Title:
Nucleic acid molecules and other molecules associated with the sucrose pathway
Kind Code:
A1


Abstract:
The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, nucleic acid sequences from maize and soybean plants associated with the sucrose pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.



Inventors:
Cheikh, Nordine (Manchester, MO, US)
Fisher, Dane K. (O'Fallon, MO, US)
Liu, Jingdong (Ballwin, MO, US)
Application Number:
12/461353
Publication Date:
12/17/2009
Filing Date:
08/10/2009
Primary Class:
Other Classes:
47/58.1R, 536/23.6
International Classes:
A01H5/00; A01G1/00; C07H21/04; C12N15/52; C12N15/82
View Patent Images:
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Primary Examiner:
PAGE, BRENT T
Attorney, Agent or Firm:
Arnold & Porter Kaye Scholer LLP (Washington, DC, US)
Claims:
1. 1-6. (canceled)

7. A transformed plant comprising a nucleic acid molecule which comprises: (a) an exogenous promoter region which functions in a plant cell to cause the production of an mRNA molecule; which is linked to; (b) a structural nucleic acid molecule, wherein said structural nucleic acid molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814, which is linked to (c) a 3′ non-translated sequence that functions in said plant cell to cause the termination of transcription and the addition of polyadenylated ribonucleotides to said 3′ end of said mRNA molecule.

8. The transformed plant according to claim 7, wherein said nucleic acid sequence is the complement of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814.

9. The transformed plant according to claim 7, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

10. The transformed plant according to claim 9, wherein said nucleic acid sequence shares between 100% and 98% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

11. The transformed plant according to claim 10, wherein said nucleic acid sequence shares between 100% and 99% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

12. The transformed plant according to claim 11, wherein said nucleic acid sequence exhibits 100% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

13. A transformed seed comprising a transformed plant cell comprising a nucleic acid molecule which comprises: (a) an exogenous promoter region which functions in said plant cell to cause the production of an mRNA molecule; which is linked to; (b) a structural nucleic acid molecule, wherein said structural nucleic acid molecule comprises a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814, which is linked to (c) a 3′ non-translated sequence that functions in said plant cell to cause the termination of transcription and the addition of polyadenylated ribonucleotides to said 3′ end of said mRNA molecule.

14. The transformed seed according to claim 13, wherein said nucleic acid sequence is the complement of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814.

15. The transformed seed according to claim 13, wherein said exogenous promoter region functions in a seed cell.

16. The transformed seed according to claim 13, wherein said nucleic acid sequence shares between 100% and 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

17. The transformed seed according to claim 16, wherein said nucleic acid sequence shares between 100% and 98% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

18. The transformed seed according to claim 17, wherein said nucleic acid sequence shares between 100% and 99% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

19. The transformed seed according to claim 18, wherein said nucleic acid sequence exhibits 100% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

20. A method of growing a transgenic plant comprising (a) planting a transformed seed comprising a nucleic acid sequence that shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814, and (b) growing a plant from said seed.

21. A substantially purified nucleic acid molecule comprising a nucleic acid sequence, wherein said nucleic acid sequence shares between 100% and 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 and the complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

22. The substantially purified nucleic acid molecule of claim 21, wherein said nucleic acid molecule encodes a soybean protein or fragment thereof.

Description:

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No. 09/237,183 filed Jan. 26, 1999, which claims benefit under 35 U.S.C § 119(e) of U.S. Provisional Application Nos. 60/067,000 filed Nov. 24, 1997; 60/069,472 filed Dec. 9, 1997; 60/072,888 filed Jan. 27, 1998; 60/074,201 filed Feb. 10, 1998; 60/074,282 filed Feb. 10, 1998; 60/074,280 filed Feb. 10, 1998; 60/074,281 filed Feb. 10, 1998; 60/074,566 filed Feb. 12, 1998; 60/074,567 filed Feb. 12, 1998; 60/074,565 filed Feb. 12, 1998; 60/075,462 filed Feb. 19, 1998; 60/074,789 filed Feb. 19, 1998; 60/075,459 filed Feb. 19, 1998; 60/075,461 filed Feb. 19, 1998; 60/075,464 filed Feb. 19, 1998; 60/075,460 filed Feb. 19, 1998; 60/075,463 filed Feb. 19, 1998; 60/076,912 filed Mar. 6, 1998; 60/077,231 filed Mar. 9, 1998; 60/077,229 filed Mar. 9, 1998; 60/077,230 filed Mar. 9, 1998; 60/078,368 filed Mar. 18, 1998; 60/080,844 filed Apr. 7, 1998; 60/083,067 filed Apr. 27, 1998; 60/083,386 filed Apr. 29, 1998; 60/083,387 filed Apr. 29, 1998; 60/083,388 filed Apr. 29, 1998; 60/083,389 filed Apr. 29, 1998; 60/083,390 filed Apr. 29, 1998; 60/085,224 filed May 13, 1998; 60/085,223 filed May 13, 1998; 60/085,222 filed May 13, 1998; 60/086,186 filed May 21, 1998; 60/086,187 filed May 21, 1998; 60/086,185 filed May 21, 1998; 60/086,184 filed May 21, 1998; 60/086,183 filed May 21, 1998; 60/086,188 filed May 21, 1998; 60/087,422 filed Jun. 1, 1998; 60/089,524 filed Jun. 16, 1998; 60/089,810 filed Jun. 18, 1998; 60/089,814 filed Jun. 18, 1998; 60/089,793 filed Jun. 18, 1998; 60/090,170 filed Jun. 22, 1998; 60/090,928 filed Jun. 26, 1998; 60/091,035 filed Jun. 29, 1998; 60/091,405 filed Jun. 30, 1998; 60/092,036 filed Jul. 8, 1998; 60/099,667 filed Sep. 9, 1998; 60/099,670 filed Sep. 9, 1998; 60/099,697 filed Sep. 9, 1998; 60/100,674 filed Sep. 16, 1998; 60/100,673 filed Sep. 16, 1998; 60/100,672 filed Sep. 16, 1998; 60/101,131 filed Sep. 21, 1998; 60/101,132 filed Sep. 21, 1998; 60/101,130 filed Sep. 21, 1998; 60/101,508 filed Sep. 22, 1998; 60/101,344 filed Sep. 22, 1998; 60/101,347 filed Sep. 22, 1998; 60/101,343 filed Sep. 22, 1998; 60/101,707 filed Sep. 25, 1998; 60/104,126 filed Oct. 13, 1998; 60/104,128 filed Oct. 13, 1998; 60/104,127 filed Oct. 13, 1998; 60/104,124 filed Oct. 13, 1998; 60/104,123 filed Oct. 13, 1998; 60/109,018 filed Nov. 18, 1998; 60/108,996 filed Nov. 18, 1998; 60/111,981 filed Dec. 11, 1998; and 60/113,224 filed Dec. 22, 1998; and claims the benefit under 35 U.S.C. § 120 as a continuation-in-part application of U.S. application Ser. Nos. 09/119,129 filed Nov. 24, 1998; 09/210,297 filed Dec. 8, 1998; and 09/229,413 filed Jan. 12, 1999, the disclosures of which are herein incorporated by reference in their entirety.

INCORPORATION OF SEQUENCE LISTING

The sequence listing is hereby incorporated by reference in its entirety, (including the file named “SequenceListing.txt” on 3 CD-ROM copies (CRF, Copy 1 and Copy 2), which is 1,352,824 bytes in size (measured in Windows XP) and was recorded on Aug. 4, 2009, which is likewise herein incorporated by reference in its entirety.)

FIELD OF THE INVENTION

The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid sequences from plant cells, in particular, nucleic acid sequences from maize and soybean plants associated with the sucrose pathway. The invention encompasses nucleic acid molecules that encode proteins and fragments of proteins. In addition, the invention also encompasses proteins and fragments of proteins so encoded and antibodies capable of binding these proteins or fragments. The invention also relates to methods of using the nucleic acid molecules, proteins and fragments of proteins and antibodies, for example for genome mapping, gene identification and analysis, plant breeding, preparation of constructs for use in plant gene expression and transgenic plants.

BACKGROUND OF THE INVENTION

Carbon fixed during photosynthesis is either retained in the chloroplast and converted to a storage carbohydrate, for example, starch, or it is transferred to the cytosol in the form of triose phosphates and converted to sucrose. The newly synthesized sucrose in source tissues is a major transported form of reduced carbon in higher plants and can be either metabolized into other carbohydrates, stored in the vacuole or exported to other plant tissues. Plant tissues where sucrose is synthesized, such as leaves, are often referred to as ‘source’ tissues. Translocated sucrose is retained in ‘sink’ tissues (such as expanding leaves, growing seeds, flowers, roots or tubers, and fruit) and may be assimilated, or further metabolized to sustain cell maintenance or fuel growth, or be converted to alternative storage compounds (e.g., starch, fats). The relative type and size of these carbohydrate pools vary during tissue development, between different plant species, and within the same species subject to different environmental conditions. Such differences are reported to affect the yield and quality of agricultural produce.

Sucrose synthesis and catabolism are reported to be highly coordinated and regulated processes that may also be coordinately regulated with other dedicated metabolic pathways in a particular plant, plant organ or cell type. Sucrose synthesis is reported to be coordinately regulated with starch metabolism and photosynthesis in green ‘source’ plant tissues. Sucrose supply by transport mechanisms to actively growing ‘sink’ tissues is reported to be coordinated with plant development. In growing sink tissues, the supply of carbohydrate is reported to be important to other metabolic pathways and physiological processes including respiration, starch biosynthesis, cell wall biogenesis, lipid and protein biosynthesis. Sucrose synthesis and/or transport is also reported to play a role in the carbohydrate capacity that is available to growing fruits and seeds. Sucrose resynthesis during seed germination is reported to play a role in seedling vigor and agronomic stand establishment in many plant species during early plant development.

In many plant species, enzymes of pathways involved in sucrose metabolism can play a role in plant physiology and plant growth and development. Compartmentation and temporal regulation of genes and enzymes of sucrose metabolic pathways can allow multiple pathways to utilize sucrose as a common metabolite. Flux through a particular sucrose metabolic pathway can define the utilization of sucrose in any tissue or developmental stage. Sucrose and its metabolite products have been reported to play a role in gene regulation and expression of the sucrose pathway and other metabolic pathways in plants.

Reviews on sucrose metabolism in plants include Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982); Hawker, In: Biochemistry of Storage Carbohydrates in Green Plants, Dey and Dixon, eds., Academic Press, London, 1-51 (1985); Huber et al., In: Carbon Partitioning Within and Between Organisms, Pollock et al., eds., Bios Scientific, Oxford, 1-26 (1992); Stitt et al., In: Biochemistry of Plants, Vol 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987); Quick and Schaffer, In: Photoassimilate Distribution In: Plants And Crops, Zamski and Schaffer, eds., Marcel Dekker Inc., New York, 115-156 (1996), all of which are herein incorporated by reference in their entirety.

The synthesis of sucrose precursors (triose and hexose phosphates) is derived from either photosynthetic CO2 fixation or degradation of previously deposited storage reserves. One substrate for sucrose synthesis in photosynthetic tissues is three carbon sugar phosphates. These are exported from the chloroplast during photosynthesis, predominantly in the form of triose phosphates. The pool of triose phosphates, dihydroxyacetone phosphate (“DHAP”), and glyceraldehyde-3-phosphate (“GAP”), is maintained at equilibrium within the cytoplasm by triose phosphate isomerase (EC 5.3.1.1). A subsequent reaction involves an aldol condensation of DHAP and GAP, catalyzed by the enzyme fructose 1,6-bisphosphate aldolase (often called aldolase) (EC 4.1.2.13) to form fructose 1,6-bisphosphate (“F1,6BP”). Fructose-1,6-bisphosphatase (“FBPase”) (EC 3.1.3.11) catalyzes the cleavage of phosphate from the C1 carbon of fructose-1,6-bisphosphate to form fructose-6-phosphate (“F6P”). This reaction is essentially irreversible and has been reported to represent the first committed step within the pathway of sucrose synthesis. The cytosolic FBPase has been reported to be subject to allosteric regulation and may serve to coordinate the rate of sucrose synthesis with that of photosynthesis. Fructose 2,6-bisphosphate (“F2,6BP”) is reported to be a regulator of FBPase (Black et al., In: Regulation of Carbohydrate Partitioning In Photosynthetic Tissue, Heath and Preiss, eds., Waverly, Baltimore, 109-126 (1985); Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987); both of which are herein incorporated by reference in their entirety). The concentration of F2,6BP is reported to be controlled in plants by two enzymes, fructose-2,6-bisphosphatase (F2,6 Bpase) (EC 3.1.3.46) and fructose-6-phosphate,2-kinase (F6P,2K) (EC 2.7.1.105) (Stitt, Annu. Rev. Plant Physiol. Plant Mol. Biol. 41: 153-181 (1990), the entirety of which is herein incorporated by reference).

Glucose-6-phosphate (“G6P”) and glucose-1-phosphate (“G1P”) are reported to be maintained in equilibrium with the F6P pool by the action of phosphoglucoisomerase (“PGI”) (EC 5.3.1.9) and phosphoglucomutase (“PGM”) (EC 5.4.2.2), respectively. Uridine diphosphate glucose (“UDPG”) and pyrophosphate (“PPi”) are formed from uridine triphosphate (“UTP”) and G1P catalyzed by the enzyme UDPG-pyrophosphorylase (“UDPGase”) (EC 2.7.7.9). This reaction is reversible and net flux in the direction of sucrose synthesis is reported to require removal of its products, particularly PPi. A pyrophosphate-dependent proton pump, vacuolar H+-translocating-pyrophosphatase (EC 3.6.1.1), has been identified within the vacuolar membrane and has been reported to utilize pyrophosphate to sustain a proton gradient formed between these two compartments (Rea et al., Trends in Biol. Sci. 17: 348-353 (1992), the entirety of which is herein incorporated by reference).

A pyrophosphate-dependent fructose-6-phosphate phosphotransferase (“PFP”) (EC 2.7.1.90) is also present in the cytoplasm and catalyzes the reversible production of F1,6BP and Pi from F6P and PPi. One reported function of PFP is to operate in a futile cycle with the cytosolic FBPase, and function as a “pseudopyrophosphatase” recycling PPi. Uridine diphosphate glucose is then combined with F6P to form sucrose-6-phosphate (“S6P”). This reaction is catalyzed by sucrose phosphate synthase (“SPS”) (EC 2.4.1.14). Attachment of UDP to the glucose moiety activates the C1 carbon atom of UDPG, which is necessary for the subsequent formation of a glycosidic bond in sucrose. In certain organisms, SPS is capable of using adenine diphosphate glucose (“ADPG”), instead of UDPG, as a substrate. The use of nucleotide biphosphate sugars is a feature of metabolic pathways leading to the production of disaccharides and polysaccharides. SPS is reported to be subject to allosteric and covalent regulation and, in conjunction with the cytosolic FBPase, reportedly serves to coordinate the rate of sucrose synthesis with the rate of photosynthesis. The reported final reaction in the pathway is catalyzed by sucrose-6-phosphate phosphatase (“SPPase” or “SPP”) (EC 3.1.3.24), which catalyzes the hydrolysis of S6P to sucrose. It has been reported that SPS and SPPase may associate to form a multienzyme complex, that the rate of sucrose-6-phosphate synthesis by SPS is enhanced in the presence of SPP, and that the rate of sucrose-6-phosphate hydrolysis by SPP is increased in the presence of SPS (Echeverria et al., Plant Physiol. 115: 223-227 (1997), herein incorporated by reference in its entirety).

I. Sucrose Synthesis

Reviews describing fructose-1,6-bisphosphatase (“FBPase”, EC 3.1.3.11) include those by Hers and Van Shaftingen, Biochem J. 206:1-12 (1982), the entirety of which is herein incorporated by reference, and Stitt, Annu. Rev. Plant Physiol. Plant Mol. Biol. 41:153-181 (1990). Two isoforms of FBPase are reported to exist in plants. The first isoform is associated with the plastid and occurs largely in photosynthetic plastids. The second isoform, located in the cytoplasm, is reported to be involved in both gluconeogenesis and sucrose synthesis (Zimmerman et al., J. Biol. Chem. 253: 5952-5956 (1978); Stitt and Heldt, Planta 164: 179-188 (1985), both of which are hereby incorporated by reference in their entirety). FBPase catalyzes an irreversible reaction in the direction of F6P synthesis in vivo and has been reported to represent the first committed step in the pathway of sucrose synthesis. The properties of the enzyme are reported to involve the action of several regulatory metabolites (Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987)). The enzyme reportedly has a high affinity for its substrate F1,6BP, a requirement for Mg2+, a requirement for a neutral pH, is weakly inhibited (Km 2-4 μm) by adenosine monophosphate (AMP), and is strongly inhibited by the regulatory metabolite F2,6BP (Hers and Van Shaftingen, Biochem J 206: 1-12 (1982); Black et al., In: Regulation of Carbohydrate Partitioning In Photosynthetic Tissue, Heath and Preiss, eds., Waverly, Baltimore, 109-126 (1985); Huber, Annu. Rev. Plant Physiol. 37: 233-246 (1986); Stiff et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987), all of which are herein incorporated by reference in their entirety). F2,6BP is also an activator of PFP and reportedly plays a role in the regulation of gluconeogenetic and respiratory metabolism.

The concentration of F2,6BP is reportedly determined in plants by two enzymes, fructose-2,6-bisphosphatase (“F2,6BPase”) (EC 3.1.3.46) and fructose-6-phosphate,2-kinase (“F6P,2K”) (EC 2.7.1.105). A review of these enzymes is provided by Stitt, Annu. Rev. Plant Physiol. Plant Mol. Biol. 41: 153-181 (1990). Regulation of the activity of the F1,6FBPase and the rate of sucrose synthesis is reported to be, at least in part, brought about by changes in the concentration of F2,6BP.

Sucrose phosphate synthase (SPS (EC 2.4.1.14)) catalyzes a reaction that is displaced from equilibrium in vivo in the direction of S6P synthesis and is reported as an essentially irreversible reaction in vivo (Stitt et al., In: Biochemistry Of Plants, Vol. 10, Hatch and Boardman, eds., Academic Press, New York, 327-407 (1987); Lunn and Rees, Biochem. J. 267: 739-743 (1990), the entirety of which is herein incorporated by reference; U.S. Pat. No. 5,665,892, the entirety of which is herein incorporated by reference). SPS has been purified from spinach and Zea mays, and the amino acid and cDNA sequences have been published (Worrel et al., Plant Cell 3:1121-1130 (1991); Klein et al., Planta 190: 498-510 (1993); Sonnewald et al., Planta 189: 174-181 (1993), all of which are herein incorporated by reference in their entirety). The enzyme has a subunit molecular weight of 117 kDa from spinach (Klein et al., Planta 190: 498-510 (1993); Sonnewald et al., Planta 189: 174-181 (1993), both of which are herein incorporated by reference) and pea (Lunn and Rees, Phytochem. 29: 1057-1063 (1990), the entirety of which is herein incorporated by reference) and 135 kDa from Zea mays (Worrel et al., Plant Cell 3:1121-1130 (1991)). The native enzyme reportedly exists as a tetramer (Walker and Huber, Plant Physiol. 89: 518-524 (1988); Lunn and Rees, Phytochem. 29: 1057-1063 (1990); Worrel et al., Plant Cell 3:1121-1130 (1991), although dimeric molecular weights have been reported (Klein et al., Planta 190: 498-510 (1993), the entirety of which is herein incorporated by reference). Activity has been observed for SPS at both dimeric and tetrameric molecular weights (Sonnewald et al., Planta 189:174-181 (1993), the entirety of which is herein incorporated by reference).

SPS is located in the cytosol, has a neutral pH optimum, and has been detected in all plant tissues which undertake active sucrose synthesis. SPS is also reported to undertake active sucrose synthesis. An increase in abundance of the enzyme is has been reported during the development of leaves, germination of seeds and ripening of fruit. The enzyme has been reported to be subject to regulation by metabolites and is activated by G6P and is inhibited by Pi. Pi and GP6 are reported to act competitively at an allosteric site of the enzyme. In the presence of high Pi concentrations, the enzyme is phosphorylated which reduces activity of the enzyme. It has also been reported that light-induced photosynthesis increases the activity of SPS in crude extracts (Sicher and Kremer, Plant Physiol. 79: 910-912 (1984), Sicher and Kremer, Plant Physiol. 79: 695-698 (1985); Pollock and Housley, Ann. Bot. 55: 593-596 (1985), all of which are herein incorporated by reference in their entirety). In addition, it has been reported that compounds altering the phosphate status of the leaf can simulate the effects of light. Feeding leaves mannose, which sequesters phosphate by its conversion to the non-metabolized mannose-6-P, has been reported to cause activation of SPS (Stitt et al., Planta 174: 217-230 (1988), the entirety of which is herein incorporated by reference).

The phosphorylation and dephosphorylation of SPS is catalyzed by SPS-phosphatase and SPS-kinase, respectively (Huber et al., Plant Physiol. 99: 1275-1278 (1992). Hydrolysis of sucrose-6-P to sucrose is catalyzed by sucrose-6-phosphatase (SPPase or SPP) (EC 3.1.3.24). The activity of both SPS and SPP is reported to be affected by a multienzyme complex between SPS and SPP (Echeverria et al., Plant Physiol. 115: 223-227 (1997)).

Regulatory properties of SPS and FBPase are reported to coordinate the rate of sucrose synthesis with that of photosynthesis (Stitt, In: Plant Physiology, Biochemistry and Molecular Biology, Dennis and Turpin, eds., Singapore, London, 319-340 (1990), the entirety of which is herein incorporated by reference). When photosynthesis produces triose phosphate in excess of the rate of sucrose synthesis, a feed-forward activation of sucrose synthesis occurs. Triose phosphate crosses the chloroplast membrane in exchange for cytosolic Pi. Under these conditions, F6P,2-kinase activity is reduced and the inhibition of F2,6Bpase is decreased.

As cytosolic F2,6BP falls, F2,6BPase activity increases, and F6P levels increase. Hexose phosphate levels are reported to increase due to PGM and PGI, and with low Pi, activate SPS and F1,6BPase. Reduction in rate of photosynthesis must result in a deactivation of sucrose synthesis, which occurs through decreased cytosolic triose-P, increased Pi and ultimately increased F2,6BP concentration and reduced SPS activity (Stitt, Phil. Trans. R Soc. Lond. B 342: 225-233 (1993); Huber et al., Plant Physiol. 99: 1275-1278 (1992); Neuhaus et al., Planta 181: 583-592 (1990), both of which are herein incorporated by reference).

II. Metabolic Pathways of Sucrose Catabolism

Sucrose can initially be cleaved by invertases (EC 3.2.1.26) or by sucrose synthases (EC 2.4.1.13). Invertases, which are classified as acid or alkaline in pH preference (Karuppiah et al., Plant Physiol. 91: 993-998 (1989); Fahrendorf and Beck, Planta 180: 237-244 (1990); Iwatsubo et al., Biosc. Biotech. Biochem. 56: 1959-1962 (1992); Unger et al., Plant Physiol. 104: 1351-1357 (1994); Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982), all of which are herein incorporated by reference in their entirety), irreversibly cleave sucrose into glucose and fructose, both of which is usually phosphorylated for further metabolism. The invertase pathway usually is associated with rapidly growing sink tissues such as expanding leaves, expanding internodes, flower petals, and early fruit development (Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982); Huber, Plant Physiol. 91:656-662 (1989); Morris and Arthur, Phytochem. 23: 2163-2167 (1984); Hawker et al., Phytochem. 15: 1441-1443 (1976); Schäffer et al., Plant Physiol. 69: 151-155 (1987), all of which are herein incorporated by reference in their entirety).

Sucrose synthase carries out the kinetically reversible transglycosylation of sucrose and UDP into fructose and UDPG, requiring only the phosphorylation of fructose for additional metabolism. Polysaccharide biosynthesis in sink tissues may utilize a sucrose synthase mediated sucrose catabolism (Avigad, In: Encyclopedia of Plant Physiology, Vol 13A, Loewus and Tanner, eds., Springer Verlag, Heidelberg, 217-347 (1982); Doehlert et al., Plant Physiol. 86: 1013-1019 (1988); Dale and Housley Plant Physiol. 82: 7-10 (1986), all of which are herein incorporated by reference). Respiring tissues reportedly utilize either sucrose synthase or invertase metabolic pathways (Echeverria and Humphreys, Phytochem. 23: 2173-2178 (1984); Uritani and Asahi, In: The Biochemistry of Plants Vol. 2, Davies, ed., Academic Press, New York, 463-487 (1980), all of which are herein incorporated by reference in their entirety). Tissues that are undergoing respiration, starch biosynthesis, amino acid and fatty acid synthesis, rapid expansion or growth, and other cellular metabolism, can utilize several sucrose metabolic pathways which may be temporally or compartmentally regulated (Doehlert et al., Plant Physiol. 86: 1013-1019 (1988); Doehlert, Plant Physiol. 78: 560-567 (1990); Doehlert and Choury, In: Recent Advances in Phloem Transport and Assimilate Compartmentation, Bonnemain et al., eds., Ouest editions, Nantes, France, 187-195 (1991); Delmer and Stone, In: The Biochemistry of Plants, Vol. 14, Preiss, ed., Academic Press, San Diego, 373-420 (1988); Maas et al., EMBO J. 9: 3447-3452 (1990), all of which are herein incorporated by reference in their entirety).

Hexose kinases are a class of enzymes responsible for the phosphorylation of hexoses, and are classified into two groups. Hexokinase (EC 2.7.1.1) can phosphorylate either glucose or fructose, with different isoforms often unique to different tissues or plant species. Different isoforms can have affinities for different hexoses (Turner and Copeland, Plant Physiol. 68: 1123-1127 (1981), the entirety of which is herein incorporated by reference; Copeland and Turner, In: The Biochemistry of Plants, Vol. 11, Stumpf and Conn, eds., Academic Press, New York, 107-128 (1987), the entirety of which is herein incorporated by reference). Hexokinases include fructokinases (EC 2.7.1.11), which typically have specific affinities for fructose (Doehlert, Plant Physiol. 89: 1042-1048 (1989); Renz and Stitt Planta 190: 166-175 (1993), both of which are herein incorporated by reference). Fructokinases can also be specific in their affinity for nucleotides. The extent to which a fructokinase utilizes UTP may play a physiological role in how efficiently UDP can be recycled for sucrose synthase activity in a particular tissue (Huber and Akazawa, Plant Physiol. 81: 1008-1013 (1986); Xu et al., Plant Physiol. 90: 635-642 (1989), both of which are herein incorporated by reference). UDP levels for the sucrose synthase reaction may be maintained, even in the case of an ATP-specific fructokinase, by the enzyme NDP-kinase (EC 2.7.4.6).

NDP-kinase has been reported in several plant tissues (Kirkland and Turner, J. Biochem. 72: 716-720 (1959); Bryce and Nelson, Plant Physiol. 63: 312-317 (1979); Dancer et al., Plant Physiol. 92: 637-641 (1990); Yano et al., Plant Molec. Biol. 23: 1087-1090 (1993), all of which are herein incorporated by reference in their entirety). Fructokinase can be substrate inhibited by fructose. In addition, sucrose synthase can be inhibited by fructose (Doehlert, Plant Sci. 52: 153-157 (1987); Morell and Copeland, Plant Physiol. 78: 140-154 (1985), Ross and Davies, Plant Physiol. 100: 1008-1013 (1992), all of which are herein incorporated by reference in their entirety). Whereas plant tissues where sucrose is catabolized by sucrose synthase predominantly contain fructokinases (Xu et al., Plant Physiol. 90: 635-642 (1989); Kursanov et al., Soviet Plant Physiol. 37: 507-515 (1990); Ross et al., Plant Physiol. 90: 748-756 (1994)), plant tissues where sucrose is catabolized by invertase often contain hexokinases (Nakamura et al., Plant Physiol. 81: 215-220 (1991)). Tissues which have both invertase and sucrose synthase activity may contain both hexose kinases (Nakamura et al., Plant Physiol. 81: 215-220 (1991), the entirety of which is herein incorporated by reference). F6P resulting from hexose kinase activity can be further metabolized in glycolysis or used in resynthesis of sucrose by SPS. G6P resulting from hexose kinase activity can enter the pentose phosphate pathway, via G6P dehydrogenase (EC 1.1.1.49), or be converted to F6P by phosphoglucoisomerase (“PGI”) (EC 5.3.1.9) or GIP by phosphoglucomutase (“PGM”) (EC 5.4.2.2) (Rees, In: Encyclopedia of Plant Physiology Vol 18, Douce and Day, eds., Springer Verlag, Berlin, 391-417 (1985); Copeland and Turner, In: The Biochemistry of Plants Vol. 11, Stumpf and Conn, eds., Academic Press, New York, 107-128 (1987); Foster and Smith, Planta 180: 237-244 (1993), all of which are herein incorporated by reference in their entirety).

PGI and PGM are reported to be ubiquitous and reversible with commitments of G6P to either F6P or G1P resulting from fluxes in metabolites further along each pathway, i.e., depending on the cell needs for glycolysis (F6P) or starch biosynthesis (G1P) (Edwards and Rees, Phytochem. 25: 2033-2039 (1986); Kursanov et al., Soviet Plant Physiol. 37: 507-515 (1990); Tobias et al., Plant Physiol. 99: 140-145 (1992), all of which are herein incorporated by reference in their entirety). UDPG formed by sucrose synthase may be utilized directly for cellulose or callose biosynthesis via UDP-glucose dehydrogenase (EC 1.1.1.2) (Robertson et al., Phytochem. 39: 21-28 (1995), the entirety of which is herein incorporated by reference), can be used for sucrose synthesis by SPS or sucrose synthase, or for glycolysis or starch metabolism dependent on further metabolism by UDP-glucose pyrophosphorylase (EC 2.7.7.9). UDP-glucose phosphorylase has been reported to be a largely reversible enzyme (Kleczkowski, Phytochem. 37: 1507-1515 (1994), the entirety of which is herein incorporated by reference). Flux through UDP-glucose pyrophosphorylase is reported to be influenced by metabolite levels and utilization of reaction products further along in the pathways (Doehlert et al., Plant Physiol. 86: 1013-1019 (1988); Huber and Akazawa, Plant Physiol. 81: 1008-1013 (1986); Zrenner et al., Planta 190: 247-252 (1993), all of which are herein incorporated by reference in their entirety). The reversibility of PGI, PGM and UDPGPPase has been reported to provide for metabolic variability and networking in metabolism, independent of which initial enzyme cleaved sucrose.

The fate of F6P reportedly plays a role in carbohydrate metabolism. NTP-phosphofructokinase (PFK) (EC 2.7.1.11) (Copeland and Turner, In: The Biochemistry of Plants Vol. 11, Stumpf and Conn, eds., Academic Press, New York, 107-128 (1987); Dennis and Greyson, Plant Physiol. 69: 395-404 (1987); Rees, In: The Biochemistry of Plants Vol. 14, Preiss, ed., Academic Press, San Diego, 1-33 (1988), all of which are herein incorporated by reference in their entirety) is reported to irreversibly convert F6P to F16BP and is associated with glycolysis. The reverse reaction of F16BP to F6P, associated with gluconeogenesis, is essentially irreversible, and is catalyzed by FBPase (EC 3.1.3.11) (Black et al., Plant Physiol. 69: 387-394 (1987). Both reactions may be carried out in a reversible manner by a PPi-dependent fructose-6-phosphate phosphotransferase or PPi-phosphofructokinase (PFP; EC 2.7.1.90) (Black et al., Plant Physiol. 69: 387-394 (1987).

PPi-dependent fructose-6-phosphate phosphotransferase or PPi-phosphofructokinase is reported to play a role in the generation of biosynthetic intermediates (Dennis and Greyson, Plant Physiol. 69: 395-404 (1987); Tobias et al., Plant Physiol. 99: 146-152 (1992), the entirety of which is herein incorporated by reference) in addition to the cycling of PPi for UDPGPPase and ultimately UDP for sucrose synthase (Huber and Akazawa, Plant Physiol. 81: 1008-1013 (1986); Black et al., Plant Physiol. 69: 387-394 (1987); Rees, In: The Biochemistry of Plants Vol. 14, Preiss, ed., Academic Press, San Diego, 1-33 (1988), all of which are herein incorporated by reference in their entirety).

II. Expressed Sequence Tag Nucleic Acid Molecules

Expressed sequence tags, or ESTs are randomly sequenced members of a cDNA library (or complementary DNA) (McCombie et al., Nature Genetics 1:124-130 (1992); Kurata et al., Nature Genetics 8:365-372 (1994); Okubo et al., Nature Genetics 2:173-179 (1992), all of which references are incorporated herein in their entirety). The randomly selected clones comprise insets that can represent a copy of up to the full length of a mRNA transcript.

Using conventional methodologies, cDNA libraries can be constructed from the mRNA (messenger RNA) of a given tissue or organism using poly dT primers and reverse transcriptase (Efstratiadis et al., Cell 7:279-3680 (1976), the entirety of which is herein incorporated by reference; Higuchi et al., Proc. Natl. Acad. Sci. (U.S.A.) 73:3146-3150 (1976), the entirety of which is herein incorporated by reference; Maniatis et al., Cell 8:163-182 (1976) the entirety of which is herein incorporated by reference; Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference; Okayama et al., Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference; Gubler et al., Gene 25:263-269 (1983), the entirety of which is herein incorporated by reference).

Several methods may be employed to obtain full-length cDNA constructs. For example, terminal transferase can be used to add homopolymeric tails of dC residues to the free 3′ hydroxyl groups (Land et al., Nucleic Acids Res. 9:2251-2266 (1981), the entirety of which is herein incorporated by reference). This tail can then be hybridized by a poly dG oligo which can act as a primer for the synthesis of full length second strand cDNA. Okayama and Berg, Mol. Cell. Biol. 2:161-170 (1982), the entirety of which is herein incorporated by reference, report a method for obtaining full length cDNA constructs. This method has been simplified by using synthetic primer-adapters that have both homopolymeric tails for priming the synthesis of the first and second strands and restriction sites for cloning into plasmids (Coleclough et al., Gene 34:305-314 (1985), the entirety of which is herein incorporated by reference) and bacteriophage vectors (Krawinkel et al., Nucleic Acids Res. 14:1913 (1986), the entirety of which is herein incorporated by reference; Han et al., Nucleic Acids Res. 15:6304 (1987), the entirety of which is herein incorporated by reference).

These strategies have been coupled with additional strategies for isolating rare mRNA populations. For example, a typical mammalian cell contains between 10,000 and 30,000 different mRNA sequences (Davidson, Gene Activity in Early Development, 2nd ed., Academic Press, New York (1976), the entirety of which is herein incorporated by reference). The number of clones required to achieve a given probability that a low-abundance mRNA will be present in a cDNA library is N=(ln(1−P))/(ln(1−1/n)) where N is the number of clones required, P is the probability desired and 1/n is the fractional proportion of the total mRNA that is represented by a single rare mRNA (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press (1989), the entirety of which is herein incorporated by reference).

A method to enrich preparations of mRNA for sequences of interest is to fractionate by size. One such method is to fractionate by electrophoresis through an agarose gel (Pennica et al., Nature 301:214-221 (1983), the entirety of which is herein incorporated by reference). Another such method employs sucrose gradient centrifugation in the presence of an agent, such as methylmercuric hydroxide, that denatures secondary structure in RNA (Schweinfest et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:4997-5000 (1982), the entirety of which is herein incorporated by reference).

A frequently adopted method is to construct equalized or normalized cDNA libraries (Ko, Nucleic Acids Res. 18:5705-5711 (1990), the entirety of which is herein incorporated by reference; Patanjali et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:1943-1947 (1991), the entirety of which is herein incorporated by reference). Typically, the cDNA population is normalized by subtractive hybridization (Schmid et al., J. Neurochem. 48:307-312 (1987), the entirety of which is herein incorporated by reference; Fargnoli et al., Anal. Biochem. 187:364-373 (1990), the entirety of which is herein incorporated by reference; Travis et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1696-1700 (1988), the entirety of which is herein incorporated by reference; Kato, Eur. J. Neurosci. 2:704-711 (1990); and Schweinfest et al., Genet. Anal. Tech. Appl. 7:64-70 (1990), the entirety of which is herein incorporated by reference). Subtraction represents another method for reducing the population of certain sequences in the cDNA library (Swaroop et al., Nucleic Acids Res. 19:1954 (1991), the entirety of which is herein incorporated by reference).

ESTs can be sequenced by a number of methods. Two basic methods may be used for DNA sequencing, the chain termination method of Sanger et al., Proc. Natl. Acad. Sci. (U.S.A.) 74:5463-5467 (1977), the entirety of which is herein incorporated by reference and the chemical degradation method of Maxam and Gilbert, Proc. Nat. Acad. Sci. (U.S.A.) 74:560-564 (1977), the entirety of which is herein incorporated by reference. Automation and advances in technology such as the replacement of radioisotopes with fluorescence-based sequencing have reduced the effort required to sequence DNA (Craxton, Methods 2:20-26 (1991), the entirety of which is herein incorporated by reference; Ju et al., Proc. Natl. Acad. Sci. (U.S.A) 92:4347-4351 (1995), the entirety of which is herein incorporated by reference; Tabor and Richardson, Proc. Natl. Acad. Sci. (U.S.A.) 92:6339-6343 (1995), the entirety of which is herein incorporated by reference). Automated sequencers are available from, for example, Pharmacia Biotech, Inc., Piscataway, N.J. (Pharmacia ALF), LI-COR, Inc., Lincoln, Nebr. (LI-COR 4,000) and Millipore, Bedford, Mass. (Millipore BaseStation).

In addition, advances in capillary gel electrophoresis have also reduced the effort required to sequence DNA and such advances provide a rapid high resolution approach for sequencing DNA samples (Swerdlow and Gesteland, Nucleic Acids Res. 18:1415-1419 (1990); Smith, Nature 349:812-813 (1991); Luckey et al., Methods Enzymol. 218:154-172 (1993); Lu et al., J Chromatog. A. 680:497-501 (1994); Carson et al., Anal. Chem. 65:3219-3226 (1993); Huang et al., Anal. Chem. 64:2149-2154 (1992); Kheterpal et al., Electrophoresis 17:1852-1859 (1996); Quesada and Zhang, Electrophoresis 17:1841-1851 (1996); Baba, Yakugaku Zasshi 117:265-281 (1997), all of which are herein incorporated by reference in their entirety).

ESTs longer than 150 nucleotides have been found to be useful for similarity searches and mapping (Adams et al., Science 252:1651-1656 (1991), herein incorporated by reference). ESTs, which can represent copies of up to the full length transcript, may be partially or completely sequenced. Between 150-450 nucleotides of sequence information is usually generated as this is the length of sequence information that is routinely and reliably produced using single run sequence data. Typically, only single run sequence data is obtained from the cDNA library (Adams et al., Science 252:1651-1656 (1991). Automated single run sequencing typically results in an approximately 2-3% error or base ambiguity rate (Boguski et al., Nature Genetics 4:332-333 (1993), the entirety of which is herein incorporated by reference).

EST databases have been constructed or partially constructed from, for example, C. elegans (McCombrie et al., Nature Genetics 1:124-131 (1992)), human liver cell line HepG2 (Okubo et al., Nature Genetics 2:173-179 (1992)), human brain RNA (Adams et al., Science 252:1651-1656 (1991); Adams et al., Nature 355:632-635 (1992)), Arabidopsis, (Newman et al., Plant Physiol. 106:1241-1255 (1994)); and rice (Kurata et al., Nature Genetics 8:365-372 (1994)).

III. Sequence Comparisons

A characteristic feature of a DNA sequence is that it can be compared with other DNA sequences. Sequence comparisons can be undertaken by determining the similarity of the test or query sequence with sequences in publicly available or proprietary databases (“similarity analysis”) or by searching for certain motifs (“intrinsic sequence analysis”) (e.g. cis elements) (Coulson, Trends in Biotechnology 12:76-80 (1994), the entirety of which is herein incorporated by reference); Birren et al., Genome Analysis 1: Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997), the entirety of which is herein incorporated by reference).

Similarity analysis includes database search and alignment. Examples of public databases include the DNA Database of Japan (DDBJ) (available on the worldwide web at ddbj.nig.acjp); Genebank (available on the worldwide web at the ncbi website at: /Web/Search/Index.html); and the European Molecular Biology Laboratory Nucleic Acid Sequence Database (EMBL) (available on the worldwide web at ebi.ac.uldebi_docs/emb1_db/embl-db.html). Other appropriate databases include dbEST (available on the worldwide web at the ncbi website at: /dbEST/index.html), SwisProt (available on the worldwide web at ebi.ac.ulc/ebi_docs/swisprot_db/swisshome.html), PIR (available on the worldwide web at nbrt.georgetown.edu/pir), and The Institute for Genome Research (available on the worldwide web at tigr.org/tdb/tdb.html).

A number of different search algorithms have been developed, one example of which are the suite of programs referred to as BLAST programs. There are five implementations of BLAST, three designed for nucleotide sequences queries (BLASTN, BLASTX and TBLASTX) and two designed for protein sequence queries (BLASTP and TBLASTN) (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 543-559 (1997)).

BLASTN takes a nucleotide sequence (the query sequence) and its reverse complement and searches them against a nucleotide sequence database. BLASTN was designed for speed, not maximum sensitivity and may not find distantly related coding sequences. BLASTX takes a nucleotide sequence, translates it in three forward reading frames and three reverse complement reading frames and then compares the six translations against a protein sequence database. BLASTX is useful for sensitive analysis of preliminary (single-pass) sequence data and is tolerant of sequencing errors (Gish and States, Nature Genetics 3:266-272 (1993), the entirety of which is herein incorporated by reference). BLASTN and BLASTX may be used in concert for analyzing EST data (Coulson, Trends in Biotechnology 12:76-80 (1994); Birren et al., Genome Analysis 1:543-559 (1997)).

Given a coding nucleotide sequence and the protein it encodes, it is often preferable to use the protein as the query sequence to search a database because of the greatly increased sensitivity to detect more subtle relationships. This is due to the larger alphabet of proteins (20 amino acids) compared with the alphabet of nucleic acid sequences (4 bases), where it is far easier to obtain a match by chance. In addition, with nucleotide alignments, only a match (positive score) or a mismatch (negative score) is obtained, but with proteins, the presence of conservative amino acid substitutions can be taken into account. Here, a mismatch may yield a positive score if the non-identical residue has physical/chemical properties similar to the one it replaced. Various scoring matrices are used to supply the substitution scores of all possible amino acid pairs. A general purpose scoring system is the BLOSUM62 matrix (Henikoff and Henikoff, Proteins 17:49-61 (1993), the entirety of which is herein incorporated by reference), which is currently the default choice for BLAST programs. BLOSUM62 is tailored for alignments of moderately diverged sequences and thus may not yield the best results under all conditions. Altschul, J. Mol. Biol. 36:290-300 (1993), the entirety of which is herein incorporated by reference, describes a combination of three matrices to cover all contingencies. This may improve sensitivity, but at the expense of slower searches. In practice, a single BLOSUM62 matrix is often used but others (PAM40 and PAM250) may be attempted when additional analysis is necessary. Low PAM matrices are directed at detecting very strong but localized sequence similarities, whereas high PAM matrices are directed at detecting long but weak alignments between very distantly related sequences.

Homologues in other organisms are available that can be used for comparative sequence analysis. Multiple alignments are performed to study similarities and differences in a group of related sequences. CLUSTAL W is a multiple sequence alignment package that performs progressive multiple sequence alignments based on the method of Feng and Doolittle, J. Mol. Evol. 25:351-360 (1987), the entirety of which is herein incorporated by reference. Each pair of sequences is aligned and the distance between each pair is calculated; from this distance matrix, a guide tree is calculated and all of the sequences are progressively aligned based on this tree. A feature of the program is its sensitivity to the effect of gaps on the alignment; gap penalties are varied to encourage the insertion of gaps in probable loop regions instead of in the middle of structured regions. Users can specify gap penalties, choose between a number of scoring matrices, or supply their own scoring matrix for both pairwise alignments and multiple alignments. CLUSTAL W for UNIX and VMS systems is available by anonymous ftp at: ftp,e13.i,ae,ak ebi.ac.uk. Another program is MACAW (Schuler et al., Proteins Struct. Func. Genet. 9:180-190 (1991), the entirety of which is herein incorporated by reference, for which both Macintosh and Microsoft Windows versions are available. MACAW uses a graphical interface, provides a choice of several alignment algorithms and is available by anonymous ftp at the ncbi website at: ncbi.nlm.nih.gov nlm.nih.gov (directory/pub/macaw).

Sequence motifs are derived from multiple alignments and can be used to examine individual sequences or an entire database for subtle patterns. With motifs, it is sometimes possible to detect distant relationships that may not be demonstrable based on comparisons of primary sequences alone. Currently, the largest collection of sequence motifs in the world is PROSITE (Bairoch and Bucher, Nucleic Acid Research 22:3583-3589 (1994), the entirety of which is herein incorporated by reference). PROSITE may be accessed via either the ExPASy server on the World Wide Web or anonymous ftp site. Many commercial sequence analysis packages also provide search programs that use PROSITE data.

A resource for searching protein motifs is the BLOCKS E-mail server developed by Henikoff, Trends Biochem Sci. 18:267-268 (1993), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Nucleic Acid Research 19:6565-6572 (1991), the entirety of which is herein incorporated by reference; Henikoff and Henikoff, Proteins 17:49-61 (1993). BLOCKS searches a protein or nucleotide sequence against a database of protein motifs or “blocks.” Blocks are defined as short, ungapped multiple alignments that represent highly conserved protein patterns. The blocks themselves are derived from entries in PROSITE as well as other sources. Either a protein query or a nucleotide query can be submitted to the BLOCKS server; if a nucleotide sequence is submitted, the sequence is translated in all six reading frames and motifs are sought for these conceptual translations. Once the search is completed, the server will return a ranked list of significant matches, along with an alignment of the query sequence to the matched BLOCKS entries.

Conserved protein domains can be represented by two-dimensional matrices, which measure either the frequency or probability of the occurrences of each amino acid residue and deletions or insertions in each position of the domain. This type of model, when used to search against protein databases, is sensitive and usually yields more accurate results than simple motif searches. Two popular implementations of this approach are profile searches such as GCG program ProfileSearch and Hidden Markov Models (HMMs) (Krough et al., J. Mol. Biol. 235:1501-1531, (1994); Eddy, Current Opinion in Structural Biology 6:361-365, (1996), both of which are herein incorporated by reference in their entirety). In both cases, a large number of common protein domains have been converted into profiles, as present in the PROSITE library, or HHM models, as in the Pfam protein domain library (Sonnhammer et al., Proteins 28:405-420 (1997), the entirety of which is herein incorporated by reference). Pfam contains more than 500 HMM models for enzymes, transcription factors, signal transduction molecules and structural proteins. Protein databases can be queried with these profiles or HMM models, which will identify proteins containing the domain of interest. For example, HMMSW or HMMFS, two programs in a public domain package called HMMER (Sonnhammer et al., Proteins 28:405-420 (1997)) can be used.

PROSITE and BLOCKS represent collected families of protein motifs. Thus, searching these databases entails submitting a single sequence to determine whether or not that sequence is similar to the members of an established family. Programs working in the opposite direction compare a collection of sequences with individual entries in the protein databases. An example of such a program is the Motif Search Tool, or MoST (Tatusov et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:12091-12095 (1994), the entirety of which is herein incorporated by reference). On the basis of an aligned set of input sequences, a weight matrix is calculated by using one of four methods (selected by the user). A weight matrix is simply a representation, position by position of how likely a particular amino acid will appear. The calculated weight matrix is then used to search the databases. To increase sensitivity, newly found sequences are added to the original data set, the weight matrix is recalculated and the search is performed again. This procedure continues until no new sequences are found.

SUMMARY OF THE INVENTION

The present invention provides a substantially purified nucleic acid molecule that encodes a maize or a soybean enzyme or fragment thereof, wherein the maize or the soybean enzyme is selected from the group consisting of: (a) triose phosphate isomerase; (b) fructose 1,6-bisphosphate aldolase; (c) fructose 1,6-bisphosphate; (d) fructose 6-phosphate 2-kinase; (e) phosphoglucoisomerase; (f) vacuolar H+ translocating-pyrophosphatase; (g) pyrophosphate-dependent fructose-6-phosphate phosphotransferase; (h) invertase; (i) sucrose synthase; (j) hexokinase; (k) fructokinase; (l) NDP-kinase; (m) glucose-6-phosphate 1-dehydrogenase; (n) phosphoglucomutase and (o) UDP-glucose pyrophosphorylase.

The present invention also provides a substantially purified nucleic acid molecule that encodes a plant sucrose pathway enzyme or fragment thereof, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof.

The present invention also provides a substantially purified maize or soybean enzyme or fragment thereof, wherein the maize or soybean enzyme is selected from the group consisting of (a) triose phosphate isomerase; (b) fructose 1,6-bisphosphate aldolase; (c) fructose 1,6-bisphosphate; (d) fructose 6-phosphate 2-kinase; (e) phosphoglucoisomerase; (f) vacuolar H+ translocating-pyrophosphatase; (g) pyrophosphate-dependent fructose-6-phosphate phosphotransferase; (h) invertase; (i) sucrose synthase; (j) hexokinase; (k) fructokinase; (l) NDP-kinase; (m) glucose-6-phosphate 1-dehydrogenase; (n) phosphoglucomutase and (o) UDP-glucose pyrophosphorylase.

The present invention also provides a substantially purified maize or soybean sucrose pathway protein or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 2814.

The present invention also provides a substantially purified maize or soybean triose phosphate isomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707.

The present invention also provides a substantially purified maize or soybean triose phosphate isomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162.

The present invention also provides a substantially purified maize or soybean fructose 1,6-bisphosphate e enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162.

The present invention also provides a substantially purified maize or soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166.

The present invention also provides a substantially purified maize or soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166.

The present invention also provides a substantially purified maize or soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182.

The present invention also provides a substantially purified maize or soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182.

The present invention also provides a substantially purified maize or soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241.

The present invention also provides a substantially purified maize or soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241.

The present invention also provides a substantially purified maize or soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442.

The present invention also provides a substantially purified maize or soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442.

The present invention also provides a substantially purified maize or soybean invertase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254.

The present invention also provides a substantially purified maize or soybean invertase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254.

The present invention also provides a substantially purified maize or soybean sucrose synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590.

The present invention also provides a substantially purified maize or soybean sucrose synthase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590.

The present invention also provides a substantially purified maize or soybean hexokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634.

The present invention also provides a substantially purified maize or soybean hexokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634.

The present invention also provides a substantially purified maize or soybean fructokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678.

The present invention also provides a substantially purified maize or soybean fructokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678.

The present invention also provides a substantially purified maize or soybean NDP-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681.

The present invention also provides a substantially purified maize or soybean NDP-kinase e enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681.

The present invention also provides a substantially purified maize or soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689.

The present invention also provides a substantially purified maize or soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689.

The present invention also provides a substantially purified maize or soybean phosphoglucomutase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740.

The present invention also provides a substantially purified maize or soybean phosphoglucomutase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740.

The present invention also provides a substantially purified maize or soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814.

The present invention also provides a substantially purified maize or soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814.

The present invention also provides a purified antibody or fragment thereof which is capable of specifically binding to a maize or soybean enzyme or fragment thereof, wherein the maize or soybean enzyme or fragment thereof is encoded by a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of consisting of SEQ ID NO: 1 through SEQ ID NO: 2814.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean triose phosphate isomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707 and a maize or soybean triose phosphate isomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113 and a maize or soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162 and a maize or soybean fructose 1,6-bisphosphate enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166 and a maize or soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182 and a maize or soybean phosphoglucoisomerase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241 and a maize or soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442 and a maize or soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean invertase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254 and a maize or soybean invertase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean sucrose synthase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590 and a maize or soybean sucrose synthase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean hexokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634 and a maize or soybean hexokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean fructokinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678 and a maize or soybean fructokinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean NDP-kinase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681 and a maize or soybean NDP-kinase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689 and a maize or soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean phosphoglucomutase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740 and a maize or soybean phosphoglucomutase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740.

The present invention also provides a substantially purified antibody or fragment thereof, the antibody or fragment thereof capable of specifically binding to a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof encoded by a first nucleic acid molecule which specifically hybridizes to a second nucleic acid molecule, the second nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a complement of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814 and a maize or soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; (B) a structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (a) a nucleic acid sequence which encodes for triose phosphate isomerase or fragment thereof; (b) a nucleic acid sequence which encodes for fructose 1,6-bisphosphate aldolase or fragment thereof; (c) a nucleic acid sequence which encodes for fructose 1,6-bisphosphate or fragment thereof; (d) a nucleic acid sequence which encodes for fructose 6-phosphate 2-kinase or fragment thereof; (e) a nucleic acid sequence which encodes for phosphoglucoisomerase or fragment thereof; (f) a nucleic acid sequence which encodes for vacuolar H+ translocating-pyrophosphatase or fragment thereof; (g) a nucleic acid sequence which encodes for pyrophosphate-dependent fructose-6-phosphate phosphotransferase or fragment thereof; (h) a nucleic acid sequence which encodes for invertase or fragment thereof; (i) a nucleic acid sequence which encodes for sucrose synthase or fragment thereof; (j) a nucleic acid sequence which encodes for hexokinase or fragment thereof; (k) a nucleic acid sequence which encodes for fructokinase or fragment thereof; (l) a nucleic acid sequence which encodes for NDP-kinase or fragment thereof; (m) a nucleic acid sequence which encodes for glucose-6-phosphate 1-dehydrogenase or fragment thereof; (n) a nucleic acid sequence which encodes for phosphoglucomutase or fragment thereof (o) a nucleic acid sequence which encodes for UDP-glucose pyrophosphorylase or fragment thereof and (p) a nucleic acid sequence which is complementary to any of the nucleic acid sequences of (a) through (o); and (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule encodes a plant sucrose pathway enzyme or fragment thereof, the structural nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a structural nucleic acid molecule, wherein the structural nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in plant cells to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a transformed plant having a nucleic acid molecule which comprises: (A) an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule; which is linked to: (B) a transcribed nucleic acid molecule with a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to an endogenous mRNA molecule having a nucleic acid sequence selected from the group consisting of an endogenous mRNA molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof; which is linked to (C) a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of the mRNA molecule.

The present invention also provides a method for determining a level or pattern in a plant cell of an enzyme in a plant metabolic pathway comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having the nucleic acid sequence of SEQ ID NO: 1 through SEQ ID NO: 2814 or compliments thereof, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of an mRNA for the enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the enzyme in the plant metabolic pathway.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant sucrose pathway enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant sucrose pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue comprising: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof or fragment of either, with a complementary nucleic acid molecule obtained from the plant cell or plant tissue, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue permits the detection of the plant sucrose pathway enzyme; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) detecting the level or pattern of the complementary nucleic acid, wherein the detection of the complementary nucleic acid is predictive of the level or pattern of the plant sucrose pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant sucrose pathway enzyme, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or reference plant tissue with the known level or pattern of the plant sucrose pathway enzyme.

The present invention also provides a method for determining a level or pattern of a plant sucrose pathway enzyme in a plant cell or plant tissue under evaluation which comprises assaying the concentration of a molecule, whose concentration is dependent upon the expression of a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof, in comparison to the concentration of that molecule present in a reference plant cell or a reference plant tissue with a known level or pattern of the plant sucrose pathway enzyme, wherein the assayed concentration of the molecule is compared to the assayed concentration of the molecule in the reference plant cell or the reference plant tissue with the known level or pattern of the plant sucrose pathway enzyme.

The present invention provides a method of determining a mutation in a plant whose presence is predictive of a mutation affecting a level or pattern of a protein comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid, the marker nucleic acid selected from the group of marker nucleic acid molecules which specifically hybridize to a nucleic acid molecule having a nucleic acid sequence selected from the group of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant sucrose pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant sucrose pathway enzyme comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant sucrose pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method for determining a mutation in a plant whose presence is predictive of a mutation affecting the level or pattern of a plant sucrose pathway enzyme comprising the steps: (A) incubating, under conditions permitting nucleic acid hybridization, a marker nucleic acid molecule, the marker nucleic acid molecule comprising a nucleic acid molecule that is linked to a gene, the gene specifically hybridizes to a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof and a complementary nucleic acid molecule obtained from the plant, wherein nucleic acid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant permits the detection of a polymorphism whose presence is predictive of a mutation affecting the level or pattern of the plant sucrose pathway enzyme in the plant; (B) permitting hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant; and (C) detecting the presence of the polymorphism, wherein the detection of the polymorphism is predictive of the mutation.

The present invention also provides a method of producing a plant containing an overexpressed protein comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region has a nucleic acid sequence selected from group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing an overexpressed plant sucrose enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant sucrose pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing an overexpressed plant sucrose pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof, wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in overexpression of the plant sucrose pathway enzyme protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing reduced levels of a plant sucrose pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814; wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant sucrose pathway enzyme protein; and (B) growing the transformed plant.

The present invention also provides a method of producing a plant containing reduced levels of a plant sucrose pathway enzyme comprising: (A) transforming the plant with a functional nucleic acid molecule, wherein the functional nucleic acid molecule comprises a promoter region, wherein the promoter region is linked to a structural region, wherein the structural region comprises a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof, wherein the structural region is linked to a 3′ non-translated sequence that functions in the plant to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and wherein the functional nucleic acid molecule results in co-suppression of the plant sucrose pathway enzyme; and (B) growing the transformed plant.

The present invention also provides a method for reducing expression of a plant sucrose pathway enzyme in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein the transcribed strand is complementary to a nucleic acid molecule having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either and the transcribed strand is complementary to an endogenous mRNA molecule; and wherein the transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and (B) growing the transformed plant.

The present invention also provides a method for reducing expression of a plant sucrose pathway enzyme in a plant comprising: (A) transforming the plant with a nucleic acid molecule, the nucleic acid molecule having an exogenous promoter region which functions in a plant cell to cause the production of a mRNA molecule, wherein the exogenous promoter region is linked to a transcribed nucleic acid molecule having a transcribed strand and a non-transcribed strand, wherein a transcribed mRNA of the transcribed strand is complementary to a nucleic acid molecule selected from the group consisting of an endogenous mRNA molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, an endogenous mRNA molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and an endogenous mRNA molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof, and wherein the transcribed nucleic acid molecule is linked to a 3′ non-translated sequence that functions in the plant cell to cause termination of transcription and addition of polyadenylated ribonucleotides to a 3′ end of a mRNA molecule; and (B) growing the transformed plant.

The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule has a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragment of either; and (B) calculating the degree of association between the polymorphism and the plant trait.

The present invention also provides a method of determining an association between a polymorphism and a plant trait comprising: (A) hybridizing a nucleic acid molecule specific for the polymorphism to genetic material of a plant, wherein the nucleic acid molecule is selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof or fragment of either and (B) calculating the degree of association between the polymorphism and the plant trait.

The present invention also provides a method of isolating a nucleic acid that encodes a plant sucrose pathway enzyme or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragment of either with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the first nucleic acid molecule and the second nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

The present invention also provides a method of isolating a nucleic acid molecule that encodes a plant sucrose pathway enzyme or fragment thereof comprising: (A) incubating under conditions permitting nucleic acid hybridization, a first nucleic acid molecule selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof or fragment of either, with a complementary second nucleic acid molecule obtained from a plant cell or plant tissue; (B) permitting hybridization between the plant sucrose pathway nucleic acid molecule and the complementary nucleic acid molecule obtained from the plant cell or plant tissue; and (C) isolating the second nucleic acid molecule.

DETAILED DESCRIPTION OF THE INVENTION

Definitions and Agents of the Present Invention

Definitions

As used herein, a sucrose pathway enzyme is any enzyme that is associated with the synthesis or degradation of sucrose.

As used herein, a sucrose synthesis enzyme is any enzyme that is associated with the synthesis of sucrose.

As used herein, a sucrose degradation enzyme is any enzyme that is associated with the degradation of sucrose.

As used herein, triose phosphate isomerase is any enzyme that maintains at equilibrium the pool of triose phosphates, dihydroxyacetone phosphate (“DHAP”), and glyceraldehyde-3-phosphate (“GAP”) within the cytoplasm.

As used herein, fructose 1,6-bisphosphate aldolase is any enzyme that catalyzes an aldol condensation of DHAP and GAP to form fructose 1,6-bisphosphate (“F1,6BP”).

As used herein, fructose-1,6-bisphosphatase (“FBPase”) is any enzyme that catalyzes the cleavage of phosphate from the C1 carbon of fructose-1,6-bisphosphate to form fructose-6-phosphate (“F6P”).

As used herein, fructose 6-phosphate 2-kinase is any enzyme that controls the concentration of fructose 2,6-bisphosphate.

As used herein, phosphoglucoisomerase is any enzyme that maintains glucose-6-phosphate (“G6P”) and glucose-1-phosphate (“GIP”) in equilibrium with the F6P pool.

As used herein, vacuolar H+ translocating-pyrophosphatase is any enzyme that utilizes pyrophosphate to sustain a proton gradient formed within the vacuolar membrane.

As used herein, pyrophosphate-dependent fructose-6-phosphate phosphotransferase is any enzyme that catalyzes the reversible production of F1,6BP and Pi from F6P and PPi.

As used herein, invertase is any enzyme that irreversibly cleaves sucrose into glucose and fructose.

As used herein, sucrose synthase is any enzyme that carries out the kinetically reversible transglycosylation of sucrose and UDP into fructose and UDPG.

As used herein, hexokinase is any enzyme that can phosphorylate either glucose or fructose.

As used herein, fructokinase is any enzyme that typically has a specific affinity for fructose.

As used herein, NDP-kinase is any enzyme that can maintain UDP levels for sucrose synthase reactions, even in the case of an ATP-specific fructokinase.

As used herein, glucose-6-phosphate 1-dehydrogenase is any enzyme that allows G6P resulting from hexose kinase activity to enter the pentose phosphate pathway.

As used herein, UDP-glucose dehydrogenase is any enzyme that allows UDPG formed by sucrose synthase to be utilized directly for cellulose or callose biosynthesis.

As used herein, phosphoglucomutase is any enzyme that is ubiquitous and reversible with commitments of G6P to either F6P or GIP resulting from fluxes in metabolites further along each pathway.

Agents

(a) Nucleic Acid Molecules

Agents of the present invention include plant nucleic acid molecules and more preferably include maize and soybean nucleic acid molecules and more preferably include nucleic acid molecules of the maize genotypes B73 (Illinois Foundation Seeds, Champaign, Illinois U.S.A.), B73×Mol7 (Illinois Foundation Seeds, Champaign, Illinois U.S.A.), DK604 (Dekalb Genetics, Dekalb, Illinois U.S.A.), H99 (Illinois Foundation Seeds, Champaign, Illinois U.S.A.), RX601 (Asgrow Seed Company, Des Moines, Iowa), Mol7 (Illinois Foundation Seeds, Champaign, Illinois U.S.A.), and soybean types Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa), C1944 (United States Department of Agriculture (USDA) Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), FT108 (Monsoy, Brazil), Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), BW211S Null (Tohoku University, Morioka, Japan), PI507354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), Asgrow A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.), P1227687 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.), P1229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.).

A subset of the nucleic acid molecules of the present invention includes nucleic acid molecules that are marker molecules. Another subset of the nucleic acid molecules of the present invention include nucleic acid molecules that encode a protein or fragment thereof. Another subset of the nucleic acid molecules of the present invention are EST molecules.

Fragment nucleic acid molecules may encode significant portion(s) of, or indeed most of, these nucleic acid molecules. Alternatively, the fragments may comprise smaller oligonucleotides (having from about 15 to about 250 nucleotide residues and more preferably, about 15 to about 30 nucleotide residues).

As used herein, an agent, be it a naturally occurring molecule or otherwise may be “substantially purified,” if desired, such that one or more molecules that is or may be present in a naturally occurring preparation containing that molecule will have been removed or will be present at a lower concentration than that at which it would normally be found.

The agents of the present invention will preferably be “biologically active” with respect to either a structural attribute, such as the capacity of a nucleic acid to hybridize to another nucleic acid molecule, or the ability of a protein to be bound by an antibody (or to compete with another molecule for such binding). Alternatively, such an attribute may be catalytic and thus involve the capacity of the agent to mediate a chemical reaction or response.

The agents of the present invention may also be recombinant. As used herein, the term recombinant means any agent (e.g. DNA, peptide etc.), that is, or results, however indirect, from human manipulation of a nucleic acid molecule.

It is understood that the agents of the present invention may be labeled with reagents that facilitate detection of the agent (e.g. fluorescent labels, Prober et al., Science 238:336-340 (1987); Albarella et al., EP 144914; chemical labels, Sheldon et al., U.S. Pat. No. 4,582,789; Albarella et al., U.S. Pat. No. 4,563,417; modified bases, Miyoshi et al., EP 119448, all of which are hereby incorporated by reference in their entirety).

It is further understood, that the present invention provides recombinant bacterial, mammalian, microbial, insect, fungal and plant cells and viral constructs comprising the agents of the present invention (See, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells).

Nucleic acid molecules or fragments thereof of the present invention are capable of specifically hybridizing to other nucleic acid molecules under certain circumstances. As used herein, two nucleic acid molecules are said to be capable of specifically hybridizing to one another if the two molecules are capable of forming an anti-parallel, double-stranded nucleic acid structure. A nucleic acid molecule is said to be the “complement” of another nucleic acid molecule if they exhibit complete complementarity. As used herein, molecules are said to exhibit “complete complementarity” when every nucleotide of one of the molecules is complementary to a nucleotide of the other. Two molecules are said to be “minimally complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under at least conventional “low-stringency” conditions. Similarly, the molecules are said to be “complementary” if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional “high-stringency” conditions. Conventional stringency conditions are described by Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989) and by Haymes et al., Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington, D.C. (1985), the entirety of which is herein incorporated by reference. Departures from complete complementarity are therefore permissible, as long as such departures do not completely preclude the capacity of the molecules to form a double-stranded structure. Thus, in order for a nucleic acid molecule to serve as a primer or probe it need only be sufficiently complementary in sequence to be able to form a stable double-stranded structure under the particular solvent and salt concentrations employed.

Appropriate stringency conditions which promote DNA hybridization, for example, 6.0× sodium chloride/sodium citrate (SSC) at about 45° C., followed by a wash of 2.0×SSC at 50° C., are known to those skilled in the art or can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. For example, the salt concentration in the wash step can be selected from a low stringency of about 2.0×SSC at 50° C. to a high stringency of about 0.2×SSC at 50° C. In addition, the temperature in the wash step can be increased from low stringency conditions at room temperature, about 22° C., to high stringency conditions at about 65° C. Both temperature and salt may be varied, or either the temperature or the salt concentration may be held constant while the other variable is changed.

In a preferred embodiment, a nucleic acid of the present invention will specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof under moderately stringent conditions, for example at about 2.0×SSC and about 65° C.

In a particularly preferred embodiment, a nucleic acid of the present invention will include those nucleic acid molecules that specifically hybridize to one or more of the nucleic acid molecules set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof under high stringency conditions such as 0.2×SSC and about 65° C.

In one aspect of the present invention, the nucleic acid molecules of the present invention have one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In another aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 90% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 95% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In a more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 98% sequence identity with one or more of the nucleic acid sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof. In an even more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention share between 100% and 99% sequence identity with one or more of the sequences set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof.

In a further more preferred aspect of the present invention, one or more of the nucleic acid molecules of the present invention exhibit 100% sequence identity with a nucleic acid molecule present within MONN01, SATMON001 through SATMON031, SATMON033, SATMON034, SATMON˜001, SATMONN01, SATMONN04 through SATMONN006, CMz029 through CMz031, CMz033, CMz035 through CMz037, CMz039 through CMz042, CMz044 through CMz045, CMz047 through CMz050, SOYMON001 through SOYMON038, Soy51 through Soy56, Soy58 through Soy62, Soy65 through Soy66, Soy 68 through Soy73 and Soy76 through Soy77, Lib9, Lib22 through Lib25, Lib35, Lib80 through Lib81, Lib 144, Lib146, Lib147, Lib190, Lib3032 through Lib3036 and Lib3099 (Monsanto Company, St. Louis, Mo. U.S.A.).

(i) Nucleic Acid Molecules Encoding Proteins or Fragments Thereof

Nucleic acid molecules of the present invention can comprise sequences that encode a sucrose pathway protein or fragment thereof. Such proteins or fragments thereof include homologues of known proteins in other organisms.

In a preferred embodiment of the present invention, a maize or a soybean protein or fragment thereof of the present invention is a homologue of another plant protein. In another preferred embodiment of the present invention, a maize or a soybean protein or fragment thereof of the present invention is a homologue of a fungal protein. In another preferred embodiment of the present invention, a maize or a soybean protein of the present invention is a homologue of mammalian protein. In another preferred embodiment of the present invention, a maize or a soybean protein or fragment thereof of the present invention is a homologue of a bacterial protein. In another preferred embodiment of the present invention, a soybean protein or fragment thereof of the present invention is a homologue of a maize protein. In another preferred embodiment of the present invention, a maize protein homologue or fragment thereof of the present invention is a homologue of a soybean protein.

In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or a soybean protein or fragment thereof where a maize or a soybean protein exhibits a BLAST probability score of greater than 1E-12, preferably a BLAST probability score of between about 1E-30 and about 1E-12, even more preferably a BLAST probability score of greater than 1E-30 with its homologue.

In another preferred embodiment of the present invention, the nucleic acid molecule encoding a maize or a soybean protein or fragment thereof exhibits a % identity with its homologue of between about 25% and about 40%, more preferably of between about 40 and about 70%, even more preferably of between about 70% and about 90% and even more preferably between about 90% and 99%. In another preferred embodiment, of the present invention, a maize or a soybean protein or fragments thereof exhibits a % identity with its homologue of 100%.

In a preferred embodiment of the present invention, the nucleic molecule of the present invention encodes a maize or a soybean protein or fragment thereof where a maize or a soybean protein exhibits a BLAST score of greater than 120, preferably a BLAST score of between about 1450 and about 120, even more preferably a BLAST score of greater than 1450 with its homologue.

Nucleic acid molecules of the present invention also include non-maize, non-soybean homologues. Preferred non-maize and soybean homologues are selected from the group consisting of alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm and Phaseolus.

In a preferred embodiment, nucleic acid molecules having SEQ ID NO: 1 through SEQ ID NO: 2814 or complements and fragments of either can be utilized to obtain such homologues.

The degeneracy of the genetic code, which allows different nucleic acid sequences to code for the same protein or peptide, is known in the literature. (U.S. Pat. No. 4,757,006, the entirety of which is herein incorporated by reference).

In an aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or a soybean protein or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 2814 due to the degeneracy in the genetic code in that they encode the same protein but differ in nucleic acid sequence.

In another further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or a soybean protein or fragment thereof in SEQ ID NO: 1 through SEQ ID NO: 2814 due to fact that the different nucleic acid sequence encodes a protein having one or more conservative amino acid residue. Examples of conservative substitutions are set forth in Table 1. It is understood that codons capable of coding for such conservative substitutions are known in the art.

TABLE 1
Original ResidueConservative Substitutions
AlaSer
ArgLys
AsnGln; His
AspGlu
CysSer; Ala
GlnAsn
GluAsp
GlyPro
HisAsn; Gln
IleLeu; Val
LeuIle; Val
LysArg; Gln; Glu
MetLeu; Ile
PheMet; Leu; Tyr
SerThr
ThrSer
TrpTyr
TyrTrp; Phe
ValIle; Leu

In a further aspect of the present invention, one or more of the nucleic acid molecules of the present invention differ in nucleic acid sequence from those encoding a maize or a soybean protein or fragment thereof set forth in SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof due to the fact that one or more codons encoding an amino acid has been substituted for a codon that encodes a nonessential substitution of the amino acid originally encoded.

Agents of the present invention include nucleic acid molecules that encode a maize or a soybean sucrose pathway protein or fragment thereof and particularly substantially purified nucleic acid molecules selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase protein or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase protein or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase protein or fragment thereof.

Non-limiting examples of such nucleic acid molecules of the present invention are nucleic acid molecules comprising: SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof that encode for a sucrose pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707 or fragment thereof that encode for a triose phosphate isomerase protein or fragment thereof, SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113 or fragment thereof that encode for a fructose 1,6-bisphosphate aldolase protein or fragment thereof, SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162 or fragment thereof that encode for a fructose 1,6-bisphosphate protein or fragment thereof, SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166 or fragment thereof that encode for a fructose 6-phosphate 2-kinase protein or fragment thereof, SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182 or fragment thereof that encode for a phosphoglucoisomerase protein or fragment thereof, SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241 or fragment thereof that encode for a vacuolar H+ translocating-pyrophosphatase protein or fragment thereof, SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442 or fragment thereof that encode for a pyrophosphate-dependent fructose-6-phosphate phosphotransferase protein or fragment thereof, SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254 or fragment thereof that encode for an invertase protein or fragment thereof, SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590 or fragment thereof that encode for a sucrose synthase protein or fragment thereof, SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634 or fragment thereof that encode for a hexokinase protein or fragment thereof, SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678 or fragment thereof that encode for a fructokinase protein or fragment thereof, SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681 or fragment thereof that encode for a NDP-kinase protein or fragment thereof, SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689 or fragment thereof that encode for a glucose-6-phosphate 1-dehydrogenase protein or fragment thereof, SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740 or fragment thereof that encode for a phosphoglucomutase protein or fragment thereof and SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ ID NO: 2741 through SEQ ID NO: 2814 or fragment thereof that encode for an UDP-glucose pyrophosphorylase protein or fragment thereof.

A nucleic acid molecule of the present invention can also encode a homologue of a maize or a soybean triose phosphate isomerase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate aldolase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate or fragment thereof, a maize or a soybean fructose 6-phosphate 2-kinase or fragment thereof, a maize or a soybean phosphoglucoisomerase or fragment thereof, a maize or a soybean vacuolar H+ translocating-pyrophosphatase or fragment thereof, a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase or fragment thereof, a maize or a soybean invertase or fragment thereof, a maize or a soybean sucrose synthase or fragment thereof, a maize or a soybean hexokinase or fragment thereof, a maize or a soybean fructokinase or fragment thereof, a maize or a soybean NDP-kinase or fragment thereof, a maize or a soybean glucose-6-phosphate 1-dehydrogenase or fragment thereof, a maize or a soybean phosphoglucomutase or fragment thereof and a maize or a soybean UDP-glucose pyrophosphorylase or fragment thereof. As used herein a homologue protein molecule or fragment thereof is a counterpart protein molecule or fragment thereof in a second species (e.g., maize triose phosphate isomerase protein is a homologue of soybean triose phosphate isomerase protein).

(ii) Nucleic Acid Molecule Markers and Probes

One aspect of the present invention concerns markers that include nucleic acid molecules SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either that can act as markers or other nucleic acid molecules of the present invention that can act as markers. Genetic markers of the present invention include “dominant” or “codominant” markers “Codominant markers” reveal the presence of two or more alleles (two per diploid individual) at a locus. “Dominant markers” reveal the presence of only a single allele per locus. The presence of the dominant marker phenotype (e.g., a band of DNA) is an indication that one allele is present in either the homozygous or heterozygous condition. The absence of the dominant marker phenotype (e.g. absence of a DNA band) is merely evidence that “some other” undefined allele is present. In the case of populations where individuals are predominantly homozygous and loci are predominately dimorphic, dominant and codominant markers can be equally valuable. As populations become more heterozygous and multi-allelic, codominant markers often become more informative of the genotype than dominant markers. Marker molecules can be, for example, capable of detecting polymorphisms such as single nucleotide polymorphisms (SNPs).

SNPs are single base changes in genomic DNA sequence. They occur at greater frequency and are spaced with a greater uniformly throughout a genome than other reported forms of polymorphism. The greater frequency and uniformity of SNPs means that there is greater probability that such a polymorphism will be found near or in a genetic locus of interest than would be the case for other polymorphisms. SNPs are located in protein-coding regions and noncoding regions of a genome. Some of these SNPs may result in defective or variant protein expression (e.g., as a result of mutations or defective splicing). Analysis (genotyping) of characterized SNPs can require only a plus/minus assay rather than a lengthy measurement, permitting easier automation.

SNPs can be characterized using any of a variety of methods. Such methods include the direct or indirect sequencing of the site, the use of restriction enzymes (Botstein et al., Am. J. Hum. Genet. 32:314-331 (1980), the entirety of which is herein incorporated reference; Konieczny and Ausubel, Plant J. 4:403-410 (1993), the entirety of which is herein incorporated by reference), enzymatic and chemical mismatch assays (Myers et al., Nature 313:495-498 (1985), the entirety of which is herein incorporated by reference), allele-specific PCR (Newton et al., Nucl. Acids Res. 17:2503-2516 (1989), the entirety of which is herein incorporated by reference; Wu et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:2757-2760 (1989), the entirety of which is herein incorporated by reference), ligase chain reaction (Barany, Proc. Natl. Acad. Sci. (U.S.A.) 88:189-193 (1991), the entirety of which is herein incorporated by reference), single-strand conformation polymorphism analysis (Labrune et al., Am. J. Hum. Genet. 48: 1115-1120 (1991), the entirety of which is herein incorporated by reference), primer-directed nucleotide incorporation assays (Kuppuswami et al., Proc. Natl. Acad. Sci. USA 88:1143-1147 (1991), the entirety of which is herein incorporated by reference), dideoxy fingerprinting (Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference), solid-phase ELISA-based oligonucleotide ligation assays (Nikiforov et al., Nucl. Acids Res. 22:4167-4175 (1994), the entirety of which is herein incorporated by reference), oligonucleotide fluorescence-quenching assays (Livak et al., PCR Methods Appl. 4:357-362 (1995), the entirety of which is herein incorporated by reference), 5′-nuclease allele-specific hybridization TaqMan assay (Livak et al., Nature Genet. 9:341-342 (1995), the entirety of which is herein incorporated by reference), template-directed dye-terminator incorporation (TDI) assay (Chen and Kwok, Nucl. Acids Res. 25:347-353 (1997), the entirety of which is herein incorporated by reference), allele-specific molecular beacon assay (Tyagi et al., Nature Biotech. 16: 49-53 (1998), the entirety of which is herein incorporated by reference), PinPoint assay (Haff and Smirnov, Genome Res. 7: 378-388 (1997), the entirety of which is herein incorporated by reference) and dCAPS analysis (Neff et al., Plant J 14:387-392 (1998), the entirety of which is herein incorporated by reference).

Additional markers, such as AFLP markers, RFLP markers and RAPD markers, can be utilized (Walton, Seed World 22-29 (July, 1993), the entirety of which is herein incorporated by reference; Burow and Blake, Molecular Dissection of Complex Traits, 13-29, Paterson (ed.), CRC Press, New York (1988), the entirety of which is herein incorporated by reference). DNA markers can be developed from nucleic acid molecules using restriction endonucleases, the PCR and/or DNA sequence information. RFLP markers result from single base changes or insertions/deletions. These codominant markers are highly abundant in plant genomes, have a medium level of polymorphism and are developed by a combination of restriction endonuclease digestion and Southern blotting hybridization. CAPS are similarly developed from restriction nuclease digestion but only of specific PCR products. These markers are also codominant, have a medium level of polymorphism and are highly abundant in the genome. The CAPS result from single base changes and insertions/deletions.

Another marker type, RAPDs, are developed from DNA amplification with random primers and result from single base changes and insertions/deletions in plant genomes. They are dominant markers with a medium level of polymorphisms and are highly abundant. AFLP markers require using the PCR on a subset of restriction fragments from extended adapter primers. These markers are both dominant and codominant are highly abundant in genomes and exhibit a medium level of polymorphism.

SSRs require DNA sequence information. These codominant markers result from repeat length changes, are highly polymorphic and do not exhibit as high a degree of abundance in the genome as CAPS, AFLPs and RAPDs SNPs also require DNA sequence information. These codominant markers result from single base substitutions. They are highly abundant and exhibit a medium of polymorphism (Rafalski et al., In: Nonmammalian Genomic Analysis, Birren and Lai (ed.), Academic Press, San Diego, Calif., pp. 75-134 (1996), the entirety of which is herein incorporated by reference). It is understood that a nucleic acid molecule of the present invention may be used as a marker.

A PCR probe is a nucleic acid molecule capable of initiating a polymerase activity while in a double-stranded structure to with another nucleic acid. Various methods for determining the structure of PCR probes and PCR techniques exist in the art. Computer generated searches using programs such as Primer3 (www-genome.wi.mit.edu/cgi-bin/primer/primer3.cgi), STSPipeline (www-genome.wi.mit.edu/cgi-bin/www-STS_Pipeline), or GeneUp (Pesole et al., BioTechniques 25:112-123 (1998) the entirety of which is herein incorporated by reference), for example, can be used to identify potential PCR primers.

It is understood that a fragment of one or more of the nucleic acid molecules of the present invention may be a probe and specifically a PCR probe.

(b) Protein and Peptide Molecules

A class of agents comprises one or more of the protein or fragments thereof or peptide molecules encoded by SEQ ID NO: 1 through SEQ ID NO: 2814 or one or more of the protein or fragment thereof and peptide molecules encoded by other nucleic acid agents of the present invention. As used herein, the term “protein molecule” or “peptide molecule” includes any molecule that comprises five or more amino acids. It is well known in the art that proteins may undergo modification, including post-translational modifications, such as, but not limited to, disulfide bond formation, glycosylation, phosphorylation, or oligomerization. Thus, as used herein, the term “protein molecule” or “peptide molecule” includes any protein molecule that is modified by any biological or non-biological process. The terms “amino acid” and “amino acids” refer to all naturally occurring L-amino acids. This definition is meant to include norleucine, ornithine, homocysteine and homoserine.

Non-limiting examples of the protein or fragment thereof of the present invention include a maize or a soybean sucrose pathway protein or fragment thereof; a maize or a soybean triose phosphate isomerase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate aldolase or fragment thereof, a maize or a soybean fructose 1,6-bisphosphate or fragment thereof, a maize or a soybean fructose 6-phosphate 2-kinase or fragment thereof, a maize or a soybean phosphoglucoisomerase or fragment thereof, a maize or a soybean vacuolar H+ translocating-pyrophosphatase or fragment thereof, a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase or fragment thereof, a maize or a soybean invertase or fragment thereof, a maize or a soybean sucrose synthase or fragment thereof, a maize or a soybean hexokinase or fragment thereof, a maize or a soybean fructokinase or fragment thereof, a maize or a soybean NDP-kinase or fragment thereof, a maize or a soybean glucose-6-phosphate 1-dehydrogenase or fragment thereof, a maize or a soybean phosphoglucomutase or fragment thereof and a maize or a soybean UDP-glucose pyrophosphorylase or fragment thereof.

Non-limiting examples of the protein or fragment molecules of the present invention are a sucrose pathway protein or fragment thereof encoded by: SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof that encode for a sucrose pathway protein or fragment thereof, SEQ ID NO: 1 through SEQ ID NO: 206 and SEQ ID NO: 1538 through SEQ ID NO: 1707 or fragment thereof that encode for a triose phosphate isomerase protein or fragment thereof, SEQ ID NO: 207 through SEQ ID NO: 232 and SEQ ID NO: 1708 through SEQ ID NO: 2113 or fragment thereof that encode for a fructose 1,6-bisphosphate aldolase protein or fragment thereof, SEQ ID NO: 233 through SEQ ID NO: 258 and SEQ ID NO: 2114 through SEQ ID NO: 2162 or fragment thereof that encode for a fructose 1,6-bisphosphate protein or fragment thereof, SEQ ID NO: 259 through SEQ ID NO: 275 and SEQ ID NO: 2163 through SEQ ID NO: 2166 or fragment thereof that encode for a fructose 6-phosphate 2-kinase protein or fragment thereof, SEQ ID NO: 276 through SEQ ID NO: 340 and SEQ ID NO: 2167 through SEQ ID NO: 2182 or fragment thereof that encode for a phosphoglucoisomerase protein or fragment thereof, SEQ ID NO: 341 through SEQ ID NO: 497 and SEQ ID NO: 2183 through SEQ ID NO: 2241 or fragment thereof that encode for a vacuolar H+ translocating-pyrophosphatase protein or fragment thereof, SEQ ID NO: 498 through SEQ ID NO: 507 and SEQ ID NO: 2442 or fragment thereof that encode for a pyrophosphate-dependent fructose-6-phosphate phosphotransferase protein or fragment thereof, SEQ ID NO: 508 through SEQ ID NO: 510 and SEQ ID NO: 2243 through SEQ ID NO: 2254 or fragment thereof that encode for an invertase protein or fragment thereof, SEQ ID NO: 511 through SEQ ID NO: 1086 and SEQ ID NO: 2255 through SEQ ID NO: 2590 or fragment thereof that encode for a sucrose synthase protein or fragment thereof, SEQ ID NO: 1087 through SEQ ID NO: 1135 and SEQ ID NO: 2591 through SEQ ID NO: 2634 or fragment thereof that encode for a hexokinase protein or fragment thereof, SEQ ID NO: 1136 through SEQ ID NO: 1215 and SEQ ID NO: 2635 through SEQ ID NO: 2678 or fragment thereof that encode for a fructokinase protein or fragment thereof, SEQ ID NO: 1216 through SEQ ID NO: 1251 and SEQ ID NO: 2679 through SEQ ID NO: 2681 or fragment thereof that encode for a NDP-kinase protein or fragment thereof, SEQ ID NO: 1252 through SEQ ID NO: 1254 and SEQ ID NO: 2682 through SEQ ID NO: 2689 or fragment thereof that encode for a glucose-6-phosphate 1-dehydrogenase protein or fragment thereof, SEQ ID NO: 1255 through SEQ ID NO: 1360 and SEQ ID NO: 2690 through SEQ ID NO: 2740 or fragment thereof that encode for a phosphoglucomutase protein or fragment thereof and SEQ ID NO: 1361 through SEQ ID NO: 1537 and SEQ IID NO: 2741 through SEQ ID NO: 2814 or fragment thereof that encode for an UDP-glucose pyrophosphorylase protein or fragment thereof.

One or more of the protein or fragment of peptide molecules may be produced via chemical synthesis, or more preferably, by expressing in a suitable bacterial or eucaryotic host. Suitable methods for expression are described by Sambrook et al., (In: Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989)), or similar texts. For example, the protein may be expressed in, for example, Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants; Section (b) Fungal Constructs and Fungal Transformants; Section (c) Mammalian Constructs and Transformed Mammalian Cells; Section (d) Insect Constructs and Transformed Insect Cells; and Section (e) Bacterial Constructs and Transformed Bacterial Cells.

A “protein fragment” is a peptide or polypeptide molecule whose amino acid sequence comprises a subset of the amino acid sequence of that protein. A protein or fragment thereof that comprises one or more additional peptide regions not derived from that protein is a “fusion” protein. Such molecules may be derivatized to contain carbohydrate or other moieties (such as keyhole limpet hemocyanin, etc.). Fusion protein or peptide molecules of the present invention are preferably produced via recombinant means.

Another class of agents comprise protein or peptide molecules or fragments or fusions thereof encoded by SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof in which conservative, non-essential or non-relevant amino acid residues have been added, replaced or deleted. Computerized means for designing modifications in protein structure are known in the art (Dahiyat and Mayo, Science 278:82-87 (1997), the entirety of which is herein incorporated by reference).

The protein molecules of the present invention include plant homologue proteins. An example of such a homologue is a homologue protein of a non-maize or non-soybean plant species, that include but not limited to alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir, eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus etc. Particularly preferred non-maize or non-soybean for use for the isolation of homologs would include, Arabidopsis, barley, cotton, oat, oilseed rape, rice, canola, ornamentals, sugarcane, sugarbeet, tomato, potato, wheat and turf grasses. Such a homologue can be obtained by any of a variety of methods. Most preferably, as indicated above, one or more of the disclosed sequences (SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof) will be used to define a pair of primers that may be used to isolate the homologue-encoding nucleic acid molecules from any desired species. Such molecules can be expressed to yield homologues by recombinant means.

(c) Antibodies

One aspect of the present invention concerns antibodies, single-chain antigen binding molecules, or other proteins that specifically bind to one or more of the protein or peptide molecules of the present invention and their homologues, fusions or fragments. Such antibodies may be used to quantitatively or qualitatively detect the protein or peptide molecules of the present invention. As used herein, an antibody or peptide is said to “specifically bind” to a protein or peptide molecule of the present invention if such binding is not competitively inhibited by the presence of non-related molecules.

Nucleic acid molecules that encode all or part of the protein of the present invention can be expressed, via recombinant means, to yield protein or peptides that can in turn be used to elicit antibodies that are capable of binding the expressed protein or peptide. Such antibodies may be used in immunoassays for that protein. Such protein-encoding molecules, or their fragments may be a “fusion” molecule (i.e., a part of a larger nucleic acid molecule) such that, upon expression, a fusion protein is produced. It is understood that any of the nucleic acid molecules of the present invention may be expressed, via recombinant means, to yield proteins or peptides encoded by these nucleic acid molecules.

The antibodies that specifically bind proteins and protein fragments of the present invention may be polyclonal or monoclonal and may comprise intact immunoglobulins, or antigen binding portions of immunoglobulins fragments (such as (F(ab′), F(ab′)2), or single-chain immunoglobulins producible, for example, via recombinant means. It is understood that practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of antibodies (see, for example, Harlow and Lane, In: Antibodies: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988), the entirety of which is herein incorporated by reference).

Murine monoclonal antibodies are particularly preferred. BALB/c mice are preferred for this purpose, however, equivalent strains may also be used. The animals are preferably immunized with approximately 25 μg of purified protein (or fragment thereof) that has been emulsified in a suitable adjuvant (such as TiterMax adjuvant (Vaxcel, Norcross, Ga.)). Immunization is preferably conducted at two intramuscular sites, one intraperitoneal site and one subcutaneous site at the base of the tail. An additional i.v. injection of approximately 25 μg of antigen is preferably given in normal saline three weeks later. After approximately 11 days following the second injection, the mice may be bled and the blood screened for the presence of anti-protein or peptide antibodies. Preferably, a direct binding Enzyme-Linked Immunoassay (ELISA) is employed for this purpose.

More preferably, the mouse having the highest antibody titer is given a third i.v. injection of approximately 25 μg of the same protein or fragment. The splenic leukocytes from this animal may be recovered 3 days later and then permitted to fuse, most preferably, using polyethylene glycol, with cells of a suitable myeloma cell line (such as, for example, the P3X63Ag8.653 myeloma cell line). Hybridoma cells are selected by culturing the cells under “HAT” (hypoxanthine-aminopterin-thymine) selection for about one week. The resulting clones may then be screened for their capacity to produce monoclonal antibodies (“mAbs”), preferably by direct ELISA.

In one embodiment, anti-protein or peptide monoclonal antibodies are isolated using a fusion of a protein or peptide of the present invention, or conjugate of a protein or peptide of the present invention, as immunogens. Thus, for example, a group of mice can be immunized using a fusion protein emulsified in Freund's complete adjuvant (e.g. approximately 50 μg of antigen per immunization). At three week intervals, an identical amount of antigen is emulsified in Freund's incomplete adjuvant and used to immunize the animals. Ten days following the third immunization, serum samples are taken and evaluated for the presence of antibody. If antibody titers are too low, a fourth booster can be employed. Polysera capable of binding the protein or peptide can also be obtained using this method.

In a preferred procedure for obtaining monoclonal antibodies, the spleens of the above-described immunized mice are removed, disrupted and immune splenocytes are isolated over a ficoll gradient. The isolated splenocytes are fused, using polyethylene glycol with BALB/c-derived HGPRT (hypoxanthine guanine phosphoribosyl transferase) deficient P3×63xAg8.653 plasmacytoma cells. The fused cells are plated into 96 well microtiter plates and screened for hybridoma fusion cells by their capacity to grow in culture medium supplemented with hypothanthine, aminopterin and thymidine for approximately 2-3 weeks.

Hybridoma cells that arise from such incubation are preferably screened for their capacity to produce an immunoglobulin that binds to a protein of interest. An indirect ELISA may be used for this purpose. In brief, the supernatants of hybridomas are incubated in microtiter wells that contain immobilized protein. After washing, the titer of bound immunoglobulin can be determined using, for example, a goat anti-mouse antibody conjugated to horseradish peroxidase. After additional washing, the amount of immobilized enzyme is determined (for example through the use of a chromogenic substrate). Such screening is performed as quickly as possible after the identification of the hybridoma in order to ensure that a desired clone is not overgrown by non-secreting neighbor cells. Desirably, the fusion plates are screened several times since the rates of hybridoma growth vary. In a preferred sub-embodiment, a different antigenic form may be used to screen the hybridoma. Thus, for example, the splenocytes may be immunized with one immunogen, but the resulting hybridomas can be screened using a different immunogen. It is understood that any of the protein or peptide molecules of the present invention may be used to raise antibodies.

As discussed below, such antibody molecules or their fragments may be used for diagnostic purposes. Where the antibodies are intended for diagnostic purposes, it may be desirable to derivatize them, for example with a ligand group (such as biotin) or a detectable marker group (such as a fluorescent group, a radioisotope or an enzyme).

The ability to produce antibodies that bind the protein or peptide molecules of the present invention permits the identification of mimetic compounds of those molecules. A “mimetic compound” is a compound that is not that compound, or a fragment of that compound, but which nonetheless exhibits an ability to specifically bind to antibodies directed against that compound.

It is understood that any of the agents of the present invention can be substantially purified and/or be biologically active and/or recombinant.

Uses of the Agents of the Invention

Nucleic acid molecules and fragments thereof of the present invention may be employed to obtain other nucleic acid molecules from the same species (e.g., ESTs or fragment thereof from maize may be utilized to obtain other nucleic acid molecules from maize). Such nucleic acid molecules include the nucleic acid molecules that encode the complete coding sequence of a protein and promoters and flanking sequences of such molecules. In addition, such nucleic acid molecules include nucleic acid molecules that encode for other isozymes or gene family members. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from maize or soybean. Methods for forming such libraries are well known in the art.

Nucleic acid molecules and fragments thereof of the present invention may also be employed to obtain nucleic acid homologues. Such homologues include the nucleic acid molecule of other plants or other organisms (e.g., alfalfa, Arabidopsis, barley, Brassica, broccoli, cabbage, citrus, cotton, garlic, oat, oilseed rape, onion, canola, flax, an ornamental plant, pea, peanut, pepper, potato, rice, rye, sorghum, strawberry, sugarcane, sugarbeet, tomato, wheat, poplar, pine, fir; eucalyptus, apple, lettuce, lentils, grape, banana, tea, turf grasses, sunflower, oil palm, Phaseolus, etc.) including the nucleic acid molecules that encode, in whole or in part, protein homologues of other plant species or other organisms, sequences of genetic elements such as promoters and transcriptional regulatory elements. Such molecules can be readily obtained by using the above-described nucleic acid molecules or fragments thereof to screen cDNA or genomic libraries obtained from such plant species. Methods for forming such libraries are well known in the art. Such homologue molecules may differ in their nucleotide sequences from those found in one or more of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof because complete complementarity is not needed for stable hybridization. The nucleic acid molecules of the present invention therefore also include molecules that, although capable of specifically hybridizing with the nucleic acid molecules, may lack “complete complementarity.”

Any of a variety of methods may be used to obtain one or more of the above-described nucleic acid molecules (Zamechik et al., Proc. Natl. Acad. Sci. (U.S.A) 83:4143-4146 (1986), the entirety of which is herein incorporated by reference; Goodchild et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:5507-5511 (1988), the entirety of which is herein incorporated by reference; Wickstrom et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:1028-1032 (1988), the entirety of which is herein incorporated by reference; Holt et al., Molec. Cell. Biol. 8:963-973 (1988), the entirety of which is herein incorporated by reference; Gerwirtz et al., Science 242:1303-1306 (1988), the entirety of which is herein incorporated by reference; Anfossi et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:3379-3383 (1989), the entirety of which is herein incorporated by reference; Becker et al., EMBO J. 8:3685-3691 (1989); the entirety of which is herein incorporated by reference). Automated nucleic acid synthesizers may be employed for this purpose. In lieu of such synthesis, the disclosed nucleic acid molecules may be used to define a pair of primers that can be used with the polymerase chain reaction (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1.986); Erlich et al., European Patent 50,424; European Patent 84,796; European Patent 258,017; European Patent 237,362; Mullis, European Patent 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194, all of which are herein incorporated by reference in their entirety) to amplify and obtain any desired nucleic acid molecule or fragment.

Promoter sequence(s) and other genetic elements, including but not limited to transcriptional regulatory flanking sequences, associated with one or more of the disclosed nucleic acid sequences can also be obtained using the disclosed nucleic acid sequence provided herein. In one embodiment, such sequences are obtained by incubating EST nucleic acid molecules or preferably fragments thereof with members of genomic libraries (e.g. maize and soybean) and recovering clones that hybridize to the EST nucleic acid molecule or fragment thereof. In a second embodiment, methods of “chromosome walking,” or inverse PCR may be used to obtain such sequences (Frohman et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8998-9002 (1988); Ohara et al., Proc. Natl. Acad. Sci. (U.S.A) 86:5673-5677 (1989); Pang et al., Biotechniques 22:1046-1048 (1977); Huang et al., Methods Mol. Biol. 69:89-96 (1997); Huang et al., Method Mol. Biol. 67:287-294 (1997); Benkel et al., Genet. Anal. 13:123-127 (1996); Hartl et al., Methods Mol. Biol. 58:293-301 (1996), all of which are herein incorporated by reference in their entirety).

The nucleic acid molecules of the present invention may be used to isolate promoters of cell enhanced, cell specific, tissue enhanced, tissue specific, developmentally or environmentally regulated expression profiles. Isolation and functional analysis of the 5′ flanking promoter sequences of these genes from genomic libraries, for example, using genomic screening methods and PCR techniques would result in the isolation of useful promoters and transcriptional regulatory elements. These methods are known to those of skill in the art and have been described (See, for example, Birren et al., Genome Analysis: Analyzing DNA, 1, (1997), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference). Promoters obtained utilizing the nucleic acid molecules of the present invention could also be modified to affect their control characteristics. Examples of such modifications would include but are not limited to enhanced sequences as reported in Uses of the Agents of the Invention, Section (a) Plant Constructs and Plant Transformants. Such genetic elements could be used to enhance gene expression of new and existing traits for crop improvements.

In one sub-aspect, such an analysis is conducted by determining the presence and/or identity of polymorphism(s) by one or more of the nucleic acid molecules of the present invention and more preferably one or more of the EST nucleic acid molecule or fragment thereof which are associated with a phenotype, or a predisposition to that phenotype.

Any of a variety of molecules can be used to identify such polymorphism(s). In one embodiment, one or more of the EST nucleic acid molecules (or a sub-fragment thereof) may be employed as a marker nucleic acid molecule to identify such polymorphism(s). Alternatively, such polymorphisms can be detected through the use of a marker nucleic acid molecule or a marker protein that is genetically linked to (i.e., a polynucleotide that co-segregates with) such polymorphism(s).

In an alternative embodiment, such polymorphisms can be detected through the use of a marker nucleic acid molecule that is physically linked to such polymorphism(s). For this purpose, marker nucleic acid molecules comprising a nucleotide sequence of a polynucleotide located within lmb of the polymorphism(s) and more preferably within 100 kb of the polymorphism(s) and most preferably within 1 Okb of the polymorphism(s) can be employed.

The genomes of animals and plants naturally undergo spontaneous mutation in the course of their continuing evolution (Gusella, Ann. Rev. Biochem. 55:831-854 (1986)). A “polymorphism” is a variation or difference in the sequence of the gene or its flanking regions that arises in some of the members of a species. The variant sequence and the “original” sequence co-exist in the species' population. In some instances, such co-existence is in stable or quasi-stable equilibrium.

A polymorphism is thus said to be “allelic,” in that, due to the existence of the polymorphism, some members of a species may have the original sequence (i.e., the original “allele”) whereas other members may have the variant sequence (i.e., the variant “allele”). In the simplest case, only one variant sequence may exist and the polymorphism is thus said to be di-allelic. In other cases, the species' population may contain multiple alleles and the polymorphism is termed tri-allelic, etc. A single gene may have multiple different unrelated polymorphisms. For example, it may have a di-allelic polymorphism at one site and a multi-allelic polymorphism at another site.

The variation that defines the polymorphism may range from a single nucleotide variation to the insertion or deletion of extended regions within a gene. In some cases, the DNA sequence variations are in regions of the genome that are characterized by short tandem repeats (STRs) that include tandem di- or tri-nucleotide repeated motifs of nucleotides. Polymorphisms characterized by such tandem repeats are referred to as “variable number tandem repeat” (“VNTR”) polymorphisms. VNTRs have been used in identity analysis (Weber, U.S. Pat. No. 5,075,217; Armour et al., FEBS Lett. 307:113-115 (1992); Jones et al., Eur. J. Haematol. 39:144-147 (1987); Horn et al., PCT Patent Application WO91/14003; Jeffreys, European Patent Application 370,719; Jeffreys, U.S. Pat. No. 5,175,082; Jeffreys et al., Amer. J. Hum. Genet. 39:11-24 (1986); Jeffreys et al., Nature 316:76-79 (1985); Gray et al., Proc. R. Acad. Soc. Lond. 243:241-253 (1991); Moore et al., Genomics 10:654-660 (1991); Jeffreys et al., Anim. Genet. 18:1-15 (1987); Hillel et al., Anim. Genet. 20:145-155 (1989); Hillel et al., Genet. 124:783-789 (1990), all of which are herein incorporated by reference in their entirety).

The detection of polymorphic sites in a sample of DNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis or other means.

The most preferred method of achieving such amplification employs the polymerase chain reaction (“PCR”) (Mullis et al., Cold Spring Harbor Symp. Quant. Biol. 51:263-273 (1986); Erlich et al., European Patent Appln. 50,424; European Patent Appln. 84,796; European Patent Application 258,017; European Patent Appln. 237,362; Mullis, European Patent Appln. 201,184; Mullis et al., U.S. Pat. No. 4,683,202; Erlich, U.S. Pat. No. 4,582,788; and Saiki et al., U.S. Pat. No. 4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

In lieu of PCR, alternative methods, such as the “Ligase Chain Reaction” (“LCR”) may be used (Barany, Proc. Natl. Acad. Sci. (U.S.A) 88:189-193 (1991), the entirety of which is herein incorporated by reference). LCR uses two pairs of oligonucleotide probes to exponentially amplify a specific target. The sequences of each pair of oligonucleotides is selected to permit the pair to hybridize to abutting sequences of the same strand of the target. Such hybridization forms a substrate for a template-dependent ligase. As with PCR, the resulting products thus serve as a template in subsequent cycles and an exponential amplification of the desired sequence is obtained.

LCR can be performed with oligonucleotides having the proximal and distal sequences of the same strand of a polymorphic site. In one embodiment, either oligonucleotide will be designed to include the actual polymorphic site of the polymorphism. In such an embodiment, the reaction conditions are selected such that the oligonucleotides can be ligated together only if the target molecule either contains or lacks the specific nucleotide that is complementary to the polymorphic site present on the oligonucleotide. Alternatively, the oligonucleotides may be selected such that they do not include the polymorphic site (see, Segev, PCT Application WO 90/01069, the entirety of which is herein incorporated by reference).

The “Oligonucleotide Ligation Assay” (“OLA”) may alternatively be employed (Landegren et al., Science 241:1077-1080 (1988), the entirety of which is herein incorporated by reference). The OLA protocol uses two oligonucleotides which are designed to be capable of hybridizing to abutting sequences of a single strand of a target. OLA, like LCR, is particularly suited for the detection of point mutations. Unlike LCR, however, OLA results in “linear” rather than exponential amplification of the target sequence.

Nickerson et al., have described a nucleic acid detection assay that combines attributes of PCR and OLA (Nickerson et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:8923-8927 (1990), the entirety of which is herein incorporated by reference). In this method, PCR is used to achieve the exponential amplification of target DNA, which is then detected using OLA. In addition to requiring multiple and separate, processing steps, one problem associated with such combinations is that they inherit all of the problems associated with PCR and OLA.

Schemes based on ligation of two (or more) oligonucleotides in the presence of nucleic acid having the sequence of the resulting “di-oligonucleotide”, thereby amplifying the di-oligonucleotide, are also known (Wu et al., Genomics 4:560-569 (1989), the entirety of which is herein incorporated by reference) and may be readily adapted to the purposes of the present invention.

Other known nucleic acid amplification procedures, such as allele-specific oligomers, branched DNA technology, transcription-based amplification systems, or isothermal amplification methods may also be used to amplify and analyze such polymorphisms (Malek et al., U.S. Pat. No. 5,130,238; Davey et al., European Patent Application 329,822; Schuster et al., U.S. Pat. No. 5,169,766; Miller et al., PCT Patent Application WO 89/06700; Kwoh et al., Proc. Natl. Acad. Sci. (U.S.A) 86:1173-1177 (1989); Gingeras et al., PCT Patent Application WO 88/10315; Walker et al., Proc. Natl. Acad. Sci. (U.S.A) 89:392-396 (1992), all of which are herein incorporated by reference in their entirety).

The identification of a polymorphism can be determined in a variety of ways. By correlating the presence or absence of it in a plant with the presence or absence of a phenotype, it is possible to predict the phenotype of that plant. If a polymorphism creates or destroys a restriction endonuclease cleavage site, or if it results in the loss or insertion of DNA (e.g., a VNTR polymorphism), it will alter the size or profile of the DNA fragments that are generated by digestion with that restriction endonuclease. As such, individuals that possess a variant sequence can be distinguished from those having the original sequence by restriction fragment analysis. Polymorphisms that can be identified in this manner are termed “restriction fragment length polymorphisms” (“RFLPs”). RFLPs have been widely used in human and plant genetic analyses (Glassberg, UK Patent Application 2135774; Skolnick et al., Cytogen. Cell Genet. 32:58-67 (1982); Botstein et al., Ann. J. Hum. Genet. 32:314-331 (1980); Fischer et al., (PCT Application WO90/13668); Uhlen, PCT Application WO90/11369).

Polymorphisms can also be identified by Single Strand Conformation Polymorphism (SSCP) analysis. SSCP is a method capable of identifying most sequence variations in a single strand of DNA, typically between 150 and 250 nucleotides in length (Elles, Methods in Molecular Medicine Molecular Diagnosis of Genetic Diseases, Humana Press (1996), the entirety of which is herein incorporated by reference); Orita et al., Genomics 5:874-879 (1989), the entirety of which is herein incorporated by reference). Under denaturing conditions a single strand of DNA will adopt a conformation that is uniquely dependent on its sequence conformation. This conformation usually will be different, even if only a single base is changed. Most conformations have been reported to alter the physical configuration or size sufficiently to be detectable by electrophoresis. A number of protocols have been described for SSCP including, but not limited to, Lee et al., Anal. Biochem. 205:289-293 (1992), the entirety of which is herein incorporated by reference; Suzuki et al., Anal. Biochem. 192:82-84 (1991), the entirety of which is herein incorporated by reference; Lo et al., Nucleic Acids Research 20:1005-1009 (1992), the entirety of which is herein incorporated by reference; Sarkar et al., Genomics 13:441-443 (1992), the entirety of which is herein incorporated by reference. It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by SSCP analysis.

Polymorphisms may also be found using a DNA fingerprinting technique called amplified fragment length polymorphism (AFLP), which is based on the selective PCR amplification of restriction fragments from a total digest of genomic DNA to profile that DNA (Vos et al., Nucleic Acids Res. 23:4407-4414 (1995), the entirety of which is herein incorporated by reference). This method allows for the specific co-amplification of high numbers of restriction fragments, which can be visualized by PCR without knowledge of the nucleic acid sequence.

AFLP employs basically three steps. Initially, a sample of genomic DNA is cut with restriction enzymes and oligonucleotide adapters are ligated to the restriction fragments of the DNA. The restriction fragments are then amplified using PCR by using the adapter and restriction sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotide flanking the restriction sites. These amplified fragments are then visualized on a denaturing polyacrylamide gel.

AFLP analysis has been performed on Salix (Beismann et al., Mol. Ecol. 6:989-993 (1997), the entirety of which is herein incorporated by reference), Acinetobacter (Janssen et al., Int. J. Syst. Bacteriol. 47:1179-1187 (1997), the entirety of which is herein incorporated by reference), Aeromonas popoffi (Huys et al., Int. J. Syst. Bacteriol. 47:1165-1171 (1997), the entirety of which is herein incorporated by reference), rice (McCouch et al., Plant Mol. Biol. 35:89-99 (1997), the entirety of which is herein incorporated by reference; Nandi et al., Mol. Gen. Genet. 255:1-8 (1997), the entirety of which is herein incorporated by reference; Cho et al., Genome 39:373-378 (1996), the entirety of which is herein incorporated by reference), barley (Hordeum vulgare)(Simons et al., Genomics 44:61-70 (1997), the entirety of which is herein incorporated by reference; Waugh et al., Mol. Gen. Genet. 255:311-321 (1997), the entirety of which is herein incorporated by reference; Qi et al., Mol. Gen. Genet. 254:330-336 (1997), the entirety of which is herein incorporated by reference; Becker et al., Mol. Gen. Genet. 249:65-73 (1995), the entirety of which is herein incorporated by reference), potato (Van der Voort et al., Mol. Gen. Genet. 255:438-447 (1997), the entirety of which is herein incorporated by reference; Meksem et al., Mol. Gen. Genet. 249:74-81 (1995), the entirety of which is herein incorporated by reference), Phytophthora infestans (Van der Lee et al., Fungal Genet. Biol. 21:278-291 (1997), the entirety of which is herein incorporated by reference), Bacillus anthracis (Keim et al., J. Bacteriol. 179:818-824 (1997), the entirety of which is herein incorporated by reference), Astragalus cremnophylax (Travis et al., Mol. EcoL 5:735-745 (1996), the entirety of which is herein incorporated by reference), Arabidopsis (Cnops et al., Mol. Gen. Genet. 253:32-41 (1996), the entirety of which is herein incorporated by reference), Escherichia coli (Lin et al., Nucleic Acids Res. 24:3649-3650 (1996), the entirety of which is herein incorporated by reference), Aeromonas (Huys et al., Int. J. Syst. Bacteriol. 46:572-580 (1996), the entirety of which is herein incorporated by reference), nematode (Folkertsma et al., Mol. Plant. Microbe Interact. 9:47-54 (1996), the entirety of which is herein incorporated by reference), tomato (Thomas et al., Plant J 8:785-794 (1995), the entirety of which is herein incorporated by reference) and human (Latorra et al., PCR Methods Appl. 3:351-358 (1994), the entirety of which is herein incorporated by reference). AFLP analysis has also been used for fingerprinting mRNA (Money et al., Nucleic Acids Res. 24:2616-2617 (1996), the entirety of which is herein incorporated by reference; Bachem et al., Plant J. 9:745-753 (1996), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acids of the present invention, may be utilized as markers or probes to detect polymorphisms by AFLP analysis or for fingerprinting RNA.

Polymorphisms may also be found using random amplified polymorphic DNA (RAPD) (Williams et al., Nucl. Acids Res. 18:6531-6535 (1990), the entirety of which is herein incorporated by reference) and cleaveable amplified polymorphic sequences (CAPS) (Lyamichev et al., Science 260:778-783 (1993), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention, may be utilized as markers or probes to detect polymorphisms by RAPD or CAPS analysis.

Through genetic mapping, a fine scale linkage map can be developed using DNA markers and, then, a genomic DNA library of large-sized fragments can be screened with molecular markers linked to the desired trait. Molecular markers are advantageous for agronomic traits that are otherwise difficult to tag, such as resistance to pathogens, insects and nematodes, tolerance to abiotic stress, quality parameters and quantitative traits such as high yield potential.

The essential requirements for marker-assisted selection in a plant breeding program are: (1) the marker(s) should co-segregate or be closely linked with the desired trait; (2) an efficient means of screening large populations for the molecular marker(s) should be available; and (3) the screening technique should have high reproducibility across laboratories and preferably be economical to use and be user-friendly.

The genetic linkage of marker molecules can be established by a gene mapping model such as, without limitation, the flanking marker model reported by Lander and Botstein, Genetics 121:185-199 (1989) and the interval mapping, based on maximum likelihood methods described by Lander and Botstein, Genetics 121:185-199 (1989) and implemented in the software package MAPMAKER/QTL (Lincoln and Lander, Mapping Genes Controlling Quantitative Traits Using MAPMAKER/QTL, Whitehead Institute for Biomedical Research, Massachusetts, (1990). Additional software includes Qgene, Version 2.23 (1996), Department of Plant Breeding and Biometry, 266 Emerson Hall, Cornell University, Ithaca, N.Y., the manual of which is herein incorporated by reference in its entirety). Use of Qgene software is a particularly preferred approach.

A maximum likelihood estimate (MLE) for the presence of a marker is calculated, together with an MLE assuming no QTL effect, to avoid false positives. A log10 of an odds ratio (LOD) is then calculated as: LOD=log10 (MLE for the presence of a QTL/MLE given no linked QTL).

The LOD score essentially indicates how much more likely the data are to have arisen assuming the presence of a QTL than in its absence. The LOD threshold value for avoiding a false positive with a given confidence, say 95%, depends on the number of markers and the length of the genome. Graphs indicating LOD thresholds are set forth in Lander and Botstein, Genetics 121:185-199 (1989) the entirety of which is herein incorporated by reference and further described by Arús and Moreno-González, Plant Breeding, Hayward et al., (eds.) Chapman & Hall, London, pp. 314-331 (1993), the entirety of which is herein incorporated by reference.

Additional models can be used. Many modifications and alternative approaches to interval mapping have been reported, including the use non-parametric methods (Kruglyak and Lander, Genetics 139:1421-1428 (1995), the entirety of which is herein incorporated by reference). Multiple regression methods or models can be also be used, in which the trait is regressed on a large number of markers (Jansen, Biometrics in Plant Breeding, van Oijen and Jansen (eds.), Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 116-124 (1994); Weber and Wricke, Advances in Plant Breeding, Blackwell, Berlin, 16 (1994), both of which is herein incorporated by reference in their entirety). Procedures combining interval mapping with regression analysis, whereby the phenotype is regressed onto a single putative QTL at a given marker interval and at the same time onto a number of markers that serve as ‘cofactors,’ have been reported by Jansen and Stam, Genetics 136:1447-1455 (1994), the entirety of which is herein incorporated by reference and Zeng, Genetics 136:1457-1468 (1994) the entirety of which is herein incorporated by reference. Generally, the use of cofactors reduces the bias and sampling error of the estimated QTL positions (Utz and Melchinger, Biometrics in Plant Breeding, van Oijen and Jansen (eds.) Proceedings of the Ninth Meeting of the Eucarpia Section Biometrics in Plant Breeding, The Netherlands, pp. 195-204 (1994), the entirety of which is herein incorporated by reference, thereby improving the precision and efficiency of QTL mapping (Zeng, Genetics 136:1457-1468 (1994)). These models can be extended to multi-environment experiments to analyze genotype-environment interactions (Jansen et al., Theo. Appl. Genet. 91:33-37 (1995), the entirety of which is herein incorporated by reference).

Selection of an appropriate mapping populations is important to map construction. The choice of appropriate mapping population depends on the type of marker systems employed (Tanksley et al., Molecular mapping plant chromosomes. Chromosome structure andfunction: Impact of new concepts, Gustafson and Appels (eds.), Plenum Press, New York, pp 157-173 (1988), the entirety of which is herein incorporated by reference). Consideration must be given to the source of parents (adapted vs. exotic) used in the mapping population. Chromosome pairing and recombination rates can be severely disturbed (suppressed) in wide crosses (adapted×exotic) and generally yield greatly reduced linkage distances. Wide crosses will usually provide segregating populations with a relatively large array of polymorphisms when compared to progeny in a narrow cross (adapted×adapted).

An F2 population is the first generation of selfing after the hybrid seed is produced. Usually a single F1 plant is selfed to generate a population segregating for all the genes in Mendelian (1:2:1) fashion. Maximum genetic information is obtained from a completely classified F2 population using a codominant marker system (Mather, Measurement of Linkage in Heredity, Methuen and Co., (1938), the entirety of which is herein incorporated by reference). In the case of dominant markers, progeny tests (e.g. F3, BCF2) are required to identify the heterozygotes, thus making it equivalent to a completely classified F2 population. However, this procedure is often prohibitive because of the cost and time involved in progeny testing. Progeny testing of F2 individuals is often used in map construction where phenotypes do not consistently reflect genotype (e.g. disease resistance) or where trait expression is controlled by a QTL. Segregation data from progeny test populations (e.g. F3 or BCF2) can be used in map construction. Marker-assisted selection can then be applied to cross progeny based on marker-trait map associations (F2, F3), where linkage groups have not been completely disassociated by recombination events (i.e., maximum disequillibrium).

Recombinant inbred lines (RIL) (genetically related lines; usually >F5, developed from continuously selfing F2 lines towards homozygosity) can be used as a mapping population. Information obtained from dominant markers can be maximized by using RIL because all loci are homozygous or nearly so. Under conditions of tight linkage (i.e., about <10% recombination), dominant and co-dominant markers evaluated in RIL populations provide more information per individual than either marker type in backcross populations (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992), the entirety of which is herein incorporated by reference). However, as the distance between markers becomes larger (i.e., loci become more independent), the information in RIL populations decreases dramatically when compared to codominant markers.

Backcross populations (e.g., generated from a cross between a successful variety (recurrent parent) and another variety (donor parent) carrying a trait not present in the former) can be utilized as a mapping population. A series of backcrosses to the recurrent parent can be made to recover most of its desirable traits. Thus a population is created consisting of individuals nearly like the recurrent parent but each individual carries varying amounts or mosaic of genomic regions from the donor parent. Backcross populations can be useful for mapping dominant markers if all loci in the recurrent parent are homozygous and the donor and recurrent parent have contrasting polymorphic marker alleles (Reiter et al., Proc. Natl. Acad. Sci. (U.S.A.) 89:1477-1481 (1992)). Information obtained from backcross populations using either codominant or dominant markers is less than that obtained from F2 populations because one, rather than two, recombinant gametes are sampled per plant. Backcross populations, however, are more informative (at low marker saturation) when compared to RILs as the distance between linked loci increases in RIL populations (i.e. about 15% recombination). Increased recombination can be beneficial for resolution of tight linkages, but may be undesirable in the construction of maps with low marker saturation.

Near-isogenic lines (NIL) created by many backcrosses to produce an array of individuals that are nearly identical in genetic composition except for the trait or genomic region under interrogation can be used as a mapping population. In mapping with NILs, only a portion of the polymorphic loci are expected to map to a selected region.

Bulk segregant analysis (BSA) is a method developed for the rapid identification of linkage between markers and traits of interest (Michelmore et al., Proc. Natl. Acad. Sci. (U.S.A.) 88:9828-9832 (1991), the entirety of which is herein incorporated by reference). In BSA, two bulked DNA samples are drawn from a segregating population originating from a single cross. These bulks contain individuals that are identical for a particular trait (resistant or susceptible to particular disease) or genomic region but arbitrary at unlinked regions (i.e. heterozygous). Regions unlinked to the target region will not differ between the bulked samples of many individuals in BSA.

It is understood that one or more of the nucleic acid molecules of the present invention may be used as molecular markers. It is also understood that one or more of the protein molecules of the present invention may be used as molecular markers.

In accordance with this aspect of the present invention, a sample nucleic acid is obtained from plants cells or tissues. Any source of nucleic acid may be used. Preferably, the nucleic acid is genomic DNA. The nucleic acid is subjected to restriction endonuclease digestion. For example, one or more nucleic acid molecule or fragment thereof of the present invention can be used as a probe in accordance with the above-described polymorphic methods. The polymorphism obtained in this approach can then be cloned to identify the mutation at the coding region which alters the protein's structure or regulatory region of the gene which affects its expression level.

In an aspect of the present invention, one or more of the nucleic molecules of the present invention are used to determine the level (i.e., the concentration of mRNA in a sample, etc.) in a plant (preferably maize or soybean) or pattern (i.e., the kinetics of expression, rate of decomposition, stability profile, etc.) of the expression of a protein encoded in part or whole by one or more of the nucleic acid molecule of the present invention (collectively, the “Expression Response” of a cell or tissue). As used herein, the Expression Response manifested by a cell or tissue is said to be “altered” if it differs from the Expression Response of cells or tissues of plants not exhibiting the phenotype. To determine whether a Expression Response is altered, the Expression Response manifested by the cell or tissue of the plant exhibiting the phenotype is compared with that of a similar cell or tissue sample of a plant not exhibiting the phenotype. As will be appreciated, it is not necessary to re-determine the Expression Response of the cell or tissue sample of plants not exhibiting the phenotype each time such a comparison is made; rather, the Expression Response of a particular plant may be compared with previously obtained values of normal plants. As used herein, the phenotype of the organism is any of one or more characteristics of an organism (e.g. disease resistance, pest tolerance, environmental tolerance such as tolerance to abiotic stress, male sterility, quality improvement or yield etc.). A change in genotype or phenotype may be transient or permanent. Also as used herein, a tissue sample is any sample that comprises more than one cell. In a preferred aspect, a tissue sample comprises cells that share a common characteristic (e.g. derived from root, seed, flower, leaf, stem or pollen etc.).

In one aspect of the present invention, an evaluation can be conducted to determine whether a particular mRNA molecule is present. One or more of the nucleic acid molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention are utilized to detect the presence or quantity of the mRNA species. Such molecules are then incubated with cell or tissue extracts of a plant under conditions sufficient to permit nucleic acid hybridization. The detection of double-stranded probe-mRNA hybrid molecules is indicative of the presence of the mRNA; the amount of such hybrid formed is proportional to the amount of mRNA. Thus, such probes may be used to ascertain the level and extent of the mRNA production in a plant's cells or tissues. Such nucleic acid hybridization may be conducted under quantitative conditions (thereby providing a numerical value of the amount of the mRNA present). Alternatively, the assay may be conducted as a qualitative assay that indicates either that the mRNA is present, or that its level exceeds a user set, predefined value.

A principle of in situ hybridization is that a labeled, single-stranded nucleic acid probe will hybridize to a complementary strand of cellular DNA or RNA and, under the appropriate conditions, these molecules will form a stable hybrid. When nucleic acid hybridization is combined with histological techniques, specific DNA or RNA sequences can be identified within a single cell. An advantage of in situ hybridization over more conventional techniques for the detection of nucleic acids is that it allows an investigator to determine the precise spatial population (Angerer et al., Dev. Biol. 101:477-484 (1984), the entirety of which is herein incorporated by reference; Angerer et al., Dev. Biol. 112:157-166 (1985), the entirety of which is herein incorporated by reference; Dixon et al., EMBO J. 10:1317-1324 (1991), the entirety of which is herein incorporated by reference). In situ hybridization may be used to measure the steady-state level of RNA accumulation. It is a sensitive technique and RNA sequences present in as few as 5-10 copies per cell can be detected (Hardin et al., J. Mol. Biol. 202:417-431 (1989), the entirety of which is herein incorporated by reference). A number of protocols have been devised for in situ hybridization, each with tissue preparation, hybridization and washing conditions (Meyerowitz, Plant Mol. Biol. Rep. 5:242-250 (1987), the entirety of which is herein incorporated by reference; Cox and Goldberg, In: Plant Molecular Biology: A Practical Approach, Shaw (ed.), pp 1-35, IRL Press, Oxford (1988), the entirety of which is herein incorporated by reference; Raikhel et al., In situ RNA hybridization in plant tissues, In: Plant Molecular Biology Manual, vol. B9: 1-32, Kluwer Academic Publisher, Dordrecht, Belgium (1989), the entirety of which is herein incorporated by reference).

In situ hybridization also allows for the localization of proteins within a tissue or cell (Wilkinson, In Situ Hybridization, Oxford University Press, Oxford (1992), the entirety of which is herein incorporated by reference; Langdale, In Situ Hybridization In: The Maize Handbook, Freeling and Walbot (eds.), pp 165-179, Springer-Verlag, New York (1994), the entirety of which is herein incorporated by reference). It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the level or pattern of a sucrose pathway protein or mRNA thereof by in situ hybridization.

Fluorescent in situ hybridization allows the localization of a particular DNA sequence along a chromosome which is useful, among other uses, for gene mapping, following chromosomes in hybrid lines or detecting chromosomes with translocations, transversions or deletions. In situ hybridization has been used to identify chromosomes in several plant species (Griffor et al., Plant Mol. Biol. 17:101-109 (1991), the entirety of which is herein incorporated by reference; Gustafson et al., Proc. Natl. Acad. Sci. (U.S.A) 87:1899-1902 (1990), herein incorporated by reference; Mukai and Gill, Genome 34:448-452 (1991), the entirety of which is herein incorporated by reference; Schwarzacher and Heslop-Harrison, Genome 34:317-323 (1991); Wang et al., Jpn. J. Genet. 66:313-316 (1991), the entirety of which is herein incorporated by reference; Parra and Windle, Nature Genetics 5:17-21 (1993), the entirety of which is herein incorporated by reference). It is understood that the nucleic acid molecules of the present invention may be used as probes or markers to localize sequences along a chromosome.

Another method to localize the expression of a molecule is tissue printing. Tissue printing provides a way to screen, at the same time on the same membrane many tissue sections from different plants or different developmental stages. Tissue-printing procedures utilize films designed to immobilize proteins and nucleic acids. In essence, a freshly cut section of a tissue is pressed gently onto nitrocellulose paper, nylon membrane or polyvinylidene difluoride membrane. Such membranes are commercially available (e.g. Millipore, Bedford, Mass. U.S.A.). The contents of the cut cell transfer onto the membrane and the contents and are immobilized to the membrane. The immobilized contents form a latent print that can be visualized with appropriate probes. When a plant tissue print is made on nitrocellulose paper, the cell walls leave a physical print that makes the anatomy visible without further treatment (Varner and Taylor, Plant Physiol. 91:31-33 (1989), the entirety of which is herein incorporated by reference).

Tissue printing on substrate films is described by Daoust, Exp. Cell Res. 12:203-211 (1957), the entirety of which is herein incorporated by reference, who detected amylase, protease, ribonuclease and deoxyribonuclease in animal tissues using starch, gelatin and agar films. These techniques can be applied to plant tissues (Yomo and Taylor, Planta 112:35-43 (1973); the entirety of which is herein incorporated by reference; Harris and Chrispeels, Plant Physiol. 56:292-299 (1975), the entirety of which is herein incorporated by reference). Advances in membrane technology have increased the range of applications of Daoust's tissue-printing techniques allowing (Cassab and Varner, J. Cell. Biol. 105:2581-2588 (1987), the entirety of which is herein incorporated by reference) the histochemical localization of various plant enzymes and deoxyribonuclease on nitrocellulose paper and nylon (Spruce et al., Phytochemistry 26:2901-2903 (1987), the entirety of which is herein incorporated by reference; Barres et al., Neuron 5:527-544 (1990), the entirety of which is herein incorporated by reference; Reid and Pont-Lezica, Tissue Printing: Tools for the Study of Anatomy, Histochemistry and Gene Expression, Academic Press, New York, N.Y. (1992), the entirety of which is herein incorporated by reference; Reid et al., Plant Physiol. 93:160-165 (1990), the entirety of which is herein incorporated by reference; Ye et al., Plant J. 1: 175-183 (1991), the entirety of which is herein incorporated by reference).

It is understood that one or more of the molecules of the present invention, preferably one or more of the EST nucleic acid molecules or fragments thereof of the present invention or one or more of the antibodies of the present invention may be utilized to detect the presence or quantity of a sucrose pathway protein by tissue printing.

Further it is also understood that any of the nucleic acid molecules of the present invention may be used as marker nucleic acids and or probes in connection with methods that require probes or marker nucleic acids. As used herein, a probe is an agent that is utilized to determine an attribute or feature (e.g. presence or absence, location, correlation, etc.) of a molecule, cell, tissue or plant. As used herein, a marker nucleic acid is a nucleic acid molecule that is utilized to determine an attribute or feature (e.g. presence or absence, location, correlation, etc.) or a molecule, cell, tissue or plant.

A microarray-based method for high-throughput monitoring of plant gene expression may be utilized to measure gene-specific hybridization targets. This ‘chip’-based approach involves using microarrays of nucleic acid molecules as gene-specific hybridization targets to quantitatively measure expression of the corresponding plant genes (Schena et al., Science 270:467-470 (1995), the entirety of which is herein incorporated by reference; Shalon, Ph.D. Thesis, Stanford University (1996), the entirety of which is herein incorporated by reference). Every nucleotide in a large sequence can be queried at the same time. Hybridization can be used to efficiently analyze nucleotide sequences.

Several microarray methods have been described. One method compares the sequences to be analyzed by hybridization to a set of oligonucleotides representing all possible subsequences (Bains and Smith, J. Theor. Biol. 135:303-307 (1989), the entirety of which is herein incorporated by reference). A second method hybridizes the sample to an array of oligonucleotide or cDNA molecules. An array consisting of oligonucleotides complementary to subsequences of a target sequence can be used to determine the identity of a target sequence, measure its amount and detect differences between the target and a reference sequence. Nucleic acid molecules microarrays may also be screened with protein molecules or fragments thereof to determine nucleic acid molecules that specifically bind protein molecules or fragments thereof.

The microarray approach may be used with polypeptide targets (U.S. Pat. No. 5,445,934; U.S. Pat. No. 5,143,854; U.S. Pat. No. 5,079,600; U.S. Pat. No. 4,923,901, all of which are herein incorporated by reference in their entirety). Essentially, polypeptides are synthesized on a substrate (microarray) and these polypeptides can be screened with either protein molecules or fragments thereof or nucleic acid molecules in order to screen for either protein molecules or fragments thereof or nucleic acid molecules that specifically bind the target polypeptides. (Fodor et al., Science 251:767-773 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules or protein or fragments thereof of the present invention may be utilized in a microarray based method.

In a preferred embodiment of the present invention microarrays may be prepared that comprise nucleic acid molecules where such nucleic acid molecules encode at least one, preferably at least two, more preferably at least three or preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, preferably at least ten, preferably at least eleven, preferably at least twelve, preferably at least thirteen, preferably at least fourteen preferably at least fifteen sucrose pathway enzymes. In a preferred embodiment the nucleic acid molecules are selected from the group consisting of a nucleic acid molecule that encodes a maize or a soybean triose phosphate isomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or fragment thereof, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or fragment thereof and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or fragment thereof.

Site directed mutagenesis may be utilized to modify nucleic acid sequences, particularly as it is a technique that allows one or more of the amino acids encoded by a nucleic acid molecule to be altered (e.g. a threonine to be replaced by a methionine). Three basic methods for site directed mutagenesis are often employed. These are cassette mutagenesis (Wells et al., Gene 34:315-323 (1985), the entirety of which is herein incorporated by reference), primer extension (Gilliam et al., Gene 12:129-137 (1980), the entirety of which is herein incorporated by reference; Zoller and Smith, Methods Enzymol. 100:468-500 (1983), the entirety of which is herein incorporated by reference; Dalbadie-McFarland et al., Proc. Natl. Acad. Sci. (U.S.A.) 79:6409-6413 (1982), the entirety of which is herein incorporated by reference) and methods based upon PCR (Scharf et al., Science 233:1076-1078 (1986), the entirety of which is herein incorporated by reference; Higuchi et al., Nucleic Acids Res. 16:7351-7367 (1988), the entirety of which is herein incorporated by reference). Site directed mutagenesis approaches are also described in European Patent 0 385 962, the entirety of which is herein incorporated by reference; European Patent 0 359 472, the entirety of which is herein incorporated by reference; and PCT Patent Application WO 93/07278, the entirety of which is herein incorporated by reference.

Site directed mutagenesis strategies have been applied to plants for both in vitro as well as in vivo site directed mutagenesis (Lanz et al., J. Biol. Chem. 266:9971-9976 (1991), the entirety of which is herein incorporated by reference; Kovgan and Zhdanov, Biotekhnologiya 5:148-154; No. 207160n, Chemical Abstracts 110:225 (1989), the entirety of which is herein incorporated by reference; Ge et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:4037-4041 (1989), the entirety of which is herein incorporated by reference; Zhu et al., J. Biol. Chem. 271:18494-18498 (1996), the entirety of which is herein incorporated by reference; Chu et al., Biochemistry 33:6150-6157 (1994), the entirety of which is herein incorporated by reference; Small et al., EMBO J. 11:1291-1296 (1992), the entirety of which is herein incorporated by reference; Cho et al., Mol. Biotechnol. 8:13-16 (1997), the entirety of which is herein incorporated by reference; Kita et al., J. Biol. Chem. 271:26529-26535 (1996), the entirety of which is herein incorporated by reference, Jin et al., Mol. Microbiol. 7:555-562 (1993), the entirety of which is herein incorporated by reference; Hatfield and Vierstra, J. Biol. Chem. 267:14799-14803 (1992), the entirety of which is herein incorporated by reference; Zhao et al., Biochemistry 31:5093-5099 (1992), the entirety of which is herein incorporated by reference).

Any of the nucleic acid molecules of the present invention may either be modified by site directed mutagenesis or used as, for example, nucleic acid molecules that are used to target other nucleic acid molecules for modification. It is understood that mutants with more than one altered nucleotide can be constructed using techniques that practitioners are familiar with such as isolating restriction fragments and ligating such fragments into an expression vector (see, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989)).

Sequence-specific DNA-binding proteins play a role in the regulation of transcription. The isolation of recombinant cDNAs encoding these proteins facilitates the biochemical analysis of their structural and functional properties. Genes encoding such DNA-binding proteins have been isolated using classical genetics (Vollbrecht et al., Nature 350: 241-243 (1991), the entirety of which is herein incorporated by reference) and molecular biochemical approaches, including the screening of recombinant cDNA libraries with antibodies (Landschulz et al., Genes Dev. 2:786-800 (1988), the entirety of which is herein incorporated by reference) or DNA probes (Bodner et al., Cell 55:505-518 (1988), the entirety of which is herein incorporated by reference). In addition, an in situ screening procedure has been used and has facilitated the isolation of sequence-specific DNA-binding proteins from various plant species (Gilmartin et al., Plant Cell 4:839-849 (1992), the entirety of which is herein incorporated by reference; Schindler et al., EMBO J. 11: 1261-1273 (1992), the entirety of which is herein incorporated by reference). An in situ screening protocol does not require the purification of the protein of interest (Vinson et al., Genes Dev. 2:801-806 (1988), the entirety of which is herein incorporated by reference; Singh et al., Cell 52:415-423 (1988), the entirety of which is herein incorporated by reference).

Two steps may be employed to characterize DNA-protein interactions. The first is to identify promoter fragments that interact with DNA-binding proteins, to titrate binding activity, to determine the specificity of binding and to determine whether a given DNA-binding activity can interact with related DNA sequences (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). Electrophoretic mobility-shift assay is a widely used assay. The assay provides a rapid and sensitive method for detecting DNA-binding proteins based on the observation that the mobility of a DNA fragment through a nondenaturing, low-ionic strength polyacrylamide gel is retarded upon association with a DNA-binding protein (Fried and Crother, Nucleic Acids Res. 9:6505-6525 (1981), the entirety of which is herein incorporated by reference). When one or more specific binding activities have been identified, the exact sequence of the DNA bound by the protein may be determined. Several procedures for characterizing protein/DNA-binding sites are used, including methylation and ethylation interference assays (Maxam and Gilbert, Methods Enzymol. 65:499-560 (1980), the entirety of which is herein incorporated by reference; Wissman and Hillen, Methods Enzymol. 208:365-379 (1991), the entirety of which is herein incorporated by reference), footprinting techniques employing DNase I (Galas and Schmitz, Nucleic Acids Res. 5:3157-3170 (1978), the entirety of which is herein incorporated by reference), 1,10-phenanthroline-copper ion methods (Sigman et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference) and hydroxyl radicals methods (Dixon et al., Methods Enzymol. 208:414-433 (1991), the entirety of which is herein incorporated by reference). It is understood that one or more of the nucleic acid molecules of the present invention may be utilized to identify a protein or fragment thereof that specifically binds to a nucleic acid molecule of the present invention. It is also understood that one or more of the protein molecules or fragments thereof of the present invention may be utilized to identify a nucleic acid molecule that specifically binds to it.

A two-hybrid system is based on the fact that many cellular functions are carried out by proteins, such as transcription factors, that interact (physically) with one another. Two-hybrid systems have been used to probe the function of new proteins (Chien et al., Proc. Natl. Acad. Sci. (U.S.A) 88:9578-9582 (1991) the entirety of which is herein incorporated by reference; Durfee et al., Genes Dev. 7:555-569 (1993) the entirety of which is herein incorporated by reference; Choi et al., Cell 78:499-512 (1994), the entirety of which is herein incorporated by reference; Kranz et al., Genes Dev. 8:313-327 (1994), the entirety of which is herein incorporated by reference).

Interaction mating techniques have facilitated a number of two-hybrid studies of protein-protein interaction. Interaction mating has been used to examine interactions between small sets of tens of proteins (Finley and Brent, Proc. Natl. Acad. Sci. (U.S.A.) 91:12098-12984 (1994), the entirety of which is herein incorporated by reference), larger sets of hundreds of proteins (Bendixen et al., Nucl. Acids Res. 22:1778-1779 (1994), the entirety of which is herein incorporated by reference) and to comprehensively map proteins encoded by a small genome (Bartel et al., Nature Genetics 12:72-77 (1996), the entirety of which is herein incorporated by reference). This technique utilizes proteins fused to the DNA-binding domain and proteins fused to the activation domain. They are expressed in two different haploid yeast strains of opposite mating type and the strains are mated to determine if the two proteins interact. Mating occurs when haploid yeast strains come into contact and result in the fusion of the two haploids into a diploid yeast strain. An interaction can be determined by the activation of a two-hybrid reporter gene in the diploid strain. An advantage of this technique is that it reduces the number of yeast transformations needed to test individual interactions. It is understood that the protein-protein interactions of protein or fragments thereof of the present invention may be investigated using the two-hybrid system and that any of the nucleic acid molecules of the present invention that encode such proteins or fragments thereof may be used to transform yeast in the two-hybrid system.

(a) Plant Constructs and Plant Transformants

One or more of the nucleic acid molecules of the present invention may be used in plant transformation or transfection. Exogenous genetic material may be transferred into a plant cell and the plant cell regenerated into a whole, fertile or sterile plant. Exogenous genetic material is any genetic material, whether naturally occurring or otherwise, from any source that is capable of being inserted into any organism. Such genetic material may be transferred into either monocotyledons and dicotyledons including, but not limited to maize (pp 63-69), soybean (pp 50-60), Arabidopsis (p 45), phaseolus (pp 47-49), peanut (pp 49-50), alfalfa (p 60), wheat (pp 69-71), rice (pp 72-79), oat (pp 80-81), sorghum (p 83), rye (p 84), tritordeum (p 84), millet (p85), fescue (p 85), perennial ryegrass (p 86), sugarcane (p87), cranberry (p101), papaya (pp 101-102), banana (p 103), banana (p 103), muskmelon (p 104), apple (p 104), cucumber (p 105), dendrobium (p 109), gladiolus (p110), chrysanthemum (p 110), liliacea (p 111), cotton (pp113-114), eucalyptus (p 115), sunflower (p 118), canola (p 118), turfgrass (p 121), sugarbeet (p 122), coffee (p 122) and dioscorea (p 122), (Christou, In: Particle Bombardment for Genetic Engineering of Plants, Biotechnology Intelligence Unit. Academic Press, San Diego, Calif. (1996), the entirety of which is herein incorporated by reference).

Transfer of a nucleic acid that encodes for a protein can result in overexpression of that protein in a transformed cell or transgenic plant. One or more of the proteins or fragments thereof encoded by nucleic acid molecules of the present invention may be overexpressed in a transformed cell or transformed plant. Particularly, any of the sucrose pathway proteins or fragments thereof may be overexpressed in a transformed cell or transgenic plant. Such overexpression may be the result of transient or stable transfer of the exogenous genetic material.

Exogenous genetic material may be transferred into a plant cell and the plant cell by the use of a DNA vector or construct designed for such a purpose. Design of such a vector is generally within the skill of the art (See, Plant Molecular Biology: A Laboratory Manual, Clark (ed.), Springier, New York (1997), the entirety of which is herein incorporated by reference).

A construct or vector may include a plant promoter to express the protein or protein fragment of choice. A number of promoters which are active in plant cells have been described in the literature. These include the nopaline synthase (NOS) promoter (Ebert et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749 (1987), the entirety of which is herein incorporated by reference), the octopine synthase (OCS) promoter (which are carried on tumor-inducing plasmids of Agrobacterium tumefaciens), the caulimovirus promoters such as the cauliflower mosaic virus (CaMV) 19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), the entirety of which is herein incorporated by reference) and the CAMV 35S promoter (Odell et al., Nature 313:810-812 (1985), the entirety of which is herein incorporated by reference), the figwort mosaic virus 35S-promoter, the light-inducible promoter from the small subunit of ribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter (Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A) 87:4144-4148 (1990), the entirety of which is herein incorporated by reference), the R gene complex promoter (Chandler et al., The Plant Cell 1:1175-1183 (1989), the entirety of which is herein incorporated by reference) and the chlorophyll a/b binding protein gene promoter, etc. These promoters have been used to create DNA constructs which have been expressed in plants; see, e.g., PCT publication WO 84/02913, herein incorporated by reference in its entirety.

Promoters which are known or are found to cause transcription of DNA in plant cells can be used in the present invention. Such promoters may be obtained from a variety of sources such as plants and plant viruses. It is preferred that the particular promoter selected should be capable of causing sufficient expression to result in the production of an effective amount of the sucrose pathway protein to cause the desired phenotype. In addition to promoters that are known to cause transcription of DNA in plant cells, other promoters may be identified for use in the current invention by screening a plant cDNA library for genes which are selectively or preferably expressed in the target tissues or cells.

For the purpose of expression in source tissues of the plant, such as the leaf, seed, root or stem, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. For this purpose, one may choose from a number of promoters for genes with tissue- or cell-specific or -enhanced expression. Examples of such promoters reported in the literature include the chloroplast glutamine synthetase GS2 promoter from pea (Edwards et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:3459-3463 (1990), herein incorporated by reference in its entirety), the chloroplast fructose-1,6-biphosphatase (FBPase) promoter from wheat (Lloyd et al., Mol. Gen. Genet. 225:209-216 (1991), herein incorporated by reference in its entirety), the nuclear photosynthetic ST-LS1 promoter from potato (Stockhaus et al., EMBO J. 8:2445-2451 (1989), herein incorporated by reference in its entirety), the serine/threonine kinase (PAL) promoter and the glucoamylase (CHS) promoter from Arabidopsis thaliana. Also reported to be active in photosynthetically active tissues are the ribulose-1,5-bisphosphate carboxylase (RbcS) promoter from eastern larch (Larix laricina), the promoter for the cab gene, cab6, from pine (Yamamoto et al., Plant Cell Physiol; 35:773-778 (1994), herein incorporated by reference in its entirety), the promoter for the Cab-1 gene from wheat (Fejes et al., Plant Mol. Biol. 15:921-932 (1990), herein incorporated by reference in its entirety), the promoter for the CAB-1 gene from spinach (Lubberstedt et al., Plant Physiol. 104:997-1006 (1994), herein incorporated by reference in its entirety), the promoter for the cab1R gene from rice (Luan et al., Plant Cell. 4:971-981 (1992), the entirety of which is herein incorporated by reference), the pyruvate, orthophosphate dikinase (PPDK) promoter from maize (Matsuoka et al., Proc. Natl. Acad. Sci. (U.S.A.) 90: 9586-9590 (1993), herein incorporated by reference in its entirety), the promoter for the tobacco Lhcb1*2 gene (Cerdan et al., Plant Mol. Biol. 33:245-255 (1997), herein incorporated by reference in its entirety), the Arabidopsis thaliana SUC2 sucrose-H+ symporter promoter (Truernit et al., Planta. 196:564-570 (1995), herein incorporated by reference in its entirety) and the promoter for the thylakoid membrane proteins from spinach (psaD, psaF, psaE, PC, FNR, atpC, atpD, cab, rbcS). Other promoters for the chlorophyll a/b-binding proteins may also be utilized in the present invention, such as the promoters for LhcB gene and PsbP gene from white mustard (Sinapis alba; Kretsch et al., Plant Mol. Biol. 28:219-229 (1995), the entirety of which is herein incorporated by reference).

For the purpose of expression in sink tissues of the plant, such as the tuber of the potato plant, the fruit of tomato, or the seed of maize, wheat, rice and barley, it is preferred that the promoters utilized in the present invention have relatively high expression in these specific tissues. A number of promoters for genes with tuber-specific or -enhanced expression are known, including the class I patatin promoter (Bevan et al., EMBO J. 8:1899-1906 (1986); Jefferson et al., Plant Mol. Biol. 14:995-1006 (1990), both of which are herein incorporated by reference in its entirety), the promoter for the potato tuber ADPGPP genes, both the large and small subunits, the sucrose synthase promoter (Salanoubat and Belliard, Gene. 60:47-56 (1987), Salanoubat and Belliard, Gene. 84:181-185 (1989), both of which are incorporated by reference in their entirety), the promoter for the major tuber proteins including the 22 kd protein complexes and proteinase inhibitors (Hannapel, Plant Physiol. 101:703-704 (1993), herein incorporated by reference in its entirety), the promoter for the granule bound starch synthase gene (GBSS) (Visser et al., Plant Mol. Biol. 17:691-699 (1991), herein incorporated by reference in its entirety) and other class I and II patatins promoters (Koster-Topfer et al., Mol Gen Genet. 219:390-396 (1989); Mignery et al., Gene. 62:27-44 (1988), both of which are herein incorporated by reference in their entirety).

Other promoters can also be used to express a sucrose pathway protein or fragment thereof in specific tissues, such as seeds or fruits. The promoter for β-conglycinin (Chen et al. Dev. Genet. 10: 112-122 (1989), herein incorporated by reference in its entirety) or other seed-specific promoters such as the napin and phaseolin promoters, can be used. The zeins are a group of storage proteins found in maize endosperm. Genomic clones for zein genes have been isolated (Pedersen et al., Cell 29:1015-1026 (1982), herein incorporated by reference in its entirety) and the promoters from these clones, including the 15 kD, 16 kD, 19 kD, 22 kD, 27 kD and γ genes, could also be used. Other promoters known to function, for example, in maize include the promoters for the following genes: waxy, Brittle, Shrunken 2, Branching enzymes I and II, starch synthases, debranching enzymes, oleosins, glutelins and sucrose synthases. A particularly preferred promoter for maize endosperm expression is the promoter for the glutelin gene from rice, more particularly the Osgt-1 promoter (Zheng et al., Mol. Cell. Biol. 13:5829-5842 (1993), herein incorporated by reference in its entirety). Examples of promoters suitable for expression in wheat include those promoters for the ADPglucose pyrosynthase (ADPGPP) subunits, the granule bound and other starch synthase, the branching and debranching enzymes, the embryogenesis-abundant proteins, the gliadins and the glutenins. Examples of such promoters in rice include those promoters for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases and the glutelins. A particularly preferred promoter is the promoter for rice glutelin, Osgt-1. Examples of such promoters for barley include those for the ADPGPP subunits, the granule bound and other starch synthase, the branching enzymes, the debranching enzymes, sucrose synthases, the hordeins, the embryo globulins and the aleurone specific proteins.

Root specific promoters may also be used. An example of such a promoter is the promoter for the acid chitinase gene (Samac et al., Plant Mol. Biol. 25:587-596 (1994), the entirety of which is herein incorporated by reference). Expression in root tissue could also be accomplished by utilizing the root specific subdomains of the CaMV35S promoter that have been identified (Lam et al., Proc. Natl. Acad. Sci. (U.S.) 86:7890-7894 (1989), herein incorporated by reference in its entirety). Other root cell specific promoters include those reported by Conkling et al. (Conkling et al., Plant Physiol. 93:1203-1211 (1990), the entirety of which is herein incorporated by reference).

Additional promoters that may be utilized are described, for example, in U.S. Pat. Nos. 5,378,619; 5,391,725; 5,428,147; 5,447,858; 5,608,144; 5,608,144; 5,614,399; 5,633,441; 5,633,435; and 4,633,436, all of which are herein incorporated in their entirety. In addition, a tissue specific enhancer may be used (Fromm et al., The Plant Cell 1:977-984 (1989), the entirety of which is herein incorporated by reference).

Constructs or vectors may also include with the coding region of interest a nucleic acid sequence that acts, in whole or in part, to terminate transcription of that region. For example, such sequences have been isolated including the Tr7 3′ sequence and the NOS 3′ sequence (Ingelbrecht et al., The Plant Cell 1:671-680 (1989), the entirety of which is herein incorporated by reference; Bevan et al., Nucleic Acids Res. 11:369-385 (1983), the entirety of which is herein incorporated by reference), or the like.

A vector or construct may also include regulatory elements. Examples of such include the Adh intron 1 (Callis et al., Genes and Develop. 1:1183-1200 (1987), the entirety of which is herein incorporated by reference), the sucrose synthase intron (Vasil et al., Plant Physiol. 91:1575-1579 (1989), the entirety of which is herein incorporated by reference) and the TMV omega element (Gallie et al., The Plant Cell 1:301-311 (1989), the entirety of which is herein incorporated by reference). These and other regulatory elements may be included when appropriate.

A vector or construct may also include a selectable marker. Selectable markers may also be used to select for plants or plant cells that contain the exogenous genetic material. Examples of such include, but are not limited to, a neo gene (Potrykus et al., Mol. Gen. Genet. 199:183-188 (1985), the entirety of which is herein incorporated by reference) which codes for kanamycin resistance and can be selected for using kanamycin, G418, etc.; a bar gene which codes for bialaphos resistance; a mutant EPSP synthase gene (Hinchee et al., Bio/Technology 6:915-922 (1988), the entirety of which is herein incorporated by reference) which encodes glyphosate resistance; a nitrilase gene which confers resistance to bromoxynil (Stalker et al., J. Biol. Chem. 263:6310-6314 (1988), the entirety of which is herein incorporated by reference); a mutant acetolactate synthase gene (ALS) which confers imidazolinone or sulphonylurea resistance (European Patent Application 154,204 (Sep. 11, 1985), the entirety of which is herein incorporated by reference); and a methotrexate resistant DHFR gene (Thillet et al., J. Biol. Chem. 263:12500-12508 (1988), the entirety of which is herein incorporated by reference).

A vector or construct may also include a transit peptide. Incorporation of a suitable chloroplast transit peptide may also be employed (European Patent Application Publication Number 0218571, the entirety of which is herein incorporated by reference). Translational enhancers may also be incorporated as part of the vector DNA. DNA constructs could contain one or more 5′ non-translated leader sequences which may serve to enhance expression of the gene products from the resulting mRNA transcripts. Such sequences may be derived from the promoter selected to express the gene or can be specifically modified to increase translation of the mRNA. Such regions may also be obtained from viral RNAs, from suitable eukaryotic genes, or from a synthetic gene sequence. For a review of optimizing expression of transgenes, see Koziel et al., Plant Mol. Biol. 32:393-405 (1996), the entirety of which is herein incorporated by reference.

A vector or construct may also include a screenable marker. Screenable markers may be used to monitor expression. Exemplary screenable markers include a β-glucuronidase or uidA gene (GUS) which encodes an enzyme for which various chromogenic substrates are known (Jefferson, Plant Mol. Biol, Rep. 5:387-405 (1987), the entirety of which is herein incorporated by reference; Jefferson et al., EMBO J. 6:3901-3907 (1987), the entirety of which is herein incorporated by reference); an R-locus gene, which encodes a product that regulates the production of anthocyanin pigments (red color) in plant tissues (Dellaporta et al., Stadler Symposium 11:263-282 (1988), the entirety of which is herein incorporated by reference); a α-lactamase gene (Sutcliffe et al., Proc. Natl. Acad. Sci. (U.S.) 75:3737-3741 (1978), the entirety of which is herein incorporated by reference), a gene which encodes an enzyme for which various chromogenic substrates are known (e.g., PADAC, a chromogenic cephalosporin); a luciferase gene (Ow et al., Science 234:856-859 (1986), the entirety of which is herein incorporated by reference); a xylE gene (Zukowsky et al., Proc. Natl. Acad. Sci. (U.S.A.) 80:1101-1105 (1983), the entirety of which is herein incorporated by reference) which encodes a catechol diozygenase that can convert chromogenic catechols; an α-amylase gene (Ikatu et al., Bio/Technol. 8:241-242 (1990), the entirety of which is herein incorporated by reference); a tyrosinase gene (Katz et al., J. Gen. Microbiol. 129:2703-2714 (1983), the entirety of which is herein incorporated by reference) which encodes an enzyme capable of oxidizing tyrosine to DOPA and dopaquinone which in turn condenses to melanin; an α-galactosidase, which will turn a chromogenic α-galactose substrate.

Included within the terms “selectable or screenable marker genes” are also genes which encode a secretable marker whose secretion can be detected as a means of identifying or selecting for transformed cells. Examples include markers which encode a secretable antigen that can be identified by antibody interaction, or even secretable enzymes which can be detected catalytically. Secretable proteins fall into a number of classes, including small, diffusible proteins which are detectable, (e.g., by ELISA), small active enzymes which are detectable in extracellular solution (e.g., α-amylase, β-lactamase, phosphinothricin transferase), or proteins which are inserted or trapped in the cell wall (such as proteins which include a leader sequence such as that found in the expression unit of extension or tobacco PR-S). Other possible selectable and/or screenable marker genes will be apparent to those of skill in the art.

There are many methods for introducing transforming nucleic acid molecules into plant cells. Suitable methods are believed to include virtually any method by which nucleic acid molecules may be introduced into a cell, such as by Agrobacterium infection or direct delivery of nucleic acid molecules such as, for example, by PEG-mediated transformation, by electroporation or by acceleration of DNA coated particles, etc (Potrykus, Ann. Rev. Plant Physiol. Plant Mol. Biol. 42:205-225 (1991), the entirety of which is herein incorporated by reference; Vasil, Plant Mol. Biol. 25:925-937 (1994), the entirety of which is herein incorporated by reference). For example, electroporation has been used to transform maize protoplasts (Fromm et al., Nature 312:791-793 (1986), the entirety of which is herein incorporated by reference).

Other vector systems suitable for introducing transforming DNA into a host plant cell include but are not limited to binary artificial chromosome (BIBAC) vectors (Hamilton et al., Gene 200:107-116 (1997), the entirety of which is herein incorporated by reference); and transfection with RNA viral vectors (Della-Cioppa et al., Ann. N.Y. Acad. Sci. (1996), 792 (Engineering Plants for Commercial Products and Applications), 57-61, the entirety of which is herein incorporated by reference). Additional vector systems also include plant selectable YAC vectors such as those described in Mullen et al., Molecular Breeding 4:449-457 (1988), the entirety of which is herein incorporated by reference).

Technology for introduction of DNA into cells is well known to those of skill in the art. Four general methods for delivering a gene into cells have been described: (1) chemical methods (Graham and van der Eb, Virology 54:536-539 (1973), the entirety of which is herein incorporated by reference); (2) physical methods such as microinjection (Capecchi, Cell 22:479-488 (1980), the entirety of which is herein incorporated by reference), electroporation (Wong and Neumann, Biochem. Biophys. Res. Commun. 107:584-587 (1982); Fromm et al., Proc. Natl. Acad. Sci. (U.S.A) 82:5824-5828 (1985); U.S. Pat. No. 5,384,253, all of which are herein incorporated in their entirety); and the gene gun (Johnston and Tang, Methods Cell Biol. 43:353-365 (1994), the entirety of which is herein incorporated by reference); (3) viral vectors (Clapp, Clin. Perinatol. 20:155-168 (1993); Lu et al., J. Exp. Med. 178:2089-2096 (1993); Eglitis and Anderson, Biotechniques 6:608-614 (1988), all of which are herein incorporated in their entirety); and (4) receptor-mediated mechanisms (Curiel et al., Hum. Gen. Ther. 3:147-154 (1992), Wagner et al., Proc. Natl. Acad. Sci. (U.S.A) 89:6099-6103 (1992), both of which are incorporated by reference in their entirety).

Acceleration methods that may be used include, for example, microprojectile bombardment and the like. One example of a method for delivering transforming nucleic acid molecules to plant cells is microprojectile bombardment. This method has been reviewed by Yang and Christou (eds.), Particle Bombardment Technology for Gene Transfer, Oxford Press, Oxford, England (1994), the entirety of which is herein incorporated by reference). Non-biological particles (microprojectiles) that may be coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, gold, platinum and the like.

A particular advantage of microprojectile bombardment, in addition to it being an effective means of reproducibly transforming monocots, is that neither the isolation of protoplasts (Cristou et al., Plant Physiol. 87:671-674 (1988), the entirety of which is herein incorporated by reference) nor the susceptibility of Agrobacterium infection are required. An illustrative embodiment of a method for delivering DNA into maize cells by acceleration is a biolistics α-particle delivery system, which can be used to propel particles coated with DNA through a screen, such as a stainless steel or Nytex screen, onto a filter surface covered with corn cells cultured in suspension. Gordon-Kamm et al., describes the basic procedure for coating tungsten particles with DNA (Gordon-Kamm et al., Plant Cell 2:603-618 (1990), the entirety of which is herein incorporated by reference). The screen disperses the tungsten nucleic acid particles so that they are not delivered to the recipient cells in large aggregates. A particle delivery system suitable for use with the present invention is the helium acceleration PDS-1000/He gun is available from Bio-Rad Laboratories (Bio-Rad, Hercules, Calif.) (Sanford et al., Technique 3:3-16 (1991), the entirety of which is herein incorporated by reference).

For the bombardment, cells in suspension may be concentrated on filters. Filters containing the cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the gun and the cells to be bombarded.

Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the microprojectile stopping plate. If desired, one or more screens are also positioned between the acceleration device and the cells to be bombarded. Through the use of techniques set forth herein one may obtain up to 1000 or more foci of cells transiently expressing a marker gene. The number of cells in a focus which express the exogenous gene product 48 hours post-bombardment often range from one to ten and average one to three.

In bombardment transformation, one may optimize the pre-bombardment culturing conditions and the bombardment parameters to yield the maximum numbers of stable transformants. Both the physical and biological parameters for bombardment are important in this technology. Physical factors are those that involve manipulating the DNA/microprojectile precipitate or those that affect the flight and velocity of either the macro- or microprojectiles. Biological factors include all steps involved in manipulation of cells before and immediately after bombardment, the osmotic adjustment of target cells to help alleviate the trauma associated with bombardment and also the nature of the transforming DNA, such as linearized DNA or intact supercoiled plasmids. It is believed that pre-bombardment manipulations are especially important for successful transformation of immature embryos.

In another alternative embodiment, plastids can be stably transformed. Methods disclosed for plastid transformation in higher plants include the particle gun delivery of DNA containing a selectable marker and targeting of the DNA to the plastid genome through homologous recombination (Svab et al., Proc. Natl. Acad. Sci. (U.S.A) 87:8526-8530 (1990); Svab and Maliga, Proc. Natl. Acad. Sci. (U.S.A.) 90:913-917 (1993); Staub and Maliga, EMBO J. 12:601-606 (1993); U.S. Pat. Nos. 5,451,513 and 5,545,818, all of which are herein incorporated by reference in their entirety).

Accordingly, it is contemplated that one may wish to adjust various aspects of the bombardment parameters in small scale studies to fully optimize the conditions. One may particularly wish to adjust physical parameters such as gap distance, flight distance, tissue distance and helium pressure. One may also minimize the trauma reduction factors by modifying conditions which influence the physiological state of the recipient cells and which may therefore influence transformation and integration efficiencies. For example, the osmotic state, tissue hydration and the subculture stage or cell cycle of the recipient cells may be adjusted for optimum transformation. The execution of other routine adjustments will be known to those of skill in the art in light of the present disclosure.

Agrobacterium-mediated transfer is a widely applicable system for introducing genes into plant cells because the DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art. See, for example the methods described by Fraley et al., Bio/Technology 3:629-635 (1985) and Rogers et al., Methods Enzymol. 153:253-277 (1987), both of which are herein incorporated by reference in their entirety. Further, the integration of the Ti-DNA is a relatively precise process resulting in few rearrangements. The region of DNA to be transferred is defined by the border sequences and intervening DNA is usually inserted into the plant genome as described (Spielmann et al., Mol. Gen. Genet. 205:34 (1986), the entirety of which is herein incorporated by reference).

Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations as described (Klee et al., In: Plant DNA Infectious Agents, Hohn and Schell (eds.), Springer-Verlag, New York, pp. 179-203 (1985), the entirety of which is herein incorporated by reference. Moreover, technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes and are suitable for present purposes (Rogers et al., Methods Enzymol. 153:253-277 (1987)). In addition, Agrobacterium containing both armed and disarmed Ti genes can be used for the transformations. In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene transfer.

A transgenic plant formed using Agrobacterium transformation methods typically contains a single gene on one chromosome. Such transgenic plants can be referred to as being heterozygous for the added gene. More preferred is a transgenic plant that is homozygous for the added structural gene; i.e., a transgenic plant that contains two added genes, one gene at the same locus on each chromosome of a chromosome pair. A homozygous transgenic plant can be obtained by sexually mating (selfing) an independent segregant transgenic plant that contains a single added gene, germinating some of the seed produced and analyzing the resulting plants produced for the gene of interest.

It is also to be understood that two different transgenic plants can also be mated to produce offspring that contain two independently segregating added, exogenous genes. Selfing of appropriate progeny can produce plants that are homozygous for both added, exogenous genes that encode a polypeptide of interest. Back-crossing to a parental plant and out-crossing with a non-transgenic plant are also contemplated, as is vegetative propagation.

Transformation of plant protoplasts can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation and combinations of these treatments (See, for example, Potrykus et al., Mol. Gen. Genet. 205:193-200 (1986); Lorz et al., Mol. Gen. Genet. 199:178 (1985); Fromm et al., Nature 319:791 (1986); Uchimiya et al., Mol. Gen. Genet. 204:204 (1986); Marcotte et al., Nature 335:454-457 (1988), all of which are herein incorporated by reference in their entirety).

Application of these systems to different plant strains depends upon the ability to regenerate that particular plant strain from protoplasts. Illustrative methods for the regeneration of cereals from protoplasts are described (Fujimura et al., Plant Tissue Culture Letters 2:74 (1985); Toriyama et al., Theor Appl. Genet. 205:34 (1986); Yamada et al., Plant Cell Rep. 4:85 (1986); Abdullah et al., Biotechnology 4:1087 (1986), all of which are herein incorporated by reference in their entirety).

To transform plant strains that cannot be successfully regenerated from protoplasts, other ways to introduce DNA into intact cells or tissues can be utilized. For example, regeneration of cereals from immature embryos or explants can be effected as described (Vasil, Biotechnology 6:397 (1988), the entirety of which is herein incorporated by reference). In addition, “particle gun” or high-velocity microprojectile technology can be utilized (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference).

Using the latter technology, DNA is carried through the cell wall and into the cytoplasm on the surface of small metal particles as described (Klein et al., Nature 328:70 (1987); Klein et al., Proc. Natl. Acad. Sci. (U.S.A.) 85:8502-8505 (1988); McCabe et al., Bio/Technology 6:923 (1988), all of which are herein incorporated by reference in their entirety). The metal particles penetrate through several layers of cells and thus allow the transformation of cells within tissue explants.

Other methods of cell transformation can also be used and include but are not limited to introduction of DNA into plants by direct DNA transfer into pollen (Zhou et al., Methods Enzymol. 101:433 (1983); Hess et al., Intern Rev. Cytol. 107:367 (1987); Luo et al., Plant Mol. Biol. Reporter 6:165 (1988), all of which are herein incorporated by reference in their entirety), by direct injection of DNA into reproductive organs of a plant (Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference), or by direct injection of DNA into the cells of immature embryos followed by the rehydration of desiccated embryos (Neuhaus et al., Theor. Appl. Genet. 75:30 (1987), the entirety of which is herein incorporated by reference).

The regeneration, development and cultivation of plants from single plant protoplast transformants or from various transformed explants is well known in the art (Weissbach and Weissbach, In: Methods for Plant Molecular Biology, Academic Press, San Diego, Calif., (1988), the entirety of which is herein incorporated by reference). This regeneration and growth process typically includes the steps of selection of transformed cells, culturing those individualized cells through the usual stages of embryonic development through the rooted plantlet stage. Transgenic embryos and seeds are similarly regenerated. The resulting transgenic rooted shoots are thereafter planted in an appropriate plant growth medium such as soil.

The development or regeneration of plants containing the foreign, exogenous gene that encodes a protein of interest is well known in the art. Preferably, the regenerated plants are self-pollinated to provide homozygous transgenic plants. Otherwise, pollen obtained from the regenerated plants is crossed to seed-grown plants of agronomically important lines. Conversely, pollen from plants of these important lines is used to pollinate regenerated plants. A transgenic plant of the present invention containing a desired polypeptide is cultivated using methods well known to one skilled in the art.

There are a variety of methods for the regeneration of plants from plant tissue. The particular method of regeneration will depend on the starting plant tissue and the particular plant species to be regenerated.

Methods for transforming dicots, primarily by use of Agrobacterium tumefaciens and obtaining transgenic plants have been published for cotton (U.S. Pat. No. 5,004,863; U.S. Pat. No. 5,159,135; U.S. Pat. No. 5,518,908, all of which are herein incorporated by reference in their entirety); soybean (U.S. Pat. No. 5,569,834; U.S. Pat. No. 5,416,011; McCabe et. al., Biotechnology 6:923 (1988); Christou et al., Plant Physiol. 87:671-674 (1988); all of which are herein incorporated by reference in their entirety); Brassica (U.S. Pat. No. 5,463,174, the entirety of which is herein incorporated by reference); peanut (Cheng et al., Plant Cell Rep. 15:653-657 (1996), McKently et al., Plant Cell Rep. 14:699-703 (1995), all of which are herein incorporated by reference in their entirety); papaya; and pea (Grant et al., Plant Cell Rep. 15:254-258 (1995), the entirety of which is herein incorporated by reference).

Transformation of monocotyledons using electroporation, particle bombardment and Agrobacterium have also been reported. Transformation and plant regeneration have been achieved in asparagus (Bytebier et al., Proc. Natl. Acad. Sci. (USA) 84:5354 (1987), the entirety of which is herein incorporated by reference); barley (Wan and Lemaux, Plant Physiol 104:37 (1994), the entirety of which is herein incorporated by reference); maize (Rhodes et al., Science 240:204 (1988); Gordon-Kamm et al., Plant Cell 2:603-618 (1990); Fromm et al., Bio/Technology 8:833 (1990); Koziel et al., Bio/Technology 11:194 (1993); Armstrong et al., Crop Science 35:550-557 (1995); all of which are herein incorporated by reference in their entirety); oat (Somers et al., Bio/Technology 10:1589 (1992), the entirety of which is herein incorporated by reference); orchard grass (Horn et al., Plant Cell Rep. 7:469 (1988), the entirety of which is herein incorporated by reference); rice (Toriyama et al., Theor Appl. Genet. 205:34 (1986); Part et al., Plant Mol. Biol. 32:1135-1148 (1996); Abedinia et al., Aust. J. Plant Physiol. 24:133-141 (1997); Zhang and Wu, Theor. Appl. Genet. 76:835 (1988); Zhang et al., Plant Cell Rep. 7:379 (1988); Battraw and Hall, Plant Sci. 86:191-202 (1992); Christou et al., Bio/Technology 9:957 (1991), all of which are herein incorporated by reference in their entirety); rye (De la Pena et al., Nature 325:274 (1987), the entirety of which is herein incorporated by reference); sugarcane (Bower and Birch, Plant J 2:409 (1992), the entirety of which is herein incorporated by reference); tall fescue (Wang et al., Bio/Technology 10:691 (1992), the entirety of which is herein incorporated by reference) and wheat (Vasil et al., Bio/Technology 10:667 (1992), the entirety of which is herein incorporated by reference; U.S. Pat. No. 5,631,152, the entirety of which is herein incorporated by reference.)

Assays for gene expression based on the transient expression of cloned nucleic acid constructs have been developed by introducing the nucleic acid molecules into plant cells by polyethylene glycol treatment, electroporation, or particle bombardment (Marcotte et al., Nature 335:454-457 (1988), the entirety of which is herein incorporated by reference; Marcotte et al., Plant Cell 1:523-532 (1989), the entirety of which is herein incorporated by reference; McCarty et al., Cell 66:895-905 (1991), the entirety of which is herein incorporated by reference; Hattori et al., Genes Dev. 6:609-618 (1992), the entirety of which is herein incorporated by reference; Goff et al., EMBO J. 9:2517-2522 (1990), the entirety of which is herein incorporated by reference). Transient expression systems may be used to functionally dissect gene constructs (see generally, Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995)).

Any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a permanent or transient manner in combination with other genetic elements such as vectors, promoters, enhancers etc. Further, any of the nucleic acid molecules of the present invention may be introduced into a plant cell in a manner that allows for overexpression of the protein or fragment thereof encoded by the nucleic acid molecule.

Cosuppression is the reduction in expression levels, usually at the level of RNA, of a particular endogenous gene or gene family by the expression of a homologous sense construct that is capable of transcribing mRNA of the same strandedness as the transcript of the endogenous gene (Napoli et al., Plant Cell 2:279-289 (1990), the entirety of which is herein incorporated by reference; van der Krol et al., Plant Cell 2:291-299 (1990), the entirety of which is herein incorporated by reference). Cosuppression may result from stable transformation with a single copy nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Prolls and Meyer, Plant J. 2:465-475 (1992), the entirety of which is herein incorporated by reference) or with multiple copies of a nucleic acid molecule that is homologous to a nucleic acid sequence found with the cell (Mittlesten et al., Mol. Gen. Genet. 244:325-330 (1994), the entirety of which is herein incorporated by reference). Genes, even though different, linked to homologous promoters may result in the cosuppression of the linked genes (Vaucheret, C.R. Acad. Sci. III 316:1471-1483 (1993), the entirety of which is herein incorporated by reference).

This technique has, for example, been applied to generate white flowers from red petunia and tomatoes that do not ripen on the vine. Up to 50% of petunia transformants that contained a sense copy of the glucoamylase (CHS) gene produced white flowers or floral sectors; this was as a result of the post-transcriptional loss of mRNA encoding CHS (Flavell, Proc. Natl. Acad. Sci. (U.S.A) 91:3490-3496 (1994), the entirety of which is herein incorporated by reference); van Blokland et al., Plant J. 6:861-877 (1994), the entirety of which is herein incorporated by reference). Cosuppression may require the coordinate transcription of the transgene and the endogenous gene and can be reset by a developmental control mechanism (Jorgensen, Trends Biotechnol. 8:340-344 (1990), the entirety of which is herein incorporated by reference; Meins and Kunz, In: Gene Inactivation and Homologous Recombination in Plants, Paszkowski (ed.), pp. 335-348, Kluwer Academic, Netherlands (1994), the entirety of which is herein incorporated by reference).

It is understood that one or more of the nucleic acids of the present invention may be introduced into a plant cell and transcribed using an appropriate promoter with such transcription resulting in the cosuppression of an endogenous sucrose pathway protein.

Antisense approaches are a way of preventing or reducing gene function by targeting the genetic material (Mol et al., FEBS Lett. 268:427-430 (1990), the entirety of which is herein incorporated by reference). The objective of the antisense approach is to use a sequence complementary to the target gene to block its expression and create a mutant cell line or organism in which the level of a single chosen protein is selectively reduced or abolished. Antisense techniques have several advantages over other ‘reverse genetic’ approaches. The site of inactivation and its developmental effect can be manipulated by the choice of promoter for antisense genes or by the timing of external application or microinjection. Antisense can manipulate its specificity by selecting either unique regions of the target gene or regions where it shares homology to other related genes (Hiatt et al., In: Genetic Engineering, Setlow (ed.), Vol. 11, New York: Plenum 49-63 (1989), the entirety of which is herein incorporated by reference).

The principle of regulation by antisense RNA is that RNA that is complementary to the target mRNA is introduced into cells, resulting in specific RNA:RNA duplexes being formed by base pairing between the antisense substrate and the target mRNA (Green et al., Annu. Rev. Biochem. 55:569-597 (1986), the entirety of which is herein incorporated by reference). Under one embodiment, the process involves the introduction and expression of an antisense gene sequence. Such a sequence is one in which part or all of the normal gene sequences are placed under a promoter in inverted orientation so that the ‘wrong’ or complementary strand is transcribed into a noncoding antisense RNA that hybridizes with the target mRNA and interferes with its expression (Takayama and Inouye, Crit. Rev. Biochem. Mol. Biol. 25:155-184 (1990), the entirety of which is herein incorporated by reference). An antisense vector is constructed by standard procedures and introduced into cells by transformation, transfection, electroporation, microinjection, infection, etc. The type of transformation and choice of vector will determine whether expression is transient or stable. The promoter used for the antisense gene may influence the level, timing, tissue, specificity, or inducibility of the antisense inhibition.

It is understood that the activity of a sucrose pathway protein in a plant cell may be reduced or depressed by growing a transformed plant cell containing a nucleic acid molecule whose non-transcribed strand encodes a sucrose pathway protein or fragment thereof.

Antibodies have been expressed in plants (Hiatt et al., Nature 342:76-78 (1989), the entirety of which is herein incorporated by reference; Conrad and Fielder, Plant Mol. Biol. 26:1023-1030 (1994), the entirety of which is herein incorporated by reference). Cytoplamsic expression of a scFv (single-chain Fv antibodies) has been reported to delay infection by artichoke mottled crinkle virus. Transgenic plants that express antibodies directed against endogenous proteins may exhibit a physiological effect (Philips et al., EMBO J. 16:4489-4496 (1997), the entirety of which is herein incorporated by reference; Marion-Poll, Trends in Plant Science 2:447-448 (1997), the entirety of which is herein incorporated by reference). For example, expressed anti-abscisic antibodies have been reported to result in a general perturbation of seed development (Philips et al., EMBO J. 16: 4489-4496 (1997)).

Antibodies that are catalytic may also be expressed in plants (abzymes). The principle behind abzymes is that since antibodies may be raised against many molecules, this recognition ability can be directed toward generating antibodies that bind transition states to force a chemical reaction forward (Persidas, Nature Biotechnology 15:1313-1315 (1997), the entirety of which is herein incorporated by reference; Baca et al., Ann. Rev. Biophys. Biomol. Struct. 26:461-493 (1997), the entirety of which is herein incorporated by reference). The catalytic abilities of abzymes may be enhanced by site directed mutagenesis. Examples of abzymes are, for example, set forth in U.S. Pat. No. 5,658,753; U.S. Pat. No. 5,632,990; U.S. Pat. No. 5,631,137; U.S. Pat. No. 5,602,015; U.S. Pat. No. 5,559,538; U.S. Pat. No. 5,576,174; U.S. Pat. No. 5,500,358; U.S. Pat. No. 5,318,897; U.S. Pat. No. 5,298,409; U.S. Pat. No. 5,258,289 and U.S. Pat. No. 5,194,585, all of which are herein incorporated in their entirety.

It is understood that any of the antibodies of the present invention may be expressed in plants and that such expression can result in a physiological effect. It is also understood that any of the expressed antibodies may be catalytic.

(b) Fungal Constructs and Fungal Transformants

The present invention also relates to a fungal recombinant vector comprising exogenous genetic material. The present invention also relates to a fungal cell comprising a fungal recombinant vector. The present invention also relates to methods for obtaining a recombinant fungal host cell comprising introducing into a fungal host cell exogenous genetic material.

Exogenous genetic material may be transferred into a fungal cell. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention. The fungal recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the fungal host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the fungal host.

The fungal vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the fungal cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the fungal host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the fungal host cell and, furthermore, may be non-encoding or encoding sequences.

For autonomous replication, the vector may further comprise an origin of replication enabling the vector to replicate autonomously in the host cell in question. Examples of origin of replications for use in a yeast host cell are the 2 micron origin of replication and the combination of CEN3 and ARS1. Any origin of replication may be used which is compatible with the fungal host cell of choice.

The fungal vectors of the present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. The selectable marker may be selected from the group including, but not limited to, amdS (acetamidase), argB (ornithine carbamoyltransferase), bar (phosphinothricin acetyltransferase), hygB (hygromycin phosphotransferase), niaD (nitrate reductase), pyrG (orotidine-5′-phosphate decarboxylase) and sC (sulfate adenyltransferase) and trpC (anthranilate synthase). Preferred for use in an Aspergillus cell are the amdS and pyrG markers of Aspergillus nidulans or Aspergillus oryzae and the bar marker of Streptomyces hygroscopicus. Furthermore, selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, the entirety of which is herein incorporated by reference. A nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the fungal host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof.

A promoter may be any nucleic acid sequence which shows transcriptional activity in the fungal host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell. Examples of suitable promoters for directing the transcription of a nucleic acid construct of the invention in a filamentous fungal host are promoters obtained from the genes encoding Aspergillus oryzae TAKA amylase, Rhizomucor miehei aspartic proteinase, Aspergillus niger neutral alpha-amylase, Aspergillus niger acid stable alpha-amylase, Aspergillus niger or Aspergillus awamori glucoamylase (glaA), Rhizomucor miehei lipase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Aspergillus nidulans acetamidase and hybrids thereof. In a yeast host, a useful promoter is the Saccharomyces cerevisiae enolase (eno-1) promoter. Particularly preferred promoters are the TAKA amylase, NA2-tpi (a hybrid of the promoters from the genes encoding Aspergillus niger neutral alpha-amylase and Aspergillus oryzae triose phosphate isomerase) and glaA promoters.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a terminator sequence at its 3′ terminus. The terminator sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any terminator which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred terminators are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase, Aspergillus niger alpha-glucosidase and Saccharomyces cerevisiae enolase.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5′ terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the fungal host cell of choice may be used in the present invention, but particularly preferred leaders are obtained from the genes encoding Aspergillus oryzae TAKA amylase and Aspergillus oryzae triose phosphate isomerase.

A polyadenylation sequence may also be operably linked to the 3′ terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the fungal host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention, but particularly preferred polyadenylation sequences are obtained from the genes encoding Aspergillus oryzae TAKA amylase, Aspergillus niger glucoamylase, Aspergillus nidulans anthranilate synthase and Aspergillus niger alpha-glucosidase.

To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed protein or fragment thereof within the cell, it is preferred that expression of the protein or fragment thereof gives rise to a product secreted outside the cell. To this end, a protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the fungal host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5′ end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof. Alternatively, the 5′ end of the coding sequence may contain a signal peptide coding region which is foreign to that portion of the coding sequence which encodes the secreted protein or fragment thereof. The foreign signal peptide may be required where the coding sequence does not normally contain a signal peptide coding region. Alternatively, the foreign signal peptide may simply replace the natural signal peptide to obtain enhanced secretion of the desired protein or fragment thereof. The foreign signal peptide coding region may be obtained from a glucoamylase or an amylase gene from an Aspergillus species, a lipase or proteinase gene from Rhizomucor miehei, the gene for the alpha-factor from Saccharomyces cerevisiae, or the calf preprochymosin gene. An effective signal peptide for fungal host cells is the Aspergillus oryzae TAKA amylase signal, Aspergillus niger neutral amylase signal, the Rhizomucor miehei aspartic proteinase signal, the Humicola lanuginosus cellulase signal, or the Rhizomucor miehei lipase signal. However, any signal peptide capable of permitting secretion of the protein or fragment thereof in a fungal host of choice may be used in the present invention.

A protein or fragment thereof encoding nucleic acid molecule of the present invention may also be linked to a propeptide coding region. A propeptide is an amino acid sequence found at the amino terminus of aproprotein or proenzyme. Cleavage of the propeptide from the proprotein yields a mature biochemically active protein. The resulting polypeptide is known as a propolypeptide or proenzyme (or a zymogen in some cases). Propolypeptides are generally inactive and can be converted to mature active polypeptides by catalytic or autocatalytic cleavage of the propeptide from the propolypeptide or proenzyme. The propeptide coding region may be native to the protein or fragment thereof or may be obtained from foreign sources. The foreign propeptide coding region may be obtained from the Saccharomyces cerevisiae alpha-factor gene or Myceliophthora thermophila laccase gene (WO 95/33836, the entirety of which is herein incorporated by reference).

The procedures used to ligate the elements described above to construct the recombinant expression vector of the present invention are well known to one skilled in the art (see, for example, Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd ed., Cold Spring Harbor, N.Y., (1989)).

The present invention also relates to recombinant fungal host cells produced by the methods of the present invention which are advantageously used with the recombinant vector of the present invention. The cell is preferably transformed with a vector comprising a nucleic acid sequence of the invention followed by integration of the vector into the host chromosome. The choice of fungal host cells will to a large extent depend upon the gene encoding the protein or fragment thereof and its source. The fuingal host cell may, for example, be a yeast cell or a filamentous fungal cell.

“Yeast” as used herein includes Ascosporogenous yeast (Endomycetales), Basidiosporogenous yeast and yeast belonging to the Fungi Imperfecti (Blastomycetes). The Ascosporogenous yeasts are divided into the families Spermophthoraceae and Saccharomycetaceae. The latter is comprised of four subfamilies, Schizosaccharomycoideae (for example, genus Schizosaccharomyces), Nadsonioideae, Lipomycoideae and Saccharomycoideae (for example, genera Pichia, Kluyveromyces and Saccharomyces). The Basidiosporogenous yeasts include the genera Leucosporidim, Rhodosporidium, Sporidiobolus, Filobasidium and Filobasidiella. Yeast belonging to the Fungi Imperfecti are divided into two families, Sporobolomycetaceae (for example, genera Sorobolomyces and Bullera) and Cryptococcaceae (for example, genus Candida). Since the classification of yeast may change in the future, for the purposes of this invention, yeast shall be defined as described in Biology and Activities of Yeast (Skinner et al., Soc. App. Bacteriol. Symposium Series No. 9, (1980), the entirety of which is herein incorporated by reference). The biology of yeast and manipulation of yeast genetics are well known in the art (see, for example, Biochemistry and Genetics of Yeast, Bacil et al. (ed.), 2nd edition, 1987; The Yeasts, Rose and Harrison (eds.), 2nd ed., (1987); and The Molecular Biology of the Yeast Saccharomyces, Strathern et al. (eds.), (1981), all of which are herein incorporated by reference in their entirety).

“Fungi” as used herein includes the phyla Ascomycota, Basidiomycota, Chytridiomycota and Zygomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK; the entirety of which is herein incorporated by reference) as well as the Oomycota (as cited in Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK) and all mitosporic fungi (Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). Representative groups of Ascomycota include, for example, Neurospora, Eupenicillium (=Penicillium), Emericella (=Aspergillus), Eurotiun (=Aspergillus) and the true yeasts listed above. Examples of Basidiomycota include mushrooms, rusts and smuts. Representative groups of Chytridiomycota include, for example, Allomyces, Blastocladiella, Coelomomyces and aquatic fungi. Representative groups of Oomycota include, for example, Saprolegniomycetous aquatic fungi (water molds) such as Achlya. Examples of mitosporic fungi include Aspergillus, Penicilliun, Candida and Alternaria. Representative groups of Zygomycota include, for example, Rhizopus and Mucor.

“Filamentous fungi” include all filamentous forms of the subdivision Eumycota and Oomycota (as defined by Hawksworth et al., In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK). The filamentous fungi are characterized by a vegetative mycelium composed of chitin, cellulose, glucan, chitosan, mannan and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon catabolism is obligately aerobic. In contrast, vegetative growth by yeasts such as Saccharomyces cerevisiae is by budding of a unicellular thallus and carbon catabolism may be fermentative.

In one embodiment, the fungal host cell is a yeast cell. In a preferred embodiment, the yeast host cell is a cell of the species of Candida, Kluyveromyces, Saccharomyces, Schizosaccharomyces, Pichia and Yarrowia. In a preferred embodiment, the yeast host cell is a Saccharomyces cerevisiae cell, a Saccharomyces carlsbergensis, Saccharomyces diastaticus cell, a Saccharomyces douglasii cell, a Saccharomyces kluyveri cell, a Saccharomyces norbensis cell, or a Saccharomyces oviformis cell. In another preferred embodiment, the yeast host cell is a Kluyveromyces lactis cell. In another preferred embodiment, the yeast host cell is a Yarrowia lipolytica cell.

In another embodiment, the fungal host cell is a filamentous fungal cell. In a preferred embodiment, the filamentous fungal host cell is a cell of the species of, but not limited to, Acremonium, Aspergillus, Fusarium, Humicola, Myceliophthora, Mucor, Neurospora, Penicillium, Thielavia, Tolypocladium and Trichoderma. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus cell. In another preferred embodiment, the filamentous fungal host cell is an Acremonium cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora cell. In another even preferred embodiment, the filamentous fungal host cell is a Mucor cell. In another preferred embodiment, the filamentous fungal host cell is a Neurospora cell. In another preferred embodiment, the filamentous fungal host cell is a Penicillium cell. In another preferred embodiment, the filamentous fungal host cell is a Thielavia cell. In another preferred embodiment, the filamentous fungal host cell is a Tolypocladiun cell. In another preferred embodiment, the filamentous fungal host cell is a Trichoderma cell. In a preferred embodiment, the filamentous fungal host cell is an Aspergillus oryzae cell, an Aspergillus niger cell, an Aspergillus foetidus cell, or an Aspergillus japonicus cell. In another preferred embodiment, the filamentous fungal host cell is a Fusarium oxysporum cell or a Fusarium graminearum cell. In another preferred embodiment, the filamentous fungal host cell is a Humicola insolens cell or a Humicola lanuginosus cell. In another preferred embodiment, the filamentous fungal host cell is a Myceliophthora thermophila cell. In a most preferred embodiment, the filamentous fungal host cell is a Mucor miehei cell. In a most preferred embodiment, the filamentous fungal host cell is a Neurospora crassa cell. In a most preferred embodiment, the filamentous fungal host cell is a Penicillium purpurogenum cell. In another most preferred embodiment, the filamentous fungal host cell is a Thielavia terrestris cell. In another most preferred embodiment, the Trichoderma cell is a Trichoderma reesei cell, a Trichoderma viride cell, a Trichoderma longibrachiatum cell, a Trichoderma harzianum cell, or a Trichoderma koningii cell. In a preferred embodiment, the fungal host cell is selected from an A. nidulans cell, an A. niger cell, an A. oryzae cell and an A. sojae cell. In a further preferred embodiment, the fungal host cell is an A. nidulans cell.

The recombinant fungal host cells of the present invention may further comprise one or more sequences which encode one or more factors that are advantageous in the expression of the protein or fragment thereof, for example, an activator (e.g., a trans-acting factor), a chaperone and a processing protease. The nucleic acids encoding one or more of these factors are preferably not operably linked to the nucleic acid encoding the protein or fragment thereof. An activator is a protein which activates transcription of a nucleic acid sequence encoding a polypeptide (Kudla et al., EMBO 9:1355-1364 (1990); Jarai and Buxton, Current Genetics 26:2238-244 (1994); Verdier, Yeast 6:271-297 (1990), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding an activator may be obtained from the genes encoding Saccharomyces cerevisiae heme activator protein 1 (hap1), Saccharomyces cerevisiae galactose metabolizing protein 4 (gal4) and Aspergillus nidulans ammonia regulation protein (areA). For further examples, see Verdier, Yeast 6:271-297 (1990); MacKenzie et al., Journal of Gen. Microbiol. 139:2295-2307 (1993), both of which are herein incorporated by reference in their entirety). A chaperone is a protein which assists another protein in folding properly (Hartl et al., TIBS 19:20-25 (1994); Bergeron et al., TIBS 19:124-128 (1994); Demolder et al., J Biotechnology 32:179-189 (1994); Craig, Science 260:1902-1903 (1993); Gething and Sambrook, Nature 355:33-45 (1992); Puig and Gilbert, J. Biol. Chem. 269:7764-7771 (1994); Wang and Tsou, FASEB Journal 7:1515-11157 (1993); Robinson et al., Bio/Technology 1:381-384 (1994), all of which are herein incorporated by reference in their entirety). The nucleic acid sequence encoding a chaperone may be obtained from the genes encoding Aspergillus oryzae protein disulphide isomerase, Saccharomyces cerevisiae calnexin, Saccharomyces cerevisiae BiP/GRP78 and Saccharomyces cerevisiae Hsp70. For further examples, see Gething and Sambrook, Nature 355:33-45 (1992); Hartl et al., TIBS 19:20-25 (1994). A processing protease is a protease that cleaves a propeptide to generate a mature biochemically active polypeptide (Enderlin and Ogrydziak, Yeast 10:67-79 (1994); Fuller et al., Proc. Natl. Acad. Sci. (U.S.A.) 86:1434-1438 (1989); Julius et al., Cell 37:1075-1089 (1984); Julius et al., Cell 32:839-852 (1983), all of which are incorporated by reference in their entirety). The nucleic acid sequence encoding a processing protease may be obtained from the genes encoding Aspergillus niger Kex2, Saccharomyces cerevisiae dipeptidylaminopeptidase, Saccharomyces cerevisiae Kex2 and Yarrowia lipolytica dibasic processing endoprotease (xpr6). Any factor that is functional in the fungal host cell of choice may be used in the present invention.

Fungal cells may be transformed by a process involving protoplast formation, transformation of the protoplasts and regeneration of the cell wall in a manner known per se. Suitable procedures for transformation of Aspergillus host cells are described in EP 238 023 and Yelton et al., Proc. Natl. Acad. Sci. (U.S.A.) 81:1470-1474 (1984), both of which are herein incorporated by reference in their entirety. A suitable method of transforming Fusarium species is described by Malardier et al., Gene 78:147-156 (1989), the entirety of which is herein incorporated by reference. Yeast may be transformed using the procedures described by Becker and Guarente, In: Abelson and Simon, (eds.), Guide to Yeast Genetics and Molecular Biology, Methods Enzymol. Volume 194, pp 182-187, Academic Press, Inc., New York; Ito et al., J Bacteriology 153:163 (1983); Hinnen et al., Proc. Natl. Acad. Sci. (U.S.A.) 75:1920 (1978), all of which are herein incorporated by reference in their entirety.

The present invention also relates to methods of producing the protein or fragment thereof comprising culturing the recombinant fungal host cells under conditions conducive for expression of the protein or fragment thereof. The fungal cells of the present invention are cultivated in a nutrient medium suitable for production of the protein or fragment thereof using methods known in the art. For example, the cell may be cultivated by shake flask cultivation, small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermentors performed in a suitable medium and under conditions allowing the protein or fragment thereof to be expressed and/or isolated. The cultivation takes place in a suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art (see, e.g., Bennett and LaSure (eds.), More Gene Manipulations in Fungi, Academic Press, CA, (1991), the entirety of which is herein incorporated by reference). Suitable media are available from commercial suppliers or may be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection, Manassas, Va.). If the protein or fragment thereof is secreted into the nutrient medium, a protein or fragment thereof can be recovered directly from the medium. If the protein or fragment thereof is not secreted, it is recovered from cell lysates.

The expressed protein or fragment thereof may be detected using methods known in the art that are specific for the particular protein or fragment. These detection methods may include the use of specific antibodies, formation of an enzyme product, or disappearance of an enzyme substrate. For example, if the protein or fragment thereof has enzymatic activity, an enzyme assay may be used. Alternatively, if polyclonal or monoclonal antibodies specific to the protein or fragment thereof are available, immunoassays may be employed using the antibodies to the protein or fragment thereof. The techniques of enzyme assay and immunoassay are well known to those skilled in the art.

The resulting protein or fragment thereof may be recovered by methods known in the arts. For example, the protein or fragment thereof may be recovered from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation. The recovered protein or fragment thereof may then be further purified by a variety of chromatographic procedures, e.g., ion exchange chromatography, gel filtration chromatography, affinity chromatography, or the like.

(c) Mammalian Constructs and Transformed Mammalian Cells

The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian host cell exogenous genetic material. The present invention also relates to a mammalian cell comprising a mammalian recombinant vector. The present invention also relates to methods for obtaining a recombinant mammalian host cell, comprising introducing into a mammalian cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

Mammalian cell lines available as hosts for expression are known in the art and include many immortalized cell lines available from the American Type Culture Collection (ATCC, Manassas, Va.), such as HeLa cells, Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK) cells and a number of other cell lines. Suitable promoters for mammalian cells are also known in the art and include viral promoters such as that from Simian Virus 40 (SV40) (Fiers et al., Nature 273:113 (1978), the entirety of which is herein incorporated by reference), Rous sarcoma virus (RSV), adenovirus (ADV) and bovine papilloma virus (BPV). Mammalian cells may also require terminator sequences and poly-A addition sequences. Enhancer sequences which increase expression may also be included and sequences which promote amplification of the gene may also be desirable (for example methotrexate resistance genes).

Vectors suitable for replication in mammalian cells may include viral replicons, or sequences which insure integration of the appropriate sequences encoding HCV epitopes into the host genome. For example, another vector used to express foreign DNA is vaccinia virus. In this case, for example, a nucleic acid molecule encoding a protein or fragment thereof is inserted into the vaccinia genome. Techniques for the insertion of foreign DNA into the vaccinia virus genome are known in the art and may utilize, for example, homologous recombination. Such heterologous DNA is generally inserted into a gene which is non-essential to the virus, for example, the thymidine kinase gene (tk), which also provides a selectable marker. Plasmid vectors that greatly facilitate the construction of recombinant viruses have been described (see, for example, Mackett et al, J. Virol. 49:857 (1984); Chakrabarti et al., Mol. Cell. Biol. 5:3403 (1985); Moss, In: Gene Transfer Vectors For Mammalian Cells (Miller and Calos, eds., Cold Spring Harbor Laboratory, N.Y., p. 10, (1987); all of which are herein incorporated by reference in their entirety). Expression of the HCV polypeptide then occurs in cells or animals which are infected with the live recombinant vaccinia virus.

The sequence to be integrated into the mammalian sequence may be introduced into the primary host by any convenient means, which includes calcium precipitated DNA, spheroplast fusion, transformation, electroporation, biolistics, lipofection, microinjection, or other convenient means. Where an amplifiable gene is being employed, the amplifiable gene may serve as the selection marker for selecting hosts into which the amplifiable gene has been introduced. Alternatively, one may include with the amplifiable gene another marker, such as a drug resistance marker, e.g. neomycin resistance (G418 in mammalian cells), hygromycin in resistance etc., or an auxotrophy marker (HIS3, TRP1, LEU2, URA3, ADE2, LYS2, etc.) for use in yeast cells.

Depending upon the nature of the modification and associated targeting construct, various techniques may be employed for identifying targeted integration. Conveniently, the DNA may be digested with one or more restriction enzymes and the fragments probed with an appropriate DNA fragment which will identify the properly sized restriction fragment associated with integration.

One may use different promoter sequences, enhancer sequences, or other sequence which will allow for enhanced levels of expression in the expression host. Thus, one may combine an enhancer from one source, a promoter region from another source, a 5′-noncoding region upstream from the initiation sucrose from the same or different source as the other sequences and the like. One may provide for an intron in the non-coding region with appropriate splice sites or for an alternative 3′-untranslated sequence or polyadenylation site. Depending upon the particular purpose of the modification, any of these sequences may be introduced, as desired.

Where selection is intended, the sequence to be integrated will have with it a marker gene, which allows for selection. The marker gene may conveniently be downstream from the target gene and may include resistance to a cytotoxic agent, e.g. antibiotics, heavy metals, or the like, resistance or susceptibility to HAT, gancyclovir, etc., complementation to an auxotrophic host, particularly by using an auxotrophic yeast as the host for the subject manipulations, or the like. The marker gene may also be on a separate DNA molecule, particularly with primary mammalian cells. Alternatively, one may screen the various transformants, due to the high efficiency of recombination in yeast, by using hybridization analysis, PCR, sequencing, or the like.

For homologous recombination, constructs can be prepared where the amplifiable gene will be flanked, normally on both sides with DNA homologous with the DNA of the target region. Depending upon the nature of the integrating DNA and the purpose of the integration, the homologous DNA will generally be within 100 kb, usually 50 kb, preferably about 25 kb, of the transcribed region of the target gene, more preferably within 2 kb of the target gene. Where modeling of the gene is intended, homology will usually be present proximal to the site of the mutation. The homologous DNA may include the 5′-upstream region outside of the transcriptional regulatory region or comprising any enhancer sequences, transcriptional initiation sequences, adjacent sequences, or the like. The homologous region may include a portion of the coding region, where the coding region may be comprised only of an open reading frame or combination of exons and introns. The homologous region may comprise all or a portion of an intron, where all or a portion of one or more exons may also be present. Alternatively, the homologous region may comprise the 3′-region, so as to comprise all or a portion of the transcriptional termination region, or the region 3′ of this region. The homologous regions may extend over all or a portion of the target gene or be outside the target gene comprising all or a portion of the transcriptional regulatory regions and/or the structural gene.

The integrating constructs may be prepared in accordance with conventional ways, where sequences may be synthesized, isolated from natural sources, manipulated, cloned, ligated, subjected to in vitro mutagenesis, primer repair, or the like. At various stages, the joined sequences may be cloned and analyzed by restriction analysis, sequencing, or the like. Usually during the preparation of a construct where various fragments are joined, the fragments, intermediate constructs and constructs will be carried on a cloning vector comprising a replication system functional in a prokaryotic host, e.g., E. Coli and a marker for selection, e.g., biocide resistance, complementation to an auxotrophic host, etc. Other functional sequences may also be present, such as polylinkers, for ease of introduction and excision of the construct or portions thereof, or the like. A large number of cloning vectors are available such as pBR322, the pUC series, etc. These constructs may then be used for integration into the primary mammalian host.

In the case of the primary mammalian host, a replicating vector may be used. Usually, such vector will have a viral replication system, such as SV40, bovine papilloma virus, adenovirus, or the like. The linear DNA sequence vector may also have a selectable marker for identifying transfected cells. Selectable markers include the neo gene, allowing for selection with G418, the herpes tk gene for selection with HAT medium, the gpt gene with mycophenolic acid, complementation of an auxotrophic host, etc.

The vector may or may not be capable of stable maintenance in the host. Where the vector is capable of stable maintenance, the cells will be screened for homologous integration of the vector into the genome of the host, where various techniques for curing the cells may be employed. Where the vector is not capable of stable maintenance, for example, where a temperature sensitive replication system is employed, one may change the temperature from the permissive temperature to the non-permissive temperature, so that the cells may be cured of the vector. In this case, only those cells having integration of the construct comprising the amplifiable gene and, when present, the selectable marker, will be able to survive selection.

Where a selectable marker is present, one may select for the presence of the targeting construct by means of the selectable marker. Where the selectable marker is not present, one may select for the presence of the construct by the amplifiable gene. For the neo gene or the herpes tk gene, one could employ a medium for growth of the transformants of about 0.1-1 mg/ml of G418 or may use HAT medium, respectively. Where DHFR is the amplifiable gene, the selective medium may include from about 0.01-0.5 μM of methotrexate or be deficient in glycine-hypoxanthine-thymidine and have dialysed serum (GHT media).

The DNA can be introduced into the expression host by a variety of techniques that include calcium phosphate/DNA co-precipitates, microinjection of DNA into the nucleus, electroporation, yeast protoplast fusion with intact cells, transfection, polycations, e.g., polybrene, polyornithine, etc., or the like. The DNA may be single or double stranded DNA, linear or circular. The various techniques for transforming mammalian cells are well known (see Keown et al., Methods Enzymol. (1989); Keown et al., Methods Enzymol. 185:527-537 (1990); Mansour et al., Nature 336:348-352, (1988); all of which are herein incorporated by reference in their entirety).

(d) Insect Constructs and Transformed Insect Cells

The present invention also relates to an insect recombinant vectors comprising exogenous genetic material. The present invention also relates to an insect cell comprising an insect recombinant vector. The present invention also relates to methods for obtaining a recombinant insect host cell, comprising introducing into an insect cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

The insect recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures and can bring about the expression of the nucleic acid sequence. The choice of a vector will typically depend on the compatibility of the vector with the insect host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the insect host. In addition, the insect vector may be an expression vector. Nucleic acid molecules can be suitably inserted into a replication vector for expression in the insect cell under a suitable promoter for insect cells. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid molecule to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for insect cell transformation generally include, but are not limited to, one or more of the following: a signal sequence, origin of replication, one or more marker genes and an inducible promoter.

The insect vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extrachromosomal element, a minichromosome, or an artificial chromosome. The vector may contain any means for assuring self-replication. Alternatively, the vector may be one which, when introduced into the insect cell, is integrated into the genome and replicated together with the chromosome(s) into which it has been integrated. For integration, the vector may rely on the nucleic acid sequence of the vector for stable integration of the vector into the genome by homologous or nonhomologous recombination. Alternatively, the vector may contain additional nucleic acid sequences for directing integration by homologous recombination into the genome of the insect host. The additional nucleic acid sequences enable the vector to be integrated into the host cell genome at a precise location(s) in the chromosome(s). To increase the likelihood of integration at a precise location, there should be preferably two nucleic acid sequences which individually contain a sufficient number of nucleic acids, preferably 400 bp to 1500 bp, more preferably 800 bp to 1000 bp, which are highly homologous with the corresponding target sequence to enhance the probability of homologous recombination. These nucleic acid sequences may be any sequence that is homologous with a target sequence in the genome of the insect host cell and, furthermore, may be non-encoding or encoding sequences.

Baculovirus expression vectors (BEVs) have become important tools for the expression of foreign genes, both for basic research and for the production of proteins with direct clinical applications in human and veterinary medicine (Doerfler, Curr. Top. Microbiol. Immunol. 131:51-68 (1968); Luckow and Summers, Bio/Technology 6:47-55 (1988a); Miller, Annual Review of Microbiol. 42:177-199 (1988); Summers, Curr. Comm. Molecular Biology, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1988); all of which are herein incorporated by reference in their entirety). BEVs are recombinant insect viruses in which the coding sequence for a chosen foreign gene has been inserted behind a baculovirus promoter in place of the viral gene, e.g., polyhedrin (Smith and Summers, U.S. Pat. No. 4,745,051, the entirety of which is incorporated herein by reference).

The use of baculovirus vectors relies upon the host cells being derived from Lepidopteran insects such as Spodoptera frugiperda or Trichoplusia ni. The preferred Spodoptera frugiperda cell line is the cell line Sf9. The Spodoptera frugiperda Sf9 cell line was obtained from American Type Culture Collection (Manassas, Va.) and is assigned accession number ATCC CRL 1711 (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Ag. Exper. Station Bulletin No. 1555 (1988), the entirety of which is herein incorporated by reference). Other insect cell systems, such as the silkworm B. mori may also be used.

The proteins expressed by the BEVs are, therefore, synthesized, modified and transported in host cells derived from Lepidopteran insects. Most of the genes that have been inserted and produced in the baculovirus expression vector system have been derived from vertebrate species. Other baculovirus genes in addition to the polyhedrin promoter may be employed to advantage in a baculovirus expression system. These include immediate-early (alpha), delayed-early (β), late (γ), or very late (delta), according to the phase of the viral infection during which they are expressed. The expression of these genes occurs sequentially, probably as the result of a “cascade” mechanism of transcriptional regulation. (Guarino and Summers, J. Virol. 57:563-571 (1986); Guarino and Summers, J. Virol. 61:2091-2099 (1987); Guarino and Summers, Virol. 162:444-451 (1988); all of which are herein incorporated by reference in their entirety).

Insect recombinant vectors are useful as intermediates for the infection or transformation of insect cell systems. For example, an insect recombinant vector containing a nucleic acid molecule encoding a baculovirus transcriptional promoter followed downstream by an insect signal DNA sequence is capable of directing the secretion of the desired biologically active protein from the insect cell. The vector may utilize a baculovirus transcriptional promoter region derived from any of the over 500 baculoviruses generally infecting insects, such as for example the Orders Lepidoptera, Diptera, Orthoptera, Coleoptera and Hymenoptera, including for example but not limited to the viral DNAs of Autographa californica MNPV, Bombyx mori NPV, Trichoplusia ni MNPV, Rachiplusia ou MNPV or Galleria mellonella MNPV, wherein said baculovirus transcriptional promoter is a baculovirus immediate-early gene IE1 or IEN promoter; an immediate-early gene in combination with a baculovirus delayed-early gene promoter region selected from the group consisting of 39K and a HindIII-k fragment delayed-early gene; or a baculovirus late gene promoter. The immediate-early or delayed-early promoters can be enhanced with transcriptional enhancer elements. The insect signal DNA sequence may code for a signal peptide of a Lepidopteran adipokinetic hormone precursor or a signal peptide of the Manduca sexta adipokinetic hormone precursor (Summers, U.S. Pat. No. 5,155,037; the entirety of which is herein incorporated by reference). Other insect signal DNA sequences include a signal peptide of the Orthoptera Schistocerca gregaria locust adipokinetic hormone precurser and the Drosophila melanogaster cuticle genes CP1, CP2, CP3 or CP4 or for an insect signal peptide having substantially a similar chemical composition and function (Summers, U.S. Pat. No. 5,155,037).

Insect cells are distinctly different from animal cells. Insects have a unique life cycle and have distinct cellular properties such as the lack of intracellular plasminogen activators in which are present in vertebrate cells. Another difference is the high expression levels of protein products ranging from 1 to greater than 500 mg/liter and the ease at which cDNA can be cloned into cells (Frasier, In Vitro Cell. Dev. Biol. 25:225 (1989); Summers and Smith, In: A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures; Texas Ag. Exper. Station Bulletin No. 1555 (1988), both of which are incorporated by reference in their entirety).

Recombinant protein expression in insect cells is achieved by viral infection or stable transformation. For viral infection, the desired gene is cloned into baculovirus at the site of the wild-type polyhedron gene (Webb and Summers, Technique 2:173 (1990); Bishop and Posse, Adv. Gene Technol. 1:55 (1990); both of which are incorporated by reference in their entirety). The polyhedron gene is a component of a protein coat in occlusions which encapsulate virus particles. Deletion or insertion in the polyhedron gene results the failure to form occlusion bodies. Occlusion negative viruses are morphologically different from occlusion positive viruses and enable one skilled in the art to identify and purify recombinant viruses.

The vectors of present invention preferably contain one or more selectable markers which permit easy selection of transformed cells. A selectable marker is a gene the product of which provides, for example biocide or viral resistance, resistance to heavy metals, prototrophy to auxotrophs and the like. Selection may be accomplished by co-transformation, e.g., as described in WO 91/17243, a nucleic acid sequence of the present invention may be operably linked to a suitable promoter sequence. The promoter sequence is a nucleic acid sequence which is recognized by the insect host cell for expression of the nucleic acid sequence. The promoter sequence contains transcription and translation control sequences which mediate the expression of the protein or fragment thereof. The promoter may be any nucleic acid sequence which shows transcriptional activity in the insect host cell of choice and may be obtained from genes encoding polypeptides either homologous or heterologous to the host cell.

For example, a nucleic acid molecule encoding a protein or fragment thereof may also be operably linked to a suitable leader sequence. A leader sequence is a nontranslated region of a mRNA which is important for translation by the fungal host. The leader sequence is operably linked to the 5′ terminus of the nucleic acid sequence encoding the protein or fragment thereof. The leader sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any leader sequence which is functional in the insect host cell of choice may be used in the present invention.

A polyadenylation sequence may also be operably linked to the 3′ terminus of the nucleic acid sequence of the present invention. The polyadenylation sequence is a sequence which when transcribed is recognized by the insect host to add polyadenosine residues to transcribed mRNA. The polyadenylation sequence may be native to the nucleic acid sequence encoding the protein or fragment thereof or may be obtained from foreign sources. Any polyadenylation sequence which is functional in the fungal host of choice may be used in the present invention.

To avoid the necessity of disrupting the cell to obtain the protein or fragment thereof and to minimize the amount of possible degradation of the expressed polypeptide within the cell, it is preferred that expression of the polypeptide gene gives rise to a product secreted outside the cell. To this end, the protein or fragment thereof of the present invention may be linked to a signal peptide linked to the amino terminus of the protein or fragment thereof. A signal peptide is an amino acid sequence which permits the secretion of the protein or fragment thereof from the insect host into the culture medium. The signal peptide may be native to the protein or fragment thereof of the invention or may be obtained from foreign sources. The 5′ end of the coding sequence of the nucleic acid sequence of the present invention may inherently contain a signal peptide coding region naturally linked in translation reading frame with the segment of the coding region which encodes the secreted protein or fragment thereof.

At present, a mode of achieving secretion of a foreign gene product in insect cells is by way of the foreign gene's native signal peptide. Because the foreign genes are usually from non-insect organisms, their signal sequences may be poorly recognized by insect cells and hence, levels of expression may be suboptimal. However, the efficiency of expression of foreign gene products seems to depend primarily on the characteristics of the foreign protein. On average, nuclear localized or non-structural proteins are most highly expressed, secreted proteins are intermediate and integral membrane proteins are the least expressed. One factor generally affecting the efficiency of the production of foreign gene products in a heterologous host system is the presence of native signal sequences (also termed presequences, targeting signals, or leader sequences) associated with the foreign gene. The signal sequence is generally coded by a DNA sequence immediately following (5′ to 3′) the translation start site of the desired foreign gene.

The expression dependence on the type of signal sequence associated with a gene product can be represented by the following example: If a foreign gene is inserted at a site downstream from the translational start site of the baculovirus polyhedrin gene so as to produce a fusion protein (containing the N-terminus of the polyhedrin structural gene), the fused gene is highly expressed. But less expression is achieved when a foreign gene is inserted in a baculovirus expression vector immediately following the transcriptional start site and totally replacing the polyhedrin structural gene.

Insertions into the region −50 to −1 significantly alter (reduce) steady state transcription which, in turn, reduces translation of the foreign gene product. Use of the pVL941 vector optimizes transcription of foreign genes to the level of the polyhedrin gene transcription. Even though the transcription of a foreign gene may be optimal, optimal translation may vary because of several factors involving processing: signal peptide recognition, mRNA and ribosome binding, glycosylation, disulfide bond formation, sugar processing, oligomerization, for example.

The properties of the insect signal peptide are expected to be more optimal for the efficiency of the translation process in insect cells than those from vertebrate proteins. This phenomenon can generally be explained by the fact that proteins secreted from cells are synthesized as precursor molecules containing hydrophobic N-terminal signal peptides. The signal peptides direct transport of the select protein to its target membrane and are then cleaved by a peptidase on the membrane, such as the endoplasmic reticulum, when the protein passes through it.

Another exemplary insect signal sequence is the sequence encoding for Drosophila cuticle proteins such as CP1, CP2, CP3 or CP4 (Summers, U.S. Pat. No. 5,278,050; the entirety of which is herein incorporated by reference). Most of a 9 kb region of the Drosophila genome containing genes for the cuticle proteins has been sequenced. Four of the five cuticle genes contains a signal peptide coding sequence interrupted by a short intervening sequence (about 60 base pairs) at a conserved site. Conserved sequences occur in the 5′ mRNA untranslated region, in the adjacent 35 base pairs of upstream flanking sequence and at −200 base pairs from the mRNA start position in each of the cuticle genes.

Standard methods of insect cell culture, cotransfection and preparation of plasmids are set forth in Summers and Smith (Summers and Smith, A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures, Texas Agricultural Experiment Station Bulletin No. 1555, Texas A&M University (1987)). Procedures for the cultivation of viruses and cells are described in Volkman and Summers, J. Virol 19:820-832 (1975) and Volkman et al., J. Virol 19:820-832 (1976); both of which are herein incorporated by reference in their entirety.

(e) Bacterial Constructs and Transformed Bacterial Cells

The present invention also relates to a bacterial recombinant vector comprising exogenous genetic material. The present invention also relates to a bacteria cell comprising a bacterial recombinant vector. The present invention also relates to methods for obtaining a recombinant bacteria host cell, comprising introducing into a bacterial host cell exogenous genetic material. In a preferred embodiment the exogenous genetic material includes a nucleic acid molecule of the present invention having a sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 2814 or complements thereof or fragments of either or other nucleic acid molecule of the present invention.

The bacterial recombinant vector may be any vector which can be conveniently subjected to recombinant DNA procedures. The choice of a vector will typically depend on the compatibility of the vector with the bacterial host cell into which the vector is to be introduced. The vector may be a linear or a closed circular plasmid. The vector system may be a single vector or plasmid or two or more vectors or plasmids which together contain the total DNA to be introduced into the genome of the bacterial host. In addition, the bacterial vector may be an expression vector. Nucleic acid molecules encoding protein homologues or fragments thereof can, for example, be suitably inserted into a replicable vector for expression in the bacterium under the control of a suitable promoter for bacteria. Many vectors are available for this purpose and selection of the appropriate vector will depend mainly on the size of the nucleic acid to be inserted into the vector and the particular host cell to be transformed with the vector. Each vector contains various components depending on its function (amplification of DNA or expression of DNA) and the particular host cell with which it is compatible. The vector components for bacterial transformation generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes and an inducible promoter.

In general, plasmid vectors containing replicon and control sequences that are derived from species compatible with the host cell are used in connection with bacterial hosts. The vector ordinarily carries a replication site, as well as marking sequences that are capable of providing phenotypic selection in transformed cells. For example, E. coli is typically transformed using pBR322, a plasmid derived from an E. coli species (see, e.g., Bolivar et al., Gene 2:95 (1977); the entirety of which is herein incorporated by reference). pBR322 contains genes for ampicillin and tetracycline resistance and thus provides easy means for identifying transformed cells. The pBR322 plasmid, or other microbial plasmid or phage, also generally contains, or is modified to contain, promoters that can be used by the microbial organism for expression of the selectable marker genes.

Nucleic acid molecules encoding protein or fragments thereof may be expressed not only directly, but also as a fusion with another polypeptide, preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature polypeptide. In general, the signal sequence may be a component of the vector, or it may be a part of the polypeptide DNA that is inserted into the vector. The heterologous signal sequence selected should be one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For bacterial host cells that do not recognize and process the native polypeptide signal sequence, the signal sequence is substituted by a bacterial signal sequence selected, for example, from the group consisting of the alkaline phosphatase, penicillinase, lpp, or heat-stable enterotoxin II leaders.

Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA and includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria.

Expression and cloning vectors also generally contain a selection gene, also termed a selectable marker. This gene encodes a protein necessary for the survival or growth of transformed host cells grown in a selective culture medium. Host cells not transformed with the vector containing the selection gene will not survive in the culture medium. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli. One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous protein homologue or fragment thereof produce a protein conferring drug resistance and thus survive the selection regimen.

The expression vector for producing a protein or fragment thereof can also contains an inducible promoter that is recognized by the host bacterial organism and is operably linked to the nucleic acid encoding, for example, the nucleic acid molecule encoding the protein homologue or fragment thereof of interest. Inducible promoters suitable for use with bacterial hosts include the β-lactamase and lactose promoter systems (Chang et al., Nature 275:615 (1978); Goeddel et al., Nature 281:544 (1979); both of which are herein incorporated by reference in their entirety), the arabinose promoter system (Guzman et al., J. Bacteriol. 174:7716-7728 (1992); the entirety of which is herein incorporated by reference), alkaline phosphatase, a tryptophan (trp) promoter system (Goeddel, Nucleic Acids Res. 8:4057 (1980); EP 36,776; both of which are herein incorporated by reference in their entirety) and hybrid promoters such as the tac promoter (deBoer et al., Proc. Natl. Acad. Sci. (USA) 80:21-25 (1983); the entirety of which is herein incorporated by reference). However, other known bacterial inducible promoters are suitable (Siebenlist et al., Cell 20:269 (1980); the entirety of which is herein incorporated by reference).

Promoters for use in bacterial systems also generally contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the polypeptide of interest. The promoter can be removed from the bacterial source DNA by restriction enzyme digestion and inserted into the vector containing the desired DNA.

Construction of suitable vectors containing one or more of the above-listed components employs standard ligation techniques. Isolated plasmids or DNA fragments are cleaved, tailored and re-ligated in the form desired to generate the plasmids required. Examples of available bacterial expression vectors include, but are not limited to, the multifunctional E. coli cloning and expression vectors such as Bluescript™ (Stratagene, La Jolla, Calif.), in which, for example, encoding an A. nidulans protein homologue or fragment thereof homologue, may be ligated into the vector in frame with sequences for the amino-terminal Met and the subsequent 7 residues of β-galactosidase so that a hybrid protein is produced; pIN vectors (Van Heeke and Schuster, J. Biol. Chem. 264:5503-5509 (1989), the entirety of which is herein incorporated by reference); and the like. pGEX vectors (Promega, Madison Wis. U.S.A.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems are designed to include heparin, thrombin or factor XA protease cleavage sites so that the cloned polypeptide of interest can be released from the GST moiety at will.

Suitable host bacteria for a bacterial vector include archaebacteria and eubacteria, especially eubacteria and most preferably Enterobacteriaceae. Examples of useful bacteria include Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsiella, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla and Paracoccus. Suitable E. coli hosts include E. coli W3110 (American Type Culture Collection (ATCC) 27,325, Manassas, Va. U.S.A.), E. coli 294 (ATCC 31,446), E. coli B and E. coli X1776 (ATCC 31,537). These examples are illustrative rather than limiting. Mutant cells of any of the above-mentioned bacteria may also be employed. It is, of course, necessary to select the appropriate bacteria taking into consideration replicability of the replicon in the cells of a bacterium. For example, E. coli, Serratia, or Salmonella species can be suitably used as the host when well known plasmids such as pBR322, pBR325, pACYC177, or pKN410 are used to supply the replicon. E. coli strain W3110 is a preferred host or parent host because it is a common host strain for recombinant DNA product fermentations. Preferably, the host cell should secrete minimal amounts of proteolytic enzymes.

Host cells are transfected and preferably transformed with the above-described vectors and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.

Numerous methods of transfection are known to the ordinarily skilled artisan, for example, calcium phosphate and electroporation. Depending on the host cell used, transformation is done using standard techniques appropriate to such cells. The calcium treatment employing calcium chloride, as described in section 1.82 of Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989), is generally used for bacterial cells that contain substantial cell-wall barriers. Another method for transformation employs polyethylene glycol/DMSO, as described in Chung and Miller (Chung and Miller, Nucleic Acids Res. 16:3580 (1988); the entirety of which is herein incorporated by reference). Yet another method is the use of the technique termed electroporation.

Bacterial cells used to produce the polypeptide of interest for purposes of this invention are cultured in suitable media in which the promoters for the nucleic acid encoding the heterologous polypeptide can be artificially induced as described generally, e.g., in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Laboratory Press, (1989). Examples of suitable media are given in U.S. Pat. Nos. 5,304,472 and 5,342,763; both of which are incorporated by reference in their entirety.

In addition to the above discussed procedures, practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of macromolecules (e.g., DNA molecules, plasmids, etc.), generation of recombinant organisms and the screening and isolating of clones, (see for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989); Mailga et al., Methods in Plant Molecular Biology, Cold Spring Harbor Press (1995), the entirety of which is herein incorporated by reference; Birren et al., Genome Analysis: Analyzing DNA, 1, Cold Spring Harbor, N.Y., the entirety of which is herein incorporated by reference).

(f) Computer Readable Media

The nucleotide sequence provided in SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof, or complement thereof, or a nucleotide sequence at least 90% identical, preferably 95%, identical even more preferably 99% or 100% identical to the sequence provided in SEQ ID NO: 1 through SEQ ID NO: 2814 or fragment thereof, or complement thereof, can be “provided” in a variety of mediums to facilitate use. Such a medium can also provide a subset thereof in a form that allows a skilled artisan to examine the sequences.

A preferred subset of nucleotide sequences are those nucleic acid sequences that encodes a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof or fragment of either.

A further preferred subset of nucleic acid sequences is where the subset of sequences is two proteins or fragments thereof, more preferably three proteins or fragments thereof and even more preferable four proteins or fragments thereof, these nucleic acid sequences are selected from the group that comprises a maize or a soybean triose phosphate isomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate aldolase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 1,6-bisphosphate enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructose 6-phosphate 2-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucoisomerase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean vacuolar H+ translocating-pyrophosphatase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean pyrophosphate-dependent fructose-6-phosphate phosphotransferase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean invertase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean sucrose synthase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean hexokinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean fructokinase enzyme or complement thereof or fragment of either f, a nucleic acid molecule that encodes a maize or a soybean NDP-kinase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean glucose-6-phosphate 1-dehydrogenase enzyme or complement thereof or fragment of either, a nucleic acid molecule that encodes a maize or a soybean phosphoglucomutase enzyme or complement thereof or fragment of either and a nucleic acid molecule that encodes a maize or a soybean UDP-glucose pyrophosphorylase enzyme or complement thereof or fragment of either.

In one application of this embodiment, a nucleotide sequence of the present invention can be recorded on computer readable media. As used herein, “computer readable media” refers to any medium that can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc, storage medium and magnetic tape: optical storage media such as CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media. A skilled artisan can readily appreciate how any of the presently known computer readable mediums can be used to create a manufacture comprising computer readable medium having recorded thereon a nucleotide sequence of the present invention.

As used herein, “recorded” refers to a process for storing information on computer readable medium. A skilled artisan can readily adopt any of the presently known methods for recording information on computer readable medium to generate media comprising the nucleotide sequence information of the present invention. A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon a nucleotide sequence of the present invention. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the nucleotide sequence information of the present invention on computer readable medium. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and Microsoft Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. A skilled artisan can readily adapt any number of data processor structuring formats (e.g. text file or database) in order to obtain computer readable medium having recorded thereon the nucleotide sequence information of the present invention.

By providing one or more of nucleotide sequences of the present invention, a skilled artisan can routinely access the sequence information for a variety of purposes. Computer software is publicly available which allows a skilled artisan to access sequence information provided in a computer readable medium. The examples which follow demonstrate how software which implements the BLAST (Altschul et al., J. Mol. Biol. 215:403-410 (1990), the entirety of which is herein incorporated by reference) and BLAZE (Brutlag et al., Comp. Chem. 17:203-207 (1993), the entirety of which is herein incorporated by reference) search algorithms on a Sybase system can be used to identify open reading frames (ORFs) within the genome that contain homology to ORFs or proteins from other organisms. Such ORFs are protein-encoding fragments within the sequences of the present invention and are useful in producing commercially important proteins such as enzymes used in amino acid biosynthesis, metabolism, transcription, translation, RNA processing, nucleic acid and a protein degradation, protein modification and DNA replication, restriction, modification, recombination and repair.

The present invention further provides systems, particularly computer-based systems, which contain the sequence information described herein. Such systems are designed to identify commercially important fragments of the nucleic acid molecule of the present invention. As used herein, “a computer-based system” refers to the hardware means, software means and data storage means used to analyze the nucleotide sequence information of the present invention. The minimum hardware means of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means and data storage means. A skilled artisan can readily appreciate that any one of the currently available computer-based system are suitable for use in the present invention.

As indicated above, the computer-based systems of the present invention comprise a data storage means having stored therein a nucleotide sequence of the present invention and the necessary hardware means and software means for supporting and implementing a search means. As used herein, “data storage means” refers to memory that can store nucleotide sequence information of the present invention, or a memory access means which can access manufactures having recorded thereon the nucleotide sequence information of the present invention. As used herein, “search means” refers to one or more programs which are implemented on the computer-based system to compare a target sequence or target structural motif with the sequence information stored within the data storage means. Search means are used to identify fragments or regions of the sequence of the present invention that match a particular target sequence or target motif. A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are available can be used in the computer-based systems of the present invention. Examples of such software include, but are not limited to, MacPattern (EMBL), BLASTIN and BLASTIX (NCBIA). One of the available algorithms or implementing software packages for conducting homology searches can be adapted for use in the present computer-based systems.

The most preferred sequence length of a target sequence is from about 10 to 100 amino acids or from about 30 to 300 nucleotide residues. However, it is well recognized that during searches for commercially important fragments of the nucleic acid molecules of the present invention, such as sequence fragments involved in gene expression and protein processing, may be of shorter length.

As used herein, “a target structural motif,” or “target motif,” refers to any rationally selected sequence or combination of sequences in which the sequences the sequence(s) are chosen based on a three-dimensional configuration which is formed upon the folding of the target motif. There are a variety of target motifs known in the art. Protein target motifs include, but are not limited to, enzymatic active sites and signal sequences. Nucleic acid target motifs include, but are not limited to, promoter sequences, cis elements, hairpin structures and inducible expression elements (protein binding sequences).

Thus, the present invention further provides an input means for receiving a target sequence, a data storage means for storing the target sequences of the present invention sequence identified using a search means as described above and an output means for outputting the identified homologous sequences. A variety of structural formats for the input and output means can be used to input and output information in the computer-based systems of the present invention. A preferred format for an output means ranks fragments of the sequence of the present invention by varying degrees of homology to the target sequence or target motif. Such presentation provides a skilled artisan with a ranking of sequences which contain various amounts of the target sequence or target motif and identifies the degree of homology contained in the identified fragment.

A variety of comparing means can be used to compare a target sequence or target motif with the data storage means to identify sequence fragments sequence of the present invention. For example, implementing software which implement the BLAST and BLAZE algorithms (Altschul et al., J. Mol. Biol. 215:403-410 (1990)) can be used to identify open frames within the nucleic acid molecules of the present invention. A skilled artisan can readily recognize that any one of the publicly available homology search programs can be used as the search means for the computer-based systems of the present invention.

Having now generally described the invention, the same will be more readily understood through reference to the following examples which are provided by way of illustration and are not intended to be limiting of the present invention, unless specified.

Example 1

The MONN01 cDNA library is a normalized library generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON001 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Illinois U.S.A.) immature tassels at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON003 library is generated from maize (B73×Mol 7, Illinois Foundation Seeds, Champaign, Illinois U.S.A.) roots at the V6 developmental stage. Seeds are planted at a depth of approximately 3 cm in coil into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, the seedlings are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting at a concentration of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in approximately 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6 leaf development stage. The root system is cut from maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON004 cDNA library is generated from maize (B73×Mol7, Illinois Foundation Seeds, Champaign, Illinois U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON005 cDNA library is generated from maize (B73×Mol7, Illinois Foundation Seeds, Champaign Illinois, U.S.A.) root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON006 cDNA library is generated from maize (B73×Mol7, Illinois Foundation Seeds, Champaign Illinois, U.S.A.) total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON007 cDNA library is generated from the primary root tissue of 5 day old maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). After germination, the trays, along with the moist paper, are moved to a greenhouse where the maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles for approximately 5 days. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. The primary root tissue is collected when the seedlings are 5 days old. At this stage, the primary root (radicle) is pushed through the coleorhiza which itself is pushed through the seed coat. The primary root, which is about 2-3 cm long, is cut and immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON008 cDNA library is generated from the primary shoot (coleoptile 2-3 cm) of maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) seedlings which are approximately 5 days old. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to a greenhouse at 15 hr daytime/9 hr nighttime cycles and grown until they are 5 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 5 days old. At this stage, the primary shoot (coleoptile) is pushed through the seed coat and is about 2-3 cm long. The coleoptile is dissected away from the rest of the seedling, immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON009 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) leaves at the 8 leaf stage (V8 plant development stage). Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 8-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical, are cut at the base of the leaves. The leaves are then pooled and then immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON010 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) root tissue at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the V8 development stage. The root system is cut from this mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON011 cDNA library is generated from undeveloped maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) leaf at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The second youngest leaf which is at the base of the apical leaf of V6 stage maize plant is cut at the base and immediately transferred to liquid nitrogen containers in which the leaf is crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON012 cDNA library is generated from 2 day post germination maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark until germination (one day). Then the trays containing the seeds are moved to the greenhouse and grown at 15 hr daytime/9 hr nighttime cycles until 2 days post germination. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Tissue is collected when the seedlings are 2 days old. At the two day stage, the coleorhiza is pushed through the seed coat and the primary root (the radicle) is pierced the coleorhiza but is barely visible. Also, at this two day stage, the coleoptile is just emerging from the seed coat. The 2 days post germination seedlings are then immersed in liquid nitrogen and crushed. The harvested tissue is stored at −80° C. until preparation of total RNA.

The SATMON013 cDNA library is generated from apical maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) meristem founder at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, the plant is at the 4 leaf stage. The lead at the apex of the V4 stage maize plant is referred to as the meristem founder. This apical meristem founder is cut, immediately frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON014 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) endosperm fourteen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the maize plant ear shoots are ready for fertilization. At this stage, the ear shoots are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are pollinated and 14 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON016 library is a maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) sheath library collected at the V8 developmental stage. Seeds are planted in a depth of approximately 3 cm in solid into 2-3 inch pots containing Metro growing medium. After 2-3 weeks growth, they are transplanted into 10″ pots containing the same. Plants are watered daily before transplantation and approximately the times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plants are at the V8 stage the 5th and 6th leaves from the bottom exhibit fully developed leaf blades. At the base of these leaves, the ligule is differentiated and the leaf blade is joined to the sheath. The sheath is dissected away from the base of the leaf then the sheath is frozen in liquid nitrogen and crushed. The tissue is then stored at −80° C. until RNA preparation.

The SATMON017 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) embryo seventeen days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth the seeds are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the VI 0 stage, the ear shoots of maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold the pollen. The ear shoots are fertilized and 21 days after pollination, the ears are pulled out and the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON019 (Lib3054) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) culm (stem) at the V8 developmental stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. When the maize plant is at the V8 stage, the 5th and 6th leaves from the bottom have fully developed leaf blades. The region between the nodes of the 5th and the sixth leaves from the bottom is the region of the stem that is collected. The leaves are pulled out and the sheath is also torn away from the stem. This stem tissue is completely free of any leaf and sheath tissue. The stem tissue is then frozen in liquid nitrogen and stored at −80° C. until RNA preparation.

The SATMON020 cDNA library is from a maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) Hill Type 1′-Initiated Callus. Petri plates containing approximately 25 ml of Type II initiation media are prepared. This medium contains N6 salts and vitamins, 3% sucrose, 2.3 g/liter proline 0.1 g/liter enzymatic casein hydrolysate, 2 mg/liter 2,4- dichloro phenoxy-acetic acid (2,4, D), 15.3 mg/liter AgNO3 and 0.8% bacto agar and is adjusted to pH 6.0 before autoclaving. At 9-11 days after pollination, an ear with immature embryos measuring approximately 1-2 mm in length is chosen. The husks and silks are removed and then the ear is broken into halves and placed in an autoclaved solution of Clorox/TWEEN 20 sterilizing solution. Then the ear is rinsed with deionized water. Then each embryo is extracted from the kernel. Intact embryos are placed in contact with the medium, scutellar side up). Multiple embryos are plated on each plate and the plates are incubated in the dark at 25° C. Type II calluses are friable, can be subcultured with a spatula, frequently regenerate via somatic embryogenesis and are relatively undifferentiated. As seen in the microscope, the Tape II calluses show color ranging from translucent to light yellow and heterogeneity on with respect to embryoid structure as well as stage of embryoid development. Once Type II callus are formed, the calluses is transferred to type II callus maintenance medium without AgNO3. Every 7-10 days, the callus is subcultured. About 4 weeks after embryo isolation the callus is removed from the plates and then frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON021 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb Illinois, U.S.A.) tassel at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. As the maize plant enters the V8 stage, tassels which are 15-20 cm in length are collected and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON022 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) ear (growing silks) at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the plant is in the V8 stage. At this stage, some immature ear shoots are visible. The immature ear shoots (approximately 1 cm in length) are pulled out, frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON23 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) ear (growing silk) at the V8 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. When the tissue is harvested at the V8 stage, the length of the ear that is harvested is about 10-15 cm and the silks are just exposed (approximately 1 inch). The ear along with the silks is frozen in liquid nitrogen and then the tissue is stored at −80° C. until RNA preparation.

The SATMON024 cDNA library is generated from the immature maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) tassel at the V9 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. As a maize plant enters the V9 stage, the tassel is rapidly developing and a 37 cm tassel along with the glume, anthers and pollen is collected and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The SATMON025 cDNA library is from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) Hill Type II-Regenerated Callus. Type II callus is grown in initiation media as described for SATMON020 and then the embryoids on the surface of the Type II callus are allowed to mature and germinate. The 1-2 gm fresh weight of the soft friable type callus containing numerous embryoids are transferred to 100×15 mm petri plates containing 25 ml of regeneration media. Regeneration media consists of Murashige and Skoog (MS) basal salts, modified White's vitamins (0.2 g/liter glycine and 0.5 g/liter myo-inositoland 0.8% bacto agar (6SMSOD)). The plates are then placed in the dark after covering with parafilm. After 1 week, the plates are moved to a lighted growth chamber with 16 hr light and 8 hr dark photoperiod. Three weeks after plating the Type II callus to 6SMSOD, the callus exhibit shoot formation. The callus and the shoots are transferred to fresh 6SMSOD plates for another 2 weeks. The callus and the shoots are then transferred to petri plates with reduced sucrose (3SMSOD). Upon distinct formation of a root and shoot, the newly developed green plants are then removed out with a spatula and frozen in liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON026 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) juvenile/adult shift leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plants are at the 8-leaf development stage. Leaves are founded sequentially around the meristem over weeks of time and the older, more juvenile leaves arise earlier and in a more basal position than the younger, more adult leaves, which are in a more apical position. In a V8 plant, some leaves which are in the middle portion of the plant exhibit characteristics of both juvenile as well as adult leaves. They exhibit a yellowing color but also exhibit, in part, a green color. These leaves are termed juvenile/adult shift leaves. The juvenile/adult shift leaves (the 4th, 5th leaves from the bottom) are cut at the base, pooled and transferred to liquid nitrogen in which they are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON027 cDNA library is generated from 6 day maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) leaves. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Zea mays plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical, are all cut at the base of the leaves. All the leaves exhibit significant wilting. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON028 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) roots at the V8 developmental stage that are subject to six days water stress. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the Metro 200 growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Prior to tissue collection, when the plant is at the 8-leaf stage, water is held back for six days. The root system is cut, shaken and washed to remove soil. Root tissue is then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are then crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON029 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) seedlings at the etiolated stage. Seeds are planted on a moist filter paper on a covered tray that is kept in the dark for 4 days at approximately 70° F. Tissue is collected when the seedlings are 4 days old. By 4 days, the primary root has penetrated the coleorhiza and is about 4-5 cm and the secondary lateral roots have also made their appearance. The coleoptile has also pushed through the seed coat and is about 4-5 cm long. The seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON030 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) root tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth, they are transplanted into 10 inch pots containing the same. Plants are watered daily before transplantation and approximately 3 times a week after transplantation. Peters 15-16-17 fertilizer is applied approximately three times per week after transplanting, at a strength of 150 ppm N. Two to three times during the life time of the plant, from transplanting to flowering, a total of approximately 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 sodium vapor lamps. Tissue is collected when the maize plant is at the 4 leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is then immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON031 cDNA library is generated from the maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) leaf tissue at the V4 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is 80° F. and the nighttime temperature is 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 4-leaf development stage. The third leaf from the bottom is cut at the base and immediately frozen in liquid nitrogen and crushed. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON033 cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) embryo tissue 13 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the VI 0 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 13 days after pollination, the ears are pulled out and then the kernels are plucked cut of the ears. Each kernel is then dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the embryos are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The SATMON034 cDNA library is generated from cold stressed maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) seedlings. Seeds are planted on a moist filter paper on a covered tray that is kept on at 10° C. for 7 days. After 7 days, the temperature is shifted to 15° C. for one day until germination of the seed. Tissue is collected once the seedlings are 1 day old. At this point, the coleorhiza has just pushed out of the seed coat and the primary root is just making its appearance. The coleoptile has not yet pushed completely through the seed coat and is also just making its appearance. These 1 day old cold stressed seedlings are frozen in liquid nitrogen and crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMON˜001 (Lib36, Lib83, Lib84) cDNA library is generated from maize leaves at the V8 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V8 stage. The older more juvenile leaves in a basal position was well as the younger more adult leaves which are more apical are all cut at the base, pooled and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SATMONN01 cDNA library is generated from maize (B73, Illinois Foundation Seeds, Champaign, Illinois U.S.A.) normalized immature tassels at the V6 plant development stage normalized tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in a greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue from the maize plant is collected at the V6 stage. At that stage the tassel is an immature tassel of about 2-3 cm in length. The tassels are removed and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN04 cDNA library is generated from maize (B73×Mol7, Illinois Foundation Seeds, Champaign, Illinois U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older, more juvenile leaves, which are in a basal position, as well as the younger, more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated DATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN05 cDNA library is generated from maize (B73×Mol7, Illinois Foundation Seeds, Champaign Illinois, U.S.A.) normalized root tissue at the V6 development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The root system is cut from the mature maize plant and washed with water to free it from the soil. The tissue is immediately frozen in liquid nitrogen and the harvested tissue is then stored at −80° C. until RNA preparation. The single stranded and double stranded DNA representing approximately 1×16 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The SATMONN06 cDNA library is generated from maize (B73×Mol7, Illinois Foundation Seeds, Champaign Illinois, U.S.A.) normalized total leaf tissue at the V6 plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 6-leaf development stage. The older more juvenile leaves, which are in a basal position, as well as the younger more adult leaves, which are more apical are cut at the base of the leaves. The leaves are then pooled and immediately transferred to liquid nitrogen containers in which the pooled leaves are crushed. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The CMZ029 (SATMON036) cDNA library is generated from maize (DK604, Dekalb Genetics, Dekalb, Illinois U.S.A.) endosperm 22 days after pollination. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the green house in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. After the V10 stage, the ear shoots of the maize plant, which are ready for fertilization, are enclosed in a paper bag before silk emergent to withhold the pollen. The ear shoots are pollinated and 22 days after pollination, the ears are pulled out and then the kernels are plucked out of the ears. Each kernel is then dissected into the embryo and the endosperm and the alurone layer is removed. After dissection, the endosperms are immediately frozen in liquid nitrogen and then stored at −80° C. until RNA preparation.

The CMz030 (Lib143) cDNA library is generated from maize seedling tissue two days post germination. Seeds are planted on a moist filter paper on a covered try that is keep in the dark until germination. The trays are then moved to the bench top at 15 hr daytime/9 hr nighttime cycles for 2 days post-germination. The day time temperature is 80° F. and the nighttime temperature is 70° F. Tissue is collected when the seedlings are 2 days old. At this stage, the colehrhiza has pushed through the seed coat and the primary root (the radicle) is just piercing the colehrhiza and is barely visible. The seedlings are placed at 42° C. for 1 hour. Following the heat shock treatment, the seedlings are immersed in liquid nitrogen and crushed. The harvested tissue is stored at −80° until RNA preparation.

The CMz031 (Lib148) cDNA library is generated from maize pollen tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag to withhold pollen. Twenty-one days after pollination, prior to removing the ears, the paper bag is shaken to collect the mature pollen. The mature pollen is immediately frozen in liquid nitrogen containers and the pollen is crushed. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz033 (Lib189) cDNA library is generated from maize pooled leaf tissue. Samples are harvested from open pollinated plants. Tissue is collected from maize leaves at the anthesis stage. The leaves are collect from 10-12 plants and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz034 (Lib3060) cDNA library is generated from maize mature tissue at 40 days post pollination plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from leaves located two leaves below the ear leaf. This sample represents those genes expressed during onset and early stages of leaf senescence. The leaves are pooled and immediately transferred to liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz035 (Lib3061) cDNA library is generated from maize endosperm tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence to withhold pollen. Thirty-two days after pollination, the ears are pulled out and the kernels are removed from the cob. Each kernel is dissected into the embryo and the endosperm and the aleurone layer is removed. After dissection, the endosperms are immediately transferred to liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz036 (Lib3062) cDNA library is generated from maize husk tissue at the 8 week old plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from 8 week old plants. The husk is separated from the ear and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz037 (Lib3059) cDNA library is generated from maize pooled kernal at 12-15 days after pollination plant development stage. Sample were collected from field grown material. Whole kernals from hand pollinated (control pollination) are harvested as whole ears and immediately frozen on dry ice. Kernels from 10-12 ears were pooled and ground together in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz039 (Lib3066) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz040 (Lib3067) cDNA library is generated from maize kernel tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag before silk emergence to withhold pollen. Five to eight days after controlled pollination. The ears are pulled and the kernels removed. The kernels are immediately frozen in liquid nitrogen. The harvested kernels tissue is then stored at −80° C. until RNA preparation. This sample represents gene expressed in early kernel development, during periods of cell division, amyloplast biogenesis and early carbon flow across the material to filial tissue.

The CMz041 (Lib3068) cDNA library is generated from maize pollen germinating silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants when the ear shoots are ready for fertilization at the silk emergence stage. The emerging silks are pollinated with an excess of pollen under controlled pollination conditions in the green house. Eighteen hours after pollination the silks are removed from the ears and immediately frozen in liquid nitrogen containers. This sample represents genes expressed in both pollen and silk tissue early in pollination. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz042 (Lib3069) cDNA library is generated from maize ear tissue excessively pollinated at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+ stage plants and the ear shoots which are ready for fertilization are at the silk emergence stage. The immature ears are pollinated with an excess of pollen under controlled pollination conditions. Eighteen hours post-pollination, the ears are removed and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz044 (Lib3075) cDNA library is generated from maize microspore tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature anthers from 7 week old tassels. The immature anthers are first dissected from the 7 week old tassel with a scalpel on a glass slide covered with water. The microspores (immature pollen) are released into the water and are recovered by centrifugation. The microspore suspension is immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz045 (Lib3076) cDNA library is generated from maize immature ear megaspore tissue. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from immature ear (megaspore) obtained from 7 week old plants. The immature ears are harvested from the 7 week old plants and are approximately 2.5 to 3 cm in length. The kernels are removed from the cob immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz047 (Lib3078) cDNA library is generated from maize CO2 treated high-exposure shoot tissue at the V10+ plant development stage. RX601 maize seeds are sterilized for minute with a 10% clorox solution. The seeds are rolled in germination paper, and germinated in 0.5 mM calcium sulfate solution for two days are 30° C. The seedlings are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium at a rate of 2-3 seedlings per pot. Twenty pots are placed into a high CO2 environment (approximately 1000 ppm CO2). Twenty plants were grown under ambient greenhouse CO2 (approximately 450 ppm CO2). Plants are watered daily before transplantation and three times a week after transplantation. Peters 20-20-20 fertilizer is also lightly applied. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. At ten days post planting, the shoots from both atmosphere are frozen in liquid nitrogen and lightly ground. The roots are washed in deionized water to remove the support media and the tissue is immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz048 (Lib3079) cDNA library is generated from maize basal endosperm transfer layer tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected from V10+maize plants. The ear shoots, which are ready for fertilization, are enclosed in a paper bag prior to silk emergence, to withhold the pollen. Kernels are harvested at 12 days post-pollination and placed on wet ice for dissection. The kernels are cross sectioned laterally, dissecting just above the pedicel region, including 1-2 mm of the lower endosperm and the basal endosperm transfer region. The pedicel and lower endosperm region containing the basal endosperm transfer layer is pooled and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz049(Lib3088) cDNA library is generated from maize immature anther tissue at the 7 week old immature tassel stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is at the 7 week old immature tassel stage. At this stage, prior to anthesis, the immature anthers are green and enclosed in the staminate spikelet. The developing anthers are dissected away from the 7 week old immature tassel and immediately transferred to liquid nitrogen container. The harvested tissue is then stored at −80° C. until RNA preparation.

The CMz050 (Lib3114) cDNA library is generated from maize silk tissue at the V10+ plant development stage. Seeds are planted at a depth of approximately 3 cm into 2-3 inch peat pots containing Metro 200 growing medium. After 2-3 weeks growth they are transplanted into 10 inch pots containing the same growing medium. Plants are watered daily before transplantation and three times a week after transplantation. Peters 15-16-17 fertilizer is applied three times per week after transplanting at a strength of 150 ppm N. Two to three times during the lifetime of the plant, from transplanting to flowering, a total of 900 mg Fe is added to each pot. Maize plants are grown in the greenhouse in 15 hr day/9 hr night cycles. The daytime temperature is approximately 80° F. and the nighttime temperature is approximately 70° F. Supplemental lighting is provided by 1000 W sodium vapor lamps. Tissue is collected when the maize plant is beyond the 10-leaf development stage and the ear shoots are approximately 15-20 cm in length. The ears are pulled and silks are separated from the ears and immediately transferred to liquid nitrogen containers. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON001 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) total leaf tissue at the V4 plant development stage. Leaf tissue from 38, field grown V4 stage plants is harvested from the 4th node. Leaf tissue is removed from the plants and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON002 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue at the V4 plant development stage. Root tissue from 76, field grown V4 stage plants is harvested. The root systems is cut from the soybean plant and washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON003 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON004 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledon tissue harvested 2 day post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 2 days after the start of imbibition. The 2 days after imbibition samples are separated into 3 collections after removal of any adhering seed coat. At the 2 day stage, the hypocotyl axis is emerging from the soil. A few seedlings have cracked the soil surface and exhibited slight greening of the exposed cotyledons. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON005 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling hypocotyl axis tissue harvested 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after the start of imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post imbibition. At the 6 hours after imbibition stage, not all cotyledons have become fully hydrated and germination, or radicle protrusion, has not occurred. The seedlings are washed in water to remove soil, hypocotyl axis harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON006 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling cotyledons tissue harvest 6 hour post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. Trays are placed in an environmental chamber and grown at 12 hr daytime/12 hr nightime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Tissue is collected 6 hours after imbibition. The 6 hours after imbibition samples are separated into 3 collections after removal of any adhering seed coat. The 6 hours after imbibition sample is collected over the course of approximately 2 hours starting at 6 hours post-imbibition. At the 6 hours after imbibition, not all cotyledons have become fully hydrated and germination or radicle protrusion, have not occurred. The seedlings are washed in water to remove soil, cotyledon harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON007 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days post-flowering. Seed pods from field grown plants are harvested 25 and 35 days after flowering and the seeds extracted from the pods. Approximately 4.4 g and 19.3 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON008 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested from 25 and 35 days post-flowering plants. Total leaf tissue is harvested from field grown plants. Approximately 19 g and 29 g of leaves are harvested from the fourth node of the plant 25 and 35 days post-flowering and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON009 cDNA library is generated from soybean cutlivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) pod and seed tissue harvested 15 days post-flowering. Pods from field grown plants are harvested 15 days post-flowering. Approximately 3 g of pod tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON010 cDNA library is generated from soybean cultivar C1944 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) seed tissue harvested 40 days post-flowering. Pods from field grown plants are harvested 40 days post-flowering. Pods and seeds are separated, approximately 19 g of seed tissue is harvested and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON011 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4th node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON012 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue. Leaves from field grown plants are harvested from the fourth node 15 days post-flowering. Approximately 12 g of leaves are harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON013 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root and nodule tissue. Approximately, 28 g of root tissue from field grown plants is harvested 15 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON014 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 and 35 days after flowering. Seed pods from field grown plants are harvested 15 days after flowering and the seeds extracted from the pods. Approximately 5 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON015 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 45 and 55 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 19 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON016 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately, 61 g and 38 g of root tissue from field grown plants is harvested 25 and 35 days post-flowering is harvested. The root system is cut from the soybean plant and washed with water to free it from the soil. The tissue is placed in 14 ml polystyrene tubes and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON017 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue. Approximately 28 g of root tissue from field grown plants is harvested 45 and 55 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON018 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 45 and 55 days post-flowering. Leaves from field grown plants are harvested 45 and 55 days after flowering from the fourth node. Approximately 27 g and 33 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON019 cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Gemplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON020 cDNA is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 65 and 75 days post-flowering. Seed pods from field grown plants are harvested 45 and 55 days after flowering and the seeds extracted from the pods. Approximately 14 g and 31 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON021 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar Hartwig (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Plants are grown in tissue culture at room temperature. At approximately 6 weeks post-germination, the plants are exposed to sterilized Soybean Cyst Nematode eggs. Infection is then allowed to progress for 10 days. After the 10 day infection process, the tissue is harvested. Agar from the culture medium and nematodes are removed and the root tissue is immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON022 (Lib3030) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) partially opened flower tissue. Partially to fully opened flower tissue is harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. A total of 3 g of flower tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON023 cDNA library is generated from soybean genotype BW211S Null (Tohoku University, Morioka, Japan) seed tissue harvested 15 and 40 days post-flowering. Seed pods from field grown plants are harvested 15 and 40 days post-flowering and the seeds extracted from the pods. Approximately 0.7 g and 14.2 g of seeds are harvested from the respective seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON024 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) internode-2 tissue harvested 18 days post-imbibition. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium. The plants are grown in a greenhouse for 18 days after the start of imbibition at ambient temperature. Soil is checked and watered daily to maintain even moisture conditions. Stem tissue is harvested 18 days after the start of imbibition. The samples are divided into hypocotyl and internodes 1 through 5. The fifth internode contains some leaf bud material. Approximately 3 g of each sample is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON025 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) leaf tissue harvested 65 days post-flowering. Leaves are harvested from the fourth node of field grown plants 65 days post-flowering. Approximately 18.4 g of leaf tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

SOYMON026 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) root tissue harvested 65 and 75 days post-flowering. Approximately 27 g and 40 g of root tissue from field grown plants is harvested 65 and 75 days post-flowering. The root system is cut from the soybean plant, washed with water to free it from the soil and immediately frozen in dry-ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON027 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed tissue harvested 25 days post-flowering. Seed pods from field grown plants are harvested 25 days post-flowering and the seeds extracted from the pods. Approximately 17 g of seeds are harvested from the seed pods and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON028 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed root tissue. The plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of development, water is withheld from half of the plant collection (drought stressed population). After 3 days, half of the plants from the drought stressed condition and half of the plants from the control population are harvested. After another 3 days (6 days post drought induction) the remaining plants are harvested. A total of 27 g and 40 g of root tissue is harvested and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON029 cDNA library is generated from Soybean Cyst Nematode-resistant soybean cultivar PI07354 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) root tissue. Late fall to early winter greenhouse grown plants are exposed to Soybean Cyst Nematode eggs. At 10 days post-infection, the plants are uprooted, rinsed briefly and the roots frozen in liquid nitrogen. Approximately 20 grams of root tissue is harvested from the infected plants. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON030 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) flower bud tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. A total of 100 mg of flower buds are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON031 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) carpel and stamen tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Flower buds are removed from the plant at the pedicel. Flowers are dissected to separate petals, sepals and reproductive structures (carpels and stamens). A total of 300 mg of carpel and stamen tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON032 cDNA library is prepared from the Asgrow cultivar A4922 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry soybean seed meristem tissue. Surface sterilized seeds are germinated in liquid media for 24 hours. The seed axis is then excised from the barely germinating seed, placed on tissue culture media and incubated overnight at 20° C. in the dark. The supportive tissue is removed from the explant prior to harvest. Approximately 570 mg of tissue is harvested and frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON033 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heat-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to an incubator set at 40° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at −80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance. Total RNA and poly A+ RNA is prepared from equal amounts of pooled tissue.

The SOYMON034 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) cold-shocked seedling tissue without cotyledons. Seeds are imbibed and germinated in vermiculite for 2 days under constant illumination. After 48 hours, the seedlings are transferred to a cold room set at 5° C. under constant illumination. After 30, 60 and 180 minutes seedlings are harvested and dissected. A portion of the seedling consisting of the root, hypocotyl and apical hook is frozen in liquid nitrogen and stored at −80° C. The seedlings after 2 days of imbibition are beginning to emerge from the vermiculite surface. The apical hooks are dark green in appearance.

The SOYMON035 cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seed coat tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are harvested from mid to nearly full maturation (seed coats are not yellowing). The entire embryo proper is removed from the seed coat sample and the seed coat tissue are harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON036 cDNA library is generated from soybean cultivars PI171451, PI227687 and PI229358 (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) insect challenged leaves. Plants from each of the three cultivars are grown in screenhouse conditions. The screenhouse is divided in half and one half of the screenhouse is infested with soybean looper and the other half infested with velvetbean caterpillar. A single leaf is taken from each of the representative plants at 3 different time points, 11 days after infestation, 2 weeks after infestation and 5 weeks after infestation and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation. Total RNA and poly A+RNA is isolated from pooled tissue consisting of equal quantities of all 18 samples (3 genotypes×3 sample times×2 insect genotypes).

The SOYMON037 cDNA library is generated from soybean cultivar A3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) etiolated axis and radical tissue. Seeds are planted in moist vermiculite, wrapped and kept at room temperature in complete darkness until harvest. Etiolated axis and hypocotyl tissue is harvested at 2, 3 and 4 days post-planting. A total of 1 gram of each tissue type is harvested at 2, 3 and 4 days after planting and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The SOYMON038 cDNA library is generated from soybean variety Asgrow A3237 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) rehydrated dry seeds. Explants are prepared for transformation after germination of surface-sterilized seeds on solid tissue media. After 6 days, at 28° C. and 18 hours of light per day, the germinated seeds are cold shocked at 4° C. for 24 hours. Meristemic tissue and part of the hypocotyl is remove and cotyledon excised. The prepared explant is then wounded for Agrobacterium infection. The 2 grams of harvested tissue is frozen in liquid nitrogen and stored at −80° C. until RNA preparation.

The Soy51 (LIB3027) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The Soy52 (LIB3028) cDNA library is generated from normalized flower DNA. Single stranded DNA representing approximately 1×106 colony forming units of SOYMON022 harvested tissue is used as the starting material for normalization. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

The Soy53 (LIB3039) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) seedling shoot apical meristem tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Apical tissue is harvested from seedling shoot meristem tissue, 7-8 days after the start of imbibition. The apex of each seedling is dissected to include the fifth node to the apical meristem. The fifth node corresponds to the third trifoliate leaf in the very early stages of development. Stipules completely envelop the leaf primordia at this time. A total of 200 mg of apical tissue is harvested and immediately frozen in liquid nitrogen. The harvested tissue is then stored at −80° C. until RNA preparation.

The Soy54 (LIB3040) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) heart to torpedo stage embryo tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected and embryos removed from surrounding endosperm and maternal tissues. Embryos from globular to young torpedo stages (by corresponding analogy to Arabidopsis) are collected with a bias towards the middle of this spectrum. Embryos which are beginning to show asymmetric development of cotyledons are considered the upper developmental boundary for the collection and are excluded. A total of 12 mg embryo tissue is frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy55 (LIB3049) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) young seed tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Seeds are collected from very young pods (5 to 15 days after flowering). A total of 100 mg of seeds are harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy56 (LIB3029) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are not converted to double stranded form and represent a non-normalized seed pool for comparison to Soy51 cDNA libraries.

The Soy58 (LIB3050) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed root tissue subtracted from control root tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days root tissue from both drought stressed and control (watered regularly) plants are collected and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

The Soy59 (LIB3051) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) endosperm tissue. Seeds are germinated on paper towels under laboratory ambient light conditions. At 8, 10 and 14 hours after imbibition, the seed coats are harvested. The endosperm consists of a very thin layer of tissue affixed to the inside of the seed coat. The seed coat and endosperm are frozen immediately after harvest in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Soy60 (LIB3072) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed seed plus pod subtracted from control seed plus pod tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

The Soy61 (LIB3073) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). For this library's construction, the eighth fraction of the cDNA size fractionation step was used for ligation.

The Soy62 (LIB3074) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. For subtraction, target cDNA is made from the j asmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). For this library's construction, the ninth fraction of the cDNA size fractionation step was used for ligation.

The Soy65 (LIB133107) 07cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought-stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At the R3 stage of development, drought is imposed by withholding water. At 3, 4, 5 and 6 days, tissue is harvested and wilting is not obvious until the fourth day. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Soy66 (LIB3109) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) non-drought stressed abscission zone tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Plants are irrigated with 15-16-17 Peter's Mix. At 3, 4, 5 and 6 days, control abscission layer tissue is harvested. Abscission layers from reproductive organs are harvested by cutting less than one millimeter proximal and distal to the layer and immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

Soy67 (LIB3065) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

Soy68 (LIB3052) cDNA library is prepared from equal amounts tissue harvested from SOYMON007, SOYMON015 and SOYMON020 prepared tissue. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. Captured hybrids are eluted with water.

Soy69 (LIB3053) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) normalized leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4th node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation. Single stranded and double stranded DNA representing approximately 1×106 colony forming units are isolated using standard protocols. RNA, complementary to the single stranded DNA, is synthesized using the double stranded DNA as a template. Biotinylated dATP is incorporated into the RNA during the synthesis reaction. The single stranded DNA is mixed with the biotinylated RNA in a 1:10 molar ratio) and allowed to hybridize. DNA-RNA hybrids are captured on Dynabeads M280 streptavidin (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The dynabeads with captured hybrids are collected with a magnet. The non-hybridized single stranded molecules remaining after hybrid capture are converted to double stranded form and represent the primary normalized library.

Soy70 (LIB3055) cDNA library is generated from soybean cultivars Cristalina (USDA Soybean Germplasm Collection, Urbana, Ill. U.S.A.) and FT108 (Monsoy, Brazil) (tropical germ plasma) leaf tissue. Leaves are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 30 g of leaves are harvested from the 4th node of each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

Soy71 (LIB3056) cDNA library is generated from soybean cultivars Cristalina and FT108 (tropical germ plasma) root tissue. Roots are harvested from plants grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 29° C. and the nighttime temperature approximately 24° C. Soil is checked and watered daily to maintain even moisture conditions. Approximately 50 g and 56 g of roots are harvested from each of the Cristalina and FT108 cultivars and immediately frozen in dry ice. The harvested tissue is then stored at −80° C. until RNA preparation.

Soy72 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed leaf control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

Soy73 (LIB3093) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) drought stressed leaf subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in an environmental chamber under 12 hr daytime/12 hr nighttime cycles. The daytime temperature is approximately 26° C. and the nighttime temperature 21° C. and 70% relative humidity. Soil is checked and watered daily to maintain even moisture conditions. At the R3 stage of the plant drought is induced by withholding water. After 3 and 6 days seeds and pods from both drought stressed and control (watered regularly) plants are collected from the fifth and sixth node and frozen in dry-ice. The harvested tissue is stored at −80° C. until RNA preparation. For subtraction, target cDNA is made from the drought stressed tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.).

The Soy76 (Lib3106) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid and arachidonic treated seedling subtracted from control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.) in order to capture some of the smaller transcripts characteristic of antifungal proteins.

Soy77 (LIB3108) cDNA library is generated from soybean cultivar Asgrow 3244 (Asgrow Seed Company, Des Moines, Iowa U.S.A.) jasmonic acid control tissue. Seeds are planted at a depth of approximately 2 cm into 2-3 inch peat pots containing Metromix 350 medium and the plants are grown in a greenhouse. The daytime temperature is approximately 29.4° C. and the nighttime temperature 20° C. Soil is checked and watered daily to maintain even moisture conditions. At 9 days post planting, the plantlets are sprayed with either control buffer of 0.1% Tween-20 or jasmonic acid (Sigma J-2500, Sigma, St. Louis, Mo. U.S.A.) at 1 mg/ml in 0.1% Tween-20. Plants are sprayed until runoff and the soil and the stem is socked with the spraying solution. At 18 hours post application of jasmonic acid, the soybean plantlets appear growth retarded. Arachidonic treated seedlings are sprayed with 1 m/ml arachidonic acid in 0.1% Tween-20. After 18 hours, 24 hours and 48 hours post treatment, the cotyledons are removed and the remaining leaf and stem tissue above the soil is harvested and frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation. To make RNA, the three sample timepoints were combined and ground. The RNA from the arachidonic treated seedlings is isolated separately. For subtraction, target cDNA is made from the jasmonic acid treated tissue total RNA using the SMART cDNA synthesis system from Clonetech (Clonetech Laboratories, Palo Alto, Calif. U.S.A.). Driver first strand cDNA is covalently linked to Dynabeads following a protocol similar to that described in the Dynal literature (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.). The target cDNA is then heat denatured and the second strand trapped using Dynabeads oligo-dT. The target second strand cDNA is then hybridized to the driver cDNA in 400 μl 2×SSPE for two rounds of hybridization at 65° C. and 20 hours. After each hybridization, the hybridization solution is removed from the system and the hybridized target cDNA removed from the driver by heat denaturation in water. After hybridization, the remaining cDNA is trapped with Dynabeads oligo-dT. The trapped cDNA is then amplified as in previous PCR based libraries and the resulting cDNA ligated into the pSPORT vector (Invitrogen, Carlsbad Calif. U.S.A.). Fraction 10 of the size fractionated cDNA is ligated into the pSPORT vector in order to capture some of the smaller transcripts characteristic of antifungal proteins.

The Lib9 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Leaf blades were cut with sharp scissors at seven weeks after planting. The tissue was immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)25 (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib22 cDNA library is prepared from Arabidopsis thaliana Columbia ecotype, root tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems, floral buds appear, and a few flowers are starting to open. The 7-week old plants are rinsed intensively by tope water remove dirt from the roots, and blotted by paper towel. The tissues are immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA preparation.

The Lib23 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, stem tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Stems were collected seven to eight weeks after planting by cutting the stems from the base and cutting the top of the plant to remove the floral tissue. The tissue was immediately frozen in liquid nitrogen and stored at −80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)25 (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib24 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, flower bud tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Flower buds are green and unopened and harvested about seven weeks after planting. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)25 (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib25 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, open flower tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Flowers are completely opened with all parts of floral structure observable, but no siliques are appearing. The tissue was immediately frozen in liquid nitrogen and stored at −80° C. until total RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)25 (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib35 cDNA library of the present invention, was prepared from Arabidopsis thaliana Columbia ecotype leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems and floral buds appeared and a few flowers were starting to open. Leaf blades were collected by cutting with sharp scissors. The tissues were immediately frozen in liquid nitrogen and stored at −80° C. until use. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)25 (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library was normalized using a PCR-based protocol.

The Lib146 cDNA library is prepared from Arabidopsis thaliana, Columbia ecotype, immature seed tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. At approximately 7-8 weeks of age, the seeds are harvested. The seeds ranged in maturity from the smallest seeds that could be dissected from silques to just before starting to turn yellow in color. The tissue is immediately frozen in liquid nitrogen. The harvested tissue is stored at −80° C. until RNA extraction. PolyA mRNA is purified from the total RNA preparation using Dynabeads® Oligo(dT)25 (Dynal Inc., Lake Success, N.Y.), or equivalent methods. This library is normalized using a PCR-based protocol.

The Lib3032 (Lib80) cDNA libraries are generated from Brassica napus seeds harvested 30 days after pollination. The cDNA libraries are constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA is used as the starting material for cDNA synthesis, and first strand cDNA synthesis is carried out at 45° C.

The Lib3034 (Lib82) cDNA libraries are generated from Brassica napus seeds harvested 15 and 18 days after pollination. The cDNA libraries are constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA is used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

The Lib3099 cDNA library is generated by a subtraction procedure. The library contains cDNAs whose abundance is enriched in the Brassica napus 15 and 18 day after pollination seed tissues when compared to Brassica leaf tissues. The cDNA synthesis is performed on Brassica leaf RNA and Brassica RNA isolated from seeds harvested 15 and 18 days after pollination using a Smart PCR cDNA synthesis kit according to the manufacturers protocol (Clontech, Palo Alto, Calif. U.S.A.). The subtracted cDNA is generated using the Clontech PCR-Select subtraction kit according to the manufacturers protocol (Clontech, Palo Alto, Calif. U.S.A.). The subtracted cDNA was cloned into plasmid vector pCR2.1 according to the manufacturers protocol (Invitrogen, Carlsbad, Calif. U.S.A.).

The Lib3033 (Lib81) cDNA libraries are generated from the Schizochytrium species cells. The Schizochytrium species cells are grown in liquid media until saturation. The culture is centrifuged to pellet the cells, the medium is decanted off, and pellet immediately frozen in liquid nitrogen. Wax esters are produced under such dark, anaerobic, rich-medium conditions. High wax production by the cultures is verified by microscopy (fluorescein staining of wax bodies) and by lipid extraction/TLC/GC. The harvested cells are stored at −80° C. until RNA preparation. RNA is prepared from the frozen Euglena cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

The Lib47 cDNA library is generated from Euglena gracilus strain 753 (ATTC No. 30285, ATCC Manasas, Va. U.S.A.) grown in liquid culture. A liquid culture is innoculated with 1/10 volume of a previously grown saturated culture, and the new culture for 4 days under near-anaerobic conditions (near-anaerobic cultures are not agitated, just gently swirled once a day) in the dark in 2× Beef (10 g/l bacto peptone, 4 g/l yeast extract, 2 g/l beef extract, 6 g/l glucose). The culture is then centrifuged to pellet the cells, the medium is decanted off, and pellet immediately frozen in liquid nitrogen. Wax esters are produced under such dark, anaerobic, rich-medium conditions. High wax production by the cultures is verified by microscopy (fluorescein staining of wax bodies) and by lipid extraction/TLC/GC. The harvested cells are stored at −80° C. until RNA preparation. RNA is prepared from the frozen Euglena cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45° C.

The Lib44 cDNA library is generated from Phaeodactylum tricornatum grown in modified Jones medium for 3 days. The cells were harvested by centrifugation and the resulting pellet frozen immediately in liquid nitrogen. The harvested cells are stored at −80° C. until RNA preparation. RNA is prepared from the frozen Phaeodactylum cell pellet as follows. The pellet is pulverized to a powder in liquid nitrogen using a mortar and pestle. The powder is transferred to tubes containing 6 ml each of lysis buffer (100 mM Tris, pH 8, 0.6 M NaCl, 10 mM EDTA, and 4% (w/v) SDS) and buffered phenol, vortexed, and disrupted with a Polytron. The mixture is centrifuged 20 min at 10,000×g in Corex glass tubes to separate the phases. 5 ml of the upper phase is removed, vortexed with 5 ml fresh phenol, and centrifuged. The upper phase is removed and the RNA is precipitated overnight at 4° C. by adding 1.5 volumes of 4 M LiCl. The RNA is further purified on Rneasy columns according to the manufacturers protocol (Qiagen, Valencia, Calif. U.S.A.). The cDNA library is constructed using the SuperScript Plasmid system for cDNA synthesis and plasmid cloning (Life Technologies, Gaithersburg, Md. U.S.A.) according to the manufacturers protocol with the following modification: 40 micrograms of total RNA was used as the starting material for cDNA synthesis, and first strand cDNA synthesis was carried out at 45 degrees centigrade.

The LIB3036 genomic library is generated from Mycobacterium neoaurum US52 (ATCC No. 23072, ATCC, Manasas, Va. U.S.A.) cells. Mycobacterium neoaurum US52 is a gram-positive Actinomycete bacterium. Mycobacterium neoaurum US52 is genetically related to Mycobacterium tuberculosis, but there is no reason to believe that it is a primary pathogen. It normally is saprophytic, i.e. it lives in soil and gets nutrients from decaying matter. Genomic DNA obtained from Mycobacterium neoaurum US52 is digested for various times with the restriction enzyme Sau3A. The DNA fractions are size-separated on an agarose gel, and the first fraction wherein most of the partially-digested fragments are about 10 kB is used to isolated fragments in the range of 2-3 kB. For LE33036, the 2-3 kB DNA is cloned into vector pRY401 (Invitrogen, Carlsbad, Calif. U.S.A.). The vector pZERO-2 (Invitrogen, Carlsbad, Calif. U.S.A.). is used for the construction of LIB3104.

The stored RNA is purified using Trizol reagent from Life Technologies (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.), essentially as recommended by the manufacturer. Poly A+ RNA (mRNA) is purified using magnetic oligo dT beads essentially as recommended by the manufacturer (Dynabeads, Dynal Corporation, Lake Success, New York U.S.A.).

Construction of plant cDNA libraries is well-known in the art and a number of cloning strategies exist. A number of cDNA library construction kits are commercially available. The Superscript™ Plasmid System for cDNA synthesis and Plasmid Cloning (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.) is used, following the conditions suggested by the manufacturer.

Normalized libraries are made using essentially the Soares procedure (Soares et al., Proc. Natl. Acad. Sci. (U.S.A.) 91:9228-9232 (1994), the entirety of which is herein incorporated by reference). This approach is designed to reduce the initial 10,000-fold variation in individual cDNA frequencies to achieve abundances within one order of magnitude while maintaining the overall sequence complexity of the library. In the normalization process, the prevalence of high-abundance cDNA clones decreases dramatically, clones with mid-level abundance are relatively unaffected and clones for rare transcripts are effectively increased in abundance.

Example 2

The cDNA libraries are plated on LB agar containing the appropriate antibiotics for selection and incubated at 37° for a sufficient time to allow the growth of individual colonies. Single colonies are individually placed in each well of a 96-well microtiter plates containing LB liquid including the selective antibiotics. The plates are incubated overnight at approximately 37° C. with gentle shaking to promote growth of the cultures. The plasmid DNA is isolated from each clone using Qiaprep plasmid isolation kits, using the conditions recommended by the manufacturer (Qiagen Inc., Santa Clara, Calif. U.S.A.).

Template plasmid DNA clones are used for subsequent sequencing. For sequencing, the ABI PRISM dRhodamine Terminator Cycle Sequencing Ready Reaction Kit with AmpliTaq® DNA Polymerase, FS, is used (PE Applied Biosystems, Foster City, Calif. U.S.A.).

Example 3

Nucleic acid sequences that encode for the following proteins: triose phosphate isomerase, fructose 1,6-bisphosphate aldolase, fructose 1,6-bisphosphate, fructose 6-phosphate 2-kinase, phosphoglucoisomerase, vacuolar H+ translocating-pyrophosphatase, pyrophosphate-dependent fructose-6-phosphate phosphotransferase, invertase, sucrose synthase, hexokinase, fructokinase, NDP-kinase, glucose-6-phosphate 1-dehydrogenase, phosphoglucomutase and UDP-glucose pyrophosphorylase are identified from the Monsanto EST PhytoSeq database using TBLASTN (default values) (TBLASTN compares a protein query against the six reading frames of a nucleic acid sequence). Matches found with BLAST P values equal or less than 0.001 (probability) or BLAST Score of equal or greater than 90 are classified as hits. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.

In addition, the GenBank database is searched with BLASTN and BLASTX (default values) using ESTs as queries. EST that pass the hit probability threshold of 10e−8 for the following enzymes are combined with the hits generated by using TBLASTN (described above) and classified by enzyme (see Table A below).

A cluster refers to a set of overlapping clones in the PhytoSeq database. Such an overlapping relationship among clones is designated as a “cluster” when BLAST scores from pairwise sequence comparisons of the member clones meets a predetermined minimum value or product score of 50 or more (Product Score=(BLAST SCORE×Percentage Identity)/(5×minimum [length (Seq1), length (Seq2)]))

Since clusters are formed on the basis of single-linkage relationships, it is possible for two non-overlapping clones to be members of the same cluster if, for instance, they both overlap a third clone with at least the predetermined minimum BLAST score (stringency). A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a “singleton”), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. Clones grouped in a cluster in most cases represent a contiguous sequence.

TABLE A*
Seq No.Cluster IDClone IDLibraryNCBI giMethodScoreP-value% Ident
MAIZE TRIOSE PHOSPHATE ISOMERASE
1−700019675700019675H1SATMON001g546735BLASTX1341e−1178
2−700073894700073894H1SATMON007g609261BLASTN2571e−1084
3−700167260700167260H1SATMON013g609261BLASTN6441e−4479
4−700380595700380595H1SATMON021g609261BLASTN11211e−8487
5−700449667700449667H1SATMON028g217973BLASTN2041e−1893
6−700449720700449720H2SATMON028g217973BLASTN2161e−1888
7−700570661700570661H1SATMON030g168647BLASTX1311e−1188
8−700616770700616770H1SATMON033g407525BLASTX1491e−1383
9−701170944701170944H1SATMONN05g217921BLASTX1881e−2053
1011337700337974H1SATMON020g256119BLASTN5351e−6178
1111337700027829H1SATMON003g256119BLASTN7261e−5180
12126700050046H1SATMON003g1785947BLASTN4401e−2692
13282700077320H1SATMON007g217973BLASTN6661e−10897
14282700104541H1SATMON010g217973BLASTN6311e−10697
15282700047476H1SATMON003g217973BLASTN6481e−10597
16282700211559H1SATMON016g217973BLASTN5251e−10497
17282700073553H1SATMON007g217973BLASTN9811e−10398
18282700613011H1SATMON033g217973BLASTN5521e−10298
19282700352119H1SATMON023g217973BLASTN6661e−10197
20282700088148H1SATMON011g217973BLASTN6661e−10098
21282700351626H1SATMON023g217973BLASTN4011e−9998
22282700240096H1SATMON010g217973BLASTN6661e−9897
23282700083660H1SATMON011g217973BLASTN6661e−9799
24282700208721H1SATMON016g217973BLASTN4971e−9698
25282700203144H1SATMON003g217973BLASTN5111e−9696
26282700430425H1SATMONN01g217973BLASTN6661e−9698
27282700206091H1SATMON003g217973BLASTN4971e−9497
28282700077017H1SATMON007g217973BLASTN6141e−9393
29282700618792H1SATMON034g217973BLASTN5461e−9296
30282700572532H1SATMON030g407524BLASTN12121e−9284
31282700106512H1SATMON010g217973BLASTN6321e−9197
32282700195031H1SATMON014g217973BLASTN4711e−9097
33282700168131H1SATMON013g217973BLASTN4971e−8998
34282700197039H1SATMON014g217973BLASTN5461e−8998
35282700572688H1SATMON030g169820BLASTN11141e−8985
36282700021313H1SATMON001g217973BLASTN9131e−8797
37282700452417H1SATMON028g217973BLASTN4251e−8695
38282700346119H1SATMON021g217973BLASTN4441e−8696
39282700082359H1SATMON011g217973BLASTN5421e−8693
40282700240042H1SATMON010g217973BLASTN5961e−8697
41282700030064H1SATMON003g217973BLASTN5871e−8594
42282700615185H1SATMON033g217973BLASTN4301e−8498
43282700196125H1SATMON014g217973BLASTN5811e−84100
44282700243429H1SATMON010g217973BLASTN6321e−8497
45282700474112H1SATMON025g217973BLASTN5701e−8398
46282700572282H1SATMON030g407524BLASTN8381e−8382
47282700622238H1SATMON034g169820BLASTN9171e−8086
48282700095609H1SATMON008g169820BLASTN10671e−8082
49282700218886H1SATMON011g217973BLASTN5511e−7993
50282700018688H1SATMON001g217973BLASTN10661e−7999
51282700049775H1SATMON003g217973BLASTN3621e−7891
52282700575972H1SATMON030g169820BLASTN8941e−7879
53282700215519H1SATMON016g217973BLASTN4971e−7697
54282700161120H1SATMON012g217973BLASTN6221e−7698
55282700581760H1SATMON031g217973BLASTN5331e−7590
56282700104672H1SATMON010g169820BLASTN10121e−7583
57282700346053H1SATMON021g169820BLASTN10121e−7583
58282701166592H1SATMONN04g217973BLASTN6611e−7495
59282700968667H1SATMONN04g217973BLASTN4971e−7392
60282700205627H1SATMON003g217973BLASTN6661e−7399
61282700029005H1SATMON003g169820BLASTN9791e−7285
62282700476479H1SATMON025g169820BLASTN5541e−7184
63282700050148H1SATMON003g169820BLASTN6081e−7083
64282700259846H1SATMON017g217973BLASTN2831e−6994
65282700344093H1SATMON021g169820BLASTN9341e−6983
66282700082327H1SATMON011g169820BLASTN9431e−6985
67282700020156H1SATMON001g217973BLASTN4201e−6899
68282700577714H1SATMON031g169820BLASTN9281e−6885
69282700104904H1SATMON010g169820BLASTN9131e−6784
70282700104685H1SATMON010g169820BLASTN8971e−6684
71282700053463H1SATMON009g169820BLASTN9071e−6685
72282700171639H1SATMON013g217973BLASTN4011e−6598
73282700574233H1SATMON030g169820BLASTN6511e−6583
74282700262653H1SATMON017g169820BLASTN8771e−6484
75282700456738H1SATMON029g169820BLASTN8771e−6484
76282700611806H1SATMON022g169820BLASTN8771e−6483
77282700381177H1SATMON023g169820BLASTN8841e−6484
78282700103347H1SATMON010g169820BLASTN8611e−6384
79282700103605H1SATMON010g169820BLASTN8681e−6384
80282700578536H1SATMON031g169820BLASTN8561e−6284
81282700258606H1SATMON017g169820BLASTN8071e−6183
82282700335703H1SATMON019g217973BLASTN3761e−6090
83282700351044H1SATMON023g169820BLASTN4711e−5983
84282700346364H1SATMON021g169820BLASTN8131e−5985
85282700619037H1SATMON034g169820BLASTN8141e−5984
86282700465160H1SATMON025g169820BLASTN7511e−5784
87282700235687H1SATMON010g169820BLASTN7911e−5782
88282700105645H1SATMON010g169820BLASTN7931e−5783
89282700082237H1SATMON011g169820BLASTN7931e−5784
90282700261906H1SATMON017g169820BLASTN7961e−5783
91282700456154H1SATMON029g169820BLASTN7991e−5784
92282700047696H1SATMON003g169820BLASTN5611e−5683
93282700449905H1SATMON028g169820BLASTN7881e−5684
94282700336106H1SATMON019g217973BLASTN3251e−5592
95282700381867H1SATMON023g2529386BLASTN4221e−5597
96282700051335H1SATMON003g169820BLASTN6081e−5583
97282700050988H1SATMON003g169820BLASTN7681e−5586
98282700029471H1SATMON003g169820BLASTN7721e−5584
99282700106806H1SATMON010g169820BLASTN7731e−5584
100282700071749H1SATMON007g217973BLASTN3621e−5485
101282700207607H1SATMON016g217973BLASTN3621e−5485
102282700573465H2SATMON030g169820BLASTN7531e−5486
103282700220908H1SATMON011g169820BLASTN7581e−5484
104282700467719H1SATMON025g169820BLASTN7611e−5485
105282700456018H1SATMON029g169820BLASTN7641e−5481
106282700453767H1SATMON029g217973BLASTN2961e−5294
107282700026118H1SATMON003g217973BLASTN3411e−5293
108282700026760H1SATMON003g217973BLASTN4211e−5299
109282700029525H1SATMON003g169820BLASTN7381e−5285
110282700457972H1SATMON029g169820BLASTN7231e−5185
111282700455866H1SATMON029g169820BLASTN7261e−5184
112282700165290H1SATMON013g169820BLASTN7261e−5184
113282700351190H1SATMON023g169820BLASTN6721e−5081
114282700154095H1SATMON007g169820BLASTN6961e−4984
115282700450438H1SATMON028g217973BLASTN4301e−4899
116282700044892H1SATMON004g169820BLASTN6831e−4885
117282700185095H1SATMON014g169820BLASTN6731e−4784
118282700575506H1SATMON030g169820BLASTN6801e−4783
119282700161966H1SATMON012g217973BLASTN3351e−4698
120282700343401H1SATMON021g169820BLASTN4261e−4577
121282700152354H1SATMON007g169820BLASTN6531e−4584
122282701164924H1SATMONN04g169820BLASTN3971e−4484
123282700346896H1SATMON021g169820BLASTN4961e−4284
124282700210157H1SATMON016g169820BLASTN6171e−4284
125282700383103H1SATMON024g169820BLASTN5311e−4184
126282701158829H1SATMONN04g407524BLASTN5491e−4080
127282700619883H1SATMON034g217973BLASTN3251e−3899
128282700168219H1SATMON013g169820BLASTN5401e−3683
129282700155210H1SATMON007g169820BLASTN5451e−3683
130282700334861H1SATMON019g169820BLASTN4841e−3182
131282700355663H1SATMON024g217973BLASTN2131e−3088
132282700074764H1SATMON007g546734BLASTN3871e−2784
133282700621934H1SATMON034g217973BLASTN4301e−26100
134282700802084H1SATMON036g217973BLASTN2701e−2498
1353039700620444H1SATMON034g1785947BLASTN4731e−5675
1363039700356205H1SATMON024g1785947BLASTN3321e−3272
1373039700215549H1SATMON016g414549BLASTN4431e−2672
1383039700620318H1SATMON034g556171BLASTX2141e−2579
1393039700028742H1SATMON003g556171BLASTX1561e−2086
1403039700150060H1SATMON007g556171BLASTX1811e−1789
1413039700448477H1SATMON027g556171BLASTX1371e−1285
1423039700336489H1SATMON019g556171BLASTX1261e−1081
1433414700099709H1SATMON009g609261BLASTN6001e−4984
1443414700075837H1SATMON007g609261BLASTN4941e−4184
1453414700045678H1SATMON004g609261BLASTN3401e−2973
1463414700097852H1SATMON009g609261BLASTN4361e−2784
1473414700053342H1SATMON009g609261BLASTN3461e−2573
1483414700041954H1SATMON004g609261BLASTN3401e−2482
1493414700217471H1SATMON016g609261BLASTN2651e−2171
1503414700264437H1SATMON017g609261BLASTN2311e−1769
1513414700218371H1SATMON016g609261BLASTN1561e−1068
1525593700381686H1SATMON023g609261BLASTN5341e−4489
1535593700356082H1SATMON024g609261BLASTN2461e−2490
1545593700622077H1SATMON034g609261BLASTN2921e−2086
1555593700470822H1SATMON025g609262BLASTX1341e−1179
1566525700083139H1SATMON011g256119BLASTN8801e−6476
1576525700205474H1SATMON003g169820BLASTN8491e−6277
1586991700336856H1SATMON019g609261BLASTN11311e−8585
1596991700042717H1SATMON004g609261BLASTN10281e−7685
1606991700379491H1SATMON020g609261BLASTN9951e−7481
1616991700156635H1SATMON012g609261BLASTN8771e−6484
1626991700046340H1SATMON004g609261BLASTN8521e−6284
1636991700081869H1SATMON011g609261BLASTN2661e−1480
1646991700426102H1SATMONN01g806312BLASTX1341e−1389
1657384700613626H1SATMON033g609261BLASTN9201e−8785
1667384700101506H1SATMON009g609261BLASTN11241e−8485
1677384700206445H1SATMON003g609261BLASTN9871e−7379
1687384700220160H1SATMON011g609261BLASTN8781e−6485
169-L1431527LIB143-004-LIB143g217973BLASTN2901e−1393
Q1-E1-C5
170-L30613868LIB3061-017-LIB3061g217973BLASTN1821e−1370
Q1-K1-C9
171-L30623620LIB3062-034-LIB3062g609261BLASTN5991e−3974
Q1-K1-A8
172-L361705LIB36-021-LIB36g609261BLASTN2661e−1480
Q1-E1-E7
17323992LIB3062-056-LIB3062g1200507BLASTX2851e−6461
Q1-K1-F9
174282LIB3067-047-LIB3067g217973BLASTN10761e−16496
Q1-K1-H2
175282LIB3067-055-LIB3067g217973BLASTN10761e−13393
Q1-K1-G8
176282LIB3067-059-LIB3067g169820BLASTN14011e−11584
Q1-K1-D10
177282LIB3067-027-LIB3067g407524BLASTN9951e−11383
Q1-K1-B10
178282LIB189-032-LIB189g217973BLASTN6291e−11193
Q1-E1-H2
179282LIB3059-023-LIB3059g407524BLASTN14361e−11183
Q1-K1-A7
180282LIB3069-016-LIB3069g169820BLASTN13011e−10781
Q1-K1-D9
181282LIB143-006-LIB143g169820BLASTN13731e−10584
Q1-E1-A8
182282LIB3068-054-LIB3068g169820BLASTN13271e−10282
Q1-K1-C11
183282LIB3067-034-LIB3067g407524BLASTN13211e−10183
Q1-K1-B7
184282LIB143-031-LIB143g169820BLASTN13111e−10084
Q1-E1-E5
185282LIB3069-055-LIB3069g169820BLASTN10461e−9775
Q1-K1-H12
186282LIB3061-027-LIB3061g169820BLASTN9361e−9683
Q1-K1-A8
187282LIB3078-008-LIB3078g169820BLASTN12101e−9282
Q1-K1-E5
188282LIB3066-027-LIB3066g407524BLASTN11961e−9182
Q1-K1-E1
189282LIB3067-032-LIB3067g169820BLASTN11221e−8484
Q1-K1-E5
190282LIB3078-029-LIB3078g169820BLASTN8271e−8382
Q1-K1-F7
191282LIB3061-006-LIB3061g169820BLASTN10911e−8278
Q1-K1-B7
192282LIB143-048-LIB143g169820BLASTN6441e−7475
Q1-E1-F8
193282LIB3078-033-LIB3078g169820BLASTN5841e−7379
Q1-K1-B10
194282LIB3069-046-LIB3069g169820BLASTN8191e−5979
Q1-K1-C4
195282LIB3061-049-LIB3061g169820BLASTN5871e−4780
Q1-K1-H2
196282LIB143-029-LIB143g169820BLASTN6791e−4784
Q1-E1-G4
197282LIB84-027-LIB84g169820BLASTN6131e−4678
Q1-E1-E5
198282LIB3062-001-LIB3062g169820BLASTN5071e−3380
Q1-K2-F7
199282LIB3066-014-LIB3066g169820BLASTN3851e−2576
Q1-K1-H11
20029645LIB3069-014-LIB3069g168647BLASTX1311e−2734
Q1-K1-C11
20129645LIB3069-013-LIB3069g168647BLASTX1241e−2433
Q1-K1-C11
2023039LIB3062-045-LIB3062g1785947BLASTN11191e−8472
Q1-K1-F6
2035593LIB3067-045-LIB3067g609261BLASTN7021e−5875
Q1-K1-E5
2046991LIB3059-026-LIB3059g609261BLASTN14931e−11584
Q1-K1-G9
2056991LIB3078-049-LIB3078g609261BLASTN7471e−5583
Q1-K1-E4
2067384LIB3062-034-LIB3062g609261BLASTN13511e−10785
Q1-K1-A4
MAIZE FRUCTOSE 1,6-BISPHOSPHATE ALDOLASE
207−700026544700026544H1SATMON003g22144BLASTN2151e−3088
208−700073329700073329H1SATMON007g22144BLASTN5901e−8995
209−700151987700151987H1SATMON007g22144BLASTN2121e−878
210−700206575700206575H1SATMON003g22144BLASTN10091e−10996
211−700333727700333727H1SATMON019g1217893BLASTX1541e−1661
212−700429795700429795H1SATMONN01g1619605BLASTX1021e−1677
213−700804137700804137H1SATMON036g22144BLASTN7421e−5292
2141182700449930H1SATMON028g22632BLASTN8561e−6279
2151182701185559H1SATMONN06g22632BLASTN7931e−5779
2161182700203130H1SATMON003g22632BLASTN7991e−5778
2171182700083459H1SATMON011g22632BLASTN8001e−5776
2181182700465449H1SATMON025g22632BLASTN4051e−5076
2191182701165344H1SATMONN04g22632BLASTN3261e−2978
2201182700427538H1SATMONN01g438275BLASTX961e−988
22138700224356H1SATMON011g22144BLASTN12901e−9896
22238700048169H1SATMON003g22144BLASTN5281e−7298
22338700616610H1SATMON033g22144BLASTN2781e−3191
22438700355765H1SATMON024g20204BLASTX1411e−1296
2256547700194431H1SATMON014g2636513BLASTX1811e−1747
2266547700469777H1SATMON025g2636513BLASTX1741e−1648
2278494700425929H1SATMONN01g927507BLASTX671e−1189
228-L30603643LIB3060-046-LIB3060g169037BLASTX1551e−4466
Q1-K1-G7
2291182LIB3079-006-LIB3079g22632BLASTN5981e−3965
Q1-K1-H8
23028633LIB3062-015-LIB3062g1208898BLASTX1161e−2445
Q1-K1-G12
23138LIB3061-025-LIB3061g22144BLASTN8951e−13394
Q1-K1-C9
23238LIB3059-020-LIB3059g22144BLASTN7451e−5398
Q1-K1-H3
MAIZE FRUCTOSE-1,6-BISPHOSPHATASE
233−700262935700262935H1SATMON017g3041775BLASTX1841e−1894
234−700432173700432173H1SATMONN01g1790679BLASTX1231e−1656
235−700455709700455709H1SATMON029g3041776BLASTN5971e−4085
236−700573083700573083H1SATMON030g3041775BLASTX691e−1064
23712846700101851H1SATMON009g3041776BLASTN13121e−10091
23812846700101541H1SATMON009g3041776BLASTN12521e−9590
23912846700581510H1SATMON031g3041776BLASTN8721e−8290
24015627700046054H1SATMON004g21736BLASTN12131e−9291
24115627700421605H1SATMONN01g3041776BLASTN6641e−7790
24215627700445495H1SATMON027g21736BLASTN10041e−7484
24315627700042188H1SATMON004g3041776BLASTN8751e−6488
24416870700100752H1SATMON009g3041776BLASTN2571e−3375
24516870700044805H1SATMON004g3041776BLASTN1941e−1476
24616870700099217H1SATMON009g21736BLASTN2461e−959
2475480700442189H1SATMON026g3041774BLASTN5361e−5493
2488243700264654H1SATMON017g3041774BLASTN9421e−6984
2498243700479624H1SATMON034g3041774BLASTN9021e−6682
2508243700448974H1SATMON028g3041774BLASTN8761e−6484
251-L1485381LIB148-057-LIB148g440591BLASTX801e−3063
Q1-E1-E6
252-L30662839LIB3066-035-LIB3066g3041774BLASTN2151e−1577
Q1-K1-F11
253-L362913LIB36-013-LIB36g3041776BLASTN9371e−6988
Q1-E1-D10
254-L832444LIB83-005-LIB83g3041776BLASTN5751e−3793
Q1-E1-D2
25512846LIB83-008-LIB83g3041776BLASTN16101e−13592
Q1-E1-A8
25612846LIB3078-003-LIB3078g3041776BLASTN8731e−9893
Q1-K1-C7
25716870LIB3060-052-LIB3060g21736BLASTN3771e−6670
Q1-K1-D11
25826002LIB83-008-LIB83g3041776BLASTN3781e−2086
Q1-E1-B10
MAIZE FRUCTOSE-6-PHOSPHATE,2-KINASE
259−700093724700093724H1SATMON008g3170230BLASTX1231e−2153
260−700099547700099547H1SATMON009g3309582BLASTN6301e−4380
261−700100682700100682H1SATMON009g3170230BLASTX2691e−3965
262−700173085700173085H1SATMON013g2286154BLASTN11651e−88100
263−700217623700217623H1SATMON016g3170229BLASTN5931e−4073
264−700219340700219340H1SATMON011g3170230BLASTX1901e−2056
265−700265353700265353H1SATMON017g2286154BLASTN12681e−10798
266−700379777700379777H1SATMON021g3309582BLASTN9051e−6676
267−700620963700620963H1SATMON034g2286154BLASTN3761e−5285
268−701159590701159590H1SATMONN04g3309582BLASTN6821e−4873
26920094700209789H1SATMON016g2286154BLASTN10931e−9692
27020094700550375H1SATMON022g3309582BLASTN7801e−5881
27129193700021150H1SATMON001g2286154BLASTN4661e−7592
272-L30593297LIB3059-029-LIB3059g2286154BLASTN4011e−2270
Q1-K1-B3
273-L30614892LIB3061-021-LIB3061g2286154BLASTN4691e−3879
Q1-K1-G9
274-L30623700LIB3062-031-LIB3062g3170229BLASTN2301e−1070
Q1-K1-E8
27529193LIB83-007-LIB83g2286154BLASTN5951e−11390
Q1-E1-C11
MAIZE PHOSPHOGLUCOISOMERASE
276−700086021700086021H1SATMON011g1100771BLASTX2251e−2851
277−700169489700169489H1SATMON013g1100771BLASTX1521e−1359
278−700222638700222638H1SATMON011g1100771BLASTX2561e−2860
279−700445574700445574H1SATMON027g1100771BLASTX1431e−1254
280−700475232700475232H1SATMON025g596022BLASTN8451e−6190
281−700612774700612774H1SATMON033g596022BLASTN15741e−12295
28214393700222547H1SATMON011g1100771BLASTX2391e−2560
28314393700220357H1SATMON011g1100771BLASTX2181e−2368
28414393700050317H1SATMON003g1100771BLASTX1201e−2263
28514393700163544H1SATMON013g1100771BLASTX2141e−2262
28615724700207164H1SATMON017g1100771BLASTX1351e−1767
28715724700552402H1SATMON022g1100771BLASTX1351e−1160
28815724700086085H1SATMON011g1100771BLASTX1371e−1145
28920643700577051H1SATMON031g1100771BLASTX2411e−2666
29020643700201592H1SATMON003g1100771BLASTX1131e−1945
29120643700576644H1SATMON030g1100771BLASTX1131e−1743
2922351700208928H1SATMON016g1100771BLASTX2741e−4373
2932351700240758H1SATMON010g1100771BLASTX2831e−4379
2942351700352502H1SATMON023g1100771BLASTX1971e−3670
2952351700581930H1SATMON031g1100771BLASTX1641e−3472
2962351700028642H1SATMON003g1100771BLASTX2941e−3365
2972351700106092H1SATMON010g1100771BLASTX2941e−3362
2982351700082102H1SATMON011g1100771BLASTX3001e−3362
2992351700083446H1SATMON011g1100771BLASTX2741e−3065
3002351700580585H1SATMON031g1100771BLASTX1631e−2969
3012351700550608H1SATMON022g1100771BLASTX2651e−2961
3022351700106079H1SATMON010g1100771BLASTX2611e−2854
3032351700244248H1SATMON010g1100771BLASTX2381e−2567
3042351700152233H1SATMON007g1100771BLASTX1671e−2272
3052351700455043H1SATMON029g1100771BLASTX1681e−2168
3062351700615809H1SATMON033g1100771BLASTX2071e−2166
3072351701165320H1SATMONN04g1100771BLASTX1221e−1463
30832930700042996H1SATMON004g596022BLASTN4761e−9598
3094222700222539H1SATMON011g596022BLASTN11601e−87100
3104222700104023H1SATMON010g596022BLASTN10601e−84100
3114222700101580H1SATMON009g596022BLASTN8711e−7499
3124222700473395H1SATMON025g596022BLASTN3681e−4695
3134222700800179H1SATMON036g596022BLASTN2401e−11100
3148858700221523H1SATMON011g1100771BLASTX2781e−3159
315895700100965H1SATMON009g596022BLASTN16111e−12599
316895700620985H1SATMON034g596022BLASTN14181e−11498
317895700082062H1SATMON011g596022BLASTN13651e−11097
318895700573782H1SATMON030g596022BLASTN9201e−10798
319895700236138H1SATMON010g596022BLASTN13951e−107100
320895700086336H1SATMON011g596022BLASTN13701e−105100
321895700801467H1SATMON036g596022BLASTN12491e−9995
322895700801458H1SATMON036g596022BLASTN12451e−98100
323895700475024H1SATMON025g596022BLASTN11621e−9793
324895700243164H1SATMON010g596022BLASTN11051e−96100
325895700804665H1SATMON036g596022BLASTN12661e−9699
326895700021931H1SATMON001g596022BLASTN11261e−8499
327895700805540H1SATMON036g596022BLASTN7761e−5599
328895700172576H1SATMON013g596022BLASTN5711e−3898
329895700105116H1SATMON010g596022BLASTN5581e−3799
330895700472931H1SATMON025g596022BLASTN3791e−3197
33120643LIB3069-009-LIB3069g1100771BLASTX2151e−4450
Q1-K1-B3
3322351LIB3079-007-LIB3079g1100771BLASTX3041e−7772
Q1-K1-C11
33332930LIB189-001-LIB189g596022BLASTN7941e−11595
Q1-E1-E4
3344222LIB3079-001-LIB3079g596022BLASTN11321e−10189
Q1-K1-H7
335895LIB148-049-LIB148g596022BLASTN21941e−17897
Q1-E1-D6
336895LIB3066-052-LIB3066g596022BLASTN21781e−17297
Q1-K1-G8
337895LIB148-016-LIB148g596022BLASTN15671e−16199
Q1-E1-G5
338895LIB143-032-LIB143g596022BLASTN19141e−15599
Q1-E1-E10
339895LIB3061-013-LIB3061g596022BLASTN17381e−13688
Q1-K1-F7
340895LIB143-047-LIB143g596022BLASTN14901e−11988
Q1-E1-D4
MAIZE VACUOLAR H+-TRANSLOCATING-PYROPHOSPHATASE
341−700163331700163331H1SATMON0132g534915BLASTN7511e−5377
342−700171438700171438H1SATMON013g2258073BLASTN2561e−1076
343−700202576700202576H1SATMON003g2668746BLASTX2141e−2384
344−700206487700206487H1SATMON003g2570501BLASTX1741e−1786
345−700217292700217292H1SATMON016g2668746BLASTX2141e−23100
346−700240889700240889H1SATMON010g2570500BLASTN6391e−4784
347−700347658700347658H1SATMON023g2668746BLASTX2151e−2395
348−700454151700454151H1SATMON029g2668745BLASTN1721e−1090
349−700454532700454532H1SATMON029g2668745BLASTN2591e−3893
350−700552133700552133H1SATMON022g457744BLASTX1761e−1968
351−700611864700611864H1SATMON022g2668745BLASTN2031e−984
352107700622451H1SATMON034g2668745BLASTN16451e−129100
353107700571235H1SATMON030g2668745BLASTN14061e−12598
354107700266126H1SATMON017g2668745BLASTN11451e−121100
355107700621607H1SATMON034g2668745BLASTN13751e−12199
356107700345080H1SATMON021g2668745BLASTN11951e−117100
357107700624257H1SATMON034g2668745BLASTN8251e−115100
358107700030359H1SATMON003g2668745BLASTN14701e−114100
359107700214462H1SATMON016g2668745BLASTN12231e−11098
360107700356050H1SATMON024g2668745BLASTN14301e−110100
361107701181128H1SATMONN06g2668745BLASTN13681e−10598
362107700349795H1SATMON023g2668745BLASTN13701e−10595
363107700473278H1SATMON025g2668745BLASTN13551e−104100
364107700157057H1SATMON012g2668745BLASTN13451e−103100
365107700622505H1SATMON034g2668745BLASTN7621e−10096
366107700219661H1SATMON011g2668745BLASTN9421e−9899
367107700619032H1SATMON034g2668745BLASTN9891e−9896
368107700620065H1SATMON034g2668745BLASTN10691e−9894
369107700569179H1SATMON030g2668745BLASTN12331e−9798
370107700156773H1SATMON012g2668745BLASTN12761e−9799
371107700207120H1SATMON017g2668745BLASTN7401e−9699
372107700030407H1SATMON003g2668745BLASTN4801e−9598
373107700457309H1SATMON029g2668745BLASTN9791e−9599
374107700195681H1SATMON014g2668745BLASTN12461e−9599
375107700444838H1SATMON027g2668745BLASTN12491e−9596
376107700581619H1SATMON031g2668745BLASTN9431e−9496
377107700351021H1SATMON023g2668745BLASTN8531e−9192
378107700205723H1SATMON003g2668745BLASTN11381e−9195
379107700159712H1SATMON012g2668745BLASTN11991e−9194
380107700158937H1SATMON012g2668745BLASTN11321e−9096
381107700336255H1SATMON019g2668745BLASTN4891e−8594
382107700422922H1SATMONN01g2668745BLASTN6421e−8495
383107700347429H1SATMON023g2668745BLASTN8911e−8392
384107700350695H1SATMON023g2668745BLASTN9601e−8391
385107700212988H1SATMON016g2668745BLASTN9881e−8296
386107700345278H1SATMON021g2668745BLASTN9891e−8295
387107700264475H1SATMON017g2668745BLASTN10891e−8299
388107700211923H1SATMON016g2668745BLASTN9911e−8194
389107700620974H1SATMON034g2668745BLASTN9071e−8092
390107700156401H1SATMON012g2668745BLASTN10581e−7990
391107700172547H1SATMON013g2668745BLASTN10421e−7896
392107700552384H1SATMON022g2668745BLASTN9161e−7696
393107700219926H1SATMON011g2668745BLASTN10051e−75100
394107700357492H1SATMON024g2668745BLASTN6101e−7499
395107700343365H1SATMON021g2668745BLASTN8911e−7494
396107700018618H1SATMON001g2668745BLASTN10011e−7493
397107700570755H1SATMON030g2668745BLASTN8451e−7193
398107700194777H1SATMON014g2668745BLASTN9401e−69100
399107700453790H1SATMON029g2668745BLASTN9251e−6892
400107700197306H1SATMON014g2668745BLASTN9281e−6885
401107700355750H1SATMON024g2668745BLASTN3931e−6693
402107700172940H1SATMON013g2668745BLASTN9021e−6697
403107700102133H1SATMON010g2668745BLASTN8501e−62100
404107700350332H1SATMON023g2668745BLASTN5391e−5797
405107700450285H1SATMON028g2668745BLASTN7501e−53100
406107700165003H1SATMON013g2668745BLASTN5481e−5283
407107700016136H1SATMON001g2668745BLASTN5271e−5085
408107700171557H1SATMON013g2668745BLASTN7141e−5095
409107700238156H1SATMON010g2668745BLASTN7151e−5096
410107700425175H1SATMONN01g2668745BLASTN6981e−4994
411107700354402H1SATMON024g2668745BLASTN6161e−4891
412107700159204H1SATMON012g2668745BLASTN6171e−4294
413107700623602H1SATMON034g2668745BLASTN4601e−38100
414107700612844H1SATMON033g2668745BLASTN4211e−3684
415107700621062H2SATMON034g2668745BLASTN2851e−2589
416107700335685H1SATMON019g2668745BLASTN3391e−2591
41713843700334949H1SATMON019g2570500BLASTN6801e−5583
41813843700346817H1SATMON021g2570500BLASTN7051e−5483
41913843700103380H1SATMON010g2570500BLASTN7101e−5483
42013843700348280H1SATMON023g2570500BLASTN6691e−5183
42113843700453203H1SATMON028g2570500BLASTN6591e−5082
42213843700381101H1SATMON023g2570500BLASTN6211e−4782
42313843700347617H1SATMON023g2570500BLASTN5921e−4485
42413843700043259H1SATMON004g2570500BLASTN5301e−3984
42513843701184447H1SATMONN06g2570500BLASTN4811e−3578
42621076700241354H1SATMON010g166634BLASTX2011e−2058
42724066700423113H1SATMONN01g457744BLASTX1241e−2354
42824266700577157H1SATMON031g2570500BLASTN10011e−7489
4292531700099364H1SATMON009g2570500BLASTN6691e−5186
4302531700336387H1SATMON019g2570500BLASTN3891e−4785
4312531700217095H1SATMON016g2570500BLASTN4511e−3386
4322531700155869H1SATMON007g2570500BLASTN3851e−2789
4332531700575534H1SATMON030g2570500BLASTN3651e−2688
4342531700163562H1SATMON013g2570501BLASTX1451e−2494
43532364700204306H1SATMON003g2668745BLASTN4711e−2874
43632856700166756H1SATMON013g534915BLASTN7441e−5376
43732856700042535H1SATMON004g534915BLASTN6441e−4473
4383384700237775H1SATMON010g2258073BLASTN9111e−6781
4393384700342456H1SATMON021g2258073BLASTN6481e−6478
4403384700073654H1SATMON007g2668745BLASTN8601e−6378
4413384700577805H1SATMON031g2258073BLASTN8401e−6178
4423384700028881H1SATMON003g534915BLASTN8351e−6078
4433384700215076H1SATMON016g534915BLASTN8241e−5978
4443384700017479H1SATMON001g534915BLASTN7661e−5580
4453384700204495H1SATMON003g534915BLASTN3731e−5181
4463384700206347H1SATMON003g2706449BLASTN6851e−4880
4473384700351040H1SATMON023g2706449BLASTN4361e−4578
4483384700345264H1SATMON021g2706449BLASTN6161e−4282
4493384700196795H1SATMON014g2570500BLASTN5791e−3980
4503384700019241H1SATMON001g2706449BLASTN5831e−3978
4513384700018612H1SATMON001g2668745BLASTN5181e−3476
4523384700102142H1SATMON010g2668745BLASTN5391e−3478
4533384700348430H1SATMON023g534915BLASTN4891e−3078
4543384700337745H1SATMON020g2706449BLASTN4711e−2879
4553384700439515H1SATMON026g534915BLASTN4371e−2775
4563384700074977H1SATMON007g534915BLASTN4341e−2576
4573384700615213H1SATMON033g2570501BLASTX1251e−2193
4583384700074109H1SATMON007g2668746BLASTX1971e−2072
4593384700549517H1SATMON022g2668746BLASTX1721e−1775
4603384700030347H1SATMON003g2668746BLASTX1711e−1677
4613384700221176H1SATMON011g2668746BLASTX1711e−1677
4623384700433360H1SATMONN01g2668746BLASTX951e−1374
4635000700026151H1SATMON003g2903BLASTX2611e−2854
4645000700347165H1SATMON021g2624379BLASTX2231e−2451
4655000700430341H1SATMONN01g2903BLASTX1851e−1856
4665000700457781H1SATMON029g2903BLASTX1331e−1649
4675861700104993H1SATMON010g2258073BLASTN4561e−2773
4685861700203452H1SATMON003g2258073BLASTN4281e−2672
469-L1431590LIB143-006-LIB143g16347BLASTN2861e−1361
Q1-E1-C9
470-L1433414LIB143-026-LIB143g2258073BLASTN4801e−2970
Q1-E1-C3
471-L1482832LIB148-009-LIB148g2258073BLASTN10861e−8178
Q1-E1-D8
472-L30674379LIB3067-042-LIB3067g2668745BLASTN3051e−2168
Q1-K1-H8
473-L30675678LIB3067-034-LIB3067g2706449BLASTN2861e−1273
Q1-K1-E3
474107LIB3059-036-LIB3059g2668745BLASTN19651e−166100
Q1-K1-B10
475107LIB3061-035-LIB3061g2668745BLASTN9481e−13893
Q1-K1-C9
476107LIB3061-032-LIB3061g2668745BLASTN16851e−13896
Q1-K1-A12
477107LIB3062-044-LIB3062g2668745BLASTN14921e−13495
Q1-K1-F8
478107LIB3068-025-LIB3068g2668745BLASTN16871e−13296
Q1-K1-E5
479107LIB3067-022-LIB3067g2668745BLASTN15811e−12891
Q1-K1-D11
480107LIB3067-016-LIB3067g2668745BLASTN13051e−12697
Q1-K1-G4
481107LIB3067-029-LIB3067g2668745BLASTN15601e−12590
Q1-K1-C6
482107LIB189-031-LIB189g2668745BLASTN8971e−8185
Q1-E1-D3
48324066LIB3069-047-LIB3069g166634BLASTX1731e−4555
Q1-K1-C4
48424266LIB3069-006-LIB3069g2570500BLASTN7171e−5783
Q1-K1-F4
485293LIB3068-043-LIB3068g633598BLASTN5521e−3478
Q1-K1-A2
48632364LIB3066-001-LIB3066g2668745BLASTN6121e−4073
Q1-K1-B7
48732856LIB189-028-LIB189g534915BLASTN9861e−7373
Q1-E1-C4
4883384LIB143-026-LIB143g534915BLASTN12841e−9878
Q1-E1-C1
4893384LIB3068-013-LIB3068g534915BLASTN10741e−8078
Q1-K1-H2
4903384LIB3062-033-LIB3062g2668745BLASTN10091e−7576
Q1-K1-D2
4913384LIB83-002-LIB83g2706449BLASTN8201e−5978
Q1-E1-D2
4923384LIB3062-057-LIB3062g2668745BLASTN8011e−5873
Q1-K1-B7
4933384LIB3062-001-LIB3062g16347BLASTN8021e−5777
Q1-K2-H5
4943384LIB189-022-LIB189g2668745BLASTN6461e−4375
Q1-E1-D5
4953384LIB189-012-LIB189g2570501BLASTX1381e−3272
Q1-E1-F4
4965000LIB36-015-LIB36g2624379BLASTX2361e−4151
Q1-E1-D6
4975000LIB83-016-LIB83g4198BLASTN5341e−3361
Q1-E1-H7
MAIZE PYROPHOSPHATE-DEPENDENT FRUCTOSE-6-PHOSPHATE PHOSPHOTRANSFERASE
498−700208959700208959H1SATMON016g169538BLASTX1071e−1950
499−700237606700237606H1SATMON010g169538BLASTX1141e−1162
5003456700083478H1SATMON011g169538BLASTX1211e−3988
5013652700242182H1SATMON010g169538BLASTX1551e−1382
5024965700475352H1SATMON025g169538BLASTX1231e−969
5034965700550752H1SATMON022g169538BLASTX1231e−969
5045359700347441H1SATMON023g169538BLASTX1391e−1170
505-L30594734LIB3059-018-LIB3059g169538BLASTX1451e−4983
Q1-K1-H3
506-L30622375LIB3062-009-LIB3062g169538BLASTX1571e−3065
Q1-K1-B3
50732156LIB189-021-LIB189g169538BLASTX1231e−2578
Q1-E1-G8
MAIZE INVERTASES
508−700240132700240132H1SATMON010g397631BLASTX1341e−1174
5091923700574932H1SATMON030g393390BLASTX1521e−1465
5104355700379641H1SATMON021g1177601BLASTX1751e−1985
MAIZE SUCROSE SYNTHASE
511−700151470700151470H1SATMON007g1196837BLASTX1971e−2764
512−700214035700214035H1SATMON016g22485BLASTN5231e−3479
513−700262270700262270H1SATMON017g2570066BLASTN8661e−6376
514−700334686700334686H1SATMON019g1100216BLASTN4241e−3188
515−700381593700381593H1SATMON023g22485BLASTN2191e−1397
516−700404808700404808H1SATMON026g2570066BLASTN8591e−7082
517−700456905700456905H1SATMON029g22485BLASTN5281e−6490
518−700571529700571529H1SATMON030g19106BLASTX1391e−2456
519−700576567700576567H1SATMON030g22485BLASTN2851e−1492
520−700800659700800659H1SATMON036g22485BLASTN5581e−3797
521−700802941700802941H1SATMON036g22485BLASTN3161e−2997
522−701181030701181030H1SATMONN06g2606080BLASTN6691e−4672
52313723700203023H1SATMON003g2570066BLASTN8201e−6884
52413723700215119H1SATMON016g2570066BLASTN6801e−4786
52513723700473266H1SATMON025g2570066BLASTN5371e−3585
52615661700440404H1SATMON026g2570066BLASTN3641e−3674
52715661700168252H1SATMON013g16525BLASTN4331e−2780
52820925700551647H1SATMON022g2570066BLASTN3071e−3573
52920925700257052H1SATMON017g2570067BLASTX1181e−964
53020934700217752H1SATMON016g514945BLASTN13971e−10798
53120934700332156H1SATMON019g514945BLASTN5891e−9795
53230444700257522H1SATMON017g1100216BLASTN7601e−5495
53332909700264718H1SATMON017g2570066BLASTN7021e−5776
534405700091402H1SATMON011g514945BLASTN18301e−143100
535405700572549H1SATMON030g514945BLASTN16581e−12999
536405700203058H1SATMON003g22485BLASTN13601e−127100
537405700091753H1SATMON011g514945BLASTN12451e−12699
538405700090929H1SATMON011g514945BLASTN16201e−126100
539405700091711H1SATMON011g514945BLASTN16211e−12699
540405700084254H1SATMON011g514945BLASTN16001e−124100
541405700082305H1SATMON011g514945BLASTN16011e−12499
542405700048236H1SATMON003g22485BLASTN15831e−12399
543405700086713H1SATMON011g514945BLASTN15841e−12399
544405700049353H1SATMON003g514945BLASTN15861e−12399
545405700082766H1SATMON011g22485BLASTN15891e−12398
546405700086055H1SATMON011g514945BLASTN15901e−123100
547405700215105H1SATMON016g514945BLASTN15901e−123100
548405700104149H1SATMON010g22485BLASTN15941e−12398
549405700101601H1SATMON009g514945BLASTN12701e−122100
550405700206869H1SATMON003g22485BLASTN15741e−12297
551405700088163H1SATMON011g22485BLASTN15811e−12299
552405700089166H1SATMON011g514945BLASTN15651e−121100
553405700266251H1SATMON017g514945BLASTN15701e−121100
554405700332710H1SATMON019g514945BLASTN15701e−121100
555405700571106H1SATMON030g514945BLASTN12271e−12098
556405700081893H1SATMON011g514945BLASTN15501e−12098
557405700074739H1SATMON007g514945BLASTN15501e−120100
558405700095163H1SATMON008g514945BLASTN15551e−120100
559405700612766H1SATMON033g514945BLASTN8831e−11996
560405700267271H1SATMON017g514945BLASTN15351e−119100
561405700083175H1SATMON011g514945BLASTN15351e−119100
562405700088993H1SATMON011g22485BLASTN15451e−11998
563405700094087H1SATMON008g22485BLASTN15261e−11899
564405700086708H1SATMON011g514945BLASTN15291e−11897
565405700090671H1SATMON011g514945BLASTN15301e−118100
566405700209809H1SATMON016g22485BLASTN15321e−11899
567405700084625H1SATMON011g514945BLASTN15331e−11899
568405700089718H1SATMON011g514945BLASTN11201e−117100
569405700213014H1SATMON016g514945BLASTN14051e−117100
570405700086555H1SATMON011g514945BLASTN15141e−11798
571405700475892H1SATMON025g514945BLASTN15161e−11799
572405700047374H1SATMON003g22485BLASTN15161e−11799
573405700090018H1SATMON011g514945BLASTN15191e−11799
574405700076107H1SATMON007g514945BLASTN15201e−11793
575405700213105H1SATMON016g514945BLASTN9721e−11699
576405700103806H1SATMON010g514945BLASTN15031e−11699
577405700090748H1SATMON011g514945BLASTN15051e−116100
578405700052006H1SATMON003g514945BLASTN15061e−11699
579405700614963H1SATMON033g514945BLASTN9571e−11593
580405700337255H1SATMON020g22485BLASTN9951e−11597
581405700102778H1SATMON010g22485BLASTN14931e−11599
582405700405466H1SATMON029g22485BLASTN14931e−11599
583405700209634H1SATMON016g514945BLASTN14951e−115100
584405700220467H1SATMON011g514945BLASTN14951e−115100
585405700266637H1SATMON017g514945BLASTN14801e−114100
586405700267579H1SATMON017g514945BLASTN14841e−11499
587405700088475H1SATMON011g514945BLASTN14651e−113100
588405700332618H1SATMON019g514945BLASTN14661e−11399
589405700211347H1SATMON016g514945BLASTN14701e−113100
590405700477206H1SATMON025g514945BLASTN14711e−11399
591405700336768H1SATMON019g514945BLASTN14731e−11399
592405700105305H1SATMON010g22485BLASTN14731e−11399
593405700087114H1SATMON011g514945BLASTN14731e−11399
594405700105366H1SATMON010g22485BLASTN14741e−11398
595405700104831H1SATMON010g22485BLASTN8251e−11298
596405700620134H1SATMON034g22485BLASTN11791e−11292
597405700211934H1SATMON016g22485BLASTN12151e−11298
598405700096103H1SATMON008g514945BLASTN13911e−11299
599405700264979H1SATMON017g514945BLASTN14541e−11298
600405700053864H1SATMON011g514945BLASTN14551e−112100
601405700211782H1SATMON016g514945BLASTN14601e−112100
602405700102063H1SATMON010g22485BLASTN14611e−11299
603405700207024H1SATMON003g514945BLASTN8251e−111100
604405700207970H1SATMON016g514945BLASTN11861e−11198
605405700336624H1SATMON019g514945BLASTN14401e−111100
606405700104357H1SATMON010g514945BLASTN14481e−11198
607405700222053H1SATMON011g514945BLASTN14491e−11199
608405700350806H1SATMON023g514945BLASTN6601e−11099
609405700091159H1SATMON011g514945BLASTN8701e−110100
610405700081810H1SATMON011g514945BLASTN9261e−11099
611405700102954H1SATMON010g514945BLASTN9261e−11097
612405700085307H1SATMON011g514945BLASTN10351e−110100
613405700094295H1SATMON008g22485BLASTN11371e−11096
614405700089176H1SATMON011g514945BLASTN13931e−11097
615405700093643H1SATMON008g514945BLASTN14271e−11095
616405700082421H1SATMON011g514945BLASTN14301e−11098
617405700211788H1SATMON016g514945BLASTN14311e−11099
618405700026724H1SATMON003g514945BLASTN14331e−11097
619405700085275H1SATMON011g514945BLASTN14351e−110100
620405700472161H1SATMON025g514945BLASTN7551e−10999
621405700084926H1SATMON011g514945BLASTN8251e−109100
622405700084592H1SATMON011g514945BLASTN9201e−109100
623405700053811H1SATMON011g514945BLASTN12961e−10996
624405700216963H1SATMON016g514945BLASTN14151e−109100
625405700085273H1SATMON011g22485BLASTN14161e−10998
626405700082127H1SATMON011g514945BLASTN14201e−109100
627405700085731H1SATMON011g514945BLASTN14251e−109100
628405700088595H1SATMON011g22485BLASTN14261e−10999
629405700470903H1SATMON025g514945BLASTN14261e−10999
630405700265288H1SATMON017g514945BLASTN13751e−108100
631405700072245H1SATMON007g514945BLASTN14041e−10899
632405700347692H1SATMON023g514945BLASTN14051e−10898
633405700214447H1SATMON016g514945BLASTN14061e−10899
634405700476252H1SATMON025g514945BLASTN14071e−10899
635405700336746H1SATMON019g514945BLASTN14091e−10899
636405700053833H1SATMON011g514945BLASTN14101e−108100
637405700094342H1SATMON008g514945BLASTN14101e−108100
638405700202813H1SATMON003g514945BLASTN10321e−10797
639405700050589H1SATMON003g514945BLASTN10351e−107100
640405700050011H1SATMON003g514945BLASTN10781e−10799
641405700215426H1SATMON016g514945BLASTN11891e−10796
642405700472461H1SATMON025g514945BLASTN13921e−10799
643405700336684H1SATMON019g22485BLASTN13931e−10798
644405700449826H2SATMON028g514945BLASTN13951e−107100
645405700216443H1SATMON016g514945BLASTN13961e−10799
646405700240793H1SATMON010g514945BLASTN13991e−10798
647405700215985H1SATMON016g514945BLASTN14001e−107100
648405700336740H1SATMON019g514945BLASTN9151e−10699
649405700047958H1SATMON003g514945BLASTN9871e−10696
650405700085447H1SATMON011g514945BLASTN10301e−106100
651405700084978H1SATMON011g514945BLASTN11211e−10691
652405700800439H1SATMON036g22485BLASTN13791e−10699
653405700219631H1SATMON011g514945BLASTN13801e−106100
654405700220740H1SATMON011g514945BLASTN13801e−106100
655405700243367H1SATMON010g514945BLASTN13811e−10699
656405700220363H1SATMON011g514945BLASTN13871e−10699
657405700215869H1SATMON016g514945BLASTN13901e−106100
658405700216519H1SATMON016g514945BLASTN11311e−10597
659405700052206H1SATMON003g514945BLASTN12641e−10596
660405700094975H1SATMON008g514945BLASTN13681e−10599
661405700220837H1SATMON011g514945BLASTN13691e−10598
662405700221108H1SATMON011g514945BLASTN13701e−10598
663405700222850H1SATMON011g514945BLASTN13701e−105100
664405700214429H1SATMON016g514945BLASTN13731e−10599
665405700473857H1SATMON025g514945BLASTN13751e−10598
666405700213762H1SATMON016g514945BLASTN13781e−10599
667405700405254H1SATMON028g22485BLASTN12421e−10499
668405700029978H1SATMON003g22485BLASTN13241e−10497
669405700238315H1SATMON010g514945BLASTN13551e−104100
670405700241686H1SATMON010g514945BLASTN13581e−10499
671405700237721H1SATMON010g22485BLASTN13601e−104100
672405700217344H1SATMON016g514945BLASTN13601e−104100
673405700030048H1SATMON003g514945BLASTN13631e−10499
674405700211866H1SATMON016g514945BLASTN13631e−10499
675405700214860H1SATMON016g514945BLASTN13651e−104100
676405700085490H1SATMON011g514945BLASTN9001e−10398
677405700048568H1SATMON003g514945BLASTN9801e−103100
678405700381034H1SATMON023g22485BLASTN12691e−10398
679405700220930H1SATMON011g514945BLASTN13471e−10399
680405700030261H1SATMON003g514945BLASTN13531e−10398
681405700081835H1SATMON011g22485BLASTN7971e−10298
682405700205270H1SATMON003g514945BLASTN10241e−10294
683405700093612H1SATMON008g514945BLASTN10651e−10299
684405700333392H1SATMON019g514945BLASTN11081e−10297
685405700575385H1SATMON030g514945BLASTN11711e−10296
686405700241061H1SATMON010g514945BLASTN11741e−10299
687405700239916H1SATMON010g514945BLASTN12551e−102100
688405700090248H1SATMON011g514945BLASTN13341e−10298
689405700222923H1SATMON011g514945BLASTN13341e−10298
690405700216993H1SATMON016g514945BLASTN13351e−102100
691405700215984H1SATMON016g514945BLASTN13401e−102100
692405700213182H1SATMON016g514945BLASTN13401e−10298
693405700219845H1SATMON011g514945BLASTN13401e−102100
694405700237762H1SATMON010g514945BLASTN13401e−102100
695405700551043H1SATMON022g514945BLASTN13421e−10299
696405700219254H1SATMON011g514945BLASTN12521e−10199
697405700210348H1SATMON016g514945BLASTN13201e−10197
698405700215089H1SATMON016g514945BLASTN13201e−101100
699405700217251H1SATMON016g514945BLASTN13201e−101100
700405700219240H1SATMON011g514945BLASTN13201e−101100
701405700082094H1SATMON011g514945BLASTN13211e−10199
702405700219385H1SATMON011g514945BLASTN13221e−10199
703405700220052H1SATMON011g514945BLASTN13251e−101100
704405700210366H1SATMON016g514945BLASTN13291e−10193
705405700083089H1SATMON011g514945BLASTN13301e−10198
706405700340286H1SATMON020g22485BLASTN6771e−10098
707405700221062H1SATMON011g514945BLASTN8451e−10098
708405700382272H1SATMON024g22485BLASTN9581e−10096
709405700209310H1SATMON016g514945BLASTN11871e−10097
710405700052340H1SATMON003g514945BLASTN11881e−10094
711405700467851H1SATMON025g22485BLASTN12451e−10097
712405700088014H1SATMON011g514945BLASTN12701e−10098
713405700214596H1SATMON016g514945BLASTN12951e−100100
714405700157215H1SATMON012g22485BLASTN13101e−10098
715405700223892H1SATMON011g514945BLASTN13101e−100100
716405700218981H1SATMON011g514945BLASTN13101e−100100
717405700081945H1SATMON011g514945BLASTN13111e−10096
718405700217817H1SATMON016g514945BLASTN13151e−100100
719405700469042H1SATMON025g514945BLASTN5611e−9998
720405700474709H1SATMON025g514945BLASTN8011e−9999
721405700201736H1SATMON003g514945BLASTN11681e−9998
722405700223516H1SATMON011g514945BLASTN12011e−9999
723405700453941H1SATMON029g22485BLASTN12951e−9995
724405700212970H1SATMON016g514945BLASTN12971e−9999
725405700215662H1SATMON016g22485BLASTN12971e−9999
726405700802209H1SATMON036g22485BLASTN13001e−9998
727405700343716H1SATMON021g514945BLASTN13001e−99100
728405700223322H1SATMON011g514945BLASTN13001e−99100
729405700217238H1SATMON016g514945BLASTN13001e−99100
730405700195066H1SATMON014g22485BLASTN13001e−9998
731405700072395H1SATMON007g514945BLASTN13011e−9995
732405700212752H1SATMON016g22485BLASTN13051e−9998
733405700222204H1SATMON011g514945BLASTN13051e−99100
734405700550572H1SATMON022g22485BLASTN7131e−9897
735405700213879H1SATMON016g514945BLASTN8661e−9899
736405700551585H1SATMON022g514945BLASTN9161e−9899
737405700195025H1SATMON014g22485BLASTN12831e−9898
738405700800710H1SATMON036g22485BLASTN12831e−9898
739405700222985H1SATMON011g22485BLASTN12831e−9898
740405700798823H1SATMON036g22485BLASTN12841e−9898
741405700104391H1SATMON010g22485BLASTN12891e−9898
742405700466592H1SATMON025g22485BLASTN12891e−9895
743405700027037H1SATMON003g514945BLASTN9191e−9791
744405700214371H1SATMON016g514945BLASTN10331e−9795
745405700799077H1SATMON036g22485BLASTN10911e−9799
746405700467028H1SATMON025g514945BLASTN11031e−9798
747405700219393H1SATMON011g514945BLASTN12501e−9799
748405700197602H1SATMON014g22485BLASTN12731e−9797
749405700801226H1SATMON036g22485BLASTN12761e−9799
750405700216371H1SATMON016g514945BLASTN12781e−9798
751405700805695H1SATMON036g22485BLASTN12801e−9798
752405700334076H1SATMON019g514945BLASTN5031e−9698
753405700082647H1SATMON011g514945BLASTN7351e−96100
754405700458687H1SATMON029g22485BLASTN7511e−9695
755405700220750H1SATMON011g514945BLASTN11871e−9696
756405700194931H1SATMON014g22485BLASTN12621e−9699
757405700800522H1SATMON036g22485BLASTN12651e−9698
758405700244185H1SATMON010g514945BLASTN12651e−96100
759405700240785H1SATMON010g514945BLASTN12681e−9698
760405700551959H1SATMON022g514945BLASTN12701e−96100
761405700085057H1SATMON011g514945BLASTN6821e−9597
762405700332020H1SATMON019g514945BLASTN7131e−9597
763405700208841H1SATMON016g514945BLASTN8221e−9595
764405700193023H1SATMON014g22485BLASTN12481e−9598
765405700153902H1SATMON007g514945BLASTN12501e−95100
766405700196173H1SATMON014g22485BLASTN12521e−9599
767405700804187H1SATMON036g22485BLASTN12531e−9598
768405700339656H1SATMON020g22485BLASTN12571e−9599
769405700238537H1SATMON010g22485BLASTN12581e−9599
770405700551221H1SATMON022g514945BLASTN12041e−9499
771405700801807H1SATMON036g22485BLASTN12351e−94100
772405700085096H1SATMON011g514945BLASTN12351e−94100
773405700020516H1SATMON001g514945BLASTN12361e−9498
774405700193482H1SATMON014g22485BLASTN12361e−9499
775405700217793H1SATMON016g514945BLASTN12371e−9498
776405700088752H1SATMON011g514945BLASTN12401e−94100
777405700087940H1SATMON011g514945BLASTN12411e−9499
778405700089541H1SATMON011g533251BLASTN10671e−9391
779405700346461H1SATMON021g514945BLASTN11901e−9399
780405700156258H1SATMON007g514945BLASTN12251e−93100
781405700195532H1SATMON014g22485BLASTN12261e−9399
782405700196548H1SATMON014g22485BLASTN12271e−9399
783405700213282H1SATMON016g514945BLASTN12271e−9396
784405700194939H1SATMON014g22485BLASTN12281e−9398
785405700340787H1SATMON020g22485BLASTN6971e−9294
786405700105064H1SATMON010g22485BLASTN7061e−9298
787405700805132H1SATMON036g22485BLASTN10461e−9299
788405700803414H1SATMON036g22485BLASTN12111e−9299
789405700214594H1SATMON016g514945BLASTN12121e−9298
790405700215080H1SATMON016g514945BLASTN12131e−9292
791405700224219H1SATMON011g514945BLASTN12131e−9299
792405700048190H1SATMON003g514945BLASTN12181e−9292
793405700223284H1SATMON011g514945BLASTN12201e−92100
794405700152413H1SATMON007g514945BLASTN12201e−92100
795405700218003H1SATMON016g514945BLASTN12211e−9296
796405700160155H1SATMON012g22485BLASTN12221e−9299
797405700087943H1SATMON011g22485BLASTN12221e−9299
798405700474049H1SATMON025g514945BLASTN6321e−9197
799405700216994H1SATMON016g514945BLASTN10431e−9199
800405700346892H1SATMON021g514945BLASTN12101e−9196
801405700142782H1SATMON013g514945BLASTN11901e−90100
802405700244157H1SATMON010g514945BLASTN11971e−9097
803405700469243H1SATMON025g22485BLASTN7011e−8998
804405700618387H1SATMON033g514945BLASTN8531e−8993
805405700211154H1SATMON016g533251BLASTN9171e−8990
806405700081933H1SATMON011g533251BLASTN9551e−8991
807405700235229H1SATMON010g514945BLASTN9551e−8997
808405700209241H1SATMON016g514945BLASTN10761e−8998
809405700265841H1SATMON017g514945BLASTN10971e−8995
810405700193804H1SATMON014g22485BLASTN11801e−8998
811405700167742H1SATMON013g22485BLASTN11801e−89100
812405700163256H1SATMON013g514945BLASTN11821e−8997
813405700184973H1SATMON014g22485BLASTN11821e−8999
814405700171785H1SATMON013g22485BLASTN11841e−8997
815405700216915H1SATMON016g514945BLASTN11851e−89100
816405700806685H1SATMON036g22485BLASTN11861e−8999
817405700803846H1SATMON036g22485BLASTN8921e−8895
818405700218514H1SATMON011g533251BLASTN9071e−8891
819405700574674H1SATMON030g22485BLASTN9571e−8884
820405700196082H1SATMON014g22485BLASTN10541e−8894
821405700241637H1SATMON010g22485BLASTN10811e−8898
822405700801876H1SATMON036g22485BLASTN11631e−8898
823405700213443H1SATMON016g514945BLASTN11711e−8899
824405700465181H1SATMON025g22485BLASTN7971e−8792
825405700798732H1SATMON036g22485BLASTN11591e−8796
826405700153473H1SATMON007g514945BLASTN11601e−87100
827405700165496H1SATMON013g514945BLASTN11601e−8797
828405700264250H1SATMON017g514945BLASTN6431e−86100
829405700335490H1SATMON019g514945BLASTN6711e−8697
830405700575891H1SATMON030g22485BLASTN7801e−8692
831405700222931H1SATMON011g514945BLASTN11171e−8691
832405700163588H1SATMON013g514945BLASTN11401e−86100
833405700161111H1SATMON012g22485BLASTN11421e−8699
834405700016023H1SATMON001g514945BLASTN11471e−8699
835405700209043H1SATMON016g514945BLASTN6591e−8598
836405700333556H1SATMON019g514945BLASTN7831e−8589
837405700570471H1SATMON030g22485BLASTN8311e−8589
838405700171040H1SATMON013g22485BLASTN11271e−8599
839405700196146H1SATMON014g22485BLASTN11281e−8596
840405700021837H1SATMON001g514945BLASTN11301e−8598
841405700169062H1SATMON013g514945BLASTN11301e−85100
842405700218263H1SATMON016g514945BLASTN11351e−85100
843405700091281H1SATMON011g514945BLASTN11351e−85100
844405700221221H1SATMON011g22485BLASTN7351e−8498
845405700239933H1SATMON010g22485BLASTN7791e−8497
846405700193089H1SATMON014g22485BLASTN9791e−8497
847405700216767H1SATMON016g514945BLASTN10051e−8498
848405700085786H1SATMON011g514945BLASTN11171e−8499
849405700163868H1SATMON013g22485BLASTN11201e−84100
850405700167028H1SATMON013g514945BLASTN11201e−84100
851405700170416H1SATMON013g514945BLASTN11201e−84100
852405700266595H1SATMON017g514945BLASTN11201e−84100
853405700377630H1SATMON019g514945BLASTN6491e−8395
854405700207830H1SATMON016g514945BLASTN8771e−8397
855405700241726H1SATMON010g514945BLASTN11041e−8397
856405700806447H1SATMON036g22485BLASTN11061e−8393
857405700018166H1SATMON001g514945BLASTN11081e−8398
858405700083463H1SATMON011g514945BLASTN6331e−8292
859405700548890H1SATMON022g22485BLASTN7271e−8293
860405700218553H1SATMON011g22485BLASTN9791e−8295
861405700016408H1SATMON001g514945BLASTN10261e−8297
862405700569427H2SATMON030g514945BLASTN10951e−8297
863405700172546H1SATMON013g514945BLASTN11001e−82100
864405700804387H1SATMON036g22485BLASTN6681e−8197
865405700155788H1SATMON007g514945BLASTN8401e−81100
866405700807167H1SATMON036g22485BLASTN10241e−8197
867405700472356H1SATMON025g22485BLASTN10801e−8198
868405700193535H1SATMON014g22485BLASTN10821e−8199
869405700171177H1SATMON013g22485BLASTN10861e−8198
870405700799867H1SATMON036g22485BLASTN10871e−8196
871405700263716H1SATMON017g514945BLASTN10891e−8192
872405700476045H1SATMON025g22485BLASTN6081e−8088
873405700803344H1SATMON036g22485BLASTN8341e−8097
874405700168924H1SATMON013g514945BLASTN8601e−8099
875405700218569H1SATMON011g22485BLASTN9001e−8098
876405700088574H1SATMON011g514945BLASTN9001e−8086
877405700471932H1SATMON025g530978BLASTN10641e−8083
878405700020011H1SATMON001g22485BLASTN10671e−8099
879405700167511H1SATMON013g22485BLASTN10701e−80100
880405700219249H1SATMON011g514945BLASTN10701e−80100
881405700804846H1SATMON036g22485BLASTN10751e−8090
882405700150388H1SATMON007g22485BLASTN10751e−80100
883405700807395H1SATMON036g22485BLASTN5711e−7990
884405700090864H1SATMON011g514945BLASTN6301e−79100
885405700217812H1SATMON016g514945BLASTN6461e−7991
886405700203618H1SATMON003g22485BLASTN9131e−7996
887405700203302H1SATMON003g514945BLASTN10301e−79100
888405700163192H1SATMON013g22485BLASTN10561e−7997
889405700805065H1SATMON036g22485BLASTN10661e−7995
890405700086763H1SATMON011g514945BLASTN9011e−7898
891405700240070H1SATMON010g533251BLASTN9231e−7890
892405700018847H1SATMON001g22485BLASTN10451e−7898
893405700803420H1SATMON036g22485BLASTN10481e−7896
894405700799936H1SATMON036g22485BLASTN10501e−7896
895405700207637H1SATMON016g514945BLASTN8281e−7798
896405700807034H1SATMON036g22485BLASTN8081e−7691
897405700198035H1SATMON016g514945BLASTN10251e−76100
898405700169076H1SATMON013g514945BLASTN10281e−7699
899405700020564H1SATMON001g514945BLASTN10301e−7698
900405700799968H1SATMON036g22485BLASTN7011e−7599
901405700378056H1SATMON019g22485BLASTN8021e−7597
902405700219691H1SATMON011g514945BLASTN10181e−7599
903405700168945H1SATMON013g22485BLASTN8481e−7494
904405700242730H1SATMON010g514945BLASTN10061e−7499
905405700210096H1SATMON016g514945BLASTN7561e−7393
906405700333941H1SATMON019g514945BLASTN9231e−7399
907405700576645H1SATMON030g22485BLASTN9911e−7399
908405700333494H1SATMON019g514945BLASTN6011e−7291
909405700023296H1SATMON003g514945BLASTN7261e−7295
910405700802508H1SATMON036g22485BLASTN8111e−7294
911405700223382H1SATMON011g22485BLASTN8651e−7298
912405700215535H1SATMON016g514945BLASTN9421e−7296
913405700017549H1SATMON001g514945BLASTN9731e−7297
914405700799113H1SATMON036g22485BLASTN7871e−7099
915405700168696H1SATMON013g514945BLASTN9461e−6989
916405700088269H1SATMON011g514945BLASTN9461e−6993
917405700194522H1SATMON014g22485BLASTN8751e−6897
918405700203476H1SATMON003g22485BLASTN9231e−6886
919405700549205H1SATMON022g22485BLASTN3001e−6689
920405700196217H1SATMON014g22485BLASTN9071e−6696
921405700163647H1SATMON013g22485BLASTN8881e−6598
922405700804485H1SATMON036g22485BLASTN8961e−6599
923405700193074H1SATMON014g22485BLASTN6051e−6396
924405700203370H1SATMON003g514945BLASTN8571e−6298
925405700201575H1SATMON003g514945BLASTN3351e−6087
926405700378020H1SATMON019g514945BLASTN8331e−6097
927405700242865H1SATMON010g514945BLASTN8231e−5991
928405700344036H1SATMON021g514945BLASTN8251e−59100
929405700215849H1SATMON016g514945BLASTN8051e−58100
930405700443538H1SATMON027g22485BLASTN8141e−5898
931405700804448H1SATMON036g22485BLASTN7911e−5799
932405700155008H1SATMON007g22485BLASTN8021e−5798
933405700201244H1SATMON003g22485BLASTN5301e−5697
934405700616378H1SATMON033g22485BLASTN6821e−5697
935405700333357H1SATMON019g22485BLASTN7801e−5680
936405700222360H1SATMON011g514945BLASTN7771e−5592
937405700214724H1SATMON016g514945BLASTN7631e−5498
938405700571283H1SATMON030g514945BLASTN7361e−5299
939405700020194H1SATMON001g22485BLASTN4151e−5199
940405700620551H1SATMON034g22485BLASTN4731e−5195
941405700446320H1SATMON027g22485BLASTN4751e−5087
942405700241357H1SATMON010g22485BLASTN7011e−4999
943405700617094H1SATMON033g22485BLASTN6731e−4797
944405700206691H1SATMON003g514945BLASTN6801e−4790
945405700091580H1SATMON011g514945BLASTN6801e−47100
946405700574515H1SATMON030g514945BLASTN3691e−4674
947405700155148H1SATMON007g514945BLASTN3971e−4597
948405700612388H1SATMON033g514945BLASTN6251e−43100
949405700474681H1SATMON025g22485BLASTN3791e−4191
950405700800401H1SATMON036g22485BLASTN3951e−4090
951405700155657H1SATMON007g514945BLASTN5911e−4095
952405700076002H1SATMON007g514945BLASTN5751e−39100
953405700802090H1SATMON036g22485BLASTN5771e−3998
954405700170104H1SATMON013g22485BLASTN5651e−38100
955405701183763H1SATMONN06g514945BLASTN5691e−3890
956405700084688H1SATMON011g514945BLASTN3801e−3698
957405700473655H1SATMON025g22485BLASTN5301e−35100
958405700615166H1SATMON033g514945BLASTN5311e−3594
959405700085562H1SATMON011g533251BLASTN5321e−3598
960405700153049H1SATMON007g514945BLASTN5371e−3594
961405700090656H1SATMON011g514945BLASTN4891e−3498
962405700802054H1SATMON036g22485BLASTN3451e−3199
963405700802284H1SATMON036g22485BLASTN4881e−3197
964405700802312H1SATMON036g22485BLASTN2701e−30100
965405700153683H1SATMON007g22485BLASTN4611e−2998
966405700028453H1SATMON003g22485BLASTN3211e−2799
967405700089391H1SATMON011g514945BLASTN4041e−2496
968405700381969H1SATMON023g22485BLASTN3851e−2394
969405700800135H1SATMON036g22485BLASTN1801e−21100
970405700088173H1SATMON011g514945BLASTN3471e−2095
971405700202170H1SATMON003g19108BLASTX1331e−1196
972537700209929H1SATMON016g22485BLASTN14781e−11499
973537700096948H1SATMON008g22485BLASTN9111e−11399
974537700476287H1SATMON025g22485BLASTN14031e−10898
975537700803088H1SATMON036g22485BLASTN13361e−10796
976537700799436H1SATMON036g22485BLASTN13611e−10499
977537700224822H1SATMON011g22485BLASTN13001e−10396
978537700241134H1SATMON010g22485BLASTN13021e−9999
979537700803625H1SATMON036g22485BLASTN12921e−9899
980537700802549H1SATMON036g22485BLASTN12321e−9399
981537700477992H1SATMON025g22485BLASTN9431e−9297
982537700150953H1SATMON007g22485BLASTN11521e−8799
983537700205638H1SATMON003g22485BLASTN10861e−8199
984537700803732H1SATMON036g22487BLASTN3791e−7997
985537700165461H1SATMON013g22485BLASTN10641e−7998
986537700807069H1SATMON036g22485BLASTN9571e−7796
987537700800902H1SATMON036g22485BLASTN7621e−5486
988537700466671H1SATMON025g22485BLASTN5201e−4495
989537700799118H1SATMON036g22485BLASTN6261e−4399
990537700802273H1SATMON036g22485BLASTN6161e−4299
991537700804848H1SATMON036g22485BLASTN3061e−3398
9928549700103190H1SATMON010g1100216BLASTN6151e−9298
9938549700075574H1SATMON007g1100216BLASTN7011e−92100
9948549700218547H1SATMON011g514945BLASTN12081e−9199
9958549700213873H1SATMON016g1100216BLASTN6731e−9095
9968549700221147H1SATMON011g1100216BLASTN6461e−8998
9978549700207093H1SATMON003g1100216BLASTN7011e−87100
9988549700210112H1SATMON016g1100216BLASTN6151e−8498
9998549700096984H1SATMON008g514945BLASTN11111e−8399
10008549700221070H1SATMON011g1100216BLASTN6451e−8296
10018549700332046H1SATMON019g1100216BLASTN6011e−7689
10028549700150377H1SATMON007g1100216BLASTN6211e−74100
10038549700084780H1SATMON011g514945BLASTN5851e−39100
10048549700153082H1SATMON007g1100216BLASTN4951e−3689
10058549700261144H1SATMON017g1100216BLASTN3391e−3587
10068549700264112H1SATMON017g1100216BLASTN4281e−3491
10078549700473660H1SATMON025g1100216BLASTN4151e−28100
10088549700473628H1SATMON025g514945BLASTN3291e−2688
10098549700351060H1SATMON023g1100216BLASTN2911e−2291
1010-L30595280LIB3059-039-LIB3059g22485BLASTN4731e−3079
Q1-K1-A5
1011-L30612133LIB3061-024-LIB3061g22485BLASTN8491e−6180
Q1-K1-H5
1012-L30616296LIB3061-043-LIB3061g22485BLASTN4791e−9882
Q1-K1-A10
1013-L30623037LIB3062-030-LIB3062g514945BLASTN6841e−4878
Q1-K1-F12
1014-L30625289LIB3062-021-LIB3062g514945BLASTN11801e−11179
Q1-K1-C2
1015-L30663565LIB3066-053-LIB3066g530978BLASTN5681e−3676
Q1-K1-D6
1016-L30784420LIB3078-039-LIB3078g514945BLASTN4841e−4081
Q1-K1-A4
101730444LIB3069-052-LIB3069g1100216BLASTN5581e−7789
Q1-K1-F8
101832909LIB143-057-LIB143g2570066BLASTN9021e−6974
Q1-E1-F6
1019405LIB3062-021-LIB3062g514945BLASTN23681e−18899
Q1-K1-C5
1020405LIB3078-024-LIB3078g514945BLASTN23561e−18798
Q1-K1-C5
1021405LIB3059-028-LIB3059g22485BLASTN21631e−17198
Q1-K1-D5
1022405LIB3059-015-LIB3059g22485BLASTN21671e−17198
Q1-K1-E7
1023405LIB3059-044-LIB3059g514945BLASTN21701e−17198
Q1-K1-E7
1024405LIB3061-029-LIB3061g22485BLASTN20551e−17098
Q1-K1-G11
1025405LIB3059-011-LIB3059g22485BLASTN21371e−16998
Q1-K1-F5
1026405LIB3062-009-LIB3062g514945BLASTN21221e−16798
Q1-K1-D1
1027405LIB3061-011-LIB3061g22485BLASTN20911e−16598
Q1-K1-D9
1028405LIB3067-040-LIB3067g514945BLASTN19161e−16499
Q1-K1-E8
1029405LIB3062-041-LIB3062g514945BLASTN20821e−16497
Q1-K1-D4
1030405LIB3062-022-LIB3062g514945BLASTN20841e−16499
Q1-K1-C9
1031405LIB3062-033-LIB3062g514945BLASTN18541e−16195
Q1-K1-C7
1032405LIB3062-002-LIB3062g514945BLASTN18541e−16197
Q1-K2-F9
1033405LIB3059-010-LIB3059g22485BLASTN20181e−15999
Q1-K1-C9
1034405LIB3059-013-LIB3059g22485BLASTN20221e−15998
Q1-K1-B10
1035405LIB3061-020-LIB3061g22485BLASTN17711e−15897
Q1-K1-F2
1036405LIB3061-022-LIB3061g22485BLASTN19091e−15898
Q1-K1-C2
1037405LIB3062-023-LIB3062g22485BLASTN15081e−15796
Q1-K1-D10
1038405LIB3061-008-LIB3061g22485BLASTN19831e−15697
Q1-K1-H11
1039405LIB3059-024-LIB3059g22485BLASTN10511e−15499
Q1-K1-H4
1040405LIB3062-048-LIB3062g22485BLASTN11871e−15494
Q1-K1-G5
1041405LIB3061-025-LIB3061g22485BLASTN18031e−15495
Q1-K1-B1
1042405LIB3061-028-LIB3061g22485BLASTN19631e−15497
Q1-K1-C4
1043405LIB3078-057-LIB3078g514945BLASTN14121e−15392
Q1-K1-D9
1044405LIB3061-021-LIB3061g22485BLASTN14651e−15396
Q1-K1-A8
1045405LIB3061-025-LIB3061g22485BLASTN15241e−15396
Q1-K1-B5
1046405LIB3061-008-LIB3061g22485BLASTN18791e−15394
Q1-K1-C7
1047405LIB3078-039-LIB3078g514945BLASTN18531e−15196
Q1-K1-A8
1048405LIB3061-049-LIB3061g22485BLASTN18011e−15098
Q1-K1-E5
1049405LIB3062-001-LIB3062g514945BLASTN19161e−15094
Q1-K2-G2
1050405LIB3061-021-LIB3061g22485BLASTN19181e−15092
Q1-K1-G6
1051405LIB3061-039-LIB3061g22485BLASTN13611e−14996
Q1-K1-D2
1052405LIB3061-051-LIB3061g22485BLASTN17681e−14898
Q1-K1-G8
1053405LIB3061-015-LIB3061g22485BLASTN16671e−14693
Q1-K1-A12
1054405LIB3059-040-LIB3059g22485BLASTN18351e−14697
Q1-K1-H11
1055405LIB3061-002-LIB3061g22485BLASTN18451e−14489
Q1-K2-G5
1056405LIB3062-002-LIB3062g22485BLASTN16721e−14299
Q1-K2-G12
1057405LIB3059-048-LIB3059g22485BLASTN18221e−14299
Q1-K1-H5
1058405LIB3078-040-LIB3078g514945BLASTN18011e−14197
Q1-K1-F8
1059405LIB3078-001-LIB3078g22485BLASTN12461e−13995
Q1-K1-C7
1060405LIB3061-024-LIB3061g22485BLASTN13761e−13994
Q1-K1-A12
1061405LIB3061-026-LIB3061g22485BLASTN16431e−13893
Q1-K1-D3
1062405LIB3061-056-LIB3061g22485BLASTN17631e−13892
Q1-K1-D8
1063405LIB3069-041-LIB3069g514945BLASTN17581e−13797
Q1-K1-G12
1064405LIB3059-025-LIB3059g22485BLASTN15321e−13294
Q1-K1-E5
1065405LIB3061-014-LIB3061g22485BLASTN12941e−13088
Q1-K1-D4
1066405LIB3061-005-LIB3061g22485BLASTN15401e−13097
Q1-K1-C9
1067405LIB3061-016-LIB3061g22485BLASTN12511e−12985
Q1-K1-G2
1068405LIB3069-029-LIB3069g514945BLASTN16571e−12988
Q1-K1-B2
1069405LIB3078-012-LIB3078g514945BLASTN8571e−12886
Q1-K1-F7
1070405LIB3078-016-LIB3078g514945BLASTN13351e−12887
Q1-K1-D7
1071405LIB3062-049-LIB3062g514945BLASTN16091e−12888
Q1-K1-A8
1072405LIB143-006-LIB143g514945BLASTN16141e−12596
Q1-E1-G12
1073405LIB3059-024-LIB3059g22485BLASTN15291e−12383
Q1-K1-E5
1074405LIB3069-008-LIB3069g514945BLASTN10361e−11594
Q1-K1-C1
1075405LIB3059-018-LIB3059g514945BLASTN9101e−10393
Q1-K1-F11
1076405LIB3078-001-LIB3078g514945BLASTN9521e−9890
Q1-K1-E8
1077405LIB3059-017-LIB3059g22485BLASTN11701e−8892
Q1-K1-G4
1078405LIB3067-045-LIB3067g533251BLASTN9171e−8787
Q1-K1-E9
1079405LIB3062-015-LIB3062g514945BLASTN10661e−8696
Q1-K1-C1
1080405LIB3059-039-LIB3059g22485BLASTN8561e−8292
Q1-K1-A3
1081405LIB3062-024-LIB3062g514945BLASTN5481e−7988
Q1-K1-C3
1082405LIB3059-029-LIB3059g22485BLASTN9251e−7494
Q1-K1-F1
1083405LIB3059-006-LIB3059g22485BLASTN5301e−5083
Q1-K1-F4
1084405LIB3067-017-LIB3067g533251BLASTN4251e−26100
Q1-K1-C3
1085405LIB3061-028-LIB3061g19106BLASTX1181e−25100
Q1-K1-A9
1086537LIB3066-009-LIB3066g22485BLASTN13691e−12296
Q1-K1-B9
MAIZE HEXOKINASE
1087−700018381700018381H1SATMON001g1899025BLASTX1661e−1648
1088−700051079700051079H1SATMON003g1899025BLASTX841e−1150
1089−700101579700101579H1SATMON009g881521BLASTX2171e−2366
1090−700105594700105594H1SATMON010g3087888BLASTX1811e−1757
1091−700106018700106018H1SATMON010g3087888BLASTX1951e−1964
1092−700157233700157233H1SATMON012g3087888BLASTX1981e−2058
1093−700202992700202992H1SATMON003g3087888BLASTX891e−958
1094−700224204700224204H1SATMON011g1899024BLASTN5201e−3470
1095−700241273700241273H1SATMON010g3087888BLASTX1841e−1858
1096−700352183700352183H1SATMON023g1899024BLASTN4811e−3170
1097−700573814700573814H1SATMON030g1899024BLASTN5351e−3467
1098−700612458700612458H1SATMON033g619928BLASTX2291e−2661
1099−701168774701168774H1SATMONN05g619927BLASTN2521e−1062
11001195700457430H1SATMON029g3087888BLASTX1221e−1953
110113262700102942H1SATMON010g3087888BLASTX1131e−1853
11021378700456148H1SATMON029g1899025BLASTX2671e−2959
11031378700455837H1SATMON029g1899025BLASTX1661e−2160
110417305700460742H1SATMON031g619928BLASTX1311e−1557
110517305700614972H1SATMON033g1899025BLASTX1001e−853
11061842700089135H1SATMON011g619928BLASTX4051e−4970
11071842700430234H1SATMONN01g619927BLASTN4611e−2872
11081842700166122H1SATMON013g619928BLASTX1831e−1884
110924376700053677H1SATMON010g1899024BLASTN6421e−4470
111024376700152328H1SATMON007g619927BLASTN5551e−3769
111124376700623451H1SATMON034g619928BLASTX1971e−3272
111228388700089065H1SATMON011g619928BLASTX1861e−3061
11133345700072110H1SATMON007g619928BLASTX1251e−2466
11143345700472061H1SATMON025g619928BLASTX1121e−2055
11153345701173753H1SATMONN05g619928BLASTX1351e−1654
11163345700202130H1SATMON003g619928BLASTX1131e−1168
11175073700582054H1SATMON031g619928BLASTX2471e−2966
11185073700053432H1SATMON009g619928BLASTX2331e−2560
11196731700099009H1SATMON009g619927BLASTN7361e−5272
11206731700089738H1SATMON011g1899024BLASTN7001e−4970
11216731700171542H1SATMON013g619927BLASTN5301e−3574
11227565700356773H1SATMON024g1899025BLASTX1771e−1762
11239695700212172H1SATMON016g1899024BLASTN8321e−6074
11249695700212124H1SATMON016g1899024BLASTN8351e−6075
11259695700094278H1SATMON008g1899024BLASTN8191e−5974
1126-L30621307LIB3062-001-LIB3062g1899025BLASTX951e−3253
Q1-K2-G11
1127-L30782665LIB3078-007-LIB3078g3087888BLASTX1301e−3947
Q1-K1-E9
112824376LIB3069-041-LIB3069g1899024BLASTN6081e−6170
Q1-K1-E7
112928244LIB3061-004-LIB3061g687676BLASTN4991e−3065
Q1-K1-F9
113028388LIB3066-030-LIB3066g619928BLASTX2991e−6364
Q1-K1-G10
11313364LIB3078-051-LIB3078g687676BLASTN6191e−4167
Q1-K1-B3
11323364LIB3078-053-LIB3078g687676BLASTN6271e−4169
Q1-K1-C9
11333364LIB84-015-LIB84g687676BLASTN5541e−3569
Q1-E1-F7
11346731LIB3061-028-LIB3061g1899024BLASTN8311e−6070
Q1-K1-C1
11359695LIB143-065-LIB143g1899024BLASTN10961e−8273
Q1-E1-C10
MAIZE FRUCTOKINASE
1136−700106058700106058H1SATMON010g1052972BLASTN2201e−968
1137−700151135700151135H1SATMON007g297014BLASTN3511e−1875
1138−700169310700169310H1SATMON013g1052972BLASTN2731e−1259
1139−700210226700210226H1SATMON016g1052973BLASTX1881e−2468
1140−700257901700257901H1SATMON017g297015BLASTX2001e−2072
1141−700621274700621274H1SATMON034g1052973BLASTX1411e−2464
114211678700105513H1SATMON010g1052972BLASTN5801e−3964
114311678700170725H1SATMON013g1052972BLASTN4781e−3166
11442526700159958H1SATMON012g1052973BLASTX1521e−1464
11452754700102678H1SATMON010g1052972BLASTN7071e−5069
11462754700102312H1SATMON010g1052972BLASTN7011e−4969
11472754700205695H1SATMON003g1915973BLASTN6331e−4369
11482754700221511H1SATMON011g1915973BLASTN5871e−4069
11492754700469079H1SATMON025g1052972BLASTN5841e−3972
11502754701173520H1SATMONN05g1915973BLASTN3421e−3670
11512754700267332H1SATMON017g1052972BLASTN5411e−3564
11522754701164907H1SATMONN04g1052973BLASTX2801e−3357
11532754700450050H2SATMON028g1052973BLASTX1601e−3160
11542754701182860H1SATMONN06g297015BLASTX1881e−2765
11552754700467520H1SATMON025g1915974BLASTX2421e−2660
11562754700159848H1SATMON012g1052973BLASTX1971e−2463
11573287700088103H1SATMON011g2102693BLASTX2391e−4374
11583287700210913H1SATMON016g2102693BLASTX2501e−3577
11593287700167609H1SATMON013g1052973BLASTX3001e−3568
11603287700085916H1SATMON011g1052972BLASTN5531e−3564
11613287700262715H1SATMON017g1915974BLASTX2011e−3371
11623287700170179H1SATMON013g1052973BLASTX2891e−3367
11633287700615671H1SATMON033g1052972BLASTN5151e−3263
11643287700223640H1SATMON011g1052973BLASTX2191e−3167
11653287700215234H1SATMON016g1052973BLASTX1901e−3067
11663287700203946H1SATMON003g1052973BLASTX1981e−3060
11673287700028411H1SATMON003g2102693BLASTX1101e−2957
11683287700224307H1SATMON011g1052973BLASTX1591e−2987
11693287700072013H1SATMON007g1052973BLASTX1911e−2965
11703287700215669H1SATMON016g1052973BLASTX2601e−2957
11713287700353954H1SATMON024g1052973BLASTX2601e−2961
11723287700342211H1SATMON021g1052973BLASTX1371e−2867
11733287700085462H1SATMON011g297014BLASTN4661e−2862
11743287700220972H1SATMON011g1052973BLASTX1091e−2783
11753287700451141H1SATMON028g1052973BLASTX2451e−2763
11763287700087484H1SATMON011g1052972BLASTN4401e−2664
11773287700343411H1SATMON021g1052973BLASTX1631e−2567
11783287700217263H1SATMON016g1915973BLASTN3931e−2568
11793287700030665H1SATMON003g1052973BLASTX1761e−2471
11803287700343380H1SATMON021g1052973BLASTX2281e−2457
11813287701159743H2SATMONN04g1052973BLASTX1831e−2355
11823287700221543H1SATMON011g1052973BLASTX2171e−2350
11833287700333946H1SATMON019g1052973BLASTX1781e−2266
11843287700091730H1SATMON011g1052973BLASTX1711e−2164
11853287700570521H1SATMON030g1915974BLASTX981e−1858
11863287700048604H1SATMON003g1052973BLASTX881e−1554
11873287700208681H1SATMON016g1052973BLASTX1291e−1555
11883287700028328H1SATMON003g1052973BLASTX1621e−1566
11893287700220530H1SATMON011g1052973BLASTX1411e−1488
11903287700243726H1SATMON010g1052973BLASTX1531e−1468
11913287700142502H1SATMON012g1052973BLASTX1571e−1447
11923287700336537H1SATMON019g1052973BLASTX1411e−1250
11933287700205308H1SATMON003g1052973BLASTX1331e−1175
11945966700084171H1SATMON011g1052972BLASTN4481e−2666
11955966700084951H1SATMON011g2102693BLASTX2141e−2273
11965966700089353H1SATMON011g2102691BLASTX1951e−2072
11975966700220723H1SATMON011g1915974BLASTX1981e−2073
11985966700084412H1SATMON011g2102693BLASTX1791e−1976
11995966700085628H1SATMON011g2102691BLASTX1801e−1872
12005966700027982H1SATMON003g2102691BLASTX1781e−1772
12015966700106884H1SATMON010g1915974BLASTX1481e−1375
12025966700053135H1SATMON008g1915974BLASTX1311e−1173
12035966700027988H1SATMON003g1915974BLASTX1341e−1165
12045966700207083H1SATMON003g1915974BLASTX1001e−1046
12055966700158574H1SATMON012g1915974BLASTX1201e−950
12062754LIB3061-030-LIB3061g1052972BLASTN8821e−6467
Q1-K1-G12
12072754LIB3061-030-LIB3061g1052972BLASTN7511e−5268
Q1-K1-G11
12083287LIB3067-040-LIB3067g1052972BLASTN6571e−4464
Q1-K1-H10
12093287LIB84-024-LIB84g1052972BLASTN6381e−4264
Q1-E1-H7
12103287LIB3069-045-LIB3069g1052972BLASTN5921e−3861
Q1-K1-F6
12113287LIB3061-014-LIB3061g1052973BLASTX1751e−3641
Q1-K1-A3
12123287LIB3062-019-LIB3062g1052973BLASTX1541e−3068
Q1-K1-H11
12133287LIB3067-054-LIB3067g1052972BLASTN4951e−3061
Q1-K1-C9
12143287LIB3067-022-LIB3067g1052973BLASTX1411e−2768
Q1-K1-H4
12153287LIB3069-045-LIB3069g1052972BLASTN4391e−2557
Q1-K1-F2
MAIZE NDP-KINASE
1216−700575072700575072H1SATMON030g303849BLASTX741e−1389
1217−701170773701170773H1SATMONN05g1777930BLASTX1321e−3071
12182462700050003H1SATMON003g218233BLASTN6561e−5883
12192462700204789H1SATMON003g218233BLASTN7801e−5887
12202462700049819H1SATMON003g218233BLASTN7861e−5886
12212462700204211H1SATMON003g218233BLASTN7861e−5886
12222462700205742H1SATMON003g218233BLASTN7631e−5786
12232462700207611H1SATMON016g218233BLASTN7641e−5787
12242462700072505H1SATMON007g218233BLASTN7401e−5586
12252462700236468H1SATMON010g218233BLASTN7101e−5286
12262462701181270H1SATMONN06g218233BLASTN4451e−5186
12272462700573201H1SATMON030g218233BLASTN6911e−5181
12282462700452623H1SATMON028g218233BLASTN6941e−5185
12292462700351523H1SATMON023g218233BLASTN6791e−5086
12302462700042795H1SATMON004g218233BLASTN6301e−4586
12312462700445979H1SATMON027g218233BLASTN5951e−4386
12322462700201855H1SATMON003g218233BLASTN6041e−4387
12332462700573101H1SATMON030g218233BLASTN5941e−4278
12342462700049543H1SATMON003g218233BLASTN5771e−4179
12352462700432359H1SATMONN01g218233BLASTN5611e−4081
12362462701182021H1SATMONN06g218233BLASTN5611e−4085
12372462701182019H1SATMONN06g218233BLASTN5661e−4086
12382462700150928H1SATMON007g218233BLASTN5691e−4085
12392462700202824H1SATMON003g218233BLASTN3361e−3986
12402462700451056H1SATMON028g218233BLASTN5531e−3985
12412462700449958H1SATMON028g218233BLASTN5441e−3886
12422462700347592H1SATMON023g218233BLASTN4031e−3478
12432462700573195H1SATMON030g218233BLASTN2001e−2284
12442462700582836H1SATMON031g303849BLASTX1571e−1583
12452462700029459H1SATMON003g303849BLASTX1341e−1184
124627065700583429H1SATMON031g1064895BLASTX721e−1354
1247-L1482546LIB148-007-LIB148g218233BLASTN3591e−1975
Q1-E1-E6
12482462LIB3067-039-LIB3067g218233BLASTN7111e−5282
Q1-K1-B10
12492462LIB3078-001-LIB3078g218233BLASTN4881e−4985
Q1-K1-F3
12502462LIB3067-029-LIB3067g1236951BLASTX1661e−3196
Q1-K1-C3
125125174LIB189-022-LIB189g758643BLASTN4401e−2576
Q1-E1-E9
MAIZE GLUCOSE-6-PHOSPHATE 1-DEHYDROGENASE
1252−700047645700047645H1SATMON003g471345BLASTX1931e−2158
1253−700210379700210379H1SATMON016g1480344BLASTX1031e−1085
12549135700203121H1SATMON003g1166405BLASTX1081e−1078
MAIZE PHOSPHOGLUCOMUTASE
1255−700045655700045655H1SATMON004g534982BLASTX1441e−1265
1256−700053330700053330H1SATMON009g3294467BLASTX2111e−2371
1257−700102193700102193H1SATMON010g534982BLASTX1451e−1453
1258−700166982700166982H1SATMON013g2795876BLASTX1681e−1652
1259−700169540700169540H1SATMON013g534982BLASTX1801e−1761
1260−700210088700210088H1SATMON016g534982BLASTX3281e−3855
1261−700573194700573194H1SATMON030g534982BLASTX1921e−2154
1262−700616588700616588H1SATMON033g3294468BLASTN5931e−4895
1263119700574655H1SATMON030g3294466BLASTN17051e−13398
1264119700574672H1SATMON030g3294466BLASTN8201e−121100
1265119700100992H1SATMON009g3294466BLASTN15451e−11999
1266119700615409H1SATMON033g3294466BLASTN10501e−118100
1267119700210693H1SATMON016g3294468BLASTN15151e−117100
1268119700381526H1SATMON023g3294468BLASTN14901e−115100
1269119700026372H1SATMON003g3294466BLASTN14631e−11399
1270119700201578H1SATMON003g3294468BLASTN6771e−11296
1271119700101083H1SATMON009g3294468BLASTN14301e−110100
1272119700217101H1SATMON016g3294468BLASTN14201e−109100
1273119700222466H1SATMON011g3294466BLASTN9571e−10697
1274119700072492H1SATMON007g3294466BLASTN13811e−10699
1275119700043724H1SATMON004g3294468BLASTN13901e−106100
1276119700346762H1SATMON021g3294468BLASTN13331e−10294
1277119700347741H1SATMON023g3294468BLASTN13391e−10297
1278119700550792H1SATMON022g3294466BLASTN7311e−10199
1279119700380144H1SATMON021g3294466BLASTN12161e−9897
1280119700241526H1SATMON010g3294466BLASTN12851e−98100
1281119700380456H1SATMON021g3294468BLASTN6501e−9799
1282119700238734H1SATMON010g3294466BLASTN9741e−9797
1283119700083634H1SATMON011g3294468BLASTN12651e−96100
1284119700383086H1SATMON024g3294466BLASTN9611e−9496
1285119700169630H1SATMON013g3294466BLASTN12451e−94100
1286119701177766H1SATMONN05g3294466BLASTN11871e−9397
1287119700142461H1SATMON012g3294466BLASTN12311e−9399
1288119700044235H1SATMON004g3294466BLASTN11751e−89100
1289119700216921H1SATMON016g3294466BLASTN11651e−88100
1290119700333779H1SATMON019g3294466BLASTN9961e−8796
1291119700021881H1SATMON001g3294468BLASTN11201e−84100
1292119700049194H1SATMON003g3294468BLASTN9401e−8298
1293119700164477H1SATMON013g3294466BLASTN10911e−8299
1294119700169514H1SATMON013g3294468BLASTN8651e−80100
1295119700050896H1SATMON003g3294466BLASTN5911e−7694
1296119700172394H1SATMON013g3294466BLASTN10241e−7699
1297119700211437H1SATMON016g3294466BLASTN9941e−7399
1298119700084535H1SATMON011g3294468BLASTN9731e−7299
1299119700203439H1SATMON003g3294466BLASTN7651e−71100
1300119700257833H1SATMON017g3294468BLASTN6111e−6994
1301119700621831H1SATMON034g3294466BLASTN4121e−5290
1302119700354511H1SATMON024g3294468BLASTN7031e−5291
1303119700203525H1SATMON003g3294468BLASTN7081e−5099
1304119700020476H1SATMON001g3294468BLASTN6581e−4599
1305119700050562H1SATMON003g3294466BLASTN5441e−4288
1306119700613868H1SATMON033g3294466BLASTN6151e−42100
1307119700574982H1SATMON030g3294466BLASTN4731e−3597
1308119700049512H1SATMON003g3294466BLASTN2681e−2995
1309119700260372H2SATMON017g3294466BLASTN2261e−1089
131016726700082801H1SATMON011g2829893BLASTX2781e−3055
131116726700212054H1SATMON016g2829893BLASTX2201e−2353
131219462700097450H1SATMON009g1814400BLASTN3231e−2964
131319462700441165H1SATMON026g1408296BLASTX2391e−2561
131424348700379424H1SATMON020g3294466BLASTN7071e−5098
13152587700089556H1SATMON011g2829893BLASTX1171e−867
13163016700204345H1SATMON003g3294468BLASTN17841e−13998
13173016700098713H1SATMON009g3294468BLASTN15161e−11799
13183016700084751H1SATMON011g3294466BLASTN14751e−114100
13193016700351326H1SATMON023g3294468BLASTN14601e−112100
13203016700097161H1SATMON009g3294466BLASTN13081e−10998
13213016700266423H1SATMON017g3294468BLASTN10651e−10896
13223016700349605H1SATMON023g3294466BLASTN13351e−107100
13233016700350209H1SATMON023g3294468BLASTN11881e−10697
13243016700265291H1SATMON017g3294468BLASTN8731e−10098
13253016700457572H1SATMON029g3294466BLASTN12881e−9898
13263016700334810H1SATMON019g3294468BLASTN8631e−9799
13273016700194444H1SATMON014g3294466BLASTN12651e−96100
13283016700457426H1SATMON029g3294466BLASTN12361e−9498
13293016700210958H1SATMON016g3294466BLASTN11481e−9298
13303016700075135H1SATMON007g3294468BLASTN12191e−9297
13313016700152065H1SATMON007g3294466BLASTN11351e−9099
13323016700219672H1SATMON011g3294468BLASTN8231e−8999
13333016700170425H1SATMON013g3294466BLASTN11101e−83100
13343016700153495H1SATMON007g3294468BLASTN6401e−82100
13353016700348567H1SATMON023g3294468BLASTN5571e−8187
13363016700803158H1SATMON036g3294468BLASTN6301e−6085
13373016700264923H1SATMON017g3294468BLASTN3401e−5098
13383016700615715H1SATMON033g3294466BLASTN5671e−4896
13393016700027830H1SATMON003g3294468BLASTN6321e−4395
13403016700350539H1SATMON023g3294466BLASTN3331e−4196
13414562700044891H1SATMON004g3294466BLASTN6501e−4574
13424562700215538H1SATMON016g3294466BLASTN5551e−3767
13439894700220429H1SATMON011g3294468BLASTN13021e−9999
13449894700236461H1SATMON010g3294466BLASTN10541e−9097
1345-L30594453LIB3059-042-LIB3059g1814401BLASTX2901e−4958
Q1-K1-B5
1346-L30605287LIB3060-049-LIB3060g534982BLASTX1721e−3477
Q1-K1-B7
1347119LIB3059-019-LIB3059g1881692BLASTN20941e−16598
Q1-K1-H1
1348119LIB3059-031-LIB3059g1881692BLASTN19261e−15196
Q1-K1-H10
1349119LIB3069-012-LIB3069g1881692BLASTN11881e−14690
Q1-K1-F2
1350119LIB36-019-LIB36g1881692BLASTN17831e−13990
Q1-E1-A7
1351119LIB3078-023-LIB3078g1881692BLASTN8601e−12487
Q1-K1-C3
1352119LIB3067-058-LIB3067g1881692BLASTN9911e−11499
Q1-K1-G1
1353119LIB3062-048-LIB3062g1881692BLASTN11811e−10397
Q1-K1-B7
1354119LIB3069-023-LIB3069g1881692BLASTN11761e−8784
Q1-K1-G4
1355119LIB3069-025-LIB3069g1881692BLASTN6111e−6591
Q1-K1-B6
135624348LIB3066-043-LIB3066g1881692BLASTN5601e−37100
Q1-K1-F11
135724348LIB3067-048-LIB3067g1881692BLASTN5431e−3699
Q1-K1-F3
13583016LIB143-002-LIB143g2829893BLASTX2241e−5172
Q1-E1-C12
13593016LIB189-034-LIB189g2829893BLASTX2161e−4868
Q1-E1-A11
13603016LIB3069-043-LIB3069g1814401BLASTX981e−3264
Q1-K1-D5
MAIZE UDP-GLUCOSE PYROPHOSPHORYLASE
1361−700197315700197315H1SATMON014g1388021BLASTX1221e−970
1362−700203530700203530H1SATMON003g1212995BLASTN5681e−3878
1363−700267284700267284H1SATMON017g1212996BLASTX1501e−1387
1364−700336683700336683H1SATMON019g1752677BLASTX1501e−2782
1365−700342324700342324H1SATMON021g3107931BLASTX951e−1480
1366−700354856700354856H1SATMON024g1388021BLASTX1211e−2275
1367−700613858700613858H1SATMON033g1212995BLASTN7761e−5988
136814982700028996H1SATMON003g1212995BLASTN5601e−3776
136914982700155115H1SATMON007g1212995BLASTN3991e−3181
137014982700356747H1SATMON024g1388021BLASTX1661e−1576
137119537700573761H1SATMON030g1212995BLASTN9541e−7079
137219537700208049H1SATMON016g1212995BLASTN9011e−6678
137319537700086382H1SATMON011g1212995BLASTN8851e−6477
137469700091881H1SATMON011g1212995BLASTN8441e−10589
137569700624406H1SATMON034g1212995BLASTN8161e−9788
137669700211464H1SATMON016g1212995BLASTN12511e−9588
137769700099836H1SATMON009g1212995BLASTN12391e−9488
137869700084756H1SATMON011g1212995BLASTN12401e−9490
137969700076136H1SATMON007g1212995BLASTN12431e−9489
138069700073071H1SATMON007g1212995BLASTN11631e−8886
138169700614228H1SATMON033g1212995BLASTN10131e−8784
138269700379926H1SATMON021g1212995BLASTN11381e−8688
138369700089172H1SATMON011g1212995BLASTN11411e−8688
138469700265063H1SATMON017g1212995BLASTN11471e−8686
138569700085964H1SATMON011g1212995BLASTN11351e−8585
138669700282281H2SATMON023g1212995BLASTN11361e−8586
138769700429855H1SATMONN01g1212995BLASTN11141e−8489
138869700347453H1SATMON023g1212995BLASTN11171e−8487
138969700265087H1SATMON017g1212995BLASTN11201e−8487
139069700092705H1SATMON008g1212995BLASTN11221e−8487
139169700212686H1SATMON016g1212995BLASTN11231e−8491
139269700623332H1SATMON034g1212995BLASTN8001e−8386
139369700041787H1SATMON004g1212995BLASTN10911e−8291
139469700219031H1SATMON011g1212995BLASTN10931e−8289
139569700218632H1SATMON011g1212995BLASTN10861e−8190
139669700211962H1SATMON016g1212995BLASTN10861e−8186
139769700220729H1SATMON011g1212995BLASTN9161e−8084
139869700197025H1SATMON014g1212995BLASTN10631e−7989
139969700086546H1SATMON011g1212995BLASTN10491e−7886
140069700217064H1SATMON016g1212995BLASTN10511e−7888
140169700799128H1SATMON036g1212995BLASTN6181e−7788
140269700265488H1SATMON017g1212995BLASTN10301e−7784
140369700043842H1SATMON004g1212995BLASTN10351e−7787
140469700236833H1SATMON010g1212995BLASTN10351e−7787
140569700219083H1SATMON011g1212995BLASTN10361e−7788
140669700042338H1SATMON004g1212995BLASTN10371e−7787
140769700352484H1SATMON023g1212995BLASTN10381e−7785
140869700083771H1SATMON011g1212995BLASTN6131e−7691
140969700473855H1SATMON025g1212995BLASTN7551e−7685
141069700353922H1SATMON024g1212995BLASTN10241e−7685
141169700023267H1SATMON003g1212995BLASTN10071e−7589
141269700157596H1SATMON012g1212995BLASTN10081e−7587
141369700218718H1SATMON011g1212995BLASTN10121e−7586
141469700162316H1SATMON012g1212995BLASTN6261e−7480
141569700046475H1SATMON004g1212995BLASTN10031e−7485
141669700466010H1SATMON025g1212995BLASTN5581e−7382
141769700571392H1SATMON030g1212995BLASTN9851e−7385
141869700165241H1SATMON013g1212995BLASTN9871e−7385
141969700457410H1SATMON029g1212995BLASTN9881e−7387
142069700194672H1SATMON014g1212995BLASTN9631e−7186
142169700089746H1SATMON011g1212995BLASTN9641e−7183
142269700801620H1SATMON036g1212995BLASTN5361e−7091
142369700264785H1SATMON017g1212995BLASTN9521e−7084
142469700244093H1SATMON010g1212995BLASTN9541e−7085
142569700043787H1SATMON004g1212995BLASTN9571e−7085
142669700267269H1SATMON017g1212995BLASTN8671e−6985
142769700167985H1SATMON013g1212995BLASTN9401e−6989
142869700799042H1SATMON036g1212995BLASTN8121e−6889
142969700163824H1SATMON013g1212995BLASTN8881e−6586
143069700098307H1SATMON009g1212995BLASTN4611e−6381
143169700805267H1SATMON036g1212995BLASTN7341e−6388
143269700204843H1SATMON003g1212995BLASTN8541e−6288
143369700206721H1SATMON003g1212995BLASTN4611e−6181
143469700018559H1SATMON001g1212995BLASTN8471e−6185
143569700026241H1SATMON003g1212995BLASTN8471e−6187
143669700099987H1SATMON009g1212995BLASTN4611e−6081
143769700475628H1SATMON025g1212995BLASTN7501e−5980
143869700016675H1SATMON001g1212995BLASTN8141e−5986
143969700150144H1SATMON007g1212995BLASTN8221e−5986
144069700267260H1SATMON017g1212995BLASTN4611e−5880
144169700261336H1SATMON017g1212995BLASTN5641e−5882
144269700618652H1SATMON033g1212995BLASTN7301e−5878
144369700469914H1SATMON025g1212995BLASTN7351e−5889
144469700048027H1SATMON003g1212995BLASTN8071e−5883
144569700165703H1SATMON013g1212995BLASTN7961e−5785
144669700265403H1SATMON017g1212995BLASTN7971e−5778
144769700099428H1SATMON009g1212995BLASTN4741e−5688
144869700243212H1SATMON010g1212995BLASTN7791e−5684
144969700092996H1SATMON008g1212995BLASTN7891e−5684
145069700803035H1SATMON036g1212995BLASTN4361e−5480
145169700235803H1SATMON010g1212995BLASTN6881e−5479
145269700172581H1SATMON013g1212995BLASTN7541e−5479
145369700214715H1SATMON016g1212995BLASTN7621e−5486
145469700223082H1SATMON011g1212995BLASTN7641e−5484
145569700093483H1SATMON008g1212995BLASTN3571e−5188
145669700261920H1SATMON017g1212995BLASTN3631e−5182
145769700221718H1SATMON011g1212995BLASTN3631e−5183
145869700453106H1SATMON028g1212995BLASTN6701e−5182
145969700210506H1SATMON016g1212995BLASTN4611e−5085
146069700212333H1SATMON016g1212995BLASTN4431e−4983
146169700072654H1SATMON007g1212995BLASTN4431e−4979
146269700218282H1SATMON016g1212995BLASTN4521e−4985
146369700263725H1SATMON017g1212995BLASTN6621e−4980
146469700343083H1SATMON021g1212995BLASTN3881e−4880
146569700219739H1SATMON011g1212995BLASTN4431e−4881
146669700620336H1SATMON034g1212995BLASTN6211e−4888
146769700264630H1SATMON017g1212995BLASTN3771e−4780
146869700439242H1SATMON026g1212995BLASTN6481e−4783
146969700259658H1SATMON017g1212995BLASTN5111e−4579
147069700263521H1SATMON017g1212995BLASTN4611e−4479
147169700261387H1SATMON017g1212995BLASTN4611e−4480
147269700439277H1SATMON026g1212995BLASTN4611e−4384
147369700452839H1SATMON028g1212995BLASTN5441e−4377
147469700220236H1SATMON011g1212995BLASTN4481e−4084
147569700472602H1SATMON025g1212995BLASTN2541e−3881
147669700266424H1SATMON017g1212995BLASTN4991e−3780
147769700449187H1SATMON028g1212995BLASTN5401e−3681
147869700202731H1SATMON003g1212995BLASTN5431e−3679
147969700156144H2SATMON007g1212995BLASTN4411e−3576
148069700442679H1SATMON026g1212995BLASTN5331e−3580
148169700449879H2SATMON028g1212995BLASTN5351e−3581
148269700266832H1SATMON017g1212995BLASTN3461e−3477
148369700332389H1SATMON019g1212995BLASTN3821e−3484
148469700804202H1SATMON036g1212995BLASTN4361e−3476
148569700151037H1SATMON007g1212995BLASTN4431e−3479
148669700802810H1SATMON036g1212995BLASTN5251e−3485
148769700455879H1SATMON029g1212995BLASTN4481e−3272
148869700427769H1SATMONN01g1212995BLASTN4811e−3181
148969700464626H1SATMON025g1212995BLASTN3881e−3076
149069700439228H1SATMON026g1212995BLASTN4701e−3077
149169700256847H1SATMON017g1212995BLASTN2641e−2985
149269700204881H1SATMON003g1212995BLASTN4301e−2981
149369700076032H1SATMON007g1212995BLASTN2181e−2672
149469700426342H1SATMONN01g1212995BLASTN4431e−2679
149569700209062H1SATMON016g1212995BLASTN2791e−2480
149669700076988H1SATMON007g1212995BLASTN3371e−2483
149769700349778H1SATMON023g1212995BLASTN4061e−2481
149869700261886H1SATMON017g1212995BLASTN2871e−1580
149969700426642H1SATMONN01g1388021BLASTX1611e−1476
150069700155195H1SATMON007g1212995BLASTN1551e−1081
150169700211992H1SATMON016g1212996BLASTX1181e−985
1502-L1485255LIB148-053-LIB148g1212995BLASTN6911e−4880
Q1-E1-E12
1503-L30663959LIB3066-015-LIB3066g218000BLASTN2511e−965
Q1-K1-F12
150419537LIB3066-025-LIB3066g1212995BLASTN10011e−7479
Q1-K1-E5
150569LIB3059-023-LIB3059g1212995BLASTN13011e−13389
Q1-K1-C8
150669LIB3078-022-LIB3078g1212995BLASTN16561e−12986
Q1-K1-C1
150769LIB3059-037-LIB3059g1212995BLASTN16461e−12886
Q1-K1-H5
150869LIB3061-030-LIB3061g1212995BLASTN14931e−12486
Q1-K1-A12
150969LIB3061-023-LIB3061g1212995BLASTN15981e−12486
Q1-K1-A1
151069LIB3079-001-LIB3079g1212995BLASTN16001e−12483
Q1-K1-D12
151169LIB189-028-LIB189g1212995BLASTN15831e−12387
Q1-E1-E3
151269LIB3067-017-LIB3067g1212995BLASTN13641e−12088
Q1-K1-D9
151369LIB3068-007-LIB3068g1212995BLASTN15011e−11685
Q1-K1-F9
151469LIB3069-025-LIB3069g1212995BLASTN14871e−11585
Q1-K1-E9
151569LIB3069-026-LIB3069g1212995BLASTN14531e−11285
Q1-K1-E11
151669LIB3066-006-LIB3066g1212995BLASTN10771e−10783
Q1-K1-G12
151769LIB3067-027-LIB3067g1212995BLASTN14011e−10786
Q1-K1-D12
151869LIB189-010-LIB189g1212995BLASTN13681e−10585
Q1-E1-H10
151969LIB3066-015-LIB3066g1212995BLASTN12891e−10482
Q1-K1-G12
152069LIB3061-016-LIB3061g1212995BLASTN11801e−10284
Q1-K1-G11
152169LIB3059-032-LIB3059g1212995BLASTN13341e−10287
Q1-K1-G11
152269LIB3067-059-LIB3067g1212995BLASTN10901e−10085
Q1-K1-G12
152369LIB3061-049-LIB3061g1212995BLASTN12231e−9879
Q1-K1-C8
152469LIB3062-044-LIB3062g1212995BLASTN12591e−9683
Q1-K1-F2
152569LIB3061-010-LIB3061g1212995BLASTN11801e−9584
Q1-K1-F5
152669LIB3067-018-LIB3067g1212995BLASTN11271e−8982
Q1-K1-A12
152769LIB3067-030-LIB3067g1212995BLASTN11711e−8883
Q1-K1-F4
152869LIB3062-021-LIB3062g1212995BLASTN11381e−8687
Q1-K1-F10
152969LIB3061-034-LIB3061g1212995BLASTN11481e−8685
Q1-K1-D8
153069LIB3066-049-LIB3066g1212995BLASTN11341e−8583
Q1-K1-C1
153169LIB3078-002-LIB3078g1212995BLASTN8591e−7786
Q1-K1-F5
153269LIB84-011-LIB84g1212995BLASTN10201e−7683
Q1-E1-G9
153369LIB3067-043-LIB3067g1212995BLASTN5741e−5977
Q1-K1-D2
153469LIB189-003-LIB189g1212995BLASTN2471e−4077
Q1-E1-G5
153569LIB3062-008-LIB3062g1212995BLASTN5761e−3763
Q1-K1-E6
153669LIB189-016-LIB189g1212996BLASTX1561e−3078
Q1-E1-H7
153769LIB3067-007-LIB3067g1212996BLASTX1451e−2882
Q1-K1-G4
SOYBEAN TRIOSE PHOSPHATE ISOMERASE
1538−700743237700743237H1SOYMON012g407525BLASTX1731e−1791
1539−700977730700977730H1SOYMON009g602589BLASTN3731e−2071
1540−701056176701056176H1SOYMON032g806311BLASTN7521e−5374
1541−701110172701110172H1SOYMON036g806311BLASTN8011e−5778
154210244700995141H1SOYMON011g806311BLASTN4701e−3087
154310244701124548H1SOYMON037g806311BLASTN4901e−3088
154410244700739771H1SOYMON012g806311BLASTN3291e−1677
154510244700999820H1SOYMON018g806312BLASTX1471e−1384
154610244701119858H1SOYMON037g806312BLASTX1181e−972
154710535700988684H1SOYMON009g806311BLASTN9051e−6679
154810535700902425H1SOYMON027g806311BLASTN8721e−6380
15491357701069004H1SOYMON034g806311BLASTN8321e−6081
15501357701151554H1SOYMON031g806311BLASTN5681e−3882
15511357700659936H1SOYMON004g806311BLASTN5451e−3679
155216700680927H1SOYMON008g256119BLASTN10201e−8178
155316700656871H1SOYMON004g256119BLASTN9031e−6681
155416701124364H1SOYMON037g256119BLASTN8721e−6480
155516701134707H2SOYMON038g256119BLASTN8741e−6481
155616700673750H1SOYMON007g256119BLASTN7811e−6081
155716701123269H1SOYMON037g602589BLASTN8191e−5978
155816701004846H1SOYMON019g256119BLASTN8011e−5880
155916700993362H1SOYMON011g256119BLASTN8081e−5880
156016701005445H1SOYMON019g256119BLASTN6301e−5678
156116701134327H1SOYMON038g602589BLASTN7821e−5679
156216701148169H1SOYMON031g602589BLASTN5741e−5176
156316701153410H1SOYMON031g602589BLASTN4511e−5080
156416700830168H1SOYMON019g256119BLASTN7051e−5077
156516701120627H1SOYMON037g602589BLASTN7151e−5078
156616700975358H1SOYMON009g602589BLASTN6281e−4977
156716700755979H1SOYMON014g602589BLASTN6971e−4979
156816701131374H1SOYMON038g602589BLASTN7031e−4979
156916700994166H1SOYMON011g602589BLASTN5131e−4777
157016701138038H1SOYMON038g602589BLASTN6721e−4777
157116700974248H1SOYMON005g602589BLASTN6581e−4677
157216700655832H1SOYMON004g602589BLASTN6641e−4678
157316700758320H1SOYMON015g602589BLASTN4091e−4580
157416701064709H1SOYMON034g602589BLASTN4771e−4578
157516701138504H1SOYMON038g602589BLASTN5911e−4576
157616700980284H1SOYMON009g602589BLASTN6521e−4579
157716701133585H2SOYMON038g602589BLASTN6341e−4478
157816700674706H1SOYMON007g602589BLASTN6341e−4478
157916700964927H1SOYMON022g602589BLASTN6391e−4478
158016700830923H1SOYMON019g602589BLASTN6261e−4376
158116700662845H1SOYMON005g602589BLASTN6171e−4276
158216701133824H1SOYMON038g602589BLASTN6191e−4278
158316700848913H1SOYMON021g602589BLASTN6031e−4177
158416701005984H1SOYMON019g602589BLASTN6041e−4178
158516701140769H1SOYMON038g602589BLASTN6051e−4176
158616700753357H1SOYMON014g602589BLASTN3281e−4078
158716701056336H1SOYMON032g602589BLASTN3441e−4077
158816700895411H1SOYMON027g602589BLASTN5931e−4078
158916701060188H1SOYMON033g602589BLASTN2771e−3980
159016700739461H1SOYMON012g602589BLASTN5731e−3977
159116700941104H1SOYMON024g602589BLASTN5791e−3979
159216700732960H1SOYMON010g602589BLASTN5811e−3978
159316700686476H1SOYMON008g602589BLASTN5831e−3979
159416701054231H1SOYMON032g602589BLASTN5831e−3977
159516700671690H1SOYMON006g602589BLASTN5661e−3877
159616700941174H1SOYMON024g602589BLASTN5691e−3878
159716701125091H1SOYMON037g256119BLASTN3581e−3774
159816700989827H1SOYMON011g602589BLASTN5551e−3778
159916700835006H1SOYMON019g602589BLASTN5551e−3775
160016700834847H1SOYMON019g602589BLASTN5591e−3778
160116700953411H1SOYMON022g602589BLASTN3141e−3680
160216700869222H1SOYMON016g602589BLASTN5411e−3678
160316700850633H1SOYMON023g602589BLASTN5441e−3678
160416700890283H1SOYMON024g602589BLASTN3101e−3580
160516700727079H1SOYMON009g414549BLASTN3581e−3573
160616700892544H1SOYMON024g602589BLASTN4861e−3578
160716700869230H1SOYMON016g602589BLASTN5281e−3578
160816700993034H1SOYMON011g602589BLASTN5181e−3475
160916700975553H1SOYMON009g414549BLASTN5241e−3479
161016700651326H1SOYMON003g602589BLASTN3561e−3380
161116701215308H1SOYMON035g414549BLASTN4501e−3375
161216700654480H1SOYMON004g414549BLASTN5111e−3380
161316701045128H1SOYMON032g414549BLASTN5121e−3378
161416701060759H1SOYMON033g414549BLASTN5131e−3380
161516700741652H1SOYMON012g602589BLASTN4931e−3279
161616700675469H1SOYMON007g602589BLASTN4941e−3278
161716700657787H1SOYMON004g414549BLASTN4951e−3279
161816701009957H2SOYMON019g414549BLASTN4951e−3280
161916700983693H1SOYMON009g414549BLASTN4951e−3280
162016701156784H1SOYMON031g602589BLASTN4951e−3278
162116700893935H1SOYMON024g602589BLASTN4811e−3179
162216701144619H1SOYMON031g414549BLASTN4851e−3178
162316701148851H1SOYMON031g602589BLASTN4871e−3179
162416701058218H1SOYMON033g602589BLASTN4951e−3178
162516700975165H1SOYMON009g414549BLASTN4661e−3080
162616701100165H1SOYMON028g602589BLASTN4851e−3079
162716701150241H1SOYMON031g602589BLASTN4551e−2979
162816701098308H1SOYMON028g414549BLASTN4601e−2979
162916701150440H1SOYMON031g602589BLASTN4621e−2978
163016700685125H1SOYMON008g414549BLASTN4711e−2981
163116701061565H1SOYMON033g414549BLASTN4711e−2981
163216700991418H1SOYMON011g602589BLASTN3941e−2868
163316701156156H1SOYMON031g602589BLASTN4561e−2878
163416701007231H2SOYMON019g602589BLASTN4611e−2879
163516700829667H1SOYMON019g414549BLASTN3331e−2773
163616701156033H1SOYMON031g602589BLASTN4321e−2778
163716701014293H1SOYMON019g414549BLASTN4461e−2777
163816701152138H1SOYMON031g414549BLASTN4501e−2781
163916700945665H1SOYMON024g414549BLASTN4501e−2781
164016701001407H1SOYMON018g169820BLASTN2191e−2672
164116700983185H1SOYMON009g414549BLASTN4351e−2672
164216700752364H1SOYMON014g414549BLASTN4411e−2676
164316700992409H1SOYMON011g414549BLASTN4271e−2575
164416701109396H1SOYMON036g414549BLASTN4201e−2476
164516701151402H1SOYMON031g556171BLASTX1511e−2385
164616701149617H1SOYMON031g556171BLASTX1581e−2386
164716700747310H1SOYMON013g414549BLASTN4061e−2373
164816701139569H1SOYMON038g556171BLASTX1911e−2284
164916701213275H1SOYMON035g602589BLASTN2551e−2280
165016701157185H1SOYMON031g556171BLASTX1971e−2090
165116700655520H1SOYMON004g556171BLASTX1661e−1986
165216701010779H1SOYMON019g556171BLASTX1731e−1964
165316701044104H1SOYMON032g556171BLASTX1881e−1989
165416700867605H1SOYMON016g556171BLASTX1601e−1770
165516701058593H1SOYMON033g168647BLASTX1691e−1694
165616701070286H1SOYMON034g168647BLASTX1641e−1591
165716700877219H1SOYMON018g168647BLASTX1541e−1493
165816700876790H1SOYMON018g168647BLASTX1541e−1493
165916700877212H1SOYMON018g168647BLASTX1541e−1493
166016700760847H1SOYMON015g556171BLASTX1381e−1386
166116700893711H1SOYMON024g168647BLASTX1401e−1382
166216700557532H1SOYMON001g256120BLASTX1151e−1288
166316700793802H1SOYMON017g556171BLASTX1381e−1293
166416700659725H1SOYMON004g556171BLASTX1441e−1247
166516701044545H1SOYMON032g556171BLASTX1441e−1292
166616701037485H1SOYMON029g556171BLASTX1351e−1196
166716700683524H1SOYMON008g168647BLASTX1361e−1190
166816700876711H1SOYMON018g168647BLASTX1091e−1085
166916701155437H1SOYMON031g556171BLASTX1301e−1092
167028599700997892H1SOYMON018g806311BLASTN8341e−6078
167131701053174H1SOYMON032g806311BLASTN5721e−3773
167231700754467H1SOYMON014g806312BLASTX1451e−2166
167331701107430H1SOYMON036g806312BLASTX1991e−2063
167431700985855H1SOYMON009g806312BLASTX1451e−1864
167531701038167H1SOYMON029g806312BLASTX1791e−1761
167631700670393H1SOYMON006g806312BLASTX1671e−1678
167731700559280H1SOYMON001g609262BLASTX1641e−1569
167831700793048H1SOYMON017g806312BLASTX971e−1260
167931700993683H1SOYMON011g806312BLASTX1031e−1160
168031700663233H1SOYMON005g806312BLASTX1301e−1156
168131700908079H1SOYMON022g806312BLASTX1031e−1060
168231701043447H1SOYMON029g609262BLASTX1261e−1084
168331700740188H1SOYMON012g806312BLASTX1031e−860
16847466700742922H1SOYMON012g806311BLASTN4351e−2776
16857466700606255H1SOYMON008g806312BLASTX1171e−1780
168616LIB3053-005-LIB3053g602589BLASTN10001e−7477
Q1-N1-F9
168716LIB3039-035-LIB3039g602589BLASTN9791e−7278
Q1-E1-C5
168816LIB3039-031-LIB3039g256119BLASTN9111e−7180
Q1-E1-A8
168916LIB3030-003-LIB3030g602589BLASTN9491e−7078
Q1-B1-C9
169016LIB3039-023-LIB3039g602589BLASTN9131e−6778
Q1-E1-H12
169116LIB3039-047-LIB3039g602589BLASTN5661e−6575
Q1-E1-D8
169216LIB3039-052-LIB3039g602589BLASTN8901e−6577
Q1-E1-D6
169316LIB3039-051-LIB3039g602589BLASTN8551e−6278
Q1-E1-A1
169416LIB3049-009-LIB3049g602589BLASTN7831e−5678
Q1-E1-G5
169516LIB3039-009-LIB3039g602589BLASTN8051e−5678
Q1-E1-C1
169616LIB3055-006-LIB3055g256119BLASTN4811e−5478
Q1-N1-H3
169716LIB3055-013-LIB3055g256119BLASTN7691e−5479
Q1-N1-C3
169816LIB3049-034-LIB3049g602589BLASTN6261e−5176
Q1-E1-A2
169916LIB3049-022-LIB3049g602589BLASTN5191e−4378
Q1-E1-F9
170016LIB3049-030-LIB3049g602589BLASTN5721e−3877
Q1-E1-C7
170116LIB3040-035-LIB3040g556171BLASTX1751e−3382
Q1-E1-C5
170216LIB3040-005-LIB3040g169820BLASTN3241e−3376
Q1-E1-H8
170316LIB3028-025-LIB3028g602589BLASTN4641e−3378
Q1-B1-D1
170416LIB3039-022-LIB3039g602589BLASTN3571e−3273
Q1-E1-D5
170516LIB3052-001-LIB3052G414549BLASTN3271e−2973
Q1-B1-C5
170628599LIB3039-047-LIB3039G806311BLASTN11831e−9481
Q1-E1-D9
170728599LIB3039-048-LIB3039G806311BLASTN10071e−9281
Q1-E1-D12
SOYBEAN FRUCTOSE 1,6-BISPHOSPHATE ALDOLASE
1708−700565253700565253H1SOYMON002G3021337BLASTN3521e−3976
1709−700865276700865276H1SOYMON016G3021337BLASTN6291e−4376
1710−700873022700873022H1SOYMON018G3696BLASTX2111e−2670
1711−700943855700943855H1SOYMON024G20204BLASTX2021e−2086
1712−700974965700974965H1SOYMON005g3021337BLASTN2591e−1084
1713−701039850701039850H1SOYMON029g22632BLASTN4081e−2376
1714−701206840701206840H1SOYMON035g3021338BLASTX1511e−1383
171511792700654881H1SOYMON004g20204BLASTX1501e−1376
171611792700746016H1SOYMON013g3021337BLASTN2841e−1267
171712314701037190H1SOYMON029g3021337BLASTN6341e−4478
171812314701042664H1SOYMON029g3021338BLASTX1971e−2066
171916700651596H1SOYMON003g3021337BLASTN11011e−8386
172016700750439H1SOYMON013g3021337BLASTN10781e−8186
172116700649475H1SOYMON003g3021337BLASTN10821e−8184
172216700652995H1SOYMON003g3021337BLASTN10841e−8182
172316700981967H1SOYMON009g3021337BLASTN10711e−8085
172416700863243H1SOYMON023g3021337BLASTN10441e−7886
172516700558625H1SOYMON001g3021337BLASTN10411e−7784
172616700564806H1SOYMON002g3021337BLASTN10211e−7680
172716700746368H1SOYMON013g3021337BLASTN8971e−7586
172816700960290H1SOYMON022g3021337BLASTN10091e−7587
172916701055132H1SOYMON032g3021337BLASTN10111e−7586
173016701056109H1SOYMON032g3021337BLASTN10121e−7584
173116701119884H1SOYMON037g3021337BLASTN10141e−7587
173216700898149H1SOYMON027g3021337BLASTN10151e−7586
173316700661436H1SOYMON005g3021337BLASTN5961e−7483
173416701042223H1SOYMON029g3021337BLASTN9971e−7484
173516700676004H1SOYMON007g3021337BLASTN9841e−7385
173616700747718H1SOYMON013g3021337BLASTN9881e−7387
173716700751133H1SOYMON014g3021337BLASTN9891e−7386
173816701215247H1SOYMON035g3021337BLASTN9891e−7384
173916700652484H1SOYMON003g3021337BLASTN9101e−7285
174016700869785H1SOYMON016g3021337BLASTN9701e−7287
174116700981960H1SOYMON009g3021337BLASTN9701e−7287
174216700969335H1SOYMON005g3021337BLASTN9721e−7282
174316700854174H1SOYMON023g3021337BLASTN9651e−7184
174416700761638H1SOYMON015g3021337BLASTN9661e−7186
174516700984860H1SOYMON009g3021337BLASTN9671e−7184
174616701005716H1SOYMON019g3021337BLASTN9671e−7183
174716700941053H1SOYMON024g3021337BLASTN9681e−7186
174816700561358H1SOYMON002g3021337BLASTN9681e−7182
174916700564906H1SOYMON002g3021337BLASTN5621e−7082
175016700833951H1SOYMON019g3021337BLASTN9541e−7088
175116701117626H1SOYMON037g3021337BLASTN9571e−7085
175216700729103H1SOYMON009g3021337BLASTN5351e−6986
175316700670615H1SOYMON006g3021337BLASTN9361e−6983
175416701053635H1SOYMON032g3021337BLASTN9411e−6984
175516700982280H1SOYMON009g3021337BLASTN9231e−6882
175616701119874H1SOYMON037g3021337BLASTN9251e−6888
175716700758937H1SOYMON015g3021337BLASTN9261e−6887
175816701214027H1SOYMON035g3021337BLASTN9281e−6882
175916700972858H1SOYMON005g3021337BLASTN9291e−6884
176016701099780H1SOYMON028g3021337BLASTN9301e−6885
176116700829560H1SOYMON019g3021337BLASTN9321e−6885
176216700971973H1SOYMON005g3021337BLASTN5761e−6785
176316701142336H1SOYMON038g3021337BLASTN7501e−6781
176416701132605H1SOYMON038g3021337BLASTN7591e−6785
176516700969222H1SOYMON005g3021337BLASTN9131e−6784
176616700670956H1SOYMON006g3021337BLASTN9201e−6784
176716700895725H1SOYMON027g3021337BLASTN9211e−6784
176816701013771H1SOYMON019g3021337BLASTN9211e−6781
176916701055481H1SOYMON032g3021337BLASTN6541e−6680
177016700753940H1SOYMON014g3021337BLASTN8991e−6684
177116700974141H1SOYMON005g3021337BLASTN9001e−6681
177216700562408H1SOYMON002g3021337BLASTN9021e−6682
177316700685292H1SOYMON008g3021337BLASTN9031e−6683
177416700985157H1SOYMON009g3021337BLASTN9071e−6682
177516701038194H1SOYMON029g3021337BLASTN9071e−6682
177616700986633H1SOYMON009g3021337BLASTN9081e−6683
177716700564282H1SOYMON002g3021337BLASTN5171e−6583
177816700733754H1SOYMON010g3021337BLASTN6801e−6584
177916700988179H1SOYMON009g3021337BLASTN8871e−6582
178016700555591H1SOYMON001g3021337BLASTN8871e−6582
178116701206717H1SOYMON035g3021337BLASTN8881e−6581
178216700968494H1SOYMON036g3021337BLASTN8891e−6586
178316700906271H1SOYMON022g3021337BLASTN8941e−6582
178416700677674H1SOYMON007g3021337BLASTN8941e−6583
178516700970391H1SOYMON005g3021337BLASTN8961e−6583
178616700753641H1SOYMON014g3021337BLASTN8971e−6582
178716700646593H1SOYMON014g3021337BLASTN4681e−6480
178816700565615H1SOYMON002g3021337BLASTN6671e−6480
178916700746523H1SOYMON013g3021337BLASTN7441e−6483
179016700899019H1SOYMON027g3021337BLASTN8751e−6483
179116701127167H1SOYMON037g3021337BLASTN8761e−6484
179216701131053H1SOYMON038g3021337BLASTN8791e−6484
179316700670980H1SOYMON006g3021337BLASTN8811e−6483
179416701055811H1SOYMON032g3021337BLASTN8811e−6485
179516700900103H1SOYMON027g3021337BLASTN8821e−6483
179616700975609H1SOYMON009g3021337BLASTN8821e−6484
179716701102865H1SOYMON028g3021337BLASTN8831e−6485
179816701145255H1SOYMON031g3021337BLASTN5091e−6380
179916701210875H1SOYMON035g3021337BLASTN6161e−6384
180016700646664H1SOYMON014g3021337BLASTN8621e−6385
180116700897337H1SOYMON027g3021337BLASTN8651e−6386
180216700736783H1SOYMON010g3021337BLASTN8671e−6383
180316701059586H1SOYMON033g3021337BLASTN8691e−6381
180416701127063H1SOYMON037g3021337BLASTN4121e−6284
180516700556614H1SOYMON001g3021337BLASTN4751e−6286
180616700672681H1SOYMON006g3021337BLASTN8181e−6282
180716700727057H1SOYMON009g3021337BLASTN8501e−6282
180816701042141H1SOYMON029g3021337BLASTN8511e−6283
180916700561860H1SOYMON002g3021337BLASTN8541e−6281
181016700677460H1SOYMON007g3021337BLASTN8551e−6283
181116700971671H1SOYMON005g3021337BLASTN8561e−6281
181216700749578H1SOYMON013g3021337BLASTN8561e−6281
181316700672288H1SOYMON006g3021337BLASTN8601e−6281
181416701068481H1SOYMON034g3021337BLASTN8611e−6281
181516700729913H1SOYMON009g3021337BLASTN6611e−6179
181616700739449H1SOYMON012g3021337BLASTN7241e−6185
181716700830902H1SOYMON019g3021337BLASTN8141e−6183
181816700895304H1SOYMON027g3021337BLASTN8401e−6182
181916700605676H2SOYMON005g3021337BLASTN8421e−6184
182016700677453H1SOYMON007g3021337BLASTN8431e−6183
182116700983108H1SOYMON009g3021337BLASTN8431e−6181
182216700889170H1SOYMON024g3021337BLASTN8451e−6186
182316701004956H1SOYMON019g3021337BLASTN8491e−6182
182416700958213H1SOYMON022g3021337BLASTN8491e−6182
182516701129305H1SOYMON037g3021337BLASTN6591e−6085
182616701014446H1SOYMON019g3021337BLASTN6691e−6085
182716700832047H1SOYMON019g3021337BLASTN7381e−6083
182816700669966H1SOYMON006g3021337BLASTN8291e−6082
182916700758028H1SOYMON015g3021337BLASTN8291e−6081
183016700659491H1SOYMON004g3021337BLASTN8291e−6083
183116701003560H1SOYMON019g3021337BLASTN8291e−6082
183216701060964H1SOYMON033g3021337BLASTN8331e−6081
183316700548284H1SOYMON002g3021337BLASTN8341e−6082
183416700894957H1SOYMON024g3021337BLASTN8371e−6081
183516700646551H1SOYMON014g3021337BLASTN4791e−5983
183616700967633H1SOYMON032g3021337BLASTN5301e−5981
183716700754430H1SOYMON014g3021337BLASTN6541e−5985
183816700865919H1SOYMON016g3021337BLASTN8141e−5981
183916700980426H1SOYMON009g3021337BLASTN8151e−5980
184016701048203H1SOYMON032g3021337BLASTN8161e−5981
184116700846414H1SOYMON021g3021337BLASTN8191e−5981
184216700851608H1SOYMON023g3021337BLASTN8221e−5981
184316700970160H1SOYMON005g3021337BLASTN8221e−5982
184416700834462H1SOYMON019g3021337BLASTN8231e−5981
184516701206312H1SOYMON035g3021337BLASTN8231e−5985
184616700562478H1SOYMON002g3021337BLASTN4871e−5884
184716700788114H1SOYMON011g3021337BLASTN7511e−5883
184816700753792H1SOYMON014g3021337BLASTN8041e−5884
184916700837427H1SOYMON020g3021337BLASTN8051e−5886
185016700753668H1SOYMON014g3021337BLASTN8061e−5885
185116700667315H1SOYMON006g3021337BLASTN8091e−5881
185216700808315H1SOYMON024g3021337BLASTN5581e−5780
185316700670207H1SOYMON006g3021337BLASTN7911e−5787
185416700849886H1SOYMON021g3021337BLASTN7911e−5783
185516700839033H1SOYMON020g3021337BLASTN7911e−5781
185616700751117H1SOYMON014g3021337BLASTN7991e−5786
185716700851803H1SOYMON023g3021337BLASTN7991e−5786
185816700669164H1SOYMON006g3021337BLASTN8001e−5780
185916700548285H1SOYMON002g3021337BLASTN8011e−5785
186016701065620H1SOYMON034g3021337BLASTN4261e−5682
186116700727996H1SOYMON009g3021337BLASTN4681e−5679
186216700869176H1SOYMON016g3021337BLASTN7861e−5685
186316700973141H1SOYMON005g3021337BLASTN4401e−5579
186416700969555H1SOYMON005g3021337BLASTN4481e−5581
186516700866138H1SOYMON016g3021337BLASTN6411e−5586
186616700904813H1SOYMON022g3021337BLASTN6991e−5585
186716700894146H1SOYMON024g3021337BLASTN7731e−5586
186816700669945H1SOYMON006g3021337BLASTN7731e−5586
186916701060489H1SOYMON033g3021337BLASTN6641e−5485
187016701125675H1SOYMON037g3021337BLASTN7211e−5485
187116700754750H1SOYMON014g3021337BLASTN7221e−5486
187216701142770H1SOYMON038g3021337BLASTN7551e−5488
187316700731095H1SOYMON009g3021337BLASTN7551e−5487
187416700667966H1SOYMON006g3021337BLASTN7561e−5484
187516700673606H1SOYMON007g3021337BLASTN7601e−5483
187616700605289H2SOYMON003g3021337BLASTN7631e−5484
187716700965253H1SOYMON022g3021337BLASTN7631e−5486
187816700732985H1SOYMON010g3021337BLASTN7651e−5487
187916700986523H1SOYMON009g3021337BLASTN4741e−5385
188016701100040H2SOYMON028g3021337BLASTN6021e−5385
188116700895328H1SOYMON027g3021337BLASTN7421e−5383
188216701141083H1SOYMON038g3021337BLASTN7511e−5385
188316700829878H1SOYMON019g3021337BLASTN4171e−5286
188416700671825H1SOYMON006g3021337BLASTN4311e−5279
188516700755240H1SOYMON014g3021337BLASTN7311e−5288
188616701011659H1SOYMON019g3021337BLASTN7341e−5286
188716701011547H1SOYMON019g3021337BLASTN3811e−5184
188816700835614H1SOYMON019g3021337BLASTN4371e−5180
188916700671849H1SOYMON006g3021337BLASTN4711e−5187
189016700734822H1SOYMON010g3021337BLASTN4861e−5179
189116700830223H1SOYMON019g3021337BLASTN6221e−5184
189216700659970H1SOYMON004g3021337BLASTN7221e−5182
189316701101779H1SOYMON028g3021337BLASTN7281e−5186
189416700852553H1SOYMON023g3021337BLASTN4901e−5088
189516700853857H1SOYMON023g3021337BLASTN7111e−5088
189616700980358H1SOYMON009g3021337BLASTN7121e−5085
189716700672182H1SOYMON006g3021337BLASTN7141e−5089
189816700748455H1SOYMON013g3021337BLASTN3961e−4985
189916700657257H1SOYMON004g3021337BLASTN6941e−4975
190016700729301H1SOYMON009g3021337BLASTN7021e−4980
190116700726175H1SOYMON009g3021337BLASTN7041e−4980
190216700966844H1SOYMON028g3021337BLASTN4141e−4781
190316700960965H1SOYMON022g3021337BLASTN4521e−4785
190416700678326H1SOYMON007g3021337BLASTN4801e−4783
190516700751042H1SOYMON014g3021337BLASTN6751e−4787
190616700830863H1SOYMON019g3021337BLASTN3431e−4684
190716701213640H1SOYMON035g3021337BLASTN6671e−4687
190816700870215H1SOYMON016g3021337BLASTN6671e−4680
190916700658278H1SOYMON004g3021337BLASTN4251e−4487
191016700942532H1SOYMON024g3021337BLASTN5831e−4483
191116700986276H1SOYMON009g3021337BLASTN6301e−4381
191216700870216H1SOYMON016g3021337BLASTN4571e−4282
191316700899828H1SOYMON027g3021337BLASTN4641e−4283
191416700678816H1SOYMON007g3021337BLASTN6181e−4286
191516700666809H1SOYMON005g3021337BLASTN6211e−4282
191616701098073H1SOYMON028g3021337BLASTN2851e−4183
191716700669492H1SOYMON006g3021337BLASTN5041e−3983
191816700975340H1SOYMON009g3021337BLASTN5741e−3981
191916700753528H1SOYMON014g3021337BLASTN5761e−3981
192016700665923H1SOYMON005g3021337BLASTN3731e−3584
192116701038320H1SOYMON029g3021337BLASTN5181e−3484
192216700755605H1SOYMON014g3021337BLASTN4311e−3381
192316700890349H1SOYMON024g3021337BLASTN5111e−3388
192416700669817H1SOYMON006g3021337BLASTN3631e−3187
192516701097640H1SOYMON028g3021337BLASTN4761e−3067
192616700562959H1SOYMON002g3021337BLASTN4821e−3081
192716700852454H1SOYMON023g3021337BLASTN4461e−2877
192816701121443H1SOYMON037g3021337BLASTN4181e−2484
192916701118247H1SOYMON037g3021337BLASTN2801e−1885
193016700665401H1SOYMON005g927505BLASTX1721e−1694
193116700750038H1SOYMON013g3021338BLASTX1621e−1584
193216700665414H1SOYMON005g3021337BLASTN2731e−1388
193316700889072H1SOYMON024g3021338BLASTX1361e−1183
193416700727964H1SOYMON009g927505BLASTX1371e−1186
193516700680648H1SOYMON008g3021337BLASTN2261e−1073
193616701044547H1SOYMON032g927505BLASTX911e−976
193716700649174H1SOYMON003g3021338BLASTX1261e−983
193816531701120682H1SOYMON037g3021337BLASTN7161e−5077
19391701700993909H1SOYMON011g22633BLASTX1121e−3178
19401701700955490H1SOYMON022g22633BLASTX1761e−2570
19411701700682081H1SOYMON008g22633BLASTX1471e−2068
19421701700988843H1SOYMON011g22633BLASTX901e−1467
19431701700740531H1SOYMON012g22633BLASTX921e−1264
19441701700790059H2SOYMON011g22633BLASTX921e−1267
19451701700872670H1SOYMON018g169037BLASTX1441e−1290
19461701700990591H1SOYMON011g22632BLASTN1991e−1168
19471701700743120H1SOYMON012g22633BLASTX921e−968
19481701700994931H1SOYMON011g22633BLASTX921e−864
19491938700738074H1SOYMON012g927507BLASTX1341e−1190
1950239701126904H1SOYMON037g169037BLASTX2311e−2481
1951239700668532H1SOYMON006g169037BLASTX2021e−2083
1952239700943660H1SOYMON024g169037BLASTX1801e−1784
1953239701009915H2SOYMON019g169037BLASTX1801e−1784
1954239701100047H2SOYMON028g169037BLASTX1601e−1584
1955239700794458H1SOYMON017g22633BLASTX1311e−1058
1956239700738441H1SOYMON012g169037BLASTX1181e−878
19573425700984050H1SOYMON009g3021337BLASTN8741e−6480
19583425701014509H1SOYMON019g3021337BLASTN5201e−6080
19593425701138819H1SOYMON038g3021337BLASTN8151e−5980
19603425700977309H1SOYMON009g3021337BLASTN8091e−5880
19613425700984876H1SOYMON009g3021337BLASTN8131e−5880
19623425701046151H1SOYMON032g3021337BLASTN7301e−5280
19633425700889668H1SOYMON024g3021337BLASTN7371e−5281
19643425700976571H1SOYMON009g3021337BLASTN7371e−5281
19653425701045371H1SOYMON032g3021337BLASTN7161e−5079
19663425700548283H1SOYMON002g3021337BLASTN7001e−4981
19673425701103461H1SOYMON028g3021337BLASTN7051e−4981
19683425700898446H1SOYMON027g3021337BLASTN6861e−4883
19693425701006432H1SOYMON019g3021337BLASTN6881e−4883
19703425701041476H1SOYMON029g3021337BLASTN6931e−4881
19713425700568335H1SOYMON002g3021337BLASTN6781e−4782
19723425701046312H1SOYMON032g3021337BLASTN6501e−4585
19733425701050171H1SOYMON032g3021337BLASTN6501e−4585
19743425700685063H1SOYMON008g3021337BLASTN6431e−4483
19753425701010250H2SOYMON019g3021337BLASTN5421e−3686
19763425700665454H1SOYMON005g3021337BLASTN5201e−3480
19773425701043888H1SOYMON032g3021337BLASTN4951e−3285
19783425700726806H1SOYMON009g3021337BLASTN2131e−2376
1979491700997879H1SOYMON018g22632BLASTN7891e−5677
1980491700646208H1SOYMON012g22632BLASTN7331e−5276
1981491700559796H1SOYMON001g22632BLASTN7151e−5076
1982491700789784H1SOYMON011g22632BLASTN6641e−4676
1983491700683122H1SOYMON008g22632BLASTN4851e−4186
1984491701105914H1SOYMON036g22632BLASTN5041e−4173
1985491700558789H1SOYMON001g22632BLASTN6071e−4174
1986491700873051H1SOYMON018g22632BLASTN6081e−4175
1987491700684010H1SOYMON008g22632BLASTN5971e−4075
1988491700786096H2SOYMON011g22632BLASTN5761e−3975
1989491700731865H1SOYMON010g22632BLASTN5821e−3975
1990491701108111H1SOYMON036g22632BLASTN4671e−3875
1991491700740887H1SOYMON012g22632BLASTN5671e−3874
1992491700559579H1SOYMON001g22632BLASTN5721e−3875
1993491700996104H1SOYMON018g22632BLASTN4761e−3776
1994491700682145H1SOYMON008g22632BLASTN5421e−3674
1995491700737263H1SOYMON010g22632BLASTN5261e−3574
1996491700547963H1SOYMON001g22632BLASTN5271e−3573
1997491700686296H1SOYMON008g22632BLASTN5271e−3573
1998491700646072H1SOYMON011g22632BLASTN5371e−3574
1999491701106662H1SOYMON036g22632BLASTN5141e−3474
2000491700684335H1SOYMON008g22632BLASTN5161e−3474
2001491701000609H1SOYMON018g22632BLASTN5201e−3474
2002491700685658H1SOYMON008g22632BLASTN5201e−3474
2003491700875532H1SOYMON018g22632BLASTN5211e−3473
2004491700730264H1SOYMON009g22632BLASTN5021e−3374
2005491700872948H1SOYMON018g22632BLASTN5021e−3374
2006491700685813H1SOYMON008g22632BLASTN5021e−3374
2007491701104554H1SOYMON036g22632BLASTN5031e−3374
2008491700960601H1SOYMON022g22632BLASTN5031e−3374
2009491700876633H1SOYMON018g22632BLASTN5031e−3374
2010491700739662H1SOYMON012g22632BLASTN5041e−3372
2011491700685904H1SOYMON008g22632BLASTN5051e−3372
2012491700995183H1SOYMON011g22632BLASTN5131e−3373
2013491700901996H1SOYMON027g22632BLASTN5131e−3374
2014491700727070H1SOYMON009g22632BLASTN4901e−3272
2015491700685790H1SOYMON008g22632BLASTN4921e−3274
2016491700998652H1SOYMON018g22632BLASTN4941e−3272
2017491700740465H1SOYMON012g22632BLASTN4821e−3174
2018491700682621H2SOYMON008g22632BLASTN4841e−3174
2019491700874316H1SOYMON018g22632BLASTN4661e−3073
2020491700686477H1SOYMON008g22632BLASTN4731e−3073
2021491700739979H1SOYMON012g22632BLASTN4761e−3074
2022491700739416H1SOYMON012g22632BLASTN4761e−3074
2023491700685976H1SOYMON008g22632BLASTN4761e−3074
2024491700739629H1SOYMON012g22632BLASTN4861e−3070
2025491700989163H1SOYMON011g22632BLASTN4681e−2972
2026491701000555H1SOYMON018g22632BLASTN4771e−2972
2027491700872702H1SOYMON018g22632BLASTN4361e−2872
2028491701000781H1SOYMON018g22632BLASTN4601e−2873
2029491700682760H1SOYMON008g22632BLASTN4631e−2872
2030491700740390H1SOYMON012g22632BLASTN4401e−2773
2031491700685346H1SOYMON008g22632BLASTN4511e−2772
2032491700557272H1SOYMON001g22632BLASTN2501e−2678
2033491700953343H1SOYMON022g22632BLASTN3491e−2674
2034491700741960H1SOYMON012g22632BLASTN4301e−2673
2035491700680247H2SOYMON008g22632BLASTN4251e−2567
2036491700680002H2SOYMON008g22632BLASTN2411e−2472
2037491700684827H1SOYMON008g22632BLASTN3791e−2474
2038491700956353H1SOYMON022g22632BLASTN4101e−2472
2039491700787513H1SOYMON011g22632BLASTN2351e−2272
2040491700725070H1SOYMON009g22632BLASTN2411e−2271
2041491700741111H1SOYMON012g22632BLASTN3041e−2273
2042491700985308H1SOYMON009g22632BLASTN2411e−2180
2043491700738230H1SOYMON012g22632BLASTN2411e−2172
2044491700991396H1SOYMON011g22632BLASTN3501e−2172
2045491700741276H1SOYMON012g22632BLASTN3791e−2171
2046491700740223H1SOYMON012g22632BLASTN2411e−2072
2047491700738808H1SOYMON012g22632BLASTN2411e−2072
2048491700997995H1SOYMON018g22632BLASTN2411e−1981
2049491700875139H1SOYMON018g22632BLASTN2411e−1971
2050491700989713H1SOYMON011g22632BLASTN2411e−1973
2051491700958366H1SOYMON022g22632BLASTN2411e−1871
2052491700683887H1SOYMON008g22632BLASTN3441e−1870
2053491700740788H1SOYMON012g22632BLASTN3391e−1770
2054491700743058H1SOYMON012g22632BLASTN2051e−1681
2055491700996423H1SOYMON018g22632BLASTN2341e−1680
2056491700686075H1SOYMON008g22632BLASTN2411e−1671
2057491700738811H1SOYMON012g22632BLASTN1931e−1572
2058491700998312H1SOYMON018g22632BLASTN2341e−1573
2059491700681825H1SOYMON008g22632BLASTN2411e−1581
2060491701109105H1SOYMON036g22632BLASTN2901e−1469
2061491701203741H2SOYMON035g22632BLASTN2301e−1378
2062491700740785H1SOYMON012g22632BLASTN2871e−1368
2063491700738486H1SOYMON012g22632BLASTN2951e−1364
2064491700739078H1SOYMON012g22632BLASTN1781e−1273
2065491701002287H1SOYMON018g22632BLASTN2551e−1274
2066491700742470H1SOYMON012g22632BLASTN2781e−1269
2067491700743421H1SOYMON012g22632BLASTN2611e−1171
2068491700744039H1SOYMON012g22632BLASTN2651e−1169
2069491700789444H2SOYMON011g22632BLASTN1581e−1087
2070491700741074H1SOYMON012g22632BLASTN1781e−1077
2071491700998877H1SOYMON018g22632BLASTN2351e−1072
2072491700740005H1SOYMON012g22633BLASTX751e−964
2073491700872703H1SOYMON018g169037BLASTX1161e−983
2074491700743301H1SOYMON012g22632BLASTN2411e−976
2075491700875039H1SOYMON018g22632BLASTN2411e−972
2076491700742515H1SOYMON012g22632BLASTN2411e−976
2077491700990557H1SOYMON011g22632BLASTN2411e−976
2078491700743995H1SOYMON012g22632BLASTN2411e−976
2079491700743495H1SOYMON012g22632BLASTN2411e−976
2080491701001909H1SOYMON018g22632BLASTN2411e−976
2081491701001445H1SOYMON018g169037BLASTX1151e−892
2082491700554881H1SOYMON001g169037BLASTX1161e−894
2083491700954194H1SOYMON022g169037BLASTX1161e−894
2084491700996869H1SOYMON018g22632BLASTN2301e−876
2085491700897820H1SOYMON027g22632BLASTN2341e−874
2086491700742574H1SOYMON012g22632BLASTN2341e−874
2087491700684738H1SOYMON008g22632BLASTN2351e−875
20887368700739343H1SOYMON012g927507BLASTX1641e−1588
2089-GM32379LIB3051-015-LIB3051g3021337BLASTN2601e−2877
Q1-E1-B12
2090-GM8265LIB3039-048-LIB3039g3021337BLASTN4811e−2965
Q1-E1-F11
209116LIB3027-010-LIB3027g3021337BLASTN13931e−10782
Q1-B1-B7
209216LIB3039-049-LIB3039g3021337BLASTN12971e−9983
Q1-E1-B8
209316LIB3051-061-LIB3051g3021337BLASTN13031e−9984
Q1-K1-E11
209416LIB3056-009-LIB3056g3021337BLASTN11261e−9684
Q1-N1-A10
209516LIB3051-025-LIB3051g3021337BLASTN12621e−9683
Q1-K1-E11
209616LIB3056-014-LIB3056g3021337BLASTN10771e−9481
Q1-N1-E1
209716LIB3055-005-LIB3055g3021337BLASTN12271e−9384
Q1-N1-A8
209816LIB3040-045-LIB3040g3021337BLASTN12111e−9283
Q1-E1-A4
209916LIB3028-010-LIB3028g3021337BLASTN12151e−9283
Q1-B1-G9
210016LIB3056-010-LIB3056g3021337BLASTN12171e−9284
Q1-N1-G8
210116LIB3039-029-LIB3039g3021337BLASTN11281e−8585
Q1-E1-A6
210216LIB3051-014-LIB3051g3021337BLASTN7161e−8083
Q1-E1-D2
210316LIB3030-010-LIB3030g3021337BLASTN10521e−7883
Q1-B1-D7
210416LIB3051-094-LIB3051g3021337BLASTN7781e−7483
Q1-K1-A9
210516LIB3028-030-LIB3028g3021337BLASTN9531e−7085
Q1-B1-C9
210616LIB3052-004-LIB3052g3021337BLASTN8681e−6382
Q1-N1-D8
210716LIB3065-014-LIB3065g3021337BLASTN5401e−6179
Q1-N1-A3
210816LIB3050-019-LIB3050g168420BLASTX2231e−4063
Q1-K1-H1
210916LIB3051-062-LIB3051g3021337BLASTN5411e−3879
Q1-K1-B5
21103425LIB3051-067-LIB3051g3021337BLASTN10821e−8178
Q1-K1-E7
21113425LIB3050-006-LIB3050g3021337BLASTN7521e−5775
Q1-E1-G7
2112491LIB3028-011-LIB3028g22632BLASTN9111e−6775
Q1-B1-B9
2113491LIB3028-011-LIB3028g22632BLASTN8861e−6577
Q1-B1-F2
SOYBEAN FRUCTOSE-1,6-BISPHOSPHATASE
2114−700685384700685384H1SOYMON008g21244BLASTN5971e−4980
2115−700737915700737915H1SOYMON012g515746BLASTN13161e−10097
2116−700741457700741457H1SOYMON012g3041774BLASTN6921e−5880
2117−700874831700874831H1SOYMON018g515746BLASTN12951e−99100
2118−700996155700996155H1SOYMON018g3041774BLASTN6511e−4583
2119−700996632700996632H1SOYMON018g515746BLASTN5071e−5190
2120−700998027700998027H1SOYMON018g515746BLASTN6361e−6594
2121−701209548701209548H1SOYMON035g3041774BLASTN6421e−4483
212210129700870828H1SOYMON018g21244BLASTN8271e−6079
212310129700741669H1SOYMON012g21244BLASTN6571e−5380
212410348700555754H1SOYMON001g21244BLASTN4661e−2977
212510348700991527H1SOYMON011g440591BLASTX1691e−1688
212613716700898719H1SOYMON027g515746BLASTN11861e−9097
212713716700993540H1SOYMON011g515746BLASTN11791e−8998
212813716700909657H1SOYMON022g515746BLASTN5681e−5786
21291894700555054H1SOYMON001g515746BLASTN13201e−101100
21301894700685264H1SOYMON008g515746BLASTN13231e−10199
21311894700558854H1SOYMON001g515746BLASTN6951e−98100
21321894700554755H1SOYMON001g515746BLASTN7671e−9899
21331894701000504H1SOYMON018g515746BLASTN6261e−9598
21341894700738115H1SOYMON012g515746BLASTN12301e−93100
21351894700992933H1SOYMON011g515746BLASTN10741e−9198
21361894701107444H1SOYMON036g515746BLASTN12011e−9199
21371894700852823H1SOYMON023g515746BLASTN10411e−9098
21381894700733478H1SOYMON010g515746BLASTN11501e−9097
21391894701105185H1SOYMON036g515746BLASTN6411e−8789
21401894700737830H1SOYMON012g515746BLASTN10601e−87100
21411894700685110H1SOYMON008g515746BLASTN5971e−8690
21421894700968307H1SOYMON036g515746BLASTN11131e−8497
21431894700653014H1SOYMON003g515746BLASTN5871e−8290
21441894700555504H1SOYMON001g515746BLASTN6261e−8188
21451894700751540H1SOYMON014g515746BLASTN5851e−7791
21461894700901976H1SOYMON027g515746BLASTN5051e−7387
21471894700986496H1SOYMON009g515746BLASTN5591e−7390
21481894700751580H1SOYMON014g515746BLASTN5691e−7289
21491894700751532H1SOYMON014g515746BLASTN5711e−7290
21501894700990937H1SOYMON011g515746BLASTN5441e−7188
21511894700740789H1SOYMON012g515746BLASTN6301e−69100
21521894700743994H1SOYMON012g515746BLASTN9451e−69100
21531894700754374H1SOYMON014g515746BLASTN4601e−6291
21541894701001295H1SOYMON018g515746BLASTN5411e−6297
21551894701155952H1SOYMON031g515746BLASTN5681e−5183
21561894700872212H1SOYMON018g515746BLASTN6701e−47100
21571894700682196H1SOYMON008g515746BLASTN6091e−4198
21581894700738779H1SOYMON012g515746BLASTN2521e−1682
215926568700844816H1SOYMON021g21244BLASTN6491e−4578
216027512701128049H1SOYMON037g440591BLASTX1851e−1887
21617128700649846H1SOYMON003g440591BLASTX1251e−1581
216210348LIB3030-010-LIB3030g21244BLASTN4761e−2876
Q1-B1-C7
FRUCTOSE-6-PHOSPHATE,2-KINASE
2163−700730441700730441H1SOYMON009g3309583BLASTX1791e−1782
2164−700953509700953509H1SOYMON022g3170229BLASTN6741e−4775
2165−700955121700955121H1SOYMON022g3309582BLASTN3031e−1468
2166-GM28972LIB3050-012-LIB3050g3170229BLASTN10731e−8080
Q1-E1-E9
SOYBEAN PHOSPHOGLUCOISOMERASE
2167−700568558700568558H1SOYMON002g1369950BLASTX1651e−1580
2168−700845275700845275H1SOYMON021g1100771BLASTX1241e−1053
2169−700960755700960755H1SOYMON022g1100771BLASTX1531e−1452
217018663700838363H1SOYMON020g1100771BLASTX2151e−2263
217118663700838355H1SOYMON020g1100771BLASTX1551e−1481
217219355700897450H1SOYMON027g1100771BLASTX2731e−3174
217319355700744258H1SOYMON013g1100771BLASTX2071e−2969
217419355701153832H1SOYMON031g1100771BLASTX2261e−2358
217520088700856114H1SOYMON023g1100771BLASTX1761e−3375
217620088700670380H1SOYMON006g1100771BLASTX2071e−3371
217720088700788785H2SOYMON011g1100771BLASTX1201e−3274
217820088700847659H1SOYMON021g1100771BLASTX1921e−3184
217920088701136417H1SOYMON038g1100771BLASTX1691e−2766
218031255701207622H1SOYMON035g1100771BLASTX1681e−2961
218120088LIB3051-014-LIB3051g1100771BLASTX4001e−6873
Q1-E1-G3
218231255LIB3056-008-LIB3056g1100771BLASTX1881e−5262
Q1-N1-G8
SOYBEAN VACUOLAR H+-TRANSLOCATING-PYROPHOSPHATASE
2183−700660662700660662H1SOYMON004g16347BLASTN5401e−3679
2184−700793860700793860H1SOYMON017g2706449BLASTN8081e−5878
2185−700837007700837007H1SOYMON020g16347BLASTN7761e−5578
2186−700890647700890647H1SOYMON024g790474BLASTN8261e−6081
2187−700942978700942978H1SOYMON024g790478BLASTN6051e−6382
2188−700944280700944280H1SOYMON024g790479BLASTX1191e−1076
2189−700974544700974544H1SOYMON005g1103711BLASTN8541e−6283
2190−700984449700984449H1SOYMON009g1103711BLASTN2871e−1271
2191−700989248700989248H1SOYMON011g534915BLASTN2761e−1467
2192−701102931701102931H1SOYMON028g2706449BLASTN4381e−4676
2193−701106870701106870H1SOYMON036g790478BLASTN6231e−4775
2194−701122796701122796H1SOYMON037g2258074BLASTX711e−1573
2195−701132123701132123H1SOYMON038g790478BLASTN6271e−4381
2196−701136557701136557H1SOYMON038g16347BLASTN3761e−3377
219714021700973215H1SOYMON005g2668745BLASTN4351e−3980
219814021701109310H1SOYMON036g2668745BLASTN2811e−2583
219916700891764H1SOYMON024g790479BLASTX1721e−1668
220019232701061126H1SOYMON033g790474BLASTN9351e−6981
220119232700962864H1SOYMON022g790474BLASTN8741e−6482
220220872700754883H1SOYMON014g790478BLASTN8241e−5981
220320872700971147H1SOYMON005g1103711BLASTN5641e−5479
22042813700797861H1SOYMON017g16347BLASTN7311e−5279
22052813700944850H1SOYMON024g2570500BLASTN7381e−5282
22062813701056207H1SOYMON032g2570500BLASTN5561e−4680
22072813700605115H2SOYMON003g2570500BLASTN4781e−4280
22082813700897063H1SOYMON027g2570500BLASTN5961e−4080
22092813700561829H1SOYMON002g2570500BLASTN5701e−3880
22102813701204883H1SOYMON035g2668745BLASTN5451e−3677
22112813700754984H1SOYMON014g2570500BLASTN5271e−3575
22122813700854552H1SOYMON023g2570500BLASTN5361e−3579
22132813700873337H1SOYMON018g2570500BLASTN5051e−3375
22142813700873349H1SOYMON018g2570500BLASTN5061e−3375
22152813700952403H1SOYMON022g2668745BLASTN4991e−3276
22162813700846561H1SOYMON021g2570500BLASTN4881e−3175
22172813700953987H1SOYMON022g2570500BLASTN4611e−2975
22182813700568667H1SOYMON002g2570500BLASTN2961e−2479
22192813700895231H1SOYMON024g2258074BLASTX2071e−2280
22202813701101791H1SOYMON028g2668746BLASTX1471e−1377
22218040701121224H1SOYMON037g534915BLASTN2981e−1477
22228040700743066H1SOYMON012g2668746BLASTX1401e−1280
22238531701005139H1SOYMON019g2258073BLASTN8711e−6379
22248531701008308H1SOYMON019g534915BLASTN7891e−5776
22258531700559054H1SOYMON001g2570500BLASTN7901e−5777
22268531700942540H1SOYMON024g2706449BLASTN7551e−5480
22278531700790983H1SOYMON011g2258073BLASTN4311e−5277
22288531701007949H1SOYMON019g2570500BLASTN4041e−4170
22298531701123827H1SOYMON037g534915BLASTN4361e−2675
22308531701013616H1SOYMON019g534915BLASTN4311e−2578
22318531700565624H1SOYMON002g2570501BLASTX1691e−1685
22328531701121092H1SOYMON037g2570501BLASTX1101e−1560
223316LIB3040-003-LIB3040g633598BLASTN5231e−5174
Q1-E1-F6
223416LIB3051-114-LIB3051g790478BLASTN4571e−4879
Q1-K1-G5
223516LIB3039-020-LIB3039g790478BLASTN3381e−3074
Q1-E1-A2
22362813LIB3028-026-LIB3028g2570500BLASTN10291e−7780
Q1-B1-B7
22378040LIB3049-045-LIB3049g2706449BLASTN7521e−5272
Q1-E1-C3
22388040LIB3049-005-LIB3049g2570501BLASTX1541e−3261
Q1-E1-A7
22398531LIB3050-013-LIB3050g2570500BLASTN7481e−5372
Q1-E1-G8
22408531LIB3073-025-LIB3073g534915BLASTN7111e−4978
Q1-K1-D6
22418531LIB3050-012-LIB3050g2258074BLASTX931e−3174
Q1-E1-D1
SOYBEAN PYROPHOSPHATE-DEPENDENT FRUCTOSE-6-PHOSPHATE
PHOSPHOTRANSFERASE
22427899701008645H1SOYMON019g169538BLASTX1601e−1583
INVERTASES
2243−700653543700653543H1SOYMON003g1160487BLASTN5411e−5584
2244−700992760700992760H1SOYMON011g550319BLASTX1171e−1249
2245−701005703701005703H1SOYMON019g861157BLASTX2131e−2246
2246−701047324701047324H1SOYMON032g1160487BLASTN6471e−4581
2247−701130328701130328H1SOYMON037g167551BLASTX2151e−2261
224820460700658149H1SOYMON004g861157BLASTX1981e−2072
224920460701041452H1SOYMON029g402740BLASTX1051e−1376
2250-GM31611LIB3051-002-LIB3051g1160487BLASTN10331e−7777
Q1-E1-B9
2251-GM34282LIB3O51-025-LIB3051g1160487BLASTN10691e−8079
Q1-K1-C4
2252-GM34976LIB3051-031-LIB3051g1160487BLASTN7691e−6680
Q1-K1-A9
225331949LIB3051-093-LIB3051g1160487BLASTN9481e−9277
Q1-K1-B1
225431949LIB3051-054-LIB3051g1160487BLASTN9031e−9082
Q1-K2-D11
SOYBEAN SUCROSE SYNTHASE
2255−700565776700565776H1SOYMON002g3169544BLASTX891e−864
2256−700606005700606005H2SOYMON007g2570066BLASTN10691e−8089
2257−700664186700664186H1SOYMON005g2606080BLASTN4261e−6291
2258−700668119700668119H1SOYMON006g2570066BLASTN2791e−1483
2259−700668348700668348H1SOYMON006g2570066BLASTN6931e−4888
2260−700671225700671225H1SOYMON006g16525BLASTN6171e−4272
2261−700673918700673918H1SOYMON007g218332BLASTN1521e−992
2262−700726266700726266H1SOYMON009g2606080BLASTN2371e−2179
2263−700747171700747171H1SOYMON013g2606080BLASTN7351e−5289
2264−700747359700747359H1SOYMON013g218332BLASTN4471e−2878
2265−700787443700787443H2SOYMON011g22485BLASTN11711e−9598
2266−700796035700796035H1SOYMON017g2570066BLASTN10391e−7790
2267−700832792700832792H1SOYMON019g2606080BLASTN4441e−3188
2268−700836673700836673H1SOYMON020g2570066BLASTN8431e−6185
2269−700841855700841855H1SOYMON020g2570066BLASTN4251e−3584
2270−700851758700851758H1SOYMON023g2570066BLASTN2111e−1591
2271−700851991700851991H1SOYMON023g2570066BLASTN7681e−5581
2272−700852943700852943H1SOYMON023g2606080BLASTN2501e−1385
2273−700853396700853396H1SOYMON023g2570067BLASTX1451e−1365
2274−700872206700872206H1SOYMON018g1488570BLASTX2351e−2564
2275−700876641700876641H1SOYMON018g2606080BLASTN4101e−5388
2276−700890526700890526H1SOYMON024g2606080BLASTN6521e−6083
2277−700893784700893784H1SOYMON024g3169543BLASTN2171e−1182
2278−700909222700909222H1SOYMON022g2570066BLASTN4401e−4472
2279−700944438700944438H1SOYMON024g3169543BLASTN6691e−4673
2280−700945733700945733H1SOYMON024g1488569BLASTN5041e−3366
2281−700969926700969926H1SOYMON005g2570066BLASTN6741e−4772
2282−701001986701001986H1SOYMON018g1146237BLASTX1061e−945
2283−701005687701005687H1SOYMON019g2606080BLASTN5911e−4085
2284−701012195701012195H1SOYMON019g2606080BLASTN4181e−4677
2285−701046403701046403H1SOYMON032g2606080BLASTN5741e−3876
2286−701058966701058966H1SOYMON033g218332BLASTN5291e−5684
2287−701150574701150574H1SOYMON031g1041247BLASTX1551e−1474
2288−701205210701205210H1SOYMON035g218332BLASTN9811e−7285
228910445700605276H2SOYMON003g2606080BLASTN8601e−6584
229010445700832417H1SOYMON019g2606080BLASTN8761e−6482
229110445700833214H1SOYMON019g2606080BLASTN7401e−5883
229210445700832409H1SOYMON019g2606080BLASTN8001e−5784
229310445701007169H1SOYMON019g2606080BLASTN6911e−5581
229410445701005913H1SOYMON019g2606080BLASTN6801e−5283
229510445701204549H2SOYMON035g2606080BLASTN7321e−5283
229610445701208347H1SOYMON035g2606080BLASTN6561e−4983
229710445700958980H1SOYMON022g2606080BLASTN6701e−4983
229810445700988126H1SOYMON009g2606080BLASTN3241e−4778
229910445700830464H1SOYMON019g2606080BLASTN3471e−4779
230010445700763911H1SOYMON019g3169543BLASTN5171e−4775
230110445700891996H1SOYMON024g2606080BLASTN6671e−4688
230210445700725104H1SOYMON009g2606080BLASTN5771e−4581
230310445701124001H1SOYMON037g2606080BLASTN6481e−4586
230410445700833919H1SOYMON019g2606080BLASTN4961e−4179
230510445701006692H1SOYMON019g2606080BLASTN5361e−4186
230610445700905349H1SOYMON022g2606080BLASTN5851e−3975
230710445701204596H2SOYMON035g2606080BLASTN5211e−3879
230810445700958885H1SOYMON022g2606080BLASTN3511e−3681
230910445701208390H1SOYMON035g2606080BLASTN2591e−2986
231010445701003131H1SOYMON019g2606080BLASTN4421e−2676
231110445701207712H1SOYMON035g2606080BLASTN2601e−1778
231210445701215107H1SOYMON035g2606080BLASTN2601e−1488
231310445700852649H1SOYMON023g2606080BLASTN2541e−1374
231411259701063407H1SOYMON033g2570066BLASTN11001e−8287
231511259700674761H1SOYMON007g2570066BLASTN7391e−7186
231611259700839148H1SOYMON020g2570066BLASTN9191e−6787
231711259700674815H1SOYMON007g2570066BLASTN9041e−6687
231812890701103318H1SOYMON028g2570066BLASTN10051e−7486
231912890700855911H1SOYMON023g2570066BLASTN5691e−6986
232012890700850874H1SOYMON023g2570066BLASTN9371e−6990
232112890700837552H1SOYMON020g2570066BLASTN8881e−6589
232214264700677058H1SOYMON007g2606080BLASTN5781e−3999
232314264700679301H1SOYMON007g2606080BLASTN3251e−1890
232414740701214452H1SOYMON035g2570066BLASTN10721e−8089
232514740701044972H1SOYMON032g2570066BLASTN5371e−4387
232614740701040560H1SOYMON029g2570066BLASTN3021e−2475
232714740700793901H1SOYMON017g2570066BLASTN2311e−1484
232815394701136903H1SOYMON038g2606080BLASTN9361e−6981
232915394701004431H1SOYMON019g218332BLASTN9421e−6980
233015394701006153H1SOYMON019g218332BLASTN9201e−6783
233115394701138281H1SOYMON038g218332BLASTN4851e−4082
233215394701209319H1SOYMON035g3169543BLASTN5081e−3381
233316344700746372H1SOYMON013g2606080BLASTN4711e−6585
233416344700945706H1SOYMON024g2606080BLASTN6351e−6584
233517781700960671H1SOYMON022g2570066BLASTN9661e−7188
233617781700838540H1SOYMON020g2570066BLASTN5321e−6283
233720151700847184H1SOYMON021g2570066BLASTN7621e−7290
233820151700831558H1SOYMON019g2570066BLASTN9801e−7289
233922196701046171H1SOYMON032g2606080BLASTN13211e−10199
234022196701207390H1SOYMON035g2606080BLASTN12581e−9598
234125275701013025H1SOYMON019g2606080BLASTN13531e−10398
234225275700561738H1SOYMON002g2606080BLASTN9531e−8491
234325380700667735H1SOYMON006g2570066BLASTN9591e−7187
234425380701047629H1SOYMON032g2570066BLASTN7741e−5589
234526818701047072H1SOYMON032g2606080BLASTN8301e−6087
234626818700737511H1SOYMON010g3169543BLASTN6071e−5783
234731182701098655H1SOYMON028g2570066BLASTN9511e−7085
2348318701052316H1SOYMON032g2606080BLASTN15551e−120100
2349318701053115H1SOYMON032g2606080BLASTN12811e−11196
2350318700983049H1SOYMON009g2606080BLASTN14381e−11096
2351318701058416H1SOYMON033g2606080BLASTN13851e−106100
2352318701013289H1SOYMON019g2606080BLASTN13741e−10599
2353318701002784H2SOYMON019g2606080BLASTN13651e−104100
2354318700868516H1SOYMON016g2606080BLASTN11951e−103100
2355318700978851H1SOYMON009g2606080BLASTN13251e−10198
2356318701204954H1SOYMON035g2606080BLASTN7701e−100100
2357318700889102H1SOYMON024g2606080BLASTN10481e−10099
2358318701053120H1SOYMON032g218332BLASTN11091e−10090
2359318700731734H1SOYMON010g2606080BLASTN13081e−10097
2360318700972625H1SOYMON005g2606080BLASTN11201e−9899
2361318701006566H1SOYMON019g2606080BLASTN9831e−9799
2362318700952789H1SOYMON022g2606080BLASTN12761e−9797
2363318701141518H1SOYMON038g2606080BLASTN7161e−9699
2364318700653475H1SOYMON003g3169543BLASTN12621e−9687
2365318700650832H1SOYMON003g2606080BLASTN6431e−9597
2366318700678981H1SOYMON007g2606080BLASTN11421e−9596
2367318700890311H1SOYMON024g2606080BLASTN12001e−95100
2368318700892212H1SOYMON024g2606080BLASTN12501e−9597
2369318700943424H1SOYMON024g2606080BLASTN12511e−9599
2370318700833982H1SOYMON019g2606080BLASTN12551e−95100
2371318700834361H1SOYMON019g2606080BLASTN9811e−9499
2372318700746379H1SOYMON013g2606080BLASTN11081e−9496
2373318700889648H1SOYMON024g2606080BLASTN12381e−9499
2374318701054868H1SOYMON032g2606080BLASTN12431e−9495
2375318700959914H1SOYMON022g2606080BLASTN12261e−9396
2376318701011518H1SOYMON019g2606080BLASTN7051e−9299
2377318700734053H1SOYMON010g2606080BLASTN7651e−92100
2378318701005295H1SOYMON019g2606080BLASTN9621e−9293
2379318700945690H1SOYMON024g2606080BLASTN10541e−9299
2380318701118196H1SOYMON037g2606080BLASTN11001e−9295
2381318700673512H1SOYMON007g2606080BLASTN12111e−9297
2382318700852712H1SOYMON023g2606080BLASTN12151e−9298
2383318701004755H1SOYMON019g2606080BLASTN12211e−9299
2384318700677915H1SOYMON007g2606080BLASTN6851e−9199
2385318700977846H1SOYMON009g2606080BLASTN7311e−9199
2386318700831789H1SOYMON019g2606080BLASTN12041e−9197
2387318700754901H1SOYMON014g2606080BLASTN12051e−91100
2388318700666594H1SOYMON005g2606080BLASTN12101e−91100
2389318700750890H1SOYMON014g2606080BLASTN11881e−9099
2390318700890229H1SOYMON024g2606080BLASTN11951e−90100
2391318700732660H1SOYMON010g2606080BLASTN11541e−8995
2392318700764730H1SOYMON023g2606080BLASTN11811e−8999
2393318701050015H1SOYMON032g218332BLASTN11851e−8989
2394318700870180H1SOYMON016g2606080BLASTN7101e−88100
2395318701204236H2SOYMON035g2606080BLASTN9041e−8898
2396318700645782H1SOYMON010g2606080BLASTN6331e−8795
2397318700831711H1SOYMON019g2606080BLASTN10251e−8796
2398318701056026H1SOYMON032g2606080BLASTN11581e−8796
2399318700678853H1SOYMON007g2606080BLASTN11611e−8797
2400318700852424H1SOYMON023g2606080BLASTN9131e−8695
2401318701049116H1SOYMON032g3169543BLASTN11461e−8689
2402318700977788H1SOYMON009g2606080BLASTN6421e−8594
2403318700833546H1SOYMON019g2606080BLASTN11341e−8594
2404318701004915H1SOYMON019g2606080BLASTN5911e−8496
2405318700730093H1SOYMON009g2606080BLASTN7551e−8496
2406318701119060H1SOYMON037g2606080BLASTN8241e−8497
2407318700963024H1SOYMON022g2606080BLASTN11161e−8490
2408318700563532H1SOYMON002g22037BLASTN11161e−8487
2409318700755891H1SOYMON014g2606080BLASTN11171e−8494
2410318700850605H1SOYMON023g2606080BLASTN11181e−8494
2411318700888245H1SOYMON024g2606080BLASTN6431e−8398
2412318701037091H1SOYMON029g2606080BLASTN8211e−8395
2413318700673790H1SOYMON007g2606080BLASTN11041e−8395
2414318700845518H1SOYMON021g2606080BLASTN6731e−8291
2415318700854591H1SOYMON023g2606080BLASTN6061e−8195
2416318700907167H1SOYMON022g2606080BLASTN9201e−8196
2417318700978575H1SOYMON009g218332BLASTN9711e−8191
2418318700853484H1SOYMON023g2606080BLASTN10791e−8192
2419318701124012H1SOYMON037g218332BLASTN10831e−8189
2420318700835387H1SOYMON019g2606080BLASTN10871e−8196
2421318700749133H1SOYMON013g2606080BLASTN5711e−8098
2422318700727185H1SOYMON009g2606080BLASTN7301e−8098
2423318700869024H1SOYMON016g2606080BLASTN8071e−7996
2424318701013537H1SOYMON019g2606080BLASTN9291e−7987
2425318701010402H1SOYMON019g218332BLASTN10551e−7985
2426318701107955H1SOYMON036g2606080BLASTN10581e−7987
2427318700731653H1SOYMON010g2606080BLASTN5781e−7894
2428318700888950H1SOYMON024g218332BLASTN7651e−7888
2429318700894112H1SOYMON024g2606080BLASTN8421e−7898
2430318701005565H1SOYMON019g2606080BLASTN10241e−7892
2431318700548286H1SOYMON002g2606080BLASTN10451e−7888
2432318700975854H1SOYMON009g22037BLASTN10531e−7886
2433318700944525H1SOYMON024g218332BLASTN10541e−7889
2434318701061312H1SOYMON033g2606080BLASTN7731e−7787
2435318700831277H1SOYMON019g2606080BLASTN9471e−7797
2436318700788482H1SOYMON011g2606080BLASTN10381e−7789
2437318701055686H1SOYMON032g2606080BLASTN10391e−7790
2438318701054768H1SOYMON032g2606080BLASTN7861e−7688
2439318700854891H1SOYMON023g2606080BLASTN10301e−7693
2440318701215276H1SOYMON035g2606080BLASTN10301e−7690
2441318700944860H1SOYMON024g2606080BLASTN8871e−7596
2442318701010957H1SOYMON019g2606080BLASTN10111e−7587
2443318701007175H1SOYMON019g2606080BLASTN10131e−7590
2444318700725567H1SOYMON009g2606080BLASTN10131e−7593
2445318700904972H1SOYMON022g22037BLASTN10151e−7589
2446318700747391H1SOYMON013g2606080BLASTN10171e−7587
2447318700747523H1SOYMON013g22037BLASTN8361e−7486
2448318700561819H1SOYMON002g218332BLASTN9991e−7482
2449318700835961H1SOYMON019g218332BLASTN10061e−7487
2450318700562318H1SOYMON002g2606080BLASTN9861e−7384
2451318700745092H1SOYMON013g2606080BLASTN9871e−7388
2452318700832618H1SOYMON019g2606080BLASTN9751e−7287
2453318700891092H1SOYMON024g2606080BLASTN9821e−7288
2454318701119264H1SOYMON037g2606080BLASTN6901e−7189
2455318700894436H1SOYMON024g2606080BLASTN9011e−7191
2456318700894532H1SOYMON024g22037BLASTN9591e−7189
2457318700891712H1SOYMON024g22037BLASTN9601e−7189
2458318700895985H1SOYMON027g2606080BLASTN9641e−7189
2459318701203243H1SOYMON035g2606080BLASTN9691e−7188
2460318700985945H1SOYMON009g218332BLASTN7131e−7090
2461318700984768H1SOYMON009g2606080BLASTN7811e−6984
2462318700675710H1SOYMON007g2606080BLASTN7841e−6991
2463318700829561H1SOYMON019g218332BLASTN9351e−6987
2464318700964918H1SOYMON022g22037BLASTN9421e−6983
2465318701046747H1SOYMON032g2606080BLASTN4221e−6884
2466318700745512H1SOYMON013g3169543BLASTN4571e−6885
2467318700666671H1SOYMON005g218332BLASTN5061e−6887
2468318700889555H1SOYMON024g3169543BLASTN9301e−6886
2469318701147844H1SOYMON031g3169543BLASTN9321e−6886
2470318701206247H1SOYMON035g3169543BLASTN9341e−6882
2471318701103801H1SOYMON036g218332BLASTN7231e−6788
2472318700943746H1SOYMON024g218332BLASTN9131e−6786
2473318700745956H1SOYMON013g22037BLASTN9211e−6783
2474318700893512H1SOYMON024g218332BLASTN8351e−6690
2475318700897675H1SOYMON027g22037BLASTN8991e−6683
2476318700565777H1SOYMON002g2606080BLASTN5101e−6589
2477318700749851H1SOYMON013g2606080BLASTN8871e−6589
2478318700746286H1SOYMON013g2606080BLASTN8761e−6482
2479318700869142H1SOYMON016g2606080BLASTN8851e−64100
2480318700892442H1SOYMON024g2606080BLASTN8721e−6384
2481318700964153H1SOYMON022g22037BLASTN8731e−6383
2482318700898176H1SOYMON027g3169543BLASTN8731e−6384
2483318701056245H1SOYMON032g218332BLASTN5431e−6184
2484318700835360H1SOYMON019g218332BLASTN8391e−6188
2485318700749067H1SOYMON013g3169543BLASTN4731e−6086
2486318701008962H1SOYMON019g3169543BLASTN6141e−6090
2487318700980315H1SOYMON009g3169543BLASTN6551e−6084
2488318701202680H1SOYMON035g2606080BLASTN6781e−6089
2489318701202364H1SOYMON035g2606080BLASTN7111e−6085
2490318701037195H1SOYMON029g218332BLASTN4391e−5986
2491318701011681H1SOYMON019g3169543BLASTN4591e−5983
2492318700976368H1SOYMON009g218332BLASTN3631e−5885
2493318700829847H1SOYMON019g218332BLASTN3841e−5886
2494318700561920H1SOYMON002g2606080BLASTN8091e−5888
2495318701004573H1SOYMON019g2606080BLASTN8131e−5877
2496318701049462H1SOYMON032g3169543BLASTN4501e−5782
2497318700866272H1SOYMON016g3169543BLASTN4211e−5477
2498318700892632H1SOYMON024g2606080BLASTN4531e−5484
2499318701215184H1SOYMON035g218332BLASTN4641e−5488
2500318700831177H1SOYMON019g2606080BLASTN7591e−5485
2501318700835115H1SOYMON019g2606080BLASTN7621e−5481
2502318701015056H1SOYMON019g3169543BLASTN4471e−5381
2503318700675496H1SOYMON007g2606080BLASTN4651e−5395
2504318701052767H1SOYMON032g2606080BLASTN7531e−5388
2505318700833078H1SOYMON019g3169543BLASTN4141e−5284
2506318700869165H1SOYMON016g3169543BLASTN5341e−5184
2507318700831532H1SOYMON019g2606080BLASTN6551e−51100
2508318701010104H2SOYMON019g2606080BLASTN6981e−5185
2509318700890513H1SOYMON024g22037BLASTN5751e−5088
2510318700890952H1SOYMON024g2606080BLASTN7091e−5075
2511318700567301H1SOYMON002g22037BLASTN7161e−5082
2512318700945284H1SOYMON024g3169543BLASTN7011e−4975
2513318701206626H1SOYMON035g3169543BLASTN7021e−4981
2514318700748456H1SOYMON013g2606080BLASTN3841e−4877
2515318700981883H1SOYMON009g2606080BLASTN4191e−4885
2516318700942575H1SOYMON024g22037BLASTN3401e−4682
2517318700945125H1SOYMON024g2606080BLASTN4051e−4681
2518318700830469H1SOYMON019g3169543BLASTN6361e−4483
2519318700991669H1SOYMON011g218332BLASTN6301e−4383
2520318700866064H1SOYMON016g3169543BLASTN4531e−4184
2521318700866806H1SOYMON016g218332BLASTN6071e−4196
2522318700893154H1SOYMON024g2606080BLASTN5391e−3887
2523318700893118H1SOYMON024g2606080BLASTN5391e−3887
2524318701142963H2SOYMON038g218332BLASTN5691e−3890
2525318700945968H1SOYMON024g218332BLASTN5721e−3886
2526318700945788H1SOYMON024g2606080BLASTN5141e−3690
2527318700563455H1SOYMON002g2606080BLASTN4961e−3283
2528318700888936H1SOYMON024g3169543BLASTN4981e−3286
2529318701039594H1SOYMON029g22037BLASTN2541e−2884
2530318701015024H1SOYMON019g218333BLASTX651e−1466
2531318700893166H1SOYMON024g22037BLASTN2321e−885
25324258700646449H1SOYMON013g22037BLASTN5841e−3970
25334258700952838H1SOYMON022g20373BLASTN5571e−3770
25344413700902256H1SOYMON027g2606080BLASTN12151e−9997
25354413700900032H1SOYMON027g2606080BLASTN7201e−9598
25364413701006182H1SOYMON019g2606080BLASTN11791e−8999
25374413700831710H1SOYMON019g2606080BLASTN10701e−8097
25384413701008850H1SOYMON019g2606080BLASTN9991e−7499
25394413701015432H1SOYMON019g2606080BLASTN8131e−6895
25404413700987094H1SOYMON009g2606080BLASTN9281e−6884
25414413700736179H1SOYMON010g2606080BLASTN7531e−6396
25424413700890230H1SOYMON024g2606080BLASTN7981e−5795
25434413701015314H1SOYMON019g2606080BLASTN6391e−4997
25444413701052019H1SOYMON032g2606080BLASTN4481e−3795
25454748701209527H1SOYMON035g2606080BLASTN12071e−9193
25464748700561984H1SOYMON002g2606080BLASTN5421e−8194
25474748700895166H1SOYMON024g2606080BLASTN10041e−7498
25484748700843735H1SOYMON021g2606080BLASTN2271e−2093
2549869700650545H1SOYMON003g2606080BLASTN8041e−10794
2550869701205255H1SOYMON035g2606080BLASTN11351e−10198
2551869700562091H1SOYMON002g2606080BLASTN13111e−10092
2552869701213906H1SOYMON035g2606080BLASTN13001e−99100
2553869700567712H1SOYMON002g2606080BLASTN6341e−9597
2554869701010943H1SOYMON019g2606080BLASTN12361e−9499
2555869701006976H1SOYMON019g2606080BLASTN6011e−9398
2556869700752409H1SOYMON014g2606080BLASTN10801e−92100
2557869701204769H1SOYMON035g2606080BLASTN7951e−90100
2558869701042737H1SOYMON029g2606080BLASTN10581e−9099
2559869700832091H1SOYMON019g2606080BLASTN11161e−8899
2560869701049161H1SOYMON032g2606080BLASTN10531e−8696
2561869700906541H1SOYMON022g2606080BLASTN10871e−8696
2562869701008182H1SOYMON019g2606080BLASTN11111e−8692
2563869700831609H1SOYMON019g2606080BLASTN6111e−8492
2564869700834954H1SOYMON019g2606080BLASTN8351e−84100
2565869701037284H1SOYMON029g2606080BLASTN8581e−8394
2566869700561458H1SOYMON002g2606080BLASTN10191e−8393
2567869701208357H1SOYMON035g2606080BLASTN11131e−8399
2568869700747138H1SOYMON013g2606080BLASTN9851e−8093
2569869701014835H1SOYMON019g2606080BLASTN8911e−7889
2570869700956359H1SOYMON022g2606080BLASTN10521e−7896
2571869701012740H1SOYMON019g2606080BLASTN6431e−7793
2572869701042523H1SOYMON029g2606080BLASTN6671e−7495
2573869701205775H1SOYMON035g2606080BLASTN7451e−74100
2574869701049184H1SOYMON032g2606080BLASTN6001e−7295
2575869700889179H1SOYMON024g2606080BLASTN9421e−6992
2576869700963920H1SOYMON022g2606080BLASTN7181e−6690
2577869700737476H1SOYMON010g2606080BLASTN5481e−4497
2578869701044544H1SOYMON032g2606080BLASTN4621e−4396
2579869700737636H1SOYMON010g2606080BLASTN4261e−3495
25809398700837013H1SOYMON020g2570066BLASTN10251e−7688
25819398700891526H1SOYMON024g2570066BLASTN8681e−6387
258214740LIB3051-038-LIB3051g2570066BLASTN13311e−10286
Q1-K1-E10
258331182LIB3051-015-LIB3051g2570066BLASTN15401e−11988
Q1-E1-F1
2584318LIB3050-024-LIB3050g2606080BLASTN17361e−13595
Q1-K1-H5
2585318LIB3050-012-LIB3050g2606080BLASTN15641e−12598
Q1-E1-F10
2586318LIB3056-013-LIB3056g3169543BLASTN16171e−12586
Q1-N1-H11
2587318LIB3028-026-LIB3028g3169543BLASTN13931e−10784
Q1-B1-F6
2588318LIB3049-031-LIB3049g3169543BLASTN12901e−9890
Q1-E1-B6
258933428LIB3051-085-LIB3051g2570066BLASTN6791e−5386
Q1-K1-D11
2590869LIB3056-014-LIB3056g2606080BLASTN15031e−13296
Q1-N1-G8
SOYBEAN HEXOKINASE
2591−700560085700560085H1SOYMON001g1899024BLASTN4561e−2767
2592−700752579700752579H1SOYMON014g836808BLASTX1131e−854
2593−700753182700753182H1SOYMON014g619928BLASTX2341e−2563
2594−700838622700838622H1SOYMON020g619927BLASTN7671e−5578
2595−700840271700840271H1SOYMON020g619927BLASTN5251e−3467
2596−700844132700844132H1SOYMON021g619927BLASTN4741e−5177
2597−700898308700898308H1SOYMON027g619927BLASTN4641e−2972
2598−700904279700904279H1SOYMON022g881521BLASTX1291e−1067
2599−700904320700904320H1SOYMON022g1899024BLASTN6121e−4271
2600−700946357700946357H1SOYMON024g619928BLASTX1121e−1869
2601−700998007700998007H1SOYMON018g1899024BLASTN3671e−2071
2602−701097096701097096H1SOYMON028g619927BLASTN4881e−3073
2603−701102877701102877H1SOYMON028g619927BLASTN5511e−3770
2604−701103285701103285H1SOYMON028g619928BLASTX1791e−1777
2605−701105838701105838H1SOYMON036g619928BLASTX2741e−3063
2606−701138291701138291H1SOYMON038g619927BLASTN8191e−5979
260712404701065794H1SOYMON034g3087888BLASTX841e−1144
260812404701131030H1SOYMON038g1899025BLASTX1201e−945
260912693700846513H1SOYMON021g619927BLASTN4591e−2870
261012693700656744H1SOYMON004g619927BLASTN2511e−1057
261112917700906858H1SOYMON022g3087888BLASTX1831e−3280
261212917700830011H1SOYMON019g619927BLASTN4951e−3270
261312917701068501H1SOYMON034g619927BLASTN4751e−2972
261412917701153981H1SOYMON031g3087887BLASTN4401e−2669
2615222700663332H1SOYMON005g619927BLASTN7241e−5176
2616222701142003H1SOYMON038g881520BLASTN5421e−3972
2617222700657213H1SOYMON004g881520BLASTN5241e−3473
2618222700833679H1SOYMON019g1899024BLASTN4531e−2880
2619222700556060H1SOYMON001g619927BLASTN4631e−2882
262023610700984359H1SOYMON009g1899024BLASTN6111e−4273
262123610701003284H1SOYMON019g1899024BLASTN5771e−3975
262225188700760643H1SOYMON015g619927BLASTN7011e−4973
262325188701056127H1SOYMON032g1899024BLASTN6491e−4570
262427316701054167H1SOYMON032g3087888BLASTX1771e−1747
262527316701054157H1SOYMON032g3087888BLASTX1771e−1747
2626488700682650H2SOYMON008g687676BLASTN7301e−5277
2627488700849894H1SOYMON021g687676BLASTN5821e−3976
2628-GM32703LIB3051-008-LIB3051g1899024BLASTN9811e−7677
Q1-E1-C12
2629-GM9523LIB3049-003-LIB3049g619928BLASTX2031e−3764
Q1-E1-A6
263012693LIB3051-106-LIB3051g619927BLASTN4591e−3871
Q1-K1-A9
2631488LIB3040-006-LIB3040g687676BLASTN6221e−4176
Q1-E1-A12
2632488LIB3053-008-LIB3053g687676BLASTN5971e−3975
Q1-N1-C6
2633488LIB3055-008-LIB3055g687676BLASTN5591e−3675
Q1-N1-F5
2634488LIB3053-010-LIB3053g687676BLASTN5141e−3275
Q1-N1-D8
SOYBEAN FRUCTOKINASE
2635−700834049700834049H1SOYMON019g1915974BLASTX1121e−1097
2636−700905716700905716H1SOYMON022g1915973BLASTN7741e−5577
2637−700978126700978126H1SOYMON009g1915973BLASTN5651e−3877
2638−700983171700983171H1SOYMON009g1915974BLASTX961e−993
2639−701069652701069652H1SOYMON034g297014BLASTN4471e−2773
2640−701118004701118004H2SOYMON037g2102690BLASTN4401e−2673
2641−701209270701209270H1SOYMON035g1052972BLASTN6481e−4579
26421174700832430H1SOYMON019g1915973BLASTN6381e−4481
26431174701101576H1SOYMON028g1915973BLASTN5921e−4079
26441174700754333H1SOYMON014g1915973BLASTN3231e−3780
26451174701004323H1SOYMON019g297014BLASTN5601e−3780
26461174700988192H1SOYMON009g1915973BLASTN5081e−3378
26471174700646337H1SOYMON013g1915974BLASTX1531e−3079
26481174701039647H1SOYMON029g1915973BLASTN2751e−1280
264916472701155250H1SOYMON031g1915973BLASTN6421e−5078
265016472700953304H1SOYMON022g1915973BLASTN6901e−4879
265116472700725996H1SOYMON009g1915973BLASTN3621e−2873
265217936700965277H1SOYMON022g2102690BLASTN3751e−4277
265317936700746240H1SOYMON013g2102690BLASTN6061e−4174
265422120701215393H1SOYMON035g2102691BLASTX1331e−1186
265522586701009695H1SOYMON019g2102690BLASTN6961e−4976
265622586700900731H1SOYMON027g2102690BLASTN4221e−2676
265723551701053585H1SOYMON032g2102691BLASTX1201e−992
265828587701156878H1SOYMON031g2102690BLASTN4481e−3372
26593876700942858H1SOYMON024g297014BLASTN7051e−4974
26603876701063105H1SOYMON033g1052972BLASTN6791e−4773
26613876700844831H1SOYMON021g1915973BLASTN4661e−3772
26625530700733713H1SOYMON010g1915974BLASTX1561e−2681
26635530701057239H1SOYMON033g1915974BLASTX1761e−1792
26645530700985231H1SOYMON009g297014BLASTN2221e−1679
26655805701010614H1SOYMON019g1915973BLASTN9581e−7180
26665805701003106H1SOYMON019g1915973BLASTN6791e−6481
26675805700748895H1SOYMON013g1915973BLASTN4751e−5583
26685805700892801H1SOYMON024g1915973BLASTN6391e−5580
26695805700891914H1SOYMON024g1915973BLASTN6391e−5581
26705805700962529H1SOYMON022g1915973BLASTN6221e−5482
26715805700869294H1SOYMON016g1915973BLASTN7601e−5480
26725805700986530H1SOYMON009g1915973BLASTN7611e−5480
26735805700661115H1SOYMON005g1915973BLASTN6821e−4878
26745805701041987H1SOYMON029g297014BLASTN4751e−4583
26755805701006803H1SOYMON019g1915973BLASTN6071e−4180
267628587LIB3028-008-LIB3028g2102690BLASTN9001e−6668
Q1-B1-H3
26775530LIB3055-004-LIB3055g297014BLASTN6061e−3976
Q1-N1-H3
26785805LIB3065-006-LIB3065g1915973BLASTN9541e−8179
Q1-N1-F11
SOYBEAN NDP-KINASE
267933331701108520H1SOYMON036g758643BLASTN4731e−3175
268023595LIB3050-018-LIB3050g758643BLASTN2951e−1376
Q1-E1-C4
268133331LIB3040-037-LIB3040g758643BLASTN4131e−4779
Q1-E1-D6
SOYBEAN GLUCOSE-6-PHOSPHATE 1-DEHYDROGENASE
2682−700869140700869140H1SOYMON016g2829880BLASTX1641e−1544
2683−701065174701065174H1SOYMON034g603219BLASTX861e−976
2684−701130434701130434H1SOYMON037g1197385BLASTX1891e−1955
2685−701149522701149522H1SOYMON031g603219BLASTX991e−871
268626484701003905H1SOYMON019g1197385BLASTX1381e−1581
26879136701038169H1SOYMON029g603219BLASTX1391e−2173
26889136700903571H1SOYMON022g603219BLASTX1441e−2081
26899136701045122H1SOYMON032g603219BLASTX1001e−1379
SOYBEAN PHOSPHOGLUCOMUTASE
2690−700554424700554424H1SOYMON001g534982BLASTX1331e−2560
2691−700556670700556670H1SOYMON001g3294468BLASTN3551e−4374
2692−700563871700563871H1SOYMON002g2795876BLASTX1011e−1675
2693−700565101700565101H1SOYMON002g3294466BLASTN5881e−4068
2694−700566749700566749H1SOYMON002g1814400BLASTN4751e−4173
2695−700681382700681382H2SOYMON008g3294467BLASTX981e−1148
2696−700763827700763827H1SOYMON018g3192042BLASTX2571e−2960
2697−700865583700865583H1SOYMON016g3192042BLASTX1341e−1757
2698−700891379700891379H1SOYMON024g534982BLASTX1671e−1553
2699−700942816700942816H1SOYMON024g3294466BLASTN6361e−4474
2700−701004954701004954H1SOYMON019g1814400BLASTN7901e−5678
2701−701011364701011364H1SOYMON019g534982BLASTX2841e−3267
2702−701057063701057063H2SOYMON033g1814401BLASTX1211e−960
2703−701119491701119491H1SOYMON037g1814400BLASTN7621e−5476
2704−701149254701149254H1SOYMON031g534982BLASTX1471e−1952
270510032700988921H1SOYMON011g1814400BLASTN9081e−6680
270610032701136003H1SOYMON038g1814400BLASTN8421e−6178
270710032700953253H1SOYMON022g1814400BLASTN8081e−5877
270810032701103083H1SOYMON028g1814400BLASTN8131e−5878
270910131701104852H1SOYMON036g3294466BLASTN3021e−2774
271010131700970420H1SOYMON005g2829893BLASTX2401e−2656
27111180701125681H1SOYMON037g2829893BLASTX1631e−1582
27121180700559947H1SOYMON001g2829893BLASTX1631e−1582
27131180700556009H1SOYMON001g2829893BLASTX1021e−1487
271413262701006086H2SOYMON019g3294466BLASTN7341e−5275
271513262701137937H1SOYMON038g3294466BLASTN4911e−3271
271613262701004207H1SOYMON019g3294466BLASTN2711e−3075
271713262700904551H1SOYMON022g3294466BLASTN4731e−3075
271813262701014357H1SOYMON019g1814401BLASTX2101e−2183
271913262701146638H1SOYMON031g1814401BLASTX1111e−2080
272013262700833416H1SOYMON019g1814400BLASTN3741e−2074
272113262701148967H1SOYMON031g1814401BLASTX1941e−1982
272213262701156042H1SOYMON031g1814401BLASTX1821e−1867
272313262700943365H1SOYMON024g1814401BLASTX1681e−1676
272413262701105762H1SOYMON036g1814401BLASTX1651e−1583
272513262701038338H1SOYMON029g1814400BLASTN1861e−1377
272613262700645989H1SOYMON011g1814401BLASTX1331e−1178
272713262700868941H1SOYMON016g1814400BLASTN1811e−980
272819312701121150H1SOYMON037g3294468BLASTN5011e−6179
272919312700742959H1SOYMON012g3294468BLASTN4401e−4583
273019312701135418H1SOYMON038g3294468BLASTN4591e−4279
273119312700979514H2SOYMON009g1814400BLASTN6121e−4278
273219883701133631H2SOYMON038g1814400BLASTN7581e−5475
273319883700970758H1SOYMON005g1814400BLASTN7171e−5077
273419883701153416H1SOYMON031g1814400BLASTN6911e−4876
273526278701214005H1SOYMON035g534982BLASTX1181e−847
2736-GM1647LIB3028-009-LIB3028g534982BLASTX1921e−4257
Q1-B1-F3
2737-GM17162LIB3055-012-LIB3055g1814400BLASTN4911e−2962
Q1-N1-B3
273813262LIB3028-003-LIB3028g1814400BLASTN10691e−8076
Q1-B1-B11
273913262LIB3054-009-LIB3054g1814400BLASTN6121e−4073
Q1-N1-A12
274013262LIB3054-009-LIB3054g1814401BLASTX2001e−3673
Q1-N1-A5
SOYBEAN UDP-GLUCOSE PYROPHOSPHORYLASE
2741−700665357700665357H1SOYMON005g1388021BLASTX1831e−1869
2742−700674325700674325H1SOYMON007g218000BLASTN6451e−4472
2743−700835903700835903H1SOYMON019g1388021BLASTX1351e−1168
2744−700841466700841466H1SOYMON020g1388021BLASTX1151e−1456
2745−700846570700846570H1SOYMON021g3107930BLASTN4861e−3170
2746−700888547700888547H1SOYMON024g3107930BLASTN5821e−3981
2747−700973436700973436H1SOYMON005g1212996BLASTX1321e−1551
2748−700985779700985779H1SOYMON009g3107930BLASTN9581e−7183
2749−700992994700992994H1SOYMON011g1388021BLASTX1031e−1064
2750−701061122701061122H1SOYMON033g1388021BLASTX1291e−1973
2751−701063465701063465H1SOYMON033g3107930BLASTN4261e−6282
2752−701118256701118256H1SOYMON037g3107930BLASTN3781e−3183
275311810700952705H1SOYMON022g3107930BLASTN6131e−5480
275411810701060568H1SOYMON033g3107930BLASTN6521e−4581
275511810701002783H2SOYMON019g3107930BLASTN4581e−4380
275611810701202674H1SOYMON035g218000BLASTN3311e−3373
275711810700871590H1SOYMON018g218000BLASTN3261e−2776
275811810700747279H1SOYMON013g1388021BLASTX1541e−2175
275911810701014424H1SOYMON019g1388021BLASTX1311e−2084
276011810701039454H1SOYMON029g1388021BLASTX1571e−1680
276111810701054271H1SOYMON032g1388021BLASTX1541e−1571
276211810700955092H1SOYMON022g1388021BLASTX1541e−1471
276311810701107189H1SOYMON036g1388021BLASTX1551e−1472
276411810701107930H1SOYMON036g218000BLASTN3081e−1475
276511810700904384H1SOYMON022g1388021BLASTX1491e−1372
276611810700729516H1SOYMON009g1388021BLASTX1551e−1372
276711810701009325H1SOYMON019g1388021BLASTX1431e−1275
276811821701060627H1SOYMON033g218000BLASTN2531e−2674
276911821701004671H1SOYMON019g21599BLASTX1661e−2277
277011821700964889H1SOYMON022g1388021BLASTX1671e−1667
277113178700562308H1SOYMON002g3107930BLASTN11981e−9187
277213178701049018H1SOYMON032g3107930BLASTN11011e−8288
277313178701126215H1SOYMON037g3107930BLASTN10721e−8088
277413178701211745H1SOYMON035g3107930BLASTN10381e−7787
277513178700850417H1SOYMON023g3107930BLASTN10221e−7687
277613178700665292H1SOYMON005g3107930BLASTN9801e−7288
277713178700994009H1SOYMON011g3107930BLASTN9581e−7186
277813178700895203H1SOYMON024g3107930BLASTN8641e−6886
277913178701151725H1SOYMON031g3107930BLASTN8001e−6687
278013178700988803H1SOYMON011g3107930BLASTN8961e−6580
278113178700646581H1SOYMON014g3107930BLASTN4831e−6281
278213178701153726H1SOYMON031g3107930BLASTN8321e−6087
278313178701152333H1SOYMON031g3107930BLASTN6741e−5679
278413178700756960H1SOYMON015g3107930BLASTN7871e−5686
278513178700556901H1SOYMON001g218000BLASTN7721e−5584
278613178701063605H1SOYMON033g3107930BLASTN5661e−5189
278713178701212385H1SOYMON035g3107930BLASTN3901e−2378
278813178700889518H1SOYMON024g3107931BLASTX1311e−1064
278917057700740176H1SOYMON012g3107930BLASTN7981e−5781
279017057700905747H1SOYMON022g3107930BLASTN5111e−3381
27911955701059208H1SOYMON033g3107930BLASTN10341e−7785
27921955700984109H1SOYMON009g3107930BLASTN9701e−7284
27931955701209482H1SOYMON035g3107930BLASTN9311e−6884
27941955700554847H1SOYMON001g3107930BLASTN4931e−6683
27951955701150363H1SOYMON031g3107930BLASTN8981e−6684
27961955700986014H1SOYMON009g3107930BLASTN9071e−6684
27971955700564270H1SOYMON002g3107930BLASTN5011e−6584
27981955700844253H1SOYMON021g3107930BLASTN8751e−6484
27991955701140892H1SOYMON038g3107930BLASTN8791e−6483
28001955700685893H1SOYMON008g3107930BLASTN8321e−6081
28011955700789732H1SOYMON011g3107930BLASTN5541e−5784
28021955700902418H1SOYMON027g3107930BLASTN7311e−5283
28031955701128306H1SOYMON037g3107930BLASTN4661e−4682
28041955701057973H1SOYMON033g3107930BLASTN4221e−4378
280521035700946288H1SOYMON024g3107931BLASTX1751e−1772
280621035701043539H1SOYMON029g3107931BLASTX1471e−1371
280730564701063642H1SOYMON033g3107931BLASTX1811e−2575
2808-GM18453LIB3065-001-LIB3065g1212996BLASTX681e−2957
Q1-N1-H4
2809-GM32502LIB3051-013-LIB3051g3107931BLASTX2271e−4751
Q1-E1-A6
281011810LIB3030-010-LIB3030g21598BLASTN11151e−8476
Q1-B1-H12
281113178LIB3056-014-LIB3056g3107930BLASTN11451e−9984
Q1-N1-G7
28121955LIB3056-012-LIB3056g3107930BLASTN8561e−6280
Q1-N1-D4
281330564LIB3050-003-LIB3050g3107930BLASTN10781e−8174
Q1-E1-D8
281430564LIB3050-010-LIB3050g3107930BLASTN10501e−7875
Q1-E1-D6
*Table Headings
Cluster ID A cluster ID is arbitrarily assigned to all of those clones which belong to the same cluster at a given stringency and a particular clone will belong to only one cluster at a given stringency. If a cluster contains only a single clone (a “singleton”), then the cluster ID number will be negative, with an absolute value equal to the clone ID number of its single member. The cluster ID entries in the table refer to the cluster with which the particular clone in each row is associated.
Clone ID The clone ID number refers to the particular clone in the PhytoSeq database. Each clone ID entry in the table refers to the clone whose sequence is used for (1) the sequence comparison whose scores are presented and/or (2) assignment to the particular cluster which is presented. Note that a clone may be included in this table even if its sequence comparison scores fail to meet the minimum standards for similarity. In such a case, the clone is included due solely to its association with a particular cluster for which sequences of one or more other member clones possess the required level of similarity.
Library The library ID refers to the particular cDNA library from which a given clone is obtained. Each cDNA library is associated with the particular tissue(s), line(s) and developmental stage(s) from which it is isolated.
NCBI gi Each sequence in the GenBank public database is arbitrarily assigned a unique NCBI gi (National Center for Biotechnology Information GenBank Identifier) number. In this table, the NCBI gi number which is associated (in the same row) with a given clone refers to the particular GenBank sequence which is used in the sequence comparison. This entry is omitted when a clone is included solely due to its association with a particular cluster.
Method The entry in the “Method” column of the table refers to the type of BLAST search that is used for the sequence comparison. “CLUSTER” is entered when the sequence comparison scores for a given clone fail to meet the minimum values required for significant similarity. In such cases, the clone is listed in the table solely as a result of its association with a given cluster for which sequences of one or more other member clones possess the required level of similarity.
Score Each entry in the “Score” column of the table refers to the BLAST score that is generated by sequence comparison of the designated clone with the designated GenBank sequence using the designated BLAST method. This entry is omitted when a clone is included solely due to its association with a particular cluster. If the program used to determine the hit is HMMSW then the score refers to HMMSW score.
P-Value The entries in the P-Value column refer to the probability that such matches occur by chance.
% Ident The entries in the “% Ident” column of the table refer to the percentage of identically matched nucleotides (or residues) that exist along the length of that portion of the sequences which is aligned by the BLAST comparison to generate the statistical scores presented. This entry is omitted when a clone is included solely due to its association with a particular cluster.