Phosphate use efficiency

Abstract

and polypeptides encoded thereby are described, together with the use of those products for making transgenic plants with increased tolerance to pH or increased phosphorus efficiency.

Description

[0001]This application is a Continuation of co-pending application Ser. No. 11/140,347, filed on May 27, 2005, the entire contents of which are hereby incorporated by reference and for which priority is claimed under 35 U.S.C. §120.

[0002]Co-pending application Ser. No. 11/140,347 claims priority under 35 U.S.C. §119(e) on U.S. Provisional Application No(s). 60/575,309 filed on May 27, 2004, the entire contents of which are hereby incorporated by reference.

FIELD OF THE INVENTION

[0003]The present invention relates to isolated polynucleotides, polypeptides encoded thereby, and the use of those sequences for making transgenic plants with modulated pH response and phosphate use efficiency.

BACKGROUND OF THE INVENTION

[0004]Plants are constantly exposed to a variety of biotic (i.e., pathogen infection and insect herbivory) and abiotic (e.g., high pH, low phosphate) stresses. To survive these challenges, plants have developed elaborate mechanisms to perceive external signals and environmental stresses and to manifest adaptive responses with proper physiological and morphological changes (Bohnert et al., 1995). Plants exposed to low or high pH conditions typically have low yields of plant material, seeds, fruit and other edible products. Extreme soil pH conditions have a major influence on nutrient availability resulting in severe agronomic losses. Plants exposed to low pH soil conditions develop deficiencies in nutrients such as copper, molybdate, potassium, sulfur, and nitrogen. Also, plants exposed to high pH soil conditions develop iron, copper, manganese, and zinc deficiencies (FIG. 1). Phosphate deficiency is a problem in both high and low pH soil conditions. Essential mineral nutrients are required in substantial amounts to sustain plant growth and maximize plant yields.

[0005]Consequently, agricultural and horticultural entities routinely alter the rhizosphere to maximize and maintain crop yields; these frequently result in more pollution and unbalancing of the natural soil mineral balance (National Research Council. (1989) Alternative Agriculture. National Academic Press, Washington D.C.). Excessive over-liming of acid soils, for instance, has resulted in the induction of iron, manganese, copper, and zinc deficiencies; deficiencies commonly observed in calcareous soil.

[0006]It would, therefore, be of great interest and importance to be able to identify genes that confer improved phosphate efficiency characteristics to thereby enable one to create transformed plants (such as crop plants) with improved phosphate efficiency characteristics to thereby better survive low and high pH conditions.

[0007]In the field of agriculture and forestry efforts are constantly being made to produce plants with an increased growth potential in order to feed the ever-increasing world population and to guarantee the supply of reproducible raw materials. This is done conventionally through plant breeding. The breeding process is, however, both time-consuming and labor-intensive. Furthermore, appropriate breeding programs must be performed for each relevant plant species.

[0008]Progress has been made in part by the genetic manipulation of plants; that is by introducing and expressing recombinant nucleic acid molecules in plants. Such approaches have the advantage of not usually being limited to one plant species, but instead being transferable among plant species. (Zhang et al. (2004) Plant Physiol. 135:615). There is a need for generally applicable processes that improve forest or agricultural plant growth potential. Therefore, the present invention relates to a process for increasing the abiotic stress tolerance and consequently the growth potential in plants, characterized by expression of recombinant DNA molecules stably integrated into the plant genome.

SUMMARY OF THE INVENTION

[0009]The present invention, therefore, relates to isolated polynucleotides, polypeptides encoded thereby, and the use of those sequences for making transgenic plants with modulated pH tolerance or phosphate use efficiency.

[0010]The present invention also relates to processes for increasing the growth potential in plants under abnormal pH or phosphate conditions, recombinant nucleic acid molecules and polypeptides used for these processes and their uses, as well as to plants themselves.

[0011]Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

BRIEF DESCRIPTION OF THE FIGURES

[0012]FIG. 1 shows the relationship between soil pH and nutrient uptake.

[0013]FIG. 2 shows pH recovery as measured by volume of seeds collected from a plant containing cDNA 1248777 compared to pH treated and un-treated controls.

DETAILED DESCRIPTION OF THE INVENTION

1. Definitions

[0014]The following terms are utilized throughout this application: [0015]Constitutive Promoter: Promoters referred to herein as "constitutive promoters" actively promote transcription under most, but not necessarily all, environmental conditions and states of development or cell differentiation. Examples of constitutive promoters include the cauliflower mosaic virus (CaMV) 35S transcript initiation region and the 1' or 2' promoter derived from T-DNA of Agrobacterium tumefaciens, and other transcription initiation regions from various plant genes, such as the maize ubiquitin-1 promoter, known to those of skill. [0016]Domain: Domains are fingerprints or signatures that can be used to characterize protein families and/or parts of proteins. Such fingerprints or signatures can comprise conserved (1) primary sequence, (2) secondary structure, and/or (3) three-dimensional conformation. Generally, each domain has been associated with either a family of proteins or motifs. Typically, these families and/or motifs have been correlated with specific in-vitro and/or in-vivo activities. A domain can be any length, including the entirety of the sequence of a protein. Detailed descriptions of the domains, associated families and motifs, and correlated activities of the polypeptides of the instant invention are described below. Usually, the polypeptides with designated domain(s) can exhibit at least one activity that is exhibited by any polypeptide that comprises the same domain(s). [0017]Endogenous: The term "endogenous," within the context of the current invention refers to any polynucleotide, polypeptide or protein sequence which is a natural part of a cell or organisms regenerated from said cell. [0018]Exogenous: "Exogenous," as referred to within, is any polynucleotide, polypeptide or protein sequence, whether chimeric or not, that is initially or subsequently introduced into the genome of an individual host cell or the organism regenerated from said host cell by any means other than by a sexual cross. Examples of means by which this can be accomplished are described below, and include Agrobacterium-mediated transformation (of dicots--e.g. Salomon et al. EMBO J. 3:141 (1984); Herrera-Estrella et al. EMBO J. 2:987 (1983); of monocots, representative papers are those by Escudero et al., Plant J. 10:355 (1996), Ishida et al., Nature Biotechnology 14:745 (1996), May et al., Bio/Technology 13:486 (1995)), biolistic methods (Armaleo et al., Current Genetics 17:97 1990)), electroporation, in planta techniques, and the like. Such a plant containing the exogenous nucleic acid is referred to here as a T₀ for the primary transgenic plant and T₁ for the first generation. The term "exogenous" as used herein is also intended to encompass inserting a naturally found element into a non-naturally found location. [0019]Functionally Comparable Proteins: This phrase describes those proteins that have at least one characteristic in common. Such characteristics include sequence similarity, biochemical activity, transcriptional pattern similarity and phenotypic activity. Typically, the functionally comparable proteins share some sequence similarity or at least one biochemical and within this definition, homologs, orthologs and analogs are considered to be functionally comparable. In addition, functionally comparable proteins generally share at least one biochemical and/or phenotypic activity.

[0020]Functionally comparable proteins will give rise to the same characteristic to a similar, but not necessarily to the same degree. Typically, comparable proteins give the same characteristics where the quantitative measurement due to one of the comparables is at lest 20% of the other; more typically, between 30 to 40%; even more typically, between 50-60%; even more typically, 70 to 80%; even more typically between 90 to 100%. [0021]Heterologous sequences: "Heterologous sequences" are those that are not operatively linked or are not contiguous to each other in nature. For example, a promoter from corn is considered heterologous to an Arabidopsis coding region sequence. Also, a promoter from a gene encoding a growth factor from corn is considered heterologous to a sequence encoding the corn receptor for the growth factor. Regulatory element sequences, such as UTRs or 3' end termination sequences that do not originate in nature from the same gene as the coding sequence originates from, are considered heterologous to said coding sequence. Elements operatively linked in nature and contiguous to each other are not heterologous to each other. On the other hand, these same elements remain operatively linked but become heterologous if other filler sequence is placed between them. Thus, the promoter and coding sequences of a corn gene expressing an amino acid transporter are not heterologous to each other, but the promoter and coding sequence of a corn gene operatively linked in a novel manner are heterologous. [0022]High pH: "High pH" can be defined as a non-optimal and terminal alkaline pH value when a given plant can no longer make use of certain essential nutrients, such as phosphate, available in the soil. For instance, if a plant grows optimally at pH of 4.0-5.0, high pH would be any pH greater than 5. If the optimal pH were in the range of 6-6.5, high pH would be a pH greater than pH 6.5. As an example, if a corn crop under optimal pH conditions would yield 134 bushels per acre and all other conditions were held constant, a high pH tolerant variety would produce similar yields at pH 9 or above. [0023]Inducible Promoter: An "inducible promoter" in the context of the current invention refers to a promoter which is regulated under certain conditions, such as light, chemical concentration, protein concentration, conditions in an organism, cell, or organelle, etc. A typical example of an inducible promoter, which can be utilized with the polynucleotides of the present invention, is PARSK1, the promoter from the Arabidopsis gene encoding a serine-threonine kinase enzyme, and which promoter is induced by dehydration, abscissic acid and sodium chloride (Wang and Goodman, Plant 1 8:37 (1995)). Examples of environmental conditions that may affect transcription by inducible promoters include anaerobic conditions, elevated temperature, or the presence of light. [0024]Low Nitrogen: "Low nitrogen" can be defined as a quantity of nitrogen, whether in the form of ammonium or nitrate, which is insufficient to sustain normal growth and yield for a given plant. The need for nitrogen fertilizers varies considerably among plants. Further, the type of soil and the conditions in the soil have a significant impact on the ability of a plant to take up nitrogen. Supplemental nitrogen fertilizers are often added to soil or applied directly to plants to enhance their growth or appearance. Even with normal fertilizer applications, the amount of nitrogen available to a plant at any given time may be too low to support optimal growth. Hence, low nitrogen must be defined in terms of the specific plant and environment in which the plant is being grown. For example, if under a given set of conditions with a specific corn hybrid the optimal nitrogen level was 160 pounds of nitrogen fertilizer per acre and under such conditions the hybrid were able to achieve a yield of 134 bushels per acre, a low nitrogen tolerant hybrid would grow optimally and produce the same yield with at least10% less or at least 20% less or at least 30% less or at least 40% less or at least 50% less nitrogen. Further, the low nitrogen hybrid would grow better after much of the initial nitrogen had been depleted and would not require multiple applications of nitrogen. [0025]Low pH: "Low pH" can be defined as that non-optimal and terminal acidic pH value when a given plant can no longer make use of certain essential nutrients, such as potassium, available in the soil. If a plant grows optimally at pH of 4.0-5.0, low pH is any pH less than 4. If the optimal pH is in the range of 6-8, low pH would be a pH less than 6. For example, if a corn crop under optimal pH conditions would yield 134 bushels per acre and all other conditions were held constant, a low pH tolerant variety would produce similar yields at pH 5, or pH 4. [0026]Low Phosphate: "Low phosphate" can be defined as a quantity of phosphate which is insufficient to sustain normal growth and yield for a given plant. The level of phosphate required for optimal plant growth differs among plant species and depends on the condition of the soil and other environmental conditions. To determine a level of phosphate that is low, comparative experiments are needed. For example, if a corn hybrid in a particular field treated with 40 pounds of phosphate per acre would yield 134 bushels per acre and all other conditions were held constant, a low phosphate tolerant hybrid would produce similar yields at 35 or less pounds of phosphate per acre or 30 or less pounds of phosphate per acre or 25 or less pounds of phosphate per acre or 20 or less pounds of phosphate per acre. [0027]Masterpool: The "master pools" discussed in these experiments are a pool of seeds from five different transgenic plants transformed with the same exogenous gene. [0028]Misexpression: The term "misexpression" refers to an increase or a decrease in the transcription of a coding region into a complementary RNA sequence as compared to the wild-type. This term also encompasses expression of a gene or coding region for a different time period as compared to the wild-type and/or from a non-natural location within the plant genome. [0029]Percentage of sequence identity: "Percentage of sequence identity," as used herein, is determined by comparing two optimally aligned sequences over a comparison window, where the fragment of the polynucleotide or amino acid sequence in the comparison window may comprise additions or deletions (e.g., gaps or overhangs) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity. Optimal alignment of sequences for comparison may be conducted by the local homology algorithm of Smith and Waterman Add. APL. Math. 2:482 (1981), by the homology alignment algorithm of Needleman and Wunsch J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson and Lipman Proc. Natl. Acad. Sci. (USA) 85: 2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, PASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.), or by inspection. Given that two sequences have been identified for comparison, GAP and BESTFIT are preferably employed to determine their optimal alignment. Typically, the default values of 5.00 for gap weight and 0.30 for gap weight length are used. The term "substantial sequence identity" between polynucleotide or polypeptide sequences refers to polynucleotide or polypeptide comprising a sequence that has at least 80% sequence identity, preferably at least 85%, more preferably at least 90% and most preferably at least 95%, even more preferably, at least 96%, 97%, 98% or 99% sequence identity compared to a reference sequence using the programs.

[0030]Query nucleic acid and amino acid sequences were searched against subject nucleic acid or amino acid sequences residing in public or proprietary databases. Such searches were done using the Washington University Basic Local Alignment Search Tool Version 1.83 (WU-Blast2) program. The WU-Blast2 program is available on the internet from Washington University. A WU-Blast2 service for Arabidopsis can also be found on the internet. Typically the following parameters of WU-Blast2 were used: Filter options were set to "default," Output format was set to "gapped alignments," the Comparison Matrix was set to "BLOSUM62," Cutoff Score (S value) was set to "default," the Expect (E threshold) was set to "default," the Number of best alignments to show was set to "100," and the "Sort output" option was set to sort the output by "pvalue." [0031]Plant Promoter: A "plant promoter" is a promoter capable of initiating transcription in plant cells and can drive or facilitate transcription of a nucleotide sequence or fragment thereof of the instant invention. Such promoters need not be of plant origin. For example, promoters derived from plant viruses, such as the CaMV35S promoter or from Agrobacterium tumefaciens such as the T-DNA promoters, can be plant promoters. A typical example of a plant promoter of plant origin is the maize ubiquitin-1 (ubi-1) promoter known to those of skill. [0032]Specific Promoter: In the context of the current invention, "specific promoters" refers to promoters that have a high preference for being active in a specific tissue or cell and/or at a specific time during development of an organism. By "high preference" is meant at least 3-fold, preferably 5-fold, more preferably at least 10-fold still more preferably at least 20-fold, 50-fold or 100-fold increase in transcription in the desired tissue over the transcription in any other tissue. Typical examples of temporal and/or tissue specific promoters of plant origin that can be used with the polynucleotides of the present invention, are: SH-EP from Vigna mungo and EP-C1 from Phaseolus vulgaris (Yamauchi et al. (1996) Plant Mol Biol. 30(2):321-9.); RCc2 and RCc3, promoters that direct root-specific gene transcription in rice (Xu et al., Plant Mol. Biol. 27:237 (1995) and TobRB27, a root-specific promoter from tobacco (Yamamoto et al., Plant Cell 3:371 (1991)). [0033]Stringency: "Stringency" as used herein is a function of probe length, probe composition (G+C content), and salt concentration, organic solvent concentration, and temperature of hybridization or wash conditions. Stringency is typically compared by the parameter T_m, which is the temperature at which 50% of the complementary molecules in the hybridization are hybridized, in terms of a temperature differential from T_m. High stringency conditions are those providing a condition of T_m-5° C. to T_m-10° C. Medium or moderate stringency conditions are those providing T_m-20° C. to T_m-29° C. Low stringency conditions are those providing a condition of T_m-40° C. to T_m-48° C. The relationship of hybridization conditions to T_m (in ° C.) is expressed in the mathematical equation

[0033]T_m=81.5-16.6(log₁₀[Na.sup.+])+0.41(% G+C)-(600/N) (1)

where N is the length of the probe. This equation works well for probes 14 to 70 nucleotides in length that are identical to the target sequence. The equation below for T_m of DNA-DNA hybrids is useful for probes in the range of 50 to greater than 500 nucleotides, and for conditions that include an organic solvent (formamide).

T_m=81.5+16.6 log {[Na.sup.+]/(1+0.7[Na.sup.+])}±0.41(% G+C)-500/L 0.63(% formamide) (2)

where L is the length of the probe in the hybrid. (P. Tijessen, "Hybridization with Nucleic Acid Probes" in Laboratory Techniques in Biochemistry and Molecular Biology, P. C. vand der Vliet, ed., c. 1993 by Elsevier, Amsterdam.) The T_m of equation (2) is affected by the nature of the hybrid; for DNA-RNA hybrids T_m is 10-15° C. higher than calculated, for RNA-RNA hybrids T_m is 20-25° C. higher. Because the T_m decreases about 1° C. for each 1% decrease in homology when a long probe is used (Bonner et al., J. Mol. Biol. 81:123 (1973)), stringency conditions can be adjusted to favor detection of identical genes or related family members.

[0034]Equation (2) is derived assuming equilibrium and therefore, hybridizations according to the present invention are most preferably performed under conditions of probe excess and for sufficient time to achieve equilibrium. The time required to reach equilibrium can be shortened by inclusion of a hybridization accelerator such as dextran sulfate or another high volume polymer in the hybridization buffer.

[0035]Stringency can be controlled during the hybridization reaction or after hybridization has occurred by altering the salt and temperature conditions of the wash solutions used. The formulas shown above are equally valid when used to compute the stringency of a wash solution. Preferred wash solution stringencies lie within the ranges stated above; high stringency is 5-8° C. below T_m, medium or moderate stringency is 26-29° C. below T_m and low stringency is 45-48° C. below T_m. [0036]Superpool: As used in the context of the current invention, a "superpool" refers to a mixture of seed from 100 different "master pools". Thus, the superpool contains an equal amount of seed from 500 different events, but only represents 100 transgenic plants with a distinct exogenous nucleotide sequence transformed into them, because the master pools are of 5 different events with the same exogenous nucleotide sequence transformed into them. [0037]T₀: As used in the current application, the term "T₀" refers to the whole plant, explant, or callous tissue inoculated with the transformation medium. [0038]T₁: As used in the current application, the term T₁ refers to the either the progeny of the T₀ plant, in the case of whole-plant transformation, or the regenerated seedling in the case of explant or callous tissue transformation. [0039]T₂: As used in the current application, the term T₂ refers to the progeny of the T₁ plant. T₂ progeny are the result of self-fertilization or cross pollination of a T₁ plant. [0040]T₃: As used in the current application, the term T₃ refers to second generation progeny of the plant that is the direct result of a transformation experiment. T₃ progeny are the result of self-fertilization or cross pollination of a T₂ plant. [0041]Zero Nitrogen: Nitrogen is not present in any amount. [0042]Zero Phosphorus: Phosphorus is not present in any amount.

2. Important Characteristics of the Polynucleotides and Polypeptides of the Invention

[0043]The polynucleotides and polypeptides of the present invention are of interest because when they are misexpressed (i.e. when expressed at a non-natural location or in an increased or decreased amount) they produce plants with modified pH tolerance or phosphate use efficiency. "Phosphate use efficiency" is a term that includes various responses to environmental conditions that affect the amount of phosphate available to the plant. For example, under both low and high pH conditions phosphate is bound within the soil, resulting in a decrease of available phosphate for maintaining or initiating physiological processes. As used herein, modulating phosphate use efficiency is intended to encompass all of these situations as well as other environmental situations that affect the plant's ability to use and/or maintain phosphate effectively (e.g. osmotic stress, etc.).

[0044]The polynucleotides and polypeptides of the invention, as discussed below and as evidenced by the results of various experiments, are useful for modulating pH tolerance or phosphate use efficiency. These traits can be used to exploit or maximize plant products for agricultural, ornamental or forestry purposes in different environment conditions of water supply. Modulating the expression of the nucleotides and polypeptides of the present invention leads to transgenic plants that will be less sensitive to variations in pH and that require less phosphate, resulting in better yields under these types of adverse conditions. Both categories of transgenic plants lead to reduced costs for the farmer and better yield in their respective environmental conditions.

3. The Polynucleotides and Polypeptides of the Invention

[0045]The polynucleotides of the invention, and the proteins expressed thereby, are set forth in the sequences present in the Sequence Listing. Some of these sequences are functionally comparable proteins.

[0046]Functionally comparable proteins are those proteins that have at least one characteristic in common. Such characteristics can include sequence similarity, biochemical activity and phenotypic activity. Typically, the functionally comparable proteins share some sequence similarity and generally share at least one biochemical and/or phenotypic activity. For example, biochemical functionally comparable proteins are proteins that act on the same reactant to give the same product.

[0047]Another class of functionally comparable proteins is phenotypic functionally comparable proteins. The members of this class regulate the same physical characteristic, such as increased drought tolerance. Proteins can be considered phenotypic functionally comparable proteins even if the proteins give rise to the same physical characteristic, but to a different degree.

[0048]The polypeptides of the invention also include those comprising the consensus sequences described in Tables 1-5, 2-6 and 3-5. A consensus sequence defines the important conserved amino acids and/or domains within a polypeptide. Thus, all those sequences that conform to the consensus sequence are suitable for the same purpose. Polypeptides comprised of a sequence within and defined by one of the consensus sequences can be utilized for the purposes of the invention namely to make transgenic plants with improved tolerance to heat or high or low water conditions.

4. Use of the Polynucleotides and Polypeptides to Make Transgenic Plants

[0049]To use the sequences of the present invention or a combination of them or parts and/or mutants and/or fusions and/or variants of them, recombinant DNA constructs are prepared which comprise the polynucleotide sequences of the invention inserted into a vector, and which are suitable for transformation of plant cells. The construct can be made using standard recombinant DNA techniques (Sambrook et al. 1989) and can be introduced to the species of interest by Agrobacterium-mediated transformation or by other means of transformation as referenced below.

[0050]The vector backbone can be any of those typical in the art such as plasmids, viruses, artificial chromosomes, BACs, YACs and PACs and vectors of the sort described by [0051](a) BAC: Shizuya et al., Proc. Natl. Acad. Sci. USA 89: 8794-8797 (1992); Hamilton et al., Proc. Natl. Acad. Sci. USA 93: 9975-9979 (1996); [0052](b) YAC: Burke et al., Science 236:806-812 (1987); [0053](c) PAC: Sternberg N. et al., Proc Natl Acad Sci USA. January; 87(1):103-7 (1990); [0054](d) Bacteria-Yeast Shuttle Vectors: Bradshaw et al., Nucl Acids Res 23: 4850-4856 (1995); [0055](e) Lambda Phage Vectors: Replacement Vector, e.g., Frischauf et al., J. Mol Biol 170: 827-842 (1983); or Insertion vector, e.g., Huynh et al., In: Glover N M (ed) DNA Cloning: A practical Approach, Vol. 1 Oxford: IRL Press (1985); T-DNA gene fusion vectors :Walden et al., Mol Cell Biol 1: 175-194 (1990); and [0056](g) Plasmid vectors: Sambrook et al., infra.

[0057]Typically, the construct comprises a vector containing a sequence of the present invention with any desired transcriptional and/or translational regulatory sequences, such as promoters, UTRs, and 3' end termination sequences. Vectors can also include origins of replication, scaffold attachment regions (SARs), markers, homologous sequences, introns, etc. The vector may also comprise a marker gene that confers a selectable phenotype on plant cells. The marker typically encodes biocide resistance, particularly antibiotic resistance, such as resistance to kanamycin, bleomycin, hygromycin, or herbicide resistance, such as resistance to glyphosate, chlorosulfuron or phosphinotricin.

[0058]A plant promoter is used that directs transcription of the gene in all tissues of a regenerated plant and may be a constitutive promoter, such as p326 or CaMV35S. Alternatively, the plant promoter directs transcription of a sequence of the invention in a specific tissue manner (tissue-specific promoter) or is otherwise under more precise environmental control (inducible promoter). Various plant promoters, including constitutive, tissue-specific and inducible, are known to those skilled in the art and can be utilized in the present invention. Typically, preferred promoters to use in the present invention are those that are induced by heat or low water conditions Such as the RD29a promoter (Kasuga et al., Plant Cell Physiol. 45:346 (2004) and Yamaguchi-Shinozaki and Shinozaki, Mol Gen Genet. 236: 331 (1993)) or other DRE-containing (dehydration-responsive elements) promoters (Liu et al, Cell 10: 1391 (1998)). Another preferred embodiment of the present invention is the use of root specific promoters such as those present in the AtXTH17, AtXTH18, AtXTH19 and AtXTH20 genes of Arabidopsis (Vissenberg et al. (2005) Plant Cell Physiol 46:192) or guard cell specific promoters such as TGG1 or KST1 (Husebye et al. (2002) Plant Physiol 128:1180; Plesch et al. (2001) Plant J 28:455).

[0059]Alternatively, misexpression can be accomplished using a two component system, whereby the first component comprises a transgenic plant comprising a transcriptional activator operatively linked to a promoter and the second component comprises a transgenic plant comprising a sequence of the invention operatively linked to the target binding sequence/region of the transcriptional activator. The two transgenic plants are crossed and the sequence of the invention is expressed in their progeny. In another alternative, the misexpression can be accomplished by transforming the sequences of the two component system into one transgenic plant line.

[0060]Any promoter that functions in plants can be used in the first component, such as those discussed above. Suitable transcriptional activator polypeptides include, but are not limited to, those encoding HAP1 and GAL4. The binding sequence recognized and targeted by the selected transcriptional activator protein (e.g. a UAS element) is used in the second component.

Transformation

[0061]Nucleotide sequences of the invention are introduced into the genome or the cell of the appropriate host plant by a variety of techniques. These techniques for transforming a wide variety of higher plant species are well known and described in the technical and scientific literature. See, e.g. Weising et al., Ann. Rev. Genet. 22:421 (1988); and Christou, Euphytica, v. 85, n.1-3:13-27, (1995).

[0062]Processes for the transformation and regeneration of monocotyledonous and dicotyledonous plants are known to the person skilled in the art. For the introduction of DNA into a plant host cell a variety of techniques is available. These techniques include transformation of plant cells by injection (e.g. Newell, 2000), microinjection (e.g. Griesbach (1987) Plant Sci. 50 69-77), electroporation of DNA (e.g. Fromm et al. (1985) Proc. Natl Acad. Sci. USA 82:5824 and Wan and Lemaux, Plant Physiol. 104 (1994), 37-48), PEG (e.g. Paszkowski et al. (1984) EMBO J. 3:2717), use of biolistics (e.g. Klein et al. (1987) Nature 327:773), fusion of cells or protoplasts (Willmitzer, L., 1993 Transgenic plants. In: Biotechnology, A Multi-Volume Comprehensive Treatise (H. J. Rehm, G. Reed, A. Puhler, P. Stadler, eds., Vol. 2, 627-659, VCH Weinheim-New York-Basel-Cambridge), via T-DNA using Agrobacterium tumefaciens (e.g. Fraley et al. (Crit. Rev. Plant. Sci. 4, 1-46 and Fromm et al., Biotechnology 8 (1990), 833-844) or Agrobacterium rhizogenes (e.g. Cho et al. (2000) Planta 210:195-204) or other bacterial hosts (e.g. Brootghaerts et al. (2005) Nature 433:629-633), as well as further possibilities.

[0063]In addition, a number of non-stable transformation methods well known to those skilled in the art may be desirable for the present invention. Such methods include, but are not limited to, transient expression (e.g. Lincoln et al. (1998) Plant Mol. Biol. Rep. 16:1-4) and viral transfection (e.g. Lacomme et al. (2001) In "Genetically Engineered Viruses" (C. J. A. Ring and E. D. Blair, Eds). Pp. 59-99, BIOS Scientific Publishers, Ltd. Oxford, UK).

[0064]Seeds are obtained from the transformed plants and used for testing stability and inheritance. Generally, two or more generations are cultivated to ensure that the phenotypic feature is stably maintained and transmitted.

[0065]One of skill will recognize that after the expression cassette is stably incorporated in transgenic plants and confirmed to be operable, it can be introduced into other plants by sexual crossing. Any of a number of standard breeding techniques can be used, depending upon the species to be crossed.

[0066]The nucleic acids of the invention can be used to confer the trait of increased tolerance to heat and/or low water conditions, without reduction in fertility, on essentially any plant.

[0067]The nucleotide sequences according to the invention encode appropriate proteins from any organism, in particular from plants, fungi, bacteria or animals.

[0068]The process according to the invention can be applied to any plant, preferably higher plants, pertaining to the classes of Angiospermae and Gymnospermae. Plants of the subclasses of the Dicotylodenae and the Monocotyledonae are particularly suitable. Dicotyledonous plants belong to the orders of the Magniolales, Illiciales, Laurales, Piperales Aristochiales, Nymphaeales, Ranunculales, Papeverales, Sarraceniaceae, Trochodendrales, Hamamelidales, Eucomiales, Leitneriales, Myricales, Fagales, Casuarinales, Caryophyllales, Batales, Polygonales, Plumbaginales, Dilleniales, Theales, Malvales, Urticales, Lecythidales, Violales, Salicales, Capparales, Ericales, Diapensales, Ebenales, Primulales, Rosales, Fabales, Podostemales, Haloragales, Myrtales, Comales, Proteales, Santales, Rafflesiales, Celastrales, Euphorbiales, Rhanmales, Sapindales, Juglandales, Geraniales, Polygalales, Umbellales, Gentianales, Polemoniales, Lamiales, Plantaginales, Scrophulariales, Campanulales, Rubiales, Dipsacales, and Asterales. Monocotyledonous plants belong to the orders of the Alismatales, Hydrocharitales, Najadales, Triuridales, Commelinales, Eriocaulales, Restionales, Poales, Juncales, Cyperales, Typhales, Bromeliales, Zingiberales, Arecales, Cyclanthales, Pandanales, Arales, Lilliales, and Orchidales. Plants belonging to the class of the Gymnospermae are Pinales, Ginkgoales, Cycadales and Gnetales.

[0069]The method of the invention is preferably used with plants that are interesting for agriculture, horticulture, biomass for bioconversion and/or forestry. Examples are tobacco, oilseed rape, sugar beet, potato, tomato, cucumber, pepper, bean, pea, citrus fruit, apple, pear, berries, plum, melon, eggplant, cotton, soybean, sunflower, rose, poinsettia, petunia, guayule, cabbage, spinach, alfalfa, artichoke, corn, wheat, rye, barley, grasses such as switch grass or turf grass, millet, hemp, banana, poplar, eucalyptus trees, conifers.

Homologs Encompassed by the Invention

[0070]Agents of the invention include proteins comprising at least about a contiguous 10 amino acid region preferably comprising at least about a contiguous 20 amino acid region, even more preferably comprising at least about a contiguous 25, 35, 50, 75 or 100 amino acid region of a protein of the present invention. In another preferred embodiment, the proteins of the present invention include between about 10 and about 25 contiguous amino acid region, more preferably between about 20 and about 50 contiguous amino acid region, and even more preferably between about 40 and about 80 contiguous amino acid region.

[0071]Due to the degeneracy of the genetic code, different nucleotide codons may be used to code for a particular amino acid. A host cell often displays a preferred pattern of codon usage. Nucleic acid sequences are preferably constructed to utilize the codon usage pattern of the particular host cell. This generally enhances the expression of the nucleic acid sequence in a transformed host cell. Any of the above described nucleic acid and amino acid sequences may be modified to reflect the preferred codon usage of a host cell or organism in which they are contained. Modification of a nucleic acid sequence for optimal codon usage in plants is described in U.S. Pat. No. 5,689,052. Additional variations in the nucleic acid sequences may encode proteins having equivalent or superior characteristics when compared to the proteins from which they are engineered.

[0072]It is understood that certain amino acids may be substituted for other amino acids in a protein or peptide structure (and the nucleic acid sequence that codes for it) without appreciable change or loss of its biological utility or activity. The amino acid changes may be achieved by changing the codons of the nucleic acid sequence.

[0073]It is well known in the art that one or more amino acids in a native sequence can be substituted with other amino acid(s), the charge and polarity of which are similar to that of the native amino acid, i.e., a conservative amino acid substitution, resulting in a silent change. Conservative substitutes for an amino acid within the native polypeptide sequence can be selected from other members of the class to which the amino acid belongs (see below). Amino acids can be divided into the following four groups: (1) acidic (negatively charged) amino acids, such as aspartic acid and glutamic acid; (2) basic (positively charged) amino acids, such as arginine, histidine, and lysine; (3) neutral polar amino acids, such as glycine, serine, threonine, cysteine, cystine, tyrosine, asparagine, and glutamine; and (4) neutral nonpolar (hydrophobic) amino acids such as alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine.

[0074]In a further aspect of the present invention, nucleic acid molecules of the present invention can comprise sequences that differ from those encoding a protein or fragment thereof selected from the group consisting of those sequences present in the Sequence Listing due to the fact that the different nucleic acid sequence encodes a protein having one or more conservative amino acid changes.

[0075]In another aspect, biologically functional equivalents of the proteins or fragments thereof of the present invention can have about 10 or fewer conservative amino acid changes, more preferably about 7 or fewer conservative amino acid changes, and most preferably about 5 or fewer conservative amino acid changes. In a preferred embodiment, the protein has between about 5 and about 500 conservative changes, more preferably between about 10 and about 300 conservative changes, even more preferably between about 25 and about 150 conservative changes, and most preferably between about 5 and about 25 conservative changes or between 1 and about 5 conservative changes.

5. Experiments Confirming the Usefulness of the Polynucleotides and Polypeptides of the Invention

[0076]5.1 Procedures

[0077]The nucleotide sequences of the invention were identified by use of a variety of screens for pH and/or low phosphate and/or low nitrogen conditions. These screens are recognized by those skilled in the art to be predictive of nucleotide sequences that provide plants with improved tolerance to pH and/or low phosphate and/or low nitrogen conditions because they emulate the different environmental conditions that can result from increased pH and/or low phosphate and/or low nitrogen conditions. These screens generally fall into two categories (1) soil screens and (2) in vitro screens.

[0078]Soil screens have the advantage of assaying the response of the entire plant to particular conditions, such as high pH or low phosphorus. On the other hand, in vitro screens have the advantage of relying on defined media and so allow more defined manipulation of growth conditions. Each of the screens used is described in more detail below.

[0079]In general, the screens used to identify the polynucleotides and polypeptides of the invention were conducted using superpools of Arabidopsis T₂ transformed plants. The T₁ plants were transformed with a Ti plasmid containing a particular SEQ ID NO in the sense orientation relative to a constitutive promoter and harboring the plant-selectable marker gene phosphinothricin acetyltansferase (PAT), which confers herbicide resistance to transformed plants. For in vitro screens, seed from multiple superpools (1,200 T₂ seeds from each superpool) were usually tested. T₃ seed were collected from the resistant plants and retested on one or more in vitro screens. The results of the screens conducted for each SEQ ID NO can be found in the Examples below.

[0080]1. High pH

[0081]Screens for high pH resistance identify seedlings better able to thrive under nutritional deficiencies (e.g. Phosphate, Manganese, Iron, Boron) imposed by alkaline conditions.

[0082]Seeds are sterilized in 50% household bleach for 5 minutes and then washed with double distilled deionized water three times. Sterilized seed is stored in the dark at 4° C. for a minimum of 3 days before use.

[0083]High pH media is prepared by mixing 0.5 g/l MES hydrate with 1X MS+0.5% Sucrose. Prior to autoclaving pH is adjusted with 10 N KNH to the following values: pH 5.7 (control), pH 7.03, pH 8.02, pH 9.01 and pH 10.18. The media pH is retested since pH values drop after autoclaving as follows: pH 5.7→pH 5.66; pH 7.03→pH6.50; pH 8.02→pH 7.50; pH 9.01→pH 8.91; pH10.18→pH 9.91. Generally speaking, pH 9.01(pH 8.91) allows germination but no growth beyond 2 to 5 mm and no root growth. Germination does not occur at higher pH (e.g. pH 10.81).

[0084]Approximately 1200 seeds are evenly spaced per MS-sucrose plate before incubating in the vertical position at 22° C. for 14 days. Under these conditions, the plates are exposed to 12,030 LUX from above and 3,190 LUX from the bottom.

[0085]Seedlings are scored for root and shoot growth after 7 and 14 days. Putative tolerant seedlings are transferred to MS pH 5.7 for recovery for 14 days prior to transplanting in soil. Finale® spraying is done after plants are moved to soil to remove non-transgenics from the population.

[0086]DNA is isolated from each T₂ plant and used in PCR reactions using the following cycling conditions: 95° C. for 5 min, 35 cycles of (94° C. for 30 sec, then 59° C. for 30 sec, then 72° C. for 1 min), 72° C. for 8 min and 4° C. hold. Aliquots of the reaction product are analyzed on a 1.0% agarose gel stained with ethidium bromide. The DNA products are sequenced to determine which insert sequences were in each superpool candidate chosen in the screen.

[0087]T₃ Seed from those plants containing sequenced PCR products are collected and retested on high pH media. In addition, plants are tested on MS media lacking Phosphate and having a pH of 5.7.

[0088]2. Zero Phosphate

[0089]Screens for zero phosphate tolerance identify seedlings better able to thrive under a phosphate nutritional deficiency.

[0090]Seeds are sterilized in 50% household bleach for 5 minutes and then washed with double distilled deionized water three times. Sterilized seed is stored in the dark at 4° C. for a miniumum of 3 days before use.

[0091]Zero phosphate media is prepared using commercially available MS media lacking phosphate, pH 5.7.

[0092]Approximately 1200 seeds are evenly spaced per MS-P plate before incubating in the vertical position at 22° C. for 14 days. Under these conditions, the plates are exposed to 12,030 LUX from above and 3,190 LUX from the bottom.

[0093]Seedlings are scored for root and shoot growth after 7 and 14 days. Putative tolerant seedlings are transferred to MS pH 5.7 for recovery for 14 days prior to transplanting in soil. Finale® spraying is done after the plants are moved to soil to remove non-transgenics from the population.

[0094]DNA is isolated from each T₂ plant and used in PCR reactions using the following cycling conditions: 95° C. for 5 min, 35 cycles of (94° C. for 30 sec, then 59° C. for 30 sec, then 72° C. for 1 min), 72° C. for 8 min and 4° C. hold. Aliquots of the reaction product are analyzed on a 1.0% agarose gel stained with ethidium bromide. The DNA products are sequenced to determined which insert sequences were in each superpool candidate chosen in the screen.

[0095]T₃ Seed from those plants containing the sequenced PCR products are collected and retested.

[0096]3. Zero Phosphate, Zero Nitrogen

[0097]Screens for zero phosphate, zero nitrogen tolerance identify seedlings better able to thrive under a phosphate nutritional deficiency.

[0098]Seeds are sterilized in 50% household bleach for 5 minutes and then washed with double distilled deionized water three times. Sterilized seed is stored in the dark at 4° C. for a miniumum of 3 days before use.

[0099]Zero phosphate, zero nitrogen media is prepared using commercially available MS media lacking phosphate, pH 5.7.

[0100]Approximately 1200 seeds are evenly spaced per MS-P-N plate before incubating in the vertical position at 22° C. for 14 days. Under these conditions, the plates are exposed to 12,030 LUX from above and 3,190 LUX from the bottom.

[0101]Growth and overall greenness are assayed 10 days post-treatment. Seedling recovery is assessed by adding a thin layer (8.3 ml) of complete MS+P+N media, pH 5.7, softened by the addition of 0.02% agar. Media is added to the edge of the plate and slowly rotated until a thin film of +PN media is present on top of the solidified -PN media. Putative tolerant seedlings are greener and have increased growth compared to controls. Finale® spraying is done after the plants are moved to soil to remove non-transgenics from the population.

[0102]DNA is isolated from each T₂ plant and used in PCR reactions using the following cycling conditions: 95° C. for 5 min, 35 cycles of (94° C. for 30 sec, then 59° C. for 30 sec, then 72° C. for 1 min), 72° C. for 8 min and 4° C. hold. Aliquots of the reaction product are analyzed on a 1.0% agarose gel stained with ethidium bromide. The DNA products are sequenced to determined which insert sequences were in each superpool candidate chosen in the screen.

[0103]T₃ Seed from those plants containing the sequenced PCR products are collected and retested.

[0104]5.2 Results

[0105]The results of the above experiments are set forth below wherein each individual example relates to all of the experimental results for a particular polynucleotide/polypeptide if the invention.

Example 1

Ceres cDNA 12335629

[0106]Clone 40781, Ceres cDNA 12335629, encodes a full-length protein with homology to a ferredoxin thioredoxin reductase from Arabidopsis thaliana.

[0107]Ectopic expression of Ceres cDNA 12335629 under the control of the CaMV35S promoter induces the following phenotypes: [0108]Better growth and recovery after exposure to high pH conditions and [0109]Continued growth under high pH induced phosphate and iron deficiencies.Generation and Phenotypic Evaluation of T₁ Lines Containing 35S::cDNA 12335629.

[0110]Wild-type Arabidopsis Wassilewskija (WS) plants were transformed with a Ti plasmid containing cDNA 12335629 in the sense orientation relative to the 35S constitutive promoter. The T_i plasmid vector used for this construct, CRS338, contains PAT and confers herbicide resistance to transformed plants. Ten independently transformed events were selected and evaluated for their qualitative phenotype in the T_i generation. No positive or negative phenotypes were observed in the T₁ plants.

Screens of Superpools on High pH Media for pH Tolerance.

[0111]Seed from superpools of the 35S over-expression lines were evaluated for greenness and size on high pH media as described above. Once cDNA 12335629 was identified in tolerant plants, the five individual T₂ events containing this cDNA (ME03527) were screened on high pH media essentially as described above, but where the media pH is 8.5, to identify events with the tolerant phenotype.

Results:

[0112]Qualitative Analysis of the Superpool Containing 35S::clone 40781 Plants on high pH

[0113]The screen resulted in a decrease in germination and/or growth for both wildtype and superpools as compared to seeds on control media. Only one line survived transplantation to soil. The candidate was greener than controls but overall size was comparable to those of wild-type. There was no delay in flowering time or decrease in seed set in comparison to un-treated wild-type but a faster flowering time and greater seed set was apparent when compared to a recovered pH treated wild-type plant (data not shown). These results are consistent with those of the T₁ generation which displayed normal flowering time and fertility.

Qualitative and Quantitative Analysis of T₃-cDNA 12335629 on High pH.

[0114]The plants were treated with Finale® to eliminate any false-positives or any lines where the Finale® marker was suppressed. All of the Finale®-resistant candidates flowered and set seed. Finale® segregation was assessed to identify events containing a single insert segregation in a 3:1 (R:S) ratio as calculated by chi-square test. All of the events segregated for a single functional insert (Table 1-1). The transgenic plants were greener and slightly larger than the control under high pH stress.

TABLE-US-00001 TABLE 1-1 Observed and expected frequencies assuming a 3:1 ratio for high pH tolerance of cDNA 12335629 progeny under high pH (pH 8.5). α of 0.05 Probability Event Generation Observed Expected χ² of Chi-Test pH Resistant T₃ 22 29 0.926 pH Sensitive T₃ 14 7 2.778 0.054 N = 36 36 36 3.704

Qualitative and Quantitative Analysis of cDNA 12335629 Progeny on Media Lacking Phosphate

[0115]Before testing independent T₂ events, plants containing cDNA 12335629 were re-assayed for phosphate starvation tolerance by growth on media containing no phosphate as described above. After seven days only slightly more tolerance compared to controls is observed, but cDNA 12335629 seedlings are a bit larger and slightly greener than those of the control. Because the slight increase in size was particularly difficult to assess, anything lower or equal to the wild-type average of 0.42 cm was assessed to be sensitive and anything higher was assessed as tolerant. Twenty-four resistant and twelve phosphate starved sensitive seedlings were compared to Finale®]frequencies and found to have a Chi-test probability of 0.49, suggesting a positive fit (Table 1-2).

TABLE-US-00002 TABLE 1-2 Observed and expected frequencies assuming a 3:1 ratio for phosphate starvation tolerance among progeny of cDNA 12335629 media lacking phosphate (-P). α of 0.05 Probability Event Generation Observed Expected χ² of Chi-Test -P Resistant T₃ 24 27 0.333 -P Sensitive T₃ 12 9 1.333 0.25 N = 36 36 36 1.666

Qualitative and Quantitative Analysis of Individual T₂ Events of cDNA 12335629 on High pH Plate Assay.

[0116]Five individual events of cDNA 12335629 (ME03527) were analyzed for a positive phenotype under high pH conditions. All five T₂ events had wild-type germination frequencies on MS pH 5.7 plates (data not shown). All T₂ lines and recovered T₃ lines showed evidence of a single insert as determined by Chi-square analysis (Table 1-3). Seeds from each of the five independent T₂ events, were plated on pH 8.5 plates and allowed to germinate and grow for 14 days.

[0117]Four of five T₂ events of ME03527 (-02,-03,-04, and -05) had a positive high pH tolerance phenotype as defined by growth and greenness. The phenotype of1V1E03527-01 was too weak to assess as positive compared to the controls (Table 1-4). Phenotype strength varied among the four positive independent events, but all showed better growth than controls. The segregation ratios, determined by a Chi-square test, show that the segregation of the transgene is the same as observed for Finale® (Table 1-4). ME03527-02,-03,-04, and -05 had the strongest and most consistent pH tolerance phenotypes.

TABLE-US-00003 TABLE 1-3 Observed and expected frequencies assuming a 3:1 (R:S) ratio for Finale ® resistance among 35S::clone 40781 T₂ and T₃ events tested for growth under high pH conditions. α of 0.05 Event Generation Observed Expected χ² Probability of Chi-Test ME03527-01 Finale ® T₂ 16 18 0.222 Resistant ME03527-01 Finale ® T₂ 8 6 0.667 0.35 Sensitive N = 24 24 24 0.889 ME03527-02 Finale ® T₂ 28 27 0.037 Resistant ME03527-02 Finale ® T₂ 8 9 0.111 0.70 Sensitive N = 36 36 36 0.148 ME03527-03 Finale ® T₂ 17 18 0.056 Resistant ME03527-03 Finale ® T₂ 7 6 0.167 0.64 Sensitive N = 24 24 24 0.223 ME03527-04 Finale ® T₂ 27 27 0 Resistant ME03527-04 Finale ® T₂ 9 9 0 1.0 Sensitive N = 36 36 36 0 ME03527-05 Finale ® T₂ 23 27 0.593 Resistant ME03527-05 Finale ® T₂ 13 9 1.778 0.12 Sensitive N = 36 36 36 2.371 cDNA 12335629 Finale ® T₃ 22 27 0.926 Resistant cDNA 12335629 Finale ® T₃ 14 9 2.778 0.054 Sensitive N = 36 36 36 3.704

TABLE-US-00004 TABLE 1-4 Observed and expected frequencies of high pH tolerance assuming segregation of transgene is the same as observed in Finale ® resistance among 35S::clone 40781 T₂ and T₃ events that showed increased growth under high pH conditions. α of 0.05 Event Generation Observed Expected χ² Probability of Chi-Test ME03527-01 pH Resistant T₂ 15 25.5 4.324 ME03527-01 pH Sensitive T₂ 19 85.5 2.970 32E-05 N = 36 34 34 7.294 ME03527-02 pH Resistant T₂ 23 24.75 0.124 ME03527-02 pH Sensitive T₂ 10 8.25 0.371 0.48 N = 36 33 33 0.495 ME03527-03 pH Resistant T₂ 23 23.25 0.003 0.92 ME03527-03 pH Sensitive T₂ 8 7.75 0.008 N = 36 31 31 0.011 ME03527-04 pH Resistant T₂ 24 27 0.333 0.25 ME03527-04 pH Sensitive T₂ 12 9 1.000 N = 36 36 36 1.333 ME03527-05 pH Resistant T₂ 19 27 2.370 0.002 ME03527-05 pH Sensitive T₂ 17 9 7.111 N = 36 36 3 9.481 cDNA 12335629 pH T₃ 19 27 2.370 0.002 Resistant cDNA 12335629 pH T₃ 17 9 7.111 Sensitive N = 36 36 36 9.481

Table 1-5 provides the results of the consensus sequence analysis based on Ceres cDNA 13487605.

TABLE-US-00005 TABLE 1-5 ##STR00001##

Example 2

Ceres cDNA 12330185

[0118]Clone 34035, Ceres cDNA 12330185, encodes a 128 amino acid protein of unknown function (DUF423) from Arabidopsis thaliana.

[0119]Ectopic expression of Ceres cDNA 12330185 under the control of the 32449 promoter induces the following phenotypes: [0120]Increased size and greenness on nutrient deficiencies incurred by high pH conditions, [0121]Better soil recovery after exposure to high pH stress, and [0122]Better recovery after exposure to conditions lacking both phosphate and nitrogen.Generation and Phenotypic Evaluation of T₁ Lines Containing p32449::cDNA 12330185.

[0123]Wild-type Arabidopsis Wassilewskija (WS) plants were transformed with a Ti plasmid containing cDNA 12330185 in the sense orientation relative to the 32449 constitutive promoter. Promoter 32449 has broad expression throughout Arabidopsis, although at much lower expression level than CaMV35S. The T_i plasmid vector used for this construct, CRS311, contains PAT and confers herbicide resistance to transformed plants. Nine independently transformed events were selected and evaluated for their qualitative phenotype in the T₁ generation. No positive or negative phenotypes were observed in the T₁ plants.

Screens of Superpools on High pH Media for pH Tolerance.

[0124]Seed from superpools of the 32449 over-expression lines were evaluated for greenness and size on high pH media as described above. Once cDNA 12330185 was identified in tolerant plants, nine individual T₂ events containing this cDNA (ME00077) were screened on high pH media essentially as described above, but where the media pH is 8.5, to identify events with the tolerant phenotype.

Results:

[0125]Qualitative Analysis of the Superpool Containing 34449::cDNA 12330185 on High pH

[0126]The cDNA 12330185 line displayed a delayed flowering time of ˜8 days and decreased seed set in comparison to the un-treated wild-type. However cDNA 12330185 displayed a faster flowering time (˜15 days) and greater seed set when compared to the high pH grown wild-type plant.

Qualitative and Quantitative Analysis of the T₃ 32449:: cDNA 12330185 on High pH.

[0127]The cDNA 12330185 line was tested for Finale® resistance and re-assayed for continued pH tolerance. The segregation ratio of T₃ seeds from cDNA 12330185 is suggestive of a single insert, as calculated by a Chi-square test (Table 2-1). The cDNA 12330185 line was re-tested on pH 9.0 media as described and found to be tolerant to high pH when compared to controls.

TABLE-US-00006 TABLE 2-1 Chi-square analysis of progeny of cDNA 12330185 on Finale ® assuming a 3:1 ratio. Event Observed Expected χ² Probability of Chi-Test Finale ® Resistant 27 27 0 Finale ® Sensitive 9 9 0 1 N = 36 36 36 0

Qualitative and Quantitative Analysis of Phosphate and Nitrate Starvation of T₃ (cDNA 12330185) Plants.

[0128]To ascertain whether the pH tolerant phenotype is related to better survival under nutrient starvation, T₃ seeds were assayed on MS media lacking both phosphate (--P) and nitrate (--N) (pH 5.7) as described above. The cDNA 12330185 line was greener and of equal size compared to wild-type controls. Ten days after the addition of +NP media film, cDNA 12330185 seedlings recovered more quickly than wild type. Twenty-five of 36 seedlings of SP9pH1 had greater growth when compared to wild type. This increased growth frequency is suggestive of a single insert as determined by Chi-square analysis (Table 2-2).

TABLE-US-00007 TABLE 2-2 Observed and expected frequencies of no phosphate/nitrate growth assuming segregation of transgene is 3:1 (R:S) of T₃ plants of cDNA 12330185 that showed increased growth under high pH conditions. α of 0.05 Event Observed Expected χ² Probability of Chi-Test NP Resistant 25 27 0.148 0.441 NP Sensitive 11 9 0.444 N = 36 36 36 0.592

Qualitative and Quantitative Analysis of Individual T₂ Events of cDNA 12330185 on High pH.

[0129]Seeds from T₂ lines representing nine individual events and containing cDNA 12330185 (ME0077-01, 02, 03, 04, 05, 06, 07, 08, 09) were plated on pH media, pH 8.5 as described above. Plates were evaluated at 7 and 12 days post-plating (Table 2-3). All nine T₂ events had wild-type germination frequencies except for ME00077-04 (Table 2-4). This germination problem however was not observed when seedlings were plated onto high pH plates.

[0130]Six of the nine events showed tolerance to high pH as defined by growth and greenness. The strongest tolerance phenotypes were in ME00077-03 and ME00077-05. ME00077-03 and ME00077-05 both had single inserts as determined by Chi-square analysis (Table 2-3).

[0131]The pH tolerant phenotype was strongest in the cDNA 12330185 T₃ line recovered from the superpool screen. We did not do a genetic mapping of this line's insert to determine which event it represented. This line's phenotype was so strong that it allowed adjacent wild-type quadrants within same plate to grow normally after 14-days. This is most likely due to acidification of surrounding media by the pH tolerant line. ME00077-03,-05 T₂ plants also showed increased recovery during phosphate and nitrogen starvation assays (data not shown). However, the cDNA 12330185 T₃ line recovered from the superpool phenotype was stronger than that observed for lines ME00077-03 and -05 under -NP starvation recovery (as noted above).

TABLE-US-00008 TABLE 2-3 ##STR00002## ##STR00003##

TABLE-US-00009 TABLE 2-4 ##STR00004## **Germination reduction in comparison to wild-type control and other ME00077 lines

TABLE-US-00010 TABLE 2-5 Observed and expected frequencies of high pH tolerance assuming segregation of transgene is the same as observed in Finale ® segregation among progeny of 32449:: cDNA 12330185 T₂ events that showed increased growth under high pH conditions. α of 0.05 Probability Event Observed Expected χ² of Chi-Test ME00077-03 pH Resistant 26 25.5 0.009 0.84 ME00077-03 pH Sensitive 8 8.5 0.029 N = 36 34 34 0.038 ME00077-05 pH Resistant 29 26.25 0.288 0.28 ME00077-05 pH Sensitive 6 8.75 0.864 N = 36 35 35 1.152 cDNA 12330185 pH 31 27 0.592 0.124 Resistant cDNA 12330185 pH 5 9 1.778 Sensitive N = 36 36 36 2.370

Table 2-6 provides the results of the consensus sequence analysis based on Ceres cDNA 12330185.

TABLE-US-00011 TABLE 2-6 ##STR00005##

Example 3

Ceres cDNA 12482777

[0132]Clone 126592, Ceres cDNA 12482777, encodes a full-length protein that has homology to an iron/manganese superoxide dismutase from Arabidopsis thaliana.

[0133]Ectopic expression of Ceres cDNA 12482777 under the control of the CaMV35S promoter induces the following phenotypes: [0134]Increased growth under high pH induced stress [0135]Better recovery after exposure to pH stress [0136]Reduced height without a reduction in harvest index.Generation and Phenotypic Evaluation of T₁ Lines Containing 35S::cDNA 12482777.

[0137]Wild-type Arabidopsis Wassilewskija (WS) plants were transformed with a Ti plasmid containing cDNA 12482777 in the sense orientation relative to the 35S constitutive promoter. The T_i plasmid vector used for this construct, CRS338, contains PAT and confers herbicide resistance to transformed plants. Seven independently transformed events were selected and evaluated for their qualitative phenotype in the T₁ generation. No negative phenotypes were observed in the T₁ plants, although an increase in the number of branches was observed one of the events.

Screens of Superpools on High pH Media for pH Tolerance.

[0138]Seed from superpools of the 35S over-expression lines were evaluated for greenness and size on high pH media as described above. T₃ seed were also assayed for total seed yield, total tissue dry weight and harvest index as described above.

Results:

[0139]Qualitative Analysis of the Superpool Containing 35S:: cDNA 12482777 Plants on High pH

[0140]The screen identified a single event that was greener and the overall size was comparable to the controls. There was no delay in flowering time or decrease in seed set compared to un-treated wild-type. After recovery, the plant containing cDNA 12482777 had significantly better seed yield, as determined by seed volume, than controls (FIG. 2).

Qualitative and Quantitative Analysis of T₃-cDNA 12482777 on High pH.

[0141]The plants were treated with Finale® to eliminate any false-positives or any lines where the Finale® marker was suppressed. All of the Finale®-resistant candidates flowered and set seed. Finale® resistance segregation in the T₃ line suggested a segregation ratio of 1:1 (R:S) as calculated by chi-square test (Table 3-1).

[0142]The plants were greener than the pre-pH treated control. There was no tolerant effect found under low phosphate conditions (data not shown), suggesting that the tolerant response is not to the nutrient deficiencies imposed by the high pH but rather to oxidative stress induced by alkalinity.

TABLE-US-00012 TABLE 3-1 Observed and expected frequencies assuming ratio for high pH tolerance among cDNA 12335629 tested for growth under high pH (pH 9.0) assuming a 3:1 (R:S) segregation ratio. α of 0.05 Probability Event Generation Observed Expected χ² of Chi-Test cDNA T₃ 23 27 0.593 12482777 pH Resistant cDNA T₃ 13 9 1.778 0.12 12482777 pH Sensitive N = 36 36 36 2.371

Qualitative and Quantitative Analysis of Harvest Index, Seed Yield, and Plant Height of T₃ Progeny of 35S:: cDNA 12482777.

[0143]A segregating population of 17 plants containing cDNA 12482777 was analyzed for harvest index and seed yield compared to wild-type populations. Based upon stem height measurements, the transgenic population of 35S:: cDNA 12482777 (10 plants) was significantly smaller than both internal (6 plants) and external wild-type/control populations. Internal wild-types/controls were those plants segregating from the T₃ population of the 35S::cDNA 12482777 line which did not contain the insert (segregating non-transgenics). External wild-types were non-transgenic plants from an outside source which shared no lineage with the line being tested. External wild-types are added to the experiment as a process control to ensure the quality of the growth conditions. Average height for transgenic plants of cDNA 12482777 was 33.44 cm±0.78 versus 44.65 cm±0.70 for the internal wild-type controls. Despite this decrease in plant height, harvest index, as measured by seed weight/total plant weight remained unaffected, i.e., these transgenic plants still produced the same ratio of total seed weight:total plant weight (biomass) as non-transgenic controls. This result means that although the total seed yield is decreased in cDNA 12482777 lines, it still has the same seed proportionally as controls. The cDNA 12482777 plants had a harvest index of 56.96±2.99 compared to the wild-type population's harvest index of 44.92±2.67 (Table 3-2A). This increase in harvest index was significant at a P-value of 0.009 (Table 3-3A).

[0144]It is important to note that seed weight of cDNA 12482777 plants with a larger harvest index was 0.30977g±0.025 while the wild-type population had an average seed weight of 0.37155g±0.027 (Table 3-3B). cDNA 12482777 has a slightly smaller seed weight than the wild-type population but not statistically different at a P-value of 0.12 (Table 3-3B), suggesting that the harvest index of 35S:: cDNA 12482777 is comparable to, if not greater than, wild-type plants. This increase in harvest index is not due to an increase in number of branches (data not shown) as observed in the T₁ generation. Instead, the internode length between siliques is reduced compared to the internal wild-type control, suggesting that cDNA 12482777 plants have more siliques per stem length.

TABLE-US-00013 TABLE 3-2A Descriptive statistical comparison of Harvest Index between segregating T₄ populations containing cDNA 12482777. Internal Wild- Harvest Index: cDNA Transgenic Harvest Index: of type 12482777 small stature Population cDNA 12482777 Wild-type stature Population Mean 56.9582619 Mean 44.91972222 Standard Error 2.990040579 Standard Error 2.667294901 Median 56.68809524 Median 45.56319444 Standard Deviation 9.455338527 Standard Deviation 6.533511501 Sample Variance 89.40342666 Sample Variance 42.68677253 Minimum 43.41666667 Minimum 33.9375 Maximum 70.11666667 Maximum 54.36666667 Sum 569.582619 Sum 269.5183333 Count 10 Count 6 Confidence Level 6.763946869 Confidence Level 6.856488619 (95.0%) (95.0%)

TABLE-US-00014 TABLE 3-2B Descriptive statistical comparison of total seed weight (g) at time of harvest between segregating T₄ populations containing cDNA 12482777. Total Seed Weight (g) Total Seed Weight (g) Internal Wild- of: cDNA 12482777: Transgenic of: cDNA 12482777: type Small Stature Population Wild-type Stature Population Mean 0.30977 Mean 0.37155 Standard Error 0.024799382 Standard Error 0.027304014 Median 0.3017 Median 0.3796 Standard Deviation 0.078422531 Standard Deviation 0.066880902 Sample Variance 0.006150093 Sample Variance 0.004473055 Minimum 0.1956 Minimum 0.2715 Maximum 0.4207 Maximum 0.4621 Sum 3.0977 Sum 2.2293 Count 10 Count 6 Confidence Level 0.056100142 Confidence Level 0.070187087 (95.0%) (95.0%)

TABLE-US-00015 TABLE 3-4A Statistical comparison of harvest index between transgenic populations of clone 126592 and internal wild-type populations using a t-test on two samples assuming unequal variances. cDNA 1248277 Wt stature (internal wild-type population) and cDNA 12482777 small stature (transgenic population). Harvest Index: Harvest Index cDNA 12482777 cDNA 12482777 Wt stature small stature Mean 44.91972222 56.9582619 Variance 42.68677253 89.40342666 Observations 6 10 Hypothesized Mean Difference 0 df 14 t Stat -3.004493678 P (T <= t) one-tail 0.004733406 t Critical one-tail 1.76130925 P (T <= t) two-tail 0.009466812 t Critical two-tail 2.144788596

TABLE-US-00016 TABLE 3-44B Statistical comparison of seed weight between transgenic population of clone 126592 and internal wild-type populations using a t-test on two samples assuming unequal variances. cDNA 12482777 Wt stature (internal wild-type population) and cDNA 12482777 small stature (transgenic population) Seed Weight 12482777: Seed Weight 12482777: WT stature Small Stature Mean 0.37155 0.30977 Variance 0.004473055 0.006150093 Observations 6 10 Hypothesized Mean 0 Difference df 12 t Stat 1.674926201 P(T <= t) one-tail 0.059894848 t Critical one-tail 1.782286745 P(T <= t) two-tail 0.119789696 t Critical two-tail 2.178812792

Table 3-5 provides the results of the consensus sequence analysis based on Ceres cDNA 12482777.

TABLE-US-00017 TABLE 3-5 ##STR00006## ##STR00007##

Example 4

Ceres cDNA 12333678

[0145]Clone 26006, Ceres cDNA 12333678, encodes a full-length glycosyl hydrolase. Ectopic expression of Ceres cDNA 12333678 under the control of the CaMV35S promoter induces the following phenotypes: [0146]Germination on high concentrations of polyethylene glycol (PEG), mannitol and abscissic acid (ABA). [0147]Continued growth on high PEG, mannitol and ABA.Generation and Phenotypic Evaluation of T₁ Lines Containing 35S::cDNA 12333678.

[0148]Wild-type Arabidopsis Wassilewskija (WS) plants were transformed with a Ti plasmid containing cDNA 12333678 in the sense orientation relative to the CaMV35S constitutive promoter. The T, plasmid vector used for this construct, CRS338, contains PAT and confers herbicide resistance to transformed plants. Ten independently transformed events were selected and evaluated for their qualitative phenotype in the T₁ generation. No positive or negative phenotypes were observed in the T₁ plants.

Screens of Superpools on High PEG, Mannitol and ABA as Surrogate Screens for Drought Tolerance.

[0149]Seeds from 13 superpools (1,200 T₂ seeds from each superpool) from the CaMV35S or 32449 over-expression lines were tested on high pH media as described above. T₃ seeds were collected from the tolerant plants and analyzed for tolerance on all additional high pH screens.

[0150]Once cDNA 12333678 was identified in tolerant plants, the individual T₂ events containing this cDNA (ME01334) were screened on high PEG, mannitol and ABA to identify events with the resistance phenotype.

[0151]Superpools (SP) are referred to as SP1, SP2 and so on. The letter following the hyphen refers to the screen (P=PEG, M=mannitol, and A=ABA) and the number following the letter refers to a number assigned to each plant obtained from that screen on that superpool. For example, SP1-M18 is the 18^th plant isolated from a mannitol screen of Superpool 1.

Results:

[0152]Qualitative Assessment of MEO1334 on high pH.

[0153]Superpool 1 was screened on high pH media as described above. PCR analyses identified ME01334 as one of the ME lines showing high pH resistance. Testing of the second generation confirmed the inheritance of the pH resistance (data not shown).

[0154]ME01334 plants that recovered after high pH produced an exceptionally large number of seeds compared to wild-type controls. Additional testing confirmed that these plants statistically produce 30-80% more seeds than either wild-type or transgenic control plants that are recovered from this screen or transferred from regular MS media.

[0155]Table 4-1 provides the results of the consensus sequence analysis based on Ceres cDNA 12333678.

TABLE-US-00018 TABLE 4-1 ##STR00008## ##STR00009## ##STR00010## ##STR00011## ##STR00012## ##STR00013##

[0156]The invention being thus described, it will be apparent to one of ordinary skill in the art that various modifications of the materials and methods for practicing the invention can be made. Such modifications are to be considered within the scope of the invention as defined by the following claims.

[0157]Each of the references from the patent and periodical literature cited herein is hereby expressly incorporated in its entirety by such citation.

Sequence CWU 1

491618DNAArabidopsis thalianamisc_featureclone34035_inplanta_experimental_L42 1gcttaaagtc tctggtcaaa tgggaaatag tgtaagaagc aatctaagag acatcagagg 60acgacgatcg atggatcctc ggatgtggca caaagtcgcc gctatttccg gtatggctgc 120tcttggtttg ggaacttatg gtgctcatgt ctttaaacca gagaaccctt cttacaaaca 180ggtgtggcaa acggcttcac tttaccattt ggttcacact gctgctcttg tttctgctcc 240tagcaccaaa tatcccaaca tttttggtgg cttgttgact gctggaattg tagccttttc 300cggcacgtgt tatatggtag cgctgcggga ggacagaaag ttttcgacat tggcaccatt 360cggaggcttt gcgttcattg ctgcatgggc aactttactt ttctaaacaa tctcataacc 420atctatattg tcaagtttgt ggtcaagctt atcctacata tgaactcact gttttttttt 480gtttacctaa gagattgctt aataacaatt ctgtgtcgac aaccattaag catcttcctt 540tacttgttca gtttgttgct aaagggatta tgtaaatgac gaccatatta atgtaatctt 600attaccatac aatttacc 6182128PRTArabidopsis thalianamisc_featurepeptide_clone34035_inplanta_experimental_L42 2Met Gly Asn Ser Val Arg Ser Asn Leu Arg Asp Ile Arg Gly Arg Arg1 5 10 15Ser Met Asp Pro Arg Met Trp His Lys Val Ala Ala Ile Ser Gly Met 20 25 30Ala Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Val Phe Lys Pro Glu 35 40 45Asn Pro Ser Tyr Lys Gln Val Trp Gln Thr Ala Ser Leu Tyr His Leu 50 55 60Val His Thr Ala Ala Leu Val Ser Ala Pro Ser Thr Lys Tyr Pro Asn65 70 75 80Ile Phe Gly Gly Leu Leu Thr Ala Gly Ile Val Ala Phe Ser Gly Thr 85 90 95Cys Tyr Met Val Ala Leu Arg Glu Asp Arg Lys Phe Ser Thr Leu Ala 100 105 110Pro Phe Gly Gly Phe Ala Phe Ile Ala Ala Trp Ala Thr Leu Leu Phe 115 120 1253111PRTBrassica napusmisc_featureCeresClone872428 3Met Asp Pro Arg Ile Trp His Lys Val Ala Ala Val Ser Gly Met Ala1 5 10 15Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Val Phe Lys Pro Glu Asn 20 25 30Pro Ser Tyr Lys Gln Val Trp Gln Thr Ala Ser Leu Tyr His Leu Val 35 40 45His Thr Ala Ala Leu Val Ser Ala Pro Ser Thr Lys Tyr Pro Asn Ile 50 55 60Phe Gly Gly Leu Leu Thr Ala Gly Ile Val Ala Phe Ser Gly Thr Cys65 70 75 80Tyr Met Val Ala Leu Arg Glu Asp Arg Lys Phe Ser Thr Leu Ala Pro 85 90 95Phe Gly Gly Phe Ala Phe Ile Ala Ala Trp Ala Thr Leu Leu Phe 100 105 1104113PRTBrassica napusmisc_featureCeresClone972918 4Met Gly Asn Cys Val Arg Ser Asn Leu Arg Asp Leu Gly Gly Arg Arg1 5 10 15Ser Met Asp Pro Arg Ile Trp His Lys Val Ala Ala Val Ser Gly Met 20 25 30Ala Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Val Phe Lys Pro Glu 35 40 45Asn Pro Ser Tyr Lys Gln Val Trp Gln Thr Ala Ser Leu Tyr His Leu 50 55 60Val His Thr Ala Ala Leu Val Ser Ala Pro Ser Thr Lys Tyr Pro Asn65 70 75 80Ile Phe Gly Gly Leu Leu Thr Ala Gly Ile Val Ala Phe Ser Gly Thr 85 90 95Tyr Glu Tyr Ala Lys Ser Phe Val Phe Val Asn Val Val Gly Val Thr 100 105 110Trp5111PRTGlycine maxmisc_featureCeresClone566573 5Met Asp Pro Gln Leu Trp His Lys Val Ala Ala Ile Ser Gly Leu Ala1 5 10 15Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Val Phe Lys Pro Gln Asn 20 25 30Pro Ala Tyr Asn Asp Val Trp His Thr Ala Ser Leu Tyr His Leu Val 35 40 45His Thr Ala Ala Leu Val Ala Ala Pro Ile Thr Lys His Pro Asn Val 50 55 60Phe Gly Gly Leu Leu Thr Ala Gly Ile Leu Ala Phe Ser Gly Thr Cys65 70 75 80Tyr Thr Val Ala Phe Leu Glu Asp Arg Lys Tyr Ser Thr Met Ala Pro 85 90 95Phe Gly Gly Phe Ala Phe Ile Ala Ala Trp Gly Ser Leu Phe Phe 100 105 1106111PRTGlycine maxmisc_featureCeresClone588155 6Met Asp Pro Gln Val Trp His Lys Val Ala Ala Ile Ser Gly Val Ala1 5 10 15Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Val Phe Lys Pro Gln Asn 20 25 30Pro Ala Tyr Lys Asp Val Trp His Thr Ala Ser Leu Tyr His Leu Val 35 40 45His Thr Ala Ala Leu Val Ala Ala Pro Ile Thr Lys His Pro Asn Val 50 55 60Phe Gly Gly Leu Leu Thr Ala Gly Ile Leu Ala Phe Ser Gly Thr Cys65 70 75 80Tyr Thr Val Ala Phe Leu Glu Asp Arg Lys Tyr Ser Thr Met Ala Pro 85 90 95Phe Gly Gly Phe Ala Phe Ile Ala Ala Trp Gly Ser Leu Phe Phe 100 105 1107115PRTTriticum aestivummisc_featureCeresClone678257 7Met Val Met Pro Thr Asp Pro Met Leu Trp His Lys Val Ala Ala Val1 5 10 15Ser Gly Val Val Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Met Phe 20 25 30Arg Pro Gln Asn Pro Arg Tyr Lys Glu Ile Trp Gln Thr Ala Ser Leu 35 40 45Tyr His Leu Val His Thr Ala Ala Leu Leu Gly Ala Pro Met Thr Lys 50 55 60Arg Pro Asn Ile Phe Gly Gly Leu Leu Thr Thr Gly Ile Val Leu Phe65 70 75 80Ser Gly Thr Cys Tyr Thr Val Ala Tyr Leu Glu Asp Arg Lys Phe Ser 85 90 95Ser Pro Ala Pro Ile Gly Gly Phe Ala Phe Ile Ala Ala Trp Ala Ser 100 105 110Leu Leu Phe 1158115PRTZea maysmisc_featureCeresClone289088 8Met Leu Ala Ala Thr Asp Pro Met Leu Trp His Lys Val Ala Ala Val1 5 10 15Ser Gly Val Ala Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Met Phe 20 25 30Arg Pro Lys Asn Pro Ala Tyr Lys Glu Val Trp His Thr Ala Ser Leu 35 40 45Tyr His Leu Val His Thr Ala Ala Leu Leu Gly Ala Pro Ile Thr Lys 50 55 60Arg Pro Asn Val Phe Gly Gly Leu Leu Thr Ala Gly Ile Val Leu Phe65 70 75 80Ser Gly Thr Cys Tyr Thr Val Ala Tyr Leu Glu Asp Arg Lys Phe Ser 85 90 95Ser Pro Ala Pro Leu Gly Gly Phe Ala Phe Ile Ala Ala Trp Ala Ser 100 105 110Leu Leu Phe 115993PRTPsathyrostachys junceamisc_featuregi|7963694 9Met Leu Trp His Lys Val Ala Ala Val Ser Gly Val Ala Ala Leu Gly1 5 10 15Leu Gly Thr Tyr Gly Ala His Met Phe Arg Pro Gln Asn Pro Lys Tyr 20 25 30Lys Glu Ile Trp Gln Thr Ala Phe Leu Tyr His Leu Val His Thr Ala 35 40 45Ala Leu Leu Gly Ala Pro Met Thr Lys Arg Pro Asn Ile Phe Gly Gly 50 55 60Leu Leu Thr Thr Gly Ile Val Leu Phe Ser Gly Thr Cys Tyr Thr Val65 70 75 80Ala Tyr Leu Glu Asp Arg Lys Phe Ser Ser Pro Ala Pro 85 9010106PRTAgropyron cristatummisc_featuregi|7963702 10Met Val Met Pro Thr Asp Pro Met Leu Trp His Lys Val Ala Ala Val1 5 10 15Ser Gly Val Ala Ala Leu Gly Leu Gly Thr Tyr Gly Ala His Met Phe 20 25 30Arg Pro Gln Asn Pro Arg Tyr Lys Glu Ile Trp Gln Thr Ala Ser Leu 35 40 45Tyr His Leu Val His Thr Ala Ala Leu Leu Gly Ala Pro Met Thr Lys 50 55 60Arg Pro Asn Ile Phe Gly Gly Leu Leu Thr Thr Gly Ile Val Leu Phe65 70 75 80Ser Gly Thr Cys Tyr Thr Val Ala Tyr Leu Glu Asp Arg Lys Phe Ser 85 90 95Ser Pro Ala Pro Ile Gly Gly Phe Ala Phe 100 10511120PRTOryza sativa subsp. japonicamisc_featuregi|50918749 11Met Ala Ala Ala Ala Ala Met Ala Met Lys Asp Pro Ser Leu Trp His1 5 10 15Lys Val Ala Ala Ile Ser Gly Val Ala Ala Leu Gly Leu Gly Thr Tyr 20 25 30Gly Ala His Met Phe Arg Pro Lys Asn Pro Ala Tyr Lys Glu Val Trp 35 40 45His Thr Ala Ser Leu Tyr His Leu Val His Thr Ala Ala Leu Leu Gly 50 55 60Ala Pro Ile Thr Lys Arg Pro Asp Val Phe Gly Gly Leu Leu Thr Ala65 70 75 80Gly Ile Val Leu Phe Ser Gly Thr Cys Tyr Thr Val Ala Tyr Leu Glu 85 90 95Asp Arg Lys Tyr Ser Ser Thr Ala Pro Leu Gly Gly Phe Ala Phe Ile 100 105 110Ala Ala Trp Ala Ser Leu Leu Phe 115 12012689DNAArabidopsis thalianamisc_featureclone40781_inplanta_experimental_L43 12ataaacgaat ccaaatttca agaggagaag aaaaatcttc aagtccacga cgaaactttt 60catcgatctt caaattccag aaaaaactcg atgaatcttc aagctgtttc ttgtagcttc 120ggattccttt cgagtccact tggtgtcact cccagaactt cgtttcgtcg cttcgtaatc 180cgagcgaaaa cggaaccgtc ggagaaatca gtagagatta tgaggaaatt ctccgagcaa 240tatgctcgtc gctctgggac ttacttctgt gttgataaag gagttacttc agtcgttatt 300aagggtttgg ctgagcataa agattcatat ggtgcaccgc tttgcccttg cagacactat 360gatgataaag ctgctgaggt tggacaaggc ttttggaatt gtccgtgtgt tccaatgaga 420gagaggaagg agtgccattg tatgcttttc ttaactcctg ataatgattt cgctggaaaa 480gatcagacga ttacatcgga tgaaataaaa gaaactacag ctaacatgtg agagagctgg 540ttcttccatg ttcatcacct ctgttcttta ggtaaaaaaa aagagagata tgtctcgccc 600caaatgcagt cttgtacatt gataccccga gcatcttctt cgttcttctg tacaactctt 660tcactcttaa gataatattc tttagtatg 68913146PRTArabidopsis thalianamisc_featurepeptide_clone40781_inplanta_experimental_L43 13Met Asn Leu Gln Ala Val Ser Cys Ser Phe Gly Phe Leu Ser Ser Pro1 5 10 15Leu Gly Val Thr Pro Arg Thr Ser Phe Arg Arg Phe Val Ile Arg Ala 20 25 30Lys Thr Glu Pro Ser Glu Lys Ser Val Glu Ile Met Arg Lys Phe Ser 35 40 45Glu Gln Tyr Ala Arg Arg Ser Gly Thr Tyr Phe Cys Val Asp Lys Gly 50 55 60Val Thr Ser Val Val Ile Lys Gly Leu Ala Glu His Lys Asp Ser Tyr65 70 75 80Gly Ala Pro Leu Cys Pro Cys Arg His Tyr Asp Asp Lys Ala Ala Glu 85 90 95Val Gly Gln Gly Phe Trp Asn Cys Pro Cys Val Pro Met Arg Glu Arg 100 105 110Lys Glu Cys His Cys Met Leu Phe Leu Thr Pro Asp Asn Asp Phe Ala 115 120 125Gly Lys Asp Gln Thr Ile Thr Ser Asp Glu Ile Lys Glu Thr Thr Ala 130 135 140Asn Met14514148PRTSpinacia oleraceamisc_featuregi|505189 14Met Lys Ala Leu Gln Ala Ser Thr Ser Tyr Ser Phe Phe Ser Lys Ser1 5 10 15Ser Ser Ala Thr Leu Gln Arg Arg Thr His Arg Pro Gln Cys Val Ile 20 25 30Leu Ser Lys Val Glu Pro Ser Asp Lys Ser Val Glu Ile Met Arg Lys 35 40 45Phe Ser Glu Gln Tyr Ala Arg Lys Ser Gly Thr Tyr Phe Cys Val Asp 50 55 60Lys Gly Val Thr Ser Val Val Ile Lys Gly Leu Ala Glu His Lys Asp65 70 75 80Ser Leu Gly Ala Pro Leu Cys Pro Cys Arg Tyr Tyr Asp Asp Lys Ala 85 90 95Ala Glu Ala Thr Gln Gly Phe Trp Asn Cys Pro Cys Val Pro Met Arg 100 105 110Glu Arg Lys Glu Cys His Cys Met Leu Phe Leu Thr Pro Glu Asn Asp 115 120 125Phe Ala Gly Lys Asp Gln Thr Ile Gly Leu Asp Glu Ile Arg Glu Val 130 135 140Thr Ala Asn Met14515143PRTBrassica napusmisc_featureCeresClone1127455 15Met Asn Pro Gln Ala Val Ser Cys Ser Phe Gly Phe Val Ser Ala Pro1 5 10 15Leu Val Ser Pro Arg Thr Ser Arg Phe Val Val Gln Ala Lys Ser Glu 20 25 30Pro Ser Glu Xaa Ser Val Glu Ile Met Arg Lys Phe Ser Glu Gln Tyr 35 40 45Ala Arg Arg Ser Gly Thr Tyr Phe Cys Val Asp Lys Gly Val Xaa Ser 50 55 60Val Val Ile Lys Gly Leu Ala Glu His Lys Asp Ser Tyr Gly Ala Pro65 70 75 80Leu Cys Pro Cys Arg His Tyr Asp Asp Lys Ala Ala Glu Val Gly Gln 85 90 95Gly Phe Trp Asn Cys Pro Cys Val Pro Met Arg Glu Arg Lys Glu Cys 100 105 110His Cys Met Leu Phe Leu Thr Pro Asp Asn Asp Phe Ala Gly Lys Asp 115 120 125Gln Thr Ile Thr Ser Asp Glu Ile Lys Glu Thr Thr Ala His Met 130 135 14016144PRTGlycine maxmisc_featureCeresClone470939 16Met Thr Thr Gln Ala Ser Thr Phe Ala Val Ala Val Pro Ser Val Ala1 5 10 15Thr Pro Phe Arg Arg His Arg Asn Pro Phe Val Val Arg Ala Gln Ala 20 25 30Glu Pro Ser Asp Lys Ser Val Glu Ile Met Arg Lys Phe Ser Glu Gln 35 40 45Tyr Ala Arg Lys Ser Gly Thr Tyr Phe Cys Val Asp Lys Gly Val Thr 50 55 60Ser Val Val Ile Lys Gly Leu Ala Asp His Lys Asp Thr Leu Gly Ala65 70 75 80Pro Leu Cys Pro Cys Arg His Tyr Asp Asp Lys Ala Ala Glu Val Ala 85 90 95Gln Gly Phe Trp Asn Cys Pro Cys Val Pro Met Arg Glu Arg Lys Glu 100 105 110Cys His Cys Met Leu Phe Leu Thr Pro Asp Asn Asp Phe Ala Gly Asn 115 120 125Glu Gln Thr Ile Thr Leu Asp Glu Ile Lys Glu Ser Thr Ala Asn Met 130 135 14017148PRTSolanum tuberosummisc_featuregi|14275859 17Met Arg Thr Leu Gln Ala Ser Thr Ser Tyr Ser Val Gly Phe Gly Ile1 5 10 15Ser Ser Phe Ala Thr Arg Pro Lys Pro Ser Thr His Arg Cys Leu Thr 20 25 30Val Ala Lys Met Glu Pro Ser Glu Lys Ser Val Glu Ile Met Arg Lys 35 40 45Phe Ser Glu Gln Tyr Ala Arg Arg Ser Glu Thr Tyr Phe Cys Met Asp 50 55 60Lys Gly Val Thr Ser Val Val Ile Lys Gly Leu Ala Glu His Lys Asp65 70 75 80Thr Leu Gly Ala Pro Leu Cys Pro Cys Arg His Tyr Asp Asp Lys Ala 85 90 95Ala Glu Ala Gln Gln Gly Phe Trp Asn Cys Pro Cys Val Pro Met Arg 100 105 110Glu Arg Lys Glu Cys His Cys Met Leu Phe Leu Thr Pro Asp Asn Asp 115 120 125Phe Ala Gly Glu Glu Gln Thr Ile Ser Met Glu Glu Ile Lys Glu Thr 130 135 140Thr Ala Asn Met14518152PRTZea maysmisc_featureCeresClone295783 18Met Thr Ser Thr Val Thr Thr Thr Val Gly Cys Gly Gly Leu Pro Val1 5 10 15Arg Pro Leu Ser Thr Ala Thr Arg Gly Arg Pro Arg Arg Cys Ala Val 20 25 30Arg Ala Gln Ala Ala Gly Ala Asp Ala Ser Asn Asp Lys Ser Val Glu 35 40 45Val Met Arg Lys Phe Ser Glu Gln Tyr Ala Arg Arg Ser Asn Thr Phe 50 55 60Phe Cys Ala Asp Lys Thr Val Thr Ala Val Val Ile Lys Gly Leu Ala65 70 75 80Asp His Arg Asp Thr Leu Gly Ala Pro Leu Cys Pro Cys Arg His Tyr 85 90 95Asp Asp Lys Ala Ala Glu Val Ala Gln Gly Phe Trp Asn Cys Pro Cys 100 105 110Val Pro Met Arg Glu Arg Lys Glu Cys His Cys Met Leu Phe Leu Thr 115 120 125Pro Asp Asn Asp Phe Ala Gly Lys Asp Gln Val Ile Ser Phe Glu Glu 130 135 140Ile Lys Glu Ala Thr Ser Lys Phe145 15019146PRTOryza sativa subsp. japonicamisc_featuregi|50898984 19Met Met Ser Met Ala Ser Thr Thr Ala Ser Pro Phe Cys Pro Ser Pro1 5 10 15Met Pro Arg Gly Arg Lys Cys Thr Val Arg Val Gln Ala Gly Ala Ala 20 25 30Gly Ala Asp Ala Ser Asp Lys Ser Leu Glu Ile Met Arg Lys Phe Ser 35 40 45Glu Gln Tyr Ala Arg Arg Ser Asn Thr Phe Phe Cys Ser Glu Lys Ser 50 55 60Val Thr Ala Val Val Ile Lys Gly Leu Ala Asp His Lys Asp Gln Leu65 70 75

80Gly Ala Pro Leu Cys Pro Cys Arg His Tyr Asp Asp Lys Ala Ala Glu 85 90 95Val Ala Gln Gly Phe Trp Asn Cys Pro Cys Val Pro Met Arg Glu Arg 100 105 110Lys Glu Cys His Cys Met Leu Phe Leu Thr Pro Asp Asn Asp Phe Ala 115 120 125Gly Gln Asp Gln Ala Ile Thr Leu Glu Glu Ile Lys Asp Ala Thr Ser 130 135 140Lys Ile145201016DNAArabidopsis thalianamisc_featureclone126592_expected_L44 20gttgaatcaa aaataatcag taacgctttg agtgaagatg atgaatgttg cagtgacagc 60cactccctcg tctctcttgt actctcctct gcttcttcct tctcaagggc caaaccggcg 120aatgcaatgg aaaagaaacg gaaagagacg gttagggaca aaggtggctg tttccggtgt 180tatcacagct ggatttgagc tgaagccacc tccatatcct cttgatgctc tggaaccgca 240tatgagccgg gaaaccttgg attatcactg gggcaaacat cacaaaactt atgtagagaa 300cctgaacaag caaatcttag gcacggatct agatgcatta tccttggaag aagttgtgct 360tctttcatac aacaaaggca atatgcttcc tgctttcaac aacgctgcac aggcttggaa 420ccacgagttc ttctgggagt ctatccaacc tggaggtgga ggaaagccaa ctggagagct 480cctcagatta atagaaagag attttgggtc tttcgaagag tttttggaaa ggttcaagtc 540ggctgcagct tcgaattttg gttcgggttg gacatggctt gcatataagg cgaatagact 600tgacgttgca aatgccgtta atcctctccc aaaggaggaa gacaagaaac ttgttatagt 660gaagacgccc aatgcagtaa atccgctcgt atgggattat tctccacttc tcaccattga 720tacctgggag cacgcttact atctggattt tgagaaccga agagctgaat acataaatac 780attcatggaa aagcttgtgt catgggaaac tgtaagcaca aggttggaat ccgcaattgc 840tcgagcagtg caaagagaac aagaaggaac agagacagaa gatgaagaga atccagatga 900tgaagtacca gaggtctatt tagatagtga catcgatgta tctgaggttg actaaaactt 960gtgaagcaat aacattagca tcttaaatgt taattacaca gagcaaattt ttttgc 101621305PRTArabidopsis thalianamisc_featurepeptide_clone126592_expected_L44 21Met Met Asn Val Ala Val Thr Ala Thr Pro Ser Ser Leu Leu Tyr Ser1 5 10 15Pro Leu Leu Leu Pro Ser Gln Gly Pro Asn Arg Arg Met Gln Trp Lys 20 25 30Arg Asn Gly Lys Arg Arg Leu Gly Thr Lys Val Ala Val Ser Gly Val 35 40 45Ile Thr Ala Gly Phe Glu Leu Lys Pro Pro Pro Tyr Pro Leu Asp Ala 50 55 60Leu Glu Pro His Met Ser Arg Glu Thr Leu Asp Tyr His Trp Gly Lys65 70 75 80His His Lys Thr Tyr Val Glu Asn Leu Asn Lys Gln Ile Leu Gly Thr 85 90 95Asp Leu Asp Ala Leu Ser Leu Glu Glu Val Val Leu Leu Ser Tyr Asn 100 105 110Lys Gly Asn Met Leu Pro Ala Phe Asn Asn Ala Ala Gln Ala Trp Asn 115 120 125His Glu Phe Phe Trp Glu Ser Ile Gln Pro Gly Gly Gly Gly Lys Pro 130 135 140Thr Gly Glu Leu Leu Arg Leu Ile Glu Arg Asp Phe Gly Ser Phe Glu145 150 155 160Glu Phe Leu Glu Arg Phe Lys Ser Ala Ala Ala Ser Asn Phe Gly Ser 165 170 175Gly Trp Thr Trp Leu Ala Tyr Lys Ala Asn Arg Leu Asp Val Ala Asn 180 185 190Ala Val Asn Pro Leu Pro Lys Glu Glu Asp Lys Lys Leu Val Ile Val 195 200 205Lys Thr Pro Asn Ala Val Asn Pro Leu Val Trp Asp Tyr Ser Pro Leu 210 215 220Leu Thr Ile Asp Thr Trp Glu His Ala Tyr Tyr Leu Asp Phe Glu Asn225 230 235 240Arg Arg Ala Glu Tyr Ile Asn Thr Phe Met Glu Lys Leu Val Ser Trp 245 250 255Glu Thr Val Ser Thr Arg Leu Glu Ser Ala Ile Ala Arg Ala Val Gln 260 265 270Arg Glu Gln Glu Gly Thr Glu Thr Glu Asp Glu Glu Asn Pro Asp Asp 275 280 285Glu Val Pro Glu Val Tyr Leu Asp Ser Asp Ile Asp Val Ser Glu Val 290 295 300Asp305221016DNAArabidopsis thalianamisc_featureclone126592_inplanta_experimental_L44 22gttgaatcaa aaataatcag taacgctttg agtgaagatg atgaatgttg cagtgacagc 60cactccctcg tctctcttgt actctcctct gcttcttcct tctcaagggc caaaccggcg 120aatgcaatgg aaaagaaacg gaaagagacg gttagggaca aaggtggctg tttccggtgt 180tatcacagct ggatttgagc tgaagccacc tccatatcct cttgatgctc tggaaccgca 240tatgagccgg gaaaccttgg attatcactg gggcaaacat cacaaaactt atgtagagaa 300cctgaacaag caaatcttag gcacggatct agatgcatta tccttggaag aagttgtgct 360tctttcatac aacaaaggca atatgcttcc tgctttcaac aacgctgcac aggcttggaa 420ccacgagttc ttctgggagt ctatccaacc tggaggtgga ggaaagccaa ctggagagct 480cctcagatta atagaaagag attttgggtc tttcgaagag tttttggaaa ggttcaagtc 540ggctgcagct tcgaattttg gttcgggttg gacatggctt gcatataagg cgaatagact 600tgacgttgca aatgccgtta atcctctccc aaaggaggaa gacaagaaac ttgttatagt 660gaagacgccc aatgcagtaa atccgctcgt atgggattat tctccacttc tcaccattga 720tacctgggag cacgcttact atctggattt tgagaaccga agagctgaat acataaatac 780attcatggaa aagcttgtgt catgggaaac tgtaagcaca aggttggaat ccgcaattgc 840tcgagcagtg caaagagaac aagaaagaac agagacagaa gatgaagaga atccagatga 900tgaagtacca gaggtctatt tagatagtga catcgatgta tctgaggttg actaaaactt 960gtgaagcaat aacattagca tcttaaatgt taattacaca gagcaaattt ttttgc 101623305PRTArabidopsis thalianamisc_featurepeptide_clone126592_inplanta_experimental_L44 23Met Met Asn Val Ala Val Thr Ala Thr Pro Ser Ser Leu Leu Tyr Ser1 5 10 15Pro Leu Leu Leu Pro Ser Gln Gly Pro Asn Arg Arg Met Gln Trp Lys 20 25 30Arg Asn Gly Lys Arg Arg Leu Gly Thr Lys Val Ala Val Ser Gly Val 35 40 45Ile Thr Ala Gly Phe Glu Leu Lys Pro Pro Pro Tyr Pro Leu Asp Ala 50 55 60Leu Glu Pro His Met Ser Arg Glu Thr Leu Asp Tyr His Trp Gly Lys65 70 75 80His His Lys Thr Tyr Val Glu Asn Leu Asn Lys Gln Ile Leu Gly Thr 85 90 95Asp Leu Asp Ala Leu Ser Leu Glu Glu Val Val Leu Leu Ser Tyr Asn 100 105 110Lys Gly Asn Met Leu Pro Ala Phe Asn Asn Ala Ala Gln Ala Trp Asn 115 120 125His Glu Phe Phe Trp Glu Ser Ile Gln Pro Gly Gly Gly Gly Lys Pro 130 135 140Thr Gly Glu Leu Leu Arg Leu Ile Glu Arg Asp Phe Gly Ser Phe Glu145 150 155 160Glu Phe Leu Glu Arg Phe Lys Ser Ala Ala Ala Ser Asn Phe Gly Ser 165 170 175Gly Trp Thr Trp Leu Ala Tyr Lys Ala Asn Arg Leu Asp Val Ala Asn 180 185 190Ala Val Asn Pro Leu Pro Lys Glu Glu Asp Lys Lys Leu Val Ile Val 195 200 205Lys Thr Pro Asn Ala Val Asn Pro Leu Val Trp Asp Tyr Ser Pro Leu 210 215 220Leu Thr Ile Asp Thr Trp Glu His Ala Tyr Tyr Leu Asp Phe Glu Asn225 230 235 240Arg Arg Ala Glu Tyr Ile Asn Thr Phe Met Glu Lys Leu Val Ser Trp 245 250 255Glu Thr Val Ser Thr Arg Leu Glu Ser Ala Ile Ala Arg Ala Val Gln 260 265 270Arg Glu Gln Glu Arg Thr Glu Thr Glu Asp Glu Glu Asn Pro Asp Asp 275 280 285Glu Val Pro Glu Val Tyr Leu Asp Ser Asp Ile Asp Val Ser Glu Val 290 295 300Asp30524158PRTBrassica napusmisc_featureCeresClone970125 24Met Met Met Thr Thr Thr Ser Ser Leu Leu Ser Pro Cys Ser Leu Leu1 5 10 15Pro Ser Gln Gly Pro Asn Arg Gln Thr Gln Trp Lys Arg His Glu Lys 20 25 30Arg Gln Phe Ser Arg Lys Val Val Val Ser Gly Val Val Arg Ala Gly 35 40 45Phe Glu Leu Lys Pro Pro Pro Tyr Pro Leu Asp Ala Leu Glu Pro His 50 55 60Met Ser Arg Glu Thr Met Asp Tyr His Trp Gly Lys His His Arg Thr65 70 75 80Tyr Val Glu Asn Leu Asn Lys Gln Ile Leu Gly Thr Asp Leu Asp Gly 85 90 95Leu Ser Leu Glu Glu Val Val Leu Leu Ser Tyr Asn Arg Gly Asn Met 100 105 110Leu Pro Val Phe Asn Asn Ala Ala Gln Ala Trp Asn His Glu Phe Phe 115 120 125Trp Glu Ser Ile Gln Pro Gly Gly Gly Gly Lys Pro Ser Gly Asp Leu 130 135 140Leu Arg Leu Ile Glu Arg Asp Phe Gly Ser Phe Asp Asp Phe145 150 15525313PRTGlycine maxmisc_featureCeresClone624535 25Met Asn Leu Leu Ser Gln Ser Thr Ala Pro Ser Thr Ser Leu Ser Pro1 5 10 15Ser Cys Phe Leu Pro Arg His Pro His Gly Ser Thr Trp Phe Ser Ser 20 25 30Gly Thr Phe Lys Phe Leu Lys Lys Glu Ser Arg Cys Leu Arg Lys Ala 35 40 45Gly Arg Thr Lys Ile Thr Ala Lys Phe Glu Leu Lys Pro Pro Pro Tyr 50 55 60Pro Leu Ser Ala Leu Glu Pro Ile Met Ser Gln Glu Thr Leu Glu Tyr65 70 75 80His Trp Gly Lys His His Arg Thr Tyr Val Asp Asn Leu Asn Arg Gln 85 90 95Ile Asp Gly Thr Asp Leu Asp Gly Asn Ser Leu Glu Asn Thr Ile Val 100 105 110Ile Thr Tyr Asn Lys Gly Asp Ile Leu Pro Ala Phe Asn Asn Ala Ala 115 120 125Gln Ala Trp Asn His Asp Phe Phe Trp Glu Ser Met Lys Pro Gly Gly 130 135 140Gly Gly Arg Pro Ser Gly Asp Leu Leu Asn Leu Ile Glu Arg Asp Phe145 150 155 160Gly Ser Phe Glu Lys Phe Leu Asp Glu Phe Lys Thr Ala Ala Ser Thr 165 170 175Gln Phe Gly Ser Gly Trp Ala Trp Leu Ala Tyr Lys Glu Ser Arg Leu 180 185 190Asp Val Glu Asn Ala Val Asn Pro Leu Gln Ser Asp Glu Asp Lys Lys 195 200 205Leu Val Val Val Lys Thr Pro Asn Ala Val Asn Pro Leu Val Trp Asn 210 215 220Tyr Tyr His Pro Leu Leu Thr Ile Asp Val Trp Glu His Ala Tyr Phe225 230 235 240Ile Asp Phe Gln Asn Gln Arg Arg Asp Tyr Ile Ser Val Phe Met Asp 245 250 255Lys Leu Val Ser Trp Asp Ala Val Ser Ser Arg Leu Glu Gln Ala Lys 260 265 270Ala Leu Ile Lys Glu Arg Glu Arg Glu Ala Glu Arg Lys Arg Arg Glu 275 280 285Glu Glu Glu Lys Arg Thr Ser Ser Glu Ala Ile Pro Glu Ile Tyr Ser 290 295 300Asp Gly Asp Ala Asp Leu Asp Ala Glu305 31026313PRTMedicago sativamisc_featuregi|16974682 26Met Lys Leu Leu Ser Pro Ser Ala Thr Ser Ser Thr His Val Ser Ser1 5 10 15Ser Ala Phe Leu Pro Asn Val Ala Gly Phe Gln Asn Leu Gly Ser Ser 20 25 30Ser Val Thr Thr Phe Lys Phe Ser Lys Lys Gln Gly Arg Cys Ile Arg 35 40 45Arg Ala Gly Gly Thr Gln Ile Thr Ala Lys Phe Glu Leu Lys Pro Pro 50 55 60Pro Tyr Pro Leu Asn Ala Ser Glu Pro Ile Met Ser Gln Asn Thr Phe65 70 75 80Glu Tyr His Trp Gly Lys His His Arg Ala Tyr Val Asp Asn Leu Asn 85 90 95Lys Gln Ile Glu Gly Thr Asp Leu Asp Gly Lys Ser Leu Glu Glu Thr 100 105 110Ile Ile Met Ser Tyr Asn Asn Gly Asp Ile Leu Pro Ala Phe Asn Asn 115 120 125Ala Ala Gln Val Trp Asn His Asp Phe Phe Trp Glu Ser Met Lys Pro 130 135 140Gly Gly Gly Gly Lys Pro Ser Gly Glu Leu Leu Lys Leu Ile Glu Arg145 150 155 160Asp Phe Gly Ser Phe Glu Lys Phe Val Glu Gln Phe Lys Leu Ala Ala 165 170 175Ser Thr Gln Phe Gly Ser Gly Trp Ala Trp Leu Ala Tyr Lys Glu Ser 180 185 190Arg Leu Asp Val Gly Asn Ala Val Asn Pro Leu Ala Thr Glu Glu Asp 195 200 205Lys Lys Leu Val Val Leu Lys Ser Pro Asn Ala Val Asn Pro Leu Val 210 215 220Trp Asn His His His Pro Leu Leu Thr Ile Asp Val Trp Glu His Ala225 230 235 240Tyr Tyr Leu Asp Tyr Gln Asn Arg Arg Pro Glu Tyr Ile Ser Val Phe 245 250 255Met Asp Lys Leu Val Ser Trp Glu Ala Val Ser Ser Arg Leu Glu Lys 260 265 270Ala Lys Ala Val Ile Ala Glu Arg Glu Lys Glu Glu Glu Arg Lys Arg 275 280 285Arg Glu Glu Glu Glu Lys Ser Thr Thr Gly Glu Asp Thr Pro Ala Pro 290 295 300Glu Ile Phe Ala Asp Ser Asp Thr Asp305 31027220PRTZea maysmisc_featureCeresClone278210 27Met Ser Leu Gly Gln Met Met Leu Ala Ser Phe Asn Glu Gly Arg Glu1 5 10 15Gln Pro His Pro Pro Phe Phe His Ala Ala Gln Val Trp Asn His Asp 20 25 30Phe Tyr Trp Arg Ser Met Lys Pro Gly Gly Gly Gly Lys Pro Pro Glu 35 40 45Arg Leu Leu Lys Phe Ile Asn Arg Asp Phe Gly Ser Tyr Glu Gly Met 50 55 60Ile Arg Gln Phe Met Asp Ala Ala Leu Thr Gln Phe Gly Ser Gly Trp65 70 75 80Val Trp Leu Ser Tyr Lys Gly Ser Gly Leu Pro Tyr Val Lys Ser Arg 85 90 95Ser Pro Ile Pro Ser Asp Asn His Gly Arg Leu Val Ile Ser Lys Thr 100 105 110Pro Asn Ala Ile Asn Pro Leu Val Trp Gly His Ser Pro Leu Leu Ala 115 120 125Ile Asp Val Trp Glu His Ala Tyr Tyr Leu Asp Tyr Glu Asp Arg Arg 130 135 140Ala Asp Tyr Val Ser Ala Ile Leu Glu Lys Leu Val Ser Trp Glu Thr145 150 155 160Val Glu Ser Arg Leu Ala Lys Ala Val Ala Arg Ala Val Glu Arg Asp 165 170 175Glu His Leu Arg Arg Arg Ile Leu Arg Lys Gln Arg Leu Ala Gln Ala 180 185 190Asn Gly Gln Ser Arg Ala Arg Ser Arg Ala Arg Gln Gly Arg Gln Gly 195 200 205Asp Gln Glu Val Ala Arg Ser Arg Pro Val Glu Ala 210 215 220281954DNAArabidopsis thalianamisc_featurePromoter 326; Report 56 28gtgggtaaaa gtatccttct ttgtgcattt ggtattttta agcatgtaat aagaaaaacc 60aaaatagacg gctggtattt aataaaagga gactaatgta tgtatagtat atgatttgtg 120tggaatataa taaagttgta aaatatagat gtgaagcgag tatctatctt ttgactttca 180aaggtgatcg atcgtgttct ttgtgatagt tttggtcgtc ggtctacaag tcaacaacca 240ccttgaagtt ttcgcgtctc ggtttcctct tcgcatctgg tatccaatag catacatata 300ccagtgcgga aaatggcgaa gactagtggg cttgaaccat aaggtttggc cccaatacgg 360attccaaaca acaagcctag cgcagtcttt tgggatgcat aagactaaac tgtcgcagtg 420atagacgtaa gatatatcga cttgattgga atcgtctaag ctaataagtt taccttgacc 480gtttatagtt gcgtcaacgt ccttatggag attgatgccc atcaaataaa cctgaaaatc 540catcaccatg accaccataa actcccttgc tgccgctgct ttggcttgag caaggtgttt 600ccttgtaaag ctccgatctt tggataaagt gttccacttt ttgcaagtag ctctgacccc 660tctcagagat gtcaccggaa tcttagacag aacctcctct gccaaatcac ttggaagatc 720ggacaatgtc atcatttttg caggtaattt ctccttcgtt gctgctttgg cttgagcacg 780gtgcttcttt gtaaagctcc gatctttgga taagagcgga tcggaatcct ctaggaggtg 840ccagtccctt gacctattaa tttatagaag gttttagtgt attttgttcc aatttcttct 900ctaacttaac aaataacaac tgcctcatag tcatgggctt caaattttat cgcttggtgt 960atttcgttat ttgcaaggcc ttggcccatt ttgagcccaa taactaaatc tagccttttc 1020agaccggaca tgaacttcgc atattggcgt aactgtgcag ttttaccttt ttcggatcag 1080acaagatcag atttagacca cccaacaata gtcagtcata tttgacaacc taagctagcc 1140gacactacta aaaagcaaac aaaagaagaa ttctatgttg tcattttacc ggtggcaagt 1200ggacccttct ataaaagagt aaagagacag cctgtgtgtg tataatctct aattatgttc 1260accgacacaa tcacacaaac ccttctctaa tcacacaact tcttcatgat ttacgacatt 1320aattatcatt aactctttaa attcacttta catgctcaaa aatatctaat ttgcagcatt 1380aatttgagta ccgataacta ttattataat cgtcgtgatt cgcaatcttc ttcattagat 1440gctgtcaagt tgtactcgca cgcggtggtc cagtgaagca aatccaacgg tttaaaacct 1500tcttacattt ctagatctaa tctgaaccgt cagatatcta gatctcattg tctgaacaca 1560gttagatgaa actgggaatg aatctggacg aaattacgat cttacaccaa ccccctcgac 1620gagctcgtat atataaagct tatacgctcc tccttcacct tcgtactact actaccacca 1680catttcttta gctcaacctt cattactaat ctccttttaa ggtatgttca cttttcttcg 1740attcatactt tctcaagatt cctgcatttc tgtagaattt gaaccaagtg tcgatttttg 1800tttgagagaa gtgttgattt atagatctgg ttattgaatc tagattccaa tttttaattg 1860attcgagttt gttatgtgtg tttatactac ttctcattga tcttgtttga tttctctgct 1920ctgtattagg tttctttcgt gaatcagatc ggaa 1954292016DNAArabidopsis thalianamisc_featurePromoter 32449; Report 92 29gatcggcctt cttcaggtct tctctgtagc tctgttactt ctatcacagt tatcgggtat 60ttgagaaaaa agagttagct aaaatgaatt tctccatata atcatggttt actacaggtt 120tacttgattc gcgttagctt tatctgcatc caaagttttt tccatgatgt tatgtcatat 180gtgataccgt tactatgttt ataactttat acagtctggt tcactggagt ttctgtgatt 240atgttgagta catactcatt catcctttgg taactctcaa gtttaggttg tttgaattgc

300ctctgttgtg atacttattg tctattgcat caatcttcta atgcaccacc ctagactatt 360tgaacaaaga gctgtttcat tcttaaacct ctgtgtctcc ttgctaaatg gtcatgcttt 420aatgtcttca cctgtctttc tcttctatag atatgtagtc ttgctagata gttagttcta 480cagctctctt ttgtagtctt gttagagagt tagttgagat attacctctt aaaagtatcc 540ttgaacgctt tccggttatg accaatttgt tgtagctcct tgtaagtaga acttactggg 600accagcgaga cagtttatgt gaatgttcat gcttaagtgt cgaacgtatc tatctctact 660atagctctgt agtcttgtta gacagttagt tttatatctc catttttttg tagtcttgct 720agttgagata ttacctcttc tcttcaaagt atccttgaac gctcaccggt tatgaaatct 780ctacactata gctctgtagt cttgctagat agttagttct ttagctctct ttttgtagcc 840tagttcttta gctctccttt tgtagccttg ctacagagta agatgggata ttacctcctt 900gaacgctctc cggttatgac caatttgttg tagctccttg taagtagaac ttaggataga 960gtgagtcaac tttaagaaag aacctagtat gtggcataac cagattgcag gctctgtctc 1020ggctacagta acgtaactct atagctcttt gttttgttca gaaagaacca gtgattggat 1080gattcgtcct tagaaactgg acctaacaac agtcattggc tttgaaatca agccacaaca 1140atgcctatat gaaccgtcca tttcatttat ccgtttcaaa ccagcccatt acatttcgtc 1200ccattgataa ccaaaagcgg ttcaatcaga ttatgtttta attttaccaa attctttatg 1260aagtttaaat tatactcaca ttaaaaggat tattggataa tgtaaaaatt ctgaacaatt 1320actgattttg gaaaattaac aaatattctt tgaaatagaa gaaaaagcct ttttcctttt 1380gacaacaaca tataaaatca tactcccatt aaaaagattt taatgtaaaa ttctgaatat 1440aagatatttt ttacaacaac aaccaaaaat atttattttt ttcctttttt acagcaacaa 1500gaaggaaaaa cttttttttt tgtcaagaaa aggggagatt atgtaaacag ataaaacagg 1560gaaaataact aaccgaactc tcttaattaa catcttcaaa taaggaaaat tatgatccgc 1620atatttagga agatcaatgc attaaaacaa cttgcacgtg gaaagagaga ctatacgctc 1680cacacaagtt gcactaatgg tacctctcac aaaccaatca aaatactgaa taatgccaac 1740gtgtacaaat tagggtttta cctcacaacc atcgaacatt ctcgaaacat tttaaacagc 1800ctggcgccat agatctaaac tctcatcgac caatttttga ccgtccgatg gaaactctag 1860cctcaaccca aaactctata taaagaaatc ttttccttcg ttattgctta ccaaatacaa 1920accctagccg ccttattcgt cttcttcgtt ctctagtttt ttcctcagtc tctgttctta 1980gatcccttgt agtttccaaa tcttccgata aggcct 201630870DNAArabidopsis thalianamisc_featureCeres cDNA 12333678 30aaaaagtacg aaaggaaaat atgagtgagg agaagaggaa gcaacacttc gtgctagtac 60atggtgcgtg ccacggcgca tggtgctggt acaaggttaa gcctcttctc gaggctttgg 120gccatcgtgt aaccgcctta gacctagctg cttccggtat agacacaacc aggtcaatca 180ctgacatttc tacatgtgaa caatattctg agccattgat gcagctaatg acttcattgc 240cgaatgatga gaaggttgta ctcgttggtc atagctttgg aggtttgagt ttagccttag 300ccatggataa gtttcccgat aaaatctctg tctctgtctt cgtgactgca ttcatgcccg 360acaccaaaca ctcaccatcg ttcgtcgagg aaaagtttgc aagcagcatg acaccagaag 420gatggatggg ctctgagctc gagacatatg gttcagataa ttccggcttg tctgtgttct 480tcagcaccga cttcatgaag caccgtctct accaactttc tcctgtggag gatcttgagc 540ttggattgct tctaaagagg cctagttcat tgtttattaa tgaattatcg aagatggaga 600acttttctga gaaagggtat ggatctgttc ctcgagctta cattgtgtgc aaagaggaca 660acattatctc ggaagaccat caacgatgga tgatccataa ttatccggcg aatttagtga 720ttgagatgga agagacggat catatgccaa tgttttgcaa acctcaagta ctaagtgacc 780atctattggc aatcgctgac aatttctctt aaataatatt ttgatgaaaa tgtatttgga 840gtggatacaa taaaaatgtg ttctaaatgg 87031263PRTArabidopsis thalianamisc_featurePeptide Ceres cDNA 12333678 31Met Ser Glu Glu Lys Arg Lys Gln His Phe Val Leu Val His Gly Ala1 5 10 15Cys His Gly Ala Trp Cys Trp Tyr Lys Val Lys Pro Leu Leu Glu Ala 20 25 30Leu Gly His Arg Val Thr Ala Leu Asp Leu Ala Ala Ser Gly Ile Asp 35 40 45Thr Thr Arg Ser Ile Thr Asp Ile Ser Thr Cys Glu Gln Tyr Ser Glu 50 55 60Pro Leu Met Gln Leu Met Thr Ser Leu Pro Asn Asp Glu Lys Val Val65 70 75 80Leu Val Gly His Ser Phe Gly Gly Leu Ser Leu Ala Leu Ala Met Asp 85 90 95Lys Phe Pro Asp Lys Ile Ser Val Ser Val Phe Val Thr Ala Phe Met 100 105 110Pro Asp Thr Lys His Ser Pro Ser Phe Val Glu Glu Lys Phe Ala Ser 115 120 125Ser Met Thr Pro Glu Gly Trp Met Gly Ser Glu Leu Glu Thr Tyr Gly 130 135 140Ser Asp Asn Ser Gly Leu Ser Val Phe Phe Ser Thr Asp Phe Met Lys145 150 155 160His Arg Leu Tyr Gln Leu Ser Pro Val Glu Asp Leu Glu Leu Gly Leu 165 170 175Leu Leu Lys Arg Pro Ser Ser Leu Phe Ile Asn Glu Leu Ser Lys Met 180 185 190Glu Asn Phe Ser Glu Lys Gly Tyr Gly Ser Val Pro Arg Ala Tyr Ile 195 200 205Val Cys Lys Glu Asp Asn Ile Ile Ser Glu Asp His Gln Arg Trp Met 210 215 220Ile His Asn Tyr Pro Ala Asn Leu Val Ile Glu Met Glu Glu Thr Asp225 230 235 240His Met Pro Met Phe Cys Lys Pro Gln Val Leu Ser Asp His Leu Leu 245 250 255Ala Ile Ala Asp Asn Phe Ser 26032267PRTCitrus sinensismisc_featuregi|14279437 32Met Glu Glu Val Val Gly Met Glu Glu Lys His Phe Val Leu Val His1 5 10 15Gly Val Asn His Gly Ala Trp Cys Trp Tyr Lys Leu Lys Ala Arg Leu 20 25 30Val Ala Gly Gly His Arg Val Thr Ala Val Asp Leu Ala Ala Ser Gly 35 40 45Ile Asn Met Lys Arg Ile Glu Asp Val His Thr Phe His Ala Tyr Ser 50 55 60Glu Pro Leu Met Glu Val Leu Ala Ser Leu Pro Ala Glu Glu Lys Val65 70 75 80Ile Leu Val Gly His Ser Leu Gly Gly Val Thr Leu Ala Leu Ala Gly 85 90 95Asp Lys Phe Pro His Lys Ile Ser Val Ala Val Phe Val Thr Ala Phe 100 105 110Met Pro Asp Thr Thr His Arg Pro Ser Phe Val Leu Glu Gln Tyr Ser 115 120 125Glu Lys Met Gly Lys Glu Asp Asp Ser Trp Leu Asp Thr Gln Phe Ser 130 135 140Gln Cys Asp Ala Ser Asn Pro Ser His Ile Ser Met Leu Phe Gly Arg145 150 155 160Glu Phe Leu Thr Ile Lys Ile Tyr Gln Leu Cys Pro Pro Glu Asp Leu 165 170 175Glu Leu Ala Lys Met Leu Val Arg Pro Gly Ser Met Phe Ile Asp Asn 180 185 190Leu Ser Lys Glu Ser Lys Phe Ser Asp Glu Gly Tyr Gly Ser Val Lys 195 200 205Arg Val Tyr Leu Val Cys Glu Glu Asp Ile Gly Leu Pro Lys Gln Phe 210 215 220Gln His Trp Met Ile Gln Asn Tyr Pro Val Asn Glu Val Met Glu Ile225 230 235 240Lys Gly Gly Asp His Met Ala Met Leu Ser Asp Pro Gln Lys Leu Cys 245 250 255Asp Cys Leu Ser Gln Ile Ser Leu Lys Tyr Ala 260 26533263PRTArabidopsis thalianamisc_featureCeresClone1010900 33Met Ser Glu Glu Lys Arg Lys Gln His Phe Val Leu Val His Gly Ser1 5 10 15Cys His Gly Ala Trp Cys Trp Tyr Lys Val Lys Pro Leu Leu Glu Ala 20 25 30Val Gly His Arg Val Thr Ala Val Asp Leu Ala Ala Ser Gly Ile Asp 35 40 45Thr Thr Arg Ser Ile Thr Asp Ile Pro Thr Cys Glu Gln Tyr Ser Glu 50 55 60Pro Leu Thr Lys Leu Leu Thr Ser Leu Pro Asn Asp Glu Lys Val Val65 70 75 80Leu Val Gly His Ser Phe Gly Gly Leu Asn Leu Ala Ile Ala Met Glu 85 90 95Lys Phe Pro Glu Lys Ile Ser Val Ala Val Phe Leu Thr Ala Phe Met 100 105 110Pro Asp Thr Glu His Ser Pro Ser Phe Val Leu Asp Lys Phe Gly Ser 115 120 125Asn Met Pro Gln Glu Ala Trp Met Gly Thr Glu Phe Glu Pro Tyr Gly 130 135 140Ser Asp Asn Ser Gly Leu Ser Met Phe Phe Ser Pro Asp Phe Met Lys145 150 155 160Leu Gly Leu Tyr Gln Leu Ser Pro Val Glu Asp Leu Glu Leu Gly Leu 165 170 175Leu Leu Met Arg Pro Gly Ser Leu Phe Ile Asn Asp Leu Ser Lys Met 180 185 190Lys Asn Phe Ser Asp Glu Gly Tyr Gly Ser Val Pro Arg Val Phe Ile 195 200 205Val Cys Lys Glu Asp Lys Ala Ile Pro Glu Glu Arg Gln Arg Trp Met 210 215 220Ile Asp Asn Phe Pro Val Asn Leu Val Met Glu Met Glu Glu Thr Asp225 230 235 240His Met Pro Met Phe Cys Lys Pro Gln Gln Leu Ser Asp Tyr Phe Leu 245 250 255Lys Ile Ala Asp Lys Phe Val 26034263PRTArabidopsis thalianamisc_featuregi|20196998 34Met Ser Glu Glu Lys Arg Lys Gln His Phe Val Leu Val His Gly Ser1 5 10 15Cys His Gly Ala Trp Cys Trp Tyr Lys Val Lys Pro Leu Leu Glu Ala 20 25 30Val Gly His Arg Val Thr Ala Val Asp Leu Ala Ala Ser Gly Ile Asp 35 40 45Thr Thr Arg Ser Ile Thr Asp Ile Pro Thr Cys Glu Gln Tyr Ser Glu 50 55 60Pro Leu Thr Lys Leu Leu Thr Ser Leu Pro Asn Asp Glu Lys Val Val65 70 75 80Leu Val Gly His Ser Phe Gly Gly Leu Asn Leu Ala Ile Ala Met Glu 85 90 95Lys Phe Pro Glu Lys Ile Ser Val Ala Val Phe Leu Thr Ala Phe Met 100 105 110Pro Asp Thr Glu His Ser Pro Ser Phe Val Leu Asp Lys Phe Gly Ser 115 120 125Asn Met Pro Gln Glu Ala Trp Met Gly Thr Glu Phe Glu Pro Tyr Gly 130 135 140Ser Asp Asn Ser Gly Leu Ser Met Phe Phe Ser Pro Asp Phe Met Lys145 150 155 160Leu Gly Leu Tyr Gln Leu Ser Pro Val Glu Asp Leu Glu Leu Gly Leu 165 170 175Leu Leu Met Arg Pro Gly Ser Leu Phe Ile Asn Asp Leu Ser Lys Met 180 185 190Lys Asn Phe Ser Asp Glu Gly Tyr Gly Ser Val Pro Arg Val Phe Ile 195 200 205Val Cys Lys Glu Asp Lys Ala Ile Pro Glu Glu Arg Gln Arg Trp Met 210 215 220Ile Asp Asn Phe Pro Val Asn Leu Val Met Glu Met Glu Glu Thr Asp225 230 235 240His Met Pro Met Phe Cys Lys Pro Gln Gln Leu Ser Asp Tyr Phe Leu 245 250 255Lys Ile Ala Asp Lys Phe Val 26035263PRTArabidopsis thalianamisc_featuregi|27754457 35Met Ser Glu Glu Lys Arg Lys Gln His Phe Val Leu Val His Gly Ser1 5 10 15Cys His Gly Ala Trp Cys Trp Tyr Lys Val Lys Pro Leu Leu Glu Ala 20 25 30Val Gly His Arg Val Thr Ala Val Asp Leu Ala Ala Ser Gly Ile Asp 35 40 45Thr Thr Arg Ser Ile Thr Asp Ile Pro Thr Cys Glu Gln Tyr Ser Glu 50 55 60Pro Leu Thr Lys Leu Leu Thr Ser Leu Pro Asn Asp Glu Lys Val Val65 70 75 80Leu Val Gly His Ser Phe Gly Gly Leu Asn Leu Ala Ile Ala Met Glu 85 90 95Lys Phe Pro Lys Lys Ile Ser Val Ala Val Phe Leu Thr Ala Phe Met 100 105 110Pro Asp Thr Glu His Ser Pro Ser Phe Val Leu Asp Lys Phe Gly Ser 115 120 125Asn Met Pro Gln Glu Ala Trp Met Gly Thr Glu Phe Glu Pro Tyr Gly 130 135 140Ser Asp Asn Ser Gly Leu Ser Met Phe Phe Ser Pro Asp Phe Met Lys145 150 155 160Leu Gly Leu Tyr Gln Leu Ser Pro Val Glu Asp Leu Glu Leu Gly Leu 165 170 175Leu Leu Met Arg Pro Gly Ser Leu Phe Ile Asn Asp Leu Ser Lys Met 180 185 190Lys Asn Phe Ser Asp Glu Gly Tyr Gly Ser Val Pro Arg Val Phe Ile 195 200 205Val Cys Lys Glu Asp Lys Ala Ile Pro Glu Glu Arg Gln Arg Trp Met 210 215 220Ile Asp Asn Phe Pro Val Asn Leu Val Met Glu Met Glu Glu Thr Asp225 230 235 240His Met Pro Met Phe Cys Lys Pro Gln Gln Leu Ser Asp Tyr Phe Leu 245 250 255Lys Ile Ala Asp Lys Phe Val 26036257PRTHevea brasiliensismisc_featuregi|50513520 36Met Ala Phe Ala His Phe Val Leu Ile His Thr Ile Cys His Gly Ala1 5 10 15Trp Ile Trp His Lys Leu Lys Pro Leu Leu Glu Ala Leu Gly His Lys 20 25 30Val Thr Ala Leu Asp Leu Ala Ala Ser Gly Val Asp Pro Arg Gln Ile 35 40 45Glu Glu Ile Gly Ser Phe Asp Glu Tyr Ser Glu Pro Leu Leu Thr Phe 50 55 60Leu Glu Ala Leu Pro Pro Gly Glu Lys Val Ile Leu Val Gly Glu Ser65 70 75 80Cys Gly Gly Leu Asn Ile Ala Ile Ala Ala Asp Lys Tyr Cys Glu Lys 85 90 95Ile Ala Ala Ala Val Phe His Asn Ser Val Leu Pro Asp Thr Glu His 100 105 110Cys Pro Ser Tyr Val Val Asp Lys Leu Met Glu Val Phe Pro Asp Trp 115 120 125Lys Asp Thr Thr Tyr Phe Thr Tyr Thr Lys Asp Gly Lys Glu Ile Thr 130 135 140Gly Leu Lys Leu Gly Phe Thr Leu Leu Arg Glu Asn Leu Tyr Thr Leu145 150 155 160Cys Gly Pro Glu Glu Tyr Glu Leu Ala Lys Met Leu Thr Arg Lys Gly 165 170 175Ser Leu Phe Gln Asn Ile Leu Ala Lys Arg Pro Phe Phe Thr Lys Glu 180 185 190Gly Tyr Gly Ser Ile Lys Lys Ile Tyr Val Trp Thr Asp Gln Asp Glu 195 200 205Ile Phe Leu Pro Glu Phe Gln Leu Trp Gln Ile Glu Asn Tyr Lys Pro 210 215 220Asp Lys Val Tyr Lys Val Glu Gly Gly Asp His Leu Leu Gln Leu Thr225 230 235 240Lys Thr Lys Glu Ile Ala Glu Ile Leu Gln Glu Val Ala Asp Thr Tyr 245 250 255Asn37257PRTHevea brasiliensismisc_featuregi|6435646 37Met Ala Phe Ala His Phe Val Leu Ile His Thr Ile Cys His Gly Ala1 5 10 15Trp Ile Trp His Lys Leu Lys Pro Leu Leu Glu Ala Leu Gly His Lys 20 25 30Val Thr Ala Leu Asp Leu Ala Ala Ser Gly Val Asp Pro Arg Gln Ile 35 40 45Glu Glu Ile Gly Ser Phe Asp Glu Tyr Ser Glu Pro Leu Leu Thr Phe 50 55 60Leu Glu Ala Leu Pro Pro Gly Glu Lys Val Ile Leu Val Gly Glu Ser65 70 75 80Cys Gly Gly Leu Asn Ile Ala Ile Ala Ala Asp Lys Tyr Cys Glu Lys 85 90 95Ile Ala Ala Ala Val Phe His Asn Ser Val Leu Pro Asp Thr Glu His 100 105 110Cys Pro Ser Tyr Val Val Asp Lys Leu Met Glu Val Phe Pro Asp Trp 115 120 125Lys Asp Thr Thr Tyr Phe Thr Tyr Thr Lys Asp Gly Lys Glu Ile Thr 130 135 140Gly Leu Lys Leu Gly Phe Thr Leu Leu Arg Glu Asn Leu Tyr Thr Leu145 150 155 160Cys Gly Pro Glu Glu Tyr Glu Leu Ala Lys Met Leu Thr Arg Lys Gly 165 170 175Ser Leu Phe Gln Asn Ile Leu Ala Lys Arg Pro Phe Phe Thr Lys Glu 180 185 190Gly Tyr Gly Ser Ile Lys Lys Ile Tyr Val Trp Thr Asp Gln Asp Glu 195 200 205Ile Phe Leu Pro Glu Phe Gln Leu Trp Gln Ile Glu Asn Tyr Lys Pro 210 215 220Asp Lys Val Tyr Lys Val Glu Gly Gly Asp His Lys Leu Gln Leu Thr225 230 235 240Lys Thr Lys Glu Ile Ala Glu Ile Leu Gln Glu Val Ala Asp Thr Tyr 245 250 255Asn38258PRTManihot esculentamisc_featuregi|2780225 38Met Ala Val Val Asp Phe Val Leu Ile His Thr Ile Cys His Gly Ala1 5 10 15Trp Ile Trp Tyr Lys Leu Lys Pro Val Leu Glu Ala Ala Gly His Lys 20 25 30Val Thr Ala Leu Asp Leu Ala Ala Ser Gly Val Asp Pro Arg Gln Ile 35 40 45Glu Gln Ile Asn Ser Phe Asp Glu Tyr Ser Glu Pro Leu Leu Thr Phe 50 55 60Met Glu Ser Leu Pro Gln Gly Glu Lys Val Ile Leu Val Gly Glu Ser65 70 75 80Cys Gly Gly Leu Asn Ile Ala Ile Ala Ala Asp Lys Tyr Pro Glu Lys 85 90 95Ile Ala Ala Ala Val Phe Gln Asn Ser Leu Leu Pro Asp Thr Lys His 100 105 110Lys Pro Ser Tyr Val Val Asp Lys Leu Met Glu Val Phe Pro Asp Trp 115 120 125Lys Asp Thr Glu Tyr Phe Glu Phe Ser Asn Ser Asn

Gly Glu Thr Ile 130 135 140Thr Gly Met Val Leu Gly Leu Lys Leu Met Arg Glu Asn Leu Tyr Thr145 150 155 160Ile Cys Pro Pro Glu Asp Tyr Glu Leu Ala Lys Met Leu Thr Arg Arg 165 170 175Gly Ser Leu Phe Gln Ser Ile Leu Ala Gln Arg Glu Lys Phe Thr Glu 180 185 190Lys Gly Tyr Gly Ser Ile Lys Lys Ile Tyr Val Trp Thr Gly Asp Asp 195 200 205Lys Ile Phe Leu Pro Glu Phe Gln Leu Trp Gln Ile Glu Asn Tyr Lys 210 215 220Pro Asp Leu Val Phe Arg Val Met Gly Gly Asp His Lys Leu Gln Leu225 230 235 240Thr Lys Thr Asn Glu Ile Ala Gly Ile Leu Gln Lys Val Ala Asp Ile 245 250 255Tyr Ala39258PRTCatharanthus roseusmisc_featuregi|53830670 39Met Glu Val Met Lys His Phe Val Thr Val His Gly Val Gly His Gly1 5 10 15Ala Trp Val Tyr Tyr Lys Leu Lys Pro Arg Ile Glu Ala Ala Gly His 20 25 30Arg Cys Thr Ala Val Asn Leu Ala Ala Ser Gly Ile Asn Glu Lys Lys 35 40 45Leu Glu Glu Val Arg Ser Ser Ile Asp Tyr Ala Ala Pro Leu Leu Glu 50 55 60Val Leu Asp Ser Val Pro Glu Asn Glu Lys Val Ile Leu Val Gly His65 70 75 80Ser Gly Gly Gly Met Thr Ala Ala Val Gly Met Glu Lys Phe Pro Asn 85 90 95Lys Ile Ser Leu Ala Val Phe Leu Asn Ala Ile Met Pro Asp Thr Glu 100 105 110Asn Arg Pro Ser Tyr Val Leu Glu Glu Tyr Thr Ala Lys Thr Pro Pro 115 120 125Glu Ala Trp Lys Asp Cys Gln Phe Ser Ala Tyr Gly Asp Pro Pro Ile 130 135 140Thr Ser Leu Val Cys Gly Pro Glu Phe Ile Ser Ser Thr Leu Tyr His145 150 155 160Leu Ser Pro Ile Glu Asp His Ala Leu Gly Lys Ile Leu Val Arg Pro 165 170 175Gly Ser Leu Phe Ile Glu Asp Leu Leu Lys Ala Glu Lys Phe Thr Glu 180 185 190Glu Gly Phe Gly Ser Val Pro Arg Val Tyr Val Ile Ala Ala Glu Asp 195 200 205Lys Thr Ile Pro Pro Glu Phe Gln Arg Trp Met Ile Glu Asn Asn Pro 210 215 220Val Lys Glu Val Lys Glu Ile Lys Gly Ala Asp His Met Pro Met Phe225 230 235 240Ser Lys Pro Asp Glu Leu Ser Gln Cys Leu Leu Asp Ile Ala Lys Lys 245 250 255His Ala40264PRTRauvolfia serpentinamisc_featuregi|6651393 40Met His Ser Ala Ala Asn Ala Lys Gln Gln Lys His Phe Val Leu Val1 5 10 15His Gly Gly Cys Leu Gly Ala Trp Ile Trp Tyr Lys Leu Lys Pro Leu 20 25 30Leu Glu Ser Ala Gly His Lys Val Thr Ala Val Asp Leu Ser Ala Ala 35 40 45Gly Ile Asn Pro Arg Arg Leu Asp Glu Ile His Thr Phe Arg Asp Tyr 50 55 60Ser Glu Pro Leu Met Glu Val Met Ala Ser Ile Pro Pro Asp Glu Lys65 70 75 80Val Val Leu Leu Gly His Ser Phe Gly Gly Met Ser Leu Gly Leu Ala 85 90 95Met Glu Thr Tyr Pro Glu Lys Ile Ser Val Ala Val Phe Met Ser Ala 100 105 110Met Met Pro Asp Pro Asn His Ser Leu Thr Tyr Pro Phe Glu Lys Tyr 115 120 125Asn Glu Lys Cys Pro Ala Asp Met Met Leu Asp Ser Gln Phe Ser Thr 130 135 140Tyr Gly Asn Pro Glu Asn Pro Gly Met Ser Met Ile Leu Gly Pro Gln145 150 155 160Phe Met Ala Leu Lys Met Phe Gln Asn Cys Ser Val Glu Asp Leu Glu 165 170 175Leu Ala Lys Met Leu Thr Arg Pro Gly Ser Leu Phe Phe Gln Asp Leu 180 185 190Ala Lys Ala Lys Lys Phe Ser Thr Glu Arg Tyr Gly Ser Val Lys Arg 195 200 205Ala Tyr Ile Phe Cys Asn Glu Asp Lys Ser Phe Pro Val Glu Phe Gln 210 215 220Lys Trp Phe Val Glu Ser Val Gly Ala Asp Lys Val Lys Glu Ile Lys225 230 235 240Glu Ala Asp His Met Gly Met Leu Ser Gln Pro Arg Glu Val Cys Lys 245 250 255Cys Leu Leu Asp Ile Ser Asp Ser 26041262PRTLycopersicon esculentummisc_featuregi|41814856 41Met Glu Lys Gly Asp Lys Asn His Phe Val Leu Val His Gly Ala Cys1 5 10 15His Gly Ala Trp Cys Trp Tyr Lys Val Val Thr Ile Leu Arg Ser Glu 20 25 30Gly His Lys Val Ser Val Leu Asp Met Ala Ala Ser Gly Ile Asn Pro 35 40 45Lys His Val Asp Asp Leu Asn Ser Met Ala Asp Tyr Asn Glu Pro Leu 50 55 60Met Glu Phe Met Asn Ser Leu Pro Gln Leu Glu Arg Val Val Leu Val65 70 75 80Gly His Ser Met Gly Gly Ile Asn Ile Ser Leu Ala Met Glu Lys Phe 85 90 95Pro Gln Lys Ile Val Val Ala Val Phe Val Thr Ala Phe Met Pro Gly 100 105 110Pro Asp Leu Asn Leu Val Ala Leu Gly Gln Gln Tyr Asn Gln Gln Val 115 120 125Glu Ser His Met Asp Thr Glu Phe Val Tyr Asn Asn Gly Gln Asp Lys 130 135 140Ala Pro Thr Ser Leu Val Leu Gly Pro Glu Val Leu Ala Thr Asn Phe145 150 155 160Tyr Gln Leu Ser Pro Pro Glu Asp Leu Thr Leu Ala Thr Tyr Leu Val 165 170 175Arg Pro Val Pro Leu Phe Asp Glu Ser Ile Leu Leu Ala Asn Thr Thr 180 185 190Leu Ser Lys Glu Lys Tyr Gly Ser Val His Arg Val Tyr Val Val Cys 195 200 205Asp Lys Asp Asn Val Leu Lys Glu Gln Gln Phe Gln Lys Trp Leu Ile 210 215 220Asn Asn Asn Pro Pro Asp Glu Val Gln Ile Ile His Asn Ala Asp His225 230 235 240Met Val Met Phe Ser Lys Pro Arg Asp Leu Ser Ser Cys Leu Val Met 245 250 255Ile Ser Gln Lys Tyr Tyr 26042260PRTNicotiana tabacummisc_featuregi|40549303 42Met Lys Glu Gly Lys His Phe Val Leu Val His Gly Ala Cys His Gly1 5 10 15Gly Trp Ser Trp Tyr Lys Leu Lys Pro Leu Leu Glu Ala Ala Gly His 20 25 30Lys Val Thr Ala Leu Asp Leu Ala Ala Ser Gly Thr Asp Leu Arg Lys 35 40 45Ile Glu Glu Leu Arg Thr Leu Tyr Asp Tyr Thr Leu Pro Leu Met Glu 50 55 60Leu Met Glu Ser Leu Ser Ala Asp Glu Lys Val Ile Leu Val Gly His65 70 75 80Ser Leu Gly Gly Met Asn Leu Gly Leu Ala Met Glu Lys Tyr Pro Gln 85 90 95Lys Ile Tyr Ala Ala Val Phe Leu Ala Ala Phe Met Pro Asp Ser Val 100 105 110His Asn Ser Ser Phe Val Leu Glu Gln Tyr Asn Glu Arg Thr Pro Ala 115 120 125Glu Asn Trp Leu Asp Thr Gln Phe Leu Pro Tyr Gly Ser Pro Glu Glu 130 135 140Pro Leu Thr Ser Met Phe Phe Gly Pro Lys Phe Leu Ala His Lys Leu145 150 155 160Tyr Gln Leu Cys Ser Pro Glu Asp Leu Ala Leu Ala Ser Ser Leu Val 165 170 175Arg Pro Ser Ser Leu Phe Met Glu Asp Leu Ser Lys Ala Lys Tyr Phe 180 185 190Thr Asp Glu Arg Phe Gly Ser Val Lys Arg Val Tyr Ile Val Cys Thr 195 200 205Glu Asp Lys Gly Ile Pro Glu Glu Phe Gln Arg Trp Gln Ile Asp Asn 210 215 220Ile Gly Val Thr Glu Ala Ile Glu Ile Lys Gly Ala Asp His Met Ala225 230 235 240Met Leu Cys Glu Pro Gln Lys Leu Cys Ala Ser Leu Leu Glu Ile Ala 245 250 255His Lys Tyr Asn 26043262PRTSolanum tuberosummisc_featuregi|56392765 43Met Glu Lys Gly Asn Lys Asn His Phe Val Leu Val His Gly Ala Cys1 5 10 15His Gly Ala Trp Cys Trp Tyr Lys Val Val Thr Ile Leu Arg Ser Glu 20 25 30Gly His Lys Val Ser Val Leu Asp Met Ala Ala Ser Gly Ile Asn Pro 35 40 45Lys His Val Glu Asp Leu Asn Ser Met Ala Asp Tyr Asn Glu Pro Leu 50 55 60Met Glu Phe Met Asn Ser Leu Pro Gln Gln Glu Arg Val Val Leu Val65 70 75 80Gly His Ser Met Gly Gly Ile Asn Ile Ser Leu Ala Met Glu Lys Phe 85 90 95Pro His Lys Ile Ala Val Ala Val Phe Val Ser Ala Ser Met Pro Gly 100 105 110Pro Asp Leu Asn Leu Val Ala Val Thr Gln Gln Tyr Ser Gln Gln Val 115 120 125Glu Thr Pro Met Asp Thr Glu Phe Val Tyr Asn Asn Gly Leu Asp Lys 130 135 140Gly Pro Thr Ser Val Val Leu Gly Pro Lys Val Leu Ala Thr Ile Tyr145 150 155 160Tyr Gln Phe Ser Pro Pro Glu Asp Leu Thr Leu Ala Thr Tyr Leu Val 165 170 175Arg Pro Val Pro Leu Phe Asp Glu Ser Val Leu Leu Thr Asn Thr Thr 180 185 190Leu Ser Lys Glu Lys Tyr Gly Ser Val His Arg Val Tyr Val Val Cys 195 200 205Asp Lys Asp Lys Val Leu Lys Glu Glu Gln Phe Gln Arg Trp Leu Ile 210 215 220Lys Asn Asn Pro Pro Asn Glu Val Gln Met Ile His Asp Ala Gly His225 230 235 240Met Val Met Phe Ser Lys Pro Arg Glu Leu Cys Ser Cys Leu Val Met 245 250 255Ile Ser Gln Lys Tyr His 26044266PRTTriticum aestivummisc_featureCeresClone644331 44Met Glu Ala Cys Ala Gly Gln Ala Ser Ser Ala His Ile Val Leu Val1 5 10 15His Gly Ala Cys Leu Gly Gly Trp Ser Trp Phe Lys Val Ala Thr Arg 20 25 30Leu Arg Ser Ala Gly His Arg Val Ser Thr Pro Asp Leu Ala Ala Ser 35 40 45Gly Val Asp Pro Arg Pro Leu Arg Glu Val Pro Thr Phe Arg Asp Tyr 50 55 60Thr Lys Pro Leu Leu Asp Leu Leu Glu Ser Leu Pro Ser Gly Glu Lys65 70 75 80Val Val Leu Val Gly His Ser Leu Gly Gly Val Asn Val Ala Leu Ala 85 90 95Cys Glu Leu Phe Pro Glu Lys Ile Ala Ala Ala Val Phe Val Ala Ala 100 105 110Phe Met Pro Asp His Arg Ser Pro Pro Ser Tyr Val Leu Glu Lys Phe 115 120 125Val Glu Gly Arg Thr Leu Asp Trp Met Asp Thr Glu Phe Lys Pro Gln 130 135 140Asp Pro Glu Gly Lys Leu Pro Thr Ser Met Leu Phe Gly Pro Leu Val145 150 155 160Thr Arg Ala Lys Phe Phe Gln Leu Cys Ser Pro Glu Asp Leu Thr Leu 165 170 175Gly Arg Ser Leu Met Arg Val Asn Ser Met Phe Val Asp Asp Leu Arg 180 185 190Leu Gln Pro Pro His Thr Glu Ala Arg Tyr Gly Ser Val Arg Lys Ala 195 200 205Tyr Val Val Phe Lys Asp Asp His Ala Ile Val Glu Gln Phe Gln Arg 210 215 220Trp Met Val His Asn Tyr Pro Val Asp Glu Val Met Glu Ile Asp Gly225 230 235 240Ala Asp His Met Ala Leu Leu Ser Thr Pro Thr Glu Leu Ala Arg Cys 245 250 255Leu Ala Asp Ile Ala Val Lys Tyr Ala Ala 260 26545265PRTTriticum aestivummisc_featureCeresClone936068 45Met Glu Gly Ser Ser Ser Gly Lys His Phe Ile Leu Ile His Gly Leu1 5 10 15Cys His Gly Ala Trp Cys Trp Tyr Lys Leu Val Pro Met Leu Arg Ala 20 25 30Ala Gly His Arg Val Thr Ala Leu Asp Met Ala Ala Ser Gly Ala His 35 40 45Pro Ala Arg Met Asp Glu Val Pro Ser Phe Glu Asp Tyr Ser Trp Pro 50 55 60Leu Leu Asp Ala Val Ala Ala Ala Pro Ala Gly Glu Arg Leu Val Leu65 70 75 80Val Gly His Ser Leu Gly Gly Leu Asn Ile Ala Leu Ala Met Glu Arg 85 90 95Phe Pro Arg Lys Val Ala Ala Ala Val Phe Leu Ala Ala Cys Met Pro 100 105 110Cys Val Gly Arg His Met Gly Ala Thr Thr Glu Glu Ile Met Arg Arg 115 120 125Ile Lys Pro Asp Phe Phe Met Asp Met Lys Arg Met Val Leu Asn Thr 130 135 140Ser Gln Gly Pro Arg Pro Ala Leu Val Phe Gly Pro Lys Ile Leu Ala145 150 155 160Ala Lys Leu Tyr Asp Arg Ser Ser Gly Glu Asp Gln Thr Leu Ala Thr 165 170 175Met Leu Val Arg Pro Gly Cys Gln Phe Leu Asp Asp Pro Thr Met Lys 180 185 190Asp Glu Ala Leu Leu Thr Glu Ala Lys Tyr Gly Ser Val Lys Lys Val 195 200 205Tyr Val Val Ala Met Ala Asp Ala Ser Asn Ser Glu Glu Met Gln Arg 210 215 220Trp Met Val Asp Met Ser Pro Gly Thr Glu Ala Glu Glu Ile Ala Gly225 230 235 240Ala Asp His Met Ala Met Cys Ser Lys Pro Arg Glu Leu Cys Asp Val 245 250 255Leu Leu Arg Ile Ala Asp Lys Tyr Glu 260 26546268PRTOryza sativa subsp. japonicamisc_featuregi|34907176 46Met Glu Ile Ser Ser Ser Ser Lys Lys His Phe Ile Leu Val His Gly1 5 10 15Leu Cys His Gly Ala Trp Cys Trp Tyr Arg Val Val Ala Ala Leu Arg 20 25 30Ala Ala Gly His Arg Ala Thr Ala Leu Asp Met Ala Ala Ser Gly Ala 35 40 45His Pro Ala Arg Val Asp Glu Val Gly Thr Phe Glu Glu Tyr Ser Arg 50 55 60Pro Leu Leu Asp Ala Val Ala Ala Ala Ala Ala Pro Gly Glu Arg Leu65 70 75 80Val Leu Val Gly His Ser His Gly Gly Leu Ser Val Ala Leu Ala Met 85 90 95Glu Arg Phe Pro Asp Lys Val Ala Ala Ala Val Phe Val Ala Ala Ala 100 105 110Met Pro Cys Val Gly Lys His Met Gly Val Pro Thr Glu Glu Phe Met 115 120 125Arg Arg Thr Ala Pro Glu Gly Leu Leu Met Asp Cys Glu Met Val Ala 130 135 140Ile Asn Asn Ser Gln Gly Ser Gly Val Ala Ile Asn Leu Gly Pro Thr145 150 155 160Phe Leu Ala Gln Lys Tyr Tyr Gln Gln Ser Pro Ala Glu Asp Leu Ala 165 170 175Leu Ala Lys Met Leu Val Arg Pro Gly Asn Gln Phe Met Asp Asp Pro 180 185 190Val Met Lys Asp Glu Ser Leu Leu Thr Asn Gly Asn Tyr Gly Ser Val 195 200 205Lys Lys Val Tyr Val Ile Ala Lys Ala Asp Ser Ser Ser Thr Glu Glu 210 215 220Met Gln Arg Trp Met Val Ala Met Ser Pro Gly Thr Asp Val Glu Glu225 230 235 240Ile Ala Gly Ala Asp His Ala Val Met Asn Ser Lys Pro Arg Glu Leu 245 250 255Cys Asp Ile Leu Ile Lys Ile Ala Asn Lys Tyr Glu 260 26547262PRTOryza sativa (japonica cultivar-group)misc_featuregi|57899620 47Met Glu Gly Ser Ser Ser Ser Ser Lys His Phe Ile Leu Val His Gly1 5 10 15Leu Cys His Gly Ala Trp Cys Trp Tyr Lys Val Val Thr Met Leu Arg 20 25 30Ser Glu Gly His Arg Val Thr Ala Leu Asp Leu Ala Ala Ser Gly Val 35 40 45His Pro Ala Arg Val Asp Glu Val His Ser Phe Glu Glu Tyr Ser Gln 50 55 60Pro Leu Leu Asp Ala Val Ala Glu Ala Pro Ala Gly Glu Arg Leu Ile65 70 75 80Leu Val Gly His Ser Phe Gly Gly Leu Ser Ile Ala Leu Ala Met Glu 85 90 95Arg Phe Pro Glu Lys Ile Ala Val Ala Val Phe Val Ala Ala Ala Val 100 105 110Pro Cys Val Gly Lys Arg Ile Ile Pro Glu Leu Ile Arg Glu Lys Ala 115 120 125Pro Lys Asp Met Leu Leu Asp Ser Lys Met Ile Pro Ile Asn Asn Lys 130 135 140Gln Gly Pro Gly Thr Ala Ile Leu Leu Gly Pro Asn Phe Leu Ala Glu145 150 155 160Lys Gly Tyr Pro Leu Ser Pro Ala Glu Asp Leu Thr Leu Ala Lys Leu 165

170 175Leu Val Arg Pro Thr Ser Gln Phe Val Asp Asp Pro Thr Met Lys Asp 180 185 190Asp Arg Leu Leu Thr Ser Ala Asn Tyr Gly Ser Val Lys Arg Val Cys 195 200 205Leu Met Ala Met Glu Asp Asp Leu Lys Glu Val His Arg Tyr Met Ile 210 215 220Thr Leu Ser Pro Gly Val Glu Val Glu Glu Ile Ala Gly Ala Asp His225 230 235 240Ala Val Met Cys Ser Arg Pro Arg Glu Leu Ser Asp Leu Leu Ala Lys 245 250 255Ile Gly Ser Lys Tyr Asp 26048265PRTCapsella rubellamisc_featuregi|15866583 48Met Gly Gly Asp Gly Gly Ala Glu Gln Pro Val Ile His Phe Val Phe1 5 10 15Val His Gly Ala Ser His Gly Ala Trp Cys Trp Tyr Lys Leu Thr Ser 20 25 30Leu Leu Glu Thr Ala Gly Phe Lys Thr Thr Ser Val Asp Leu Thr Gly 35 40 45Ala Gly Ile Ser Val Thr Asp Ser Asn Thr Val Leu Glu Ser Asp Gln 50 55 60Tyr Asn Arg Pro Leu Phe Ser Leu Leu Ser Asp Leu Pro Pro Ser His65 70 75 80Lys Val Ile Leu Val Gly His Ser Ile Gly Gly Gly Ser Val Thr Asp 85 90 95Ala Leu Cys Arg Phe Thr Asp Lys Ile Ser Met Ala Ile Tyr Leu Ala 100 105 110Ala Ser Met Val Lys Pro Gly Ser Val Pro Ser Pro His Val Ser Asp 115 120 125Met His Ala Asp Ala Arg Glu Glu Asn Ile Trp Glu Tyr Thr Tyr Gly 130 135 140Glu Gly Thr Asp Lys Pro Pro Thr Gly Val Ile Met Lys Gln Glu Phe145 150 155 160Leu Arg Gln Tyr Tyr Tyr Ser Gln Ser Pro Leu Glu Asp Val Ser Leu 165 170 175Ala Thr Lys Leu Leu Arg Pro Ala Pro Met Arg Ala Phe Gln Asp Leu 180 185 190Asp Lys Ser Pro Pro Asn Pro Glu Val Glu Lys Val Pro Arg Val Tyr 195 200 205Ile Lys Thr Gly Lys Asp Asn Leu Phe Ser Ser Val Arg Gln Asp Leu 210 215 220Leu Val Lys Asn Trp Pro Pro Ser Gln Phe Tyr Val Leu Glu Glu Ser225 230 235 240Asp His Ser Ala Phe Phe Ser Val Pro Thr Thr Leu Phe Val Tyr Leu 245 250 255Leu Arg Ala Val Ser Phe Leu His Lys 260 26549265PRTLycopersicon hirsutum f. glabratummisc_featuregi|56393011 49Met Glu Lys Ser Met Ser Pro Phe Val Lys Lys His Phe Val Leu Val1 5 10 15His Thr Ala Phe His Gly Ala Trp Cys Trp Tyr Lys Ile Val Ala Leu 20 25 30Met Arg Ser Ser Gly His Asn Val Thr Ala Leu Asp Leu Xaa Ala Ser 35 40 45Gly Ile Asn Pro Lys Gln Ala Leu Gln Ile Pro Asn Phe Ser Asp Tyr 50 55 60Leu Ser Pro Leu Met Glu Phe Met Ala Ser Leu Pro Ala Asn Glu Lys65 70 75 80Ile Ile Leu Val Gly His Ala Leu Gly Gly Leu Ala Ile Ser Lys Ala 85 90 95Met Glu Thr Phe Pro Glu Lys Ile Ser Val Ala Val Phe Leu Ser Gly 100 105 110Leu Met Pro Gly Pro Asn Ile Asp Ala Thr Thr Val Cys Thr Lys Ala 115 120 125Gly Ser Ala Val Leu Gly Gln Leu Asp Asn Cys Val Thr Tyr Glu Asn 130 135 140Gly Pro Thr Asn Pro Pro Thr Thr Leu Ile Ala Gly Pro Lys Phe Leu145 150 155 160Ala Thr Asn Val Tyr His Leu Ser Pro Ile Glu Asp Leu Ala Leu Ala 165 170 175Thr Ala Leu Val Arg Pro Leu Tyr Leu Tyr Leu Ala Glu Asp Ile Ser 180 185 190Lys Glu Val Val Leu Ser Ser Lys Arg Tyr Gly Ser Val Lys Arg Val 195 200 205Phe Ile Val Ala Thr Glu Asn Asp Ala Leu Lys Lys Glu Phe Leu Lys 210 215 220Leu Met Ile Glu Lys Asn Pro Pro Asp Glu Val Lys Glu Ile Glu Gly225 230 235 240Ser Asp His Val Thr Met Met Ser Lys Pro Gln Gln Leu Phe Thr Thr 245 250 255Leu Leu Ser Ile Ala Asn Lys Tyr Lys 260 265

Claims

1. An isolated nucleic acid molecule comprising:a) a nucleic acid having a nucleotide sequence which encodes an amino acid sequence exhibiting at least 85% sequence identity to any one of those sequences present in the Sequence Listing;b) a nucleic acid which is a complement of a nucleotide sequence according to paragraph (a);(c) a nucleic acid which is the reverse of the nucleotide sequence according to subparagraph (a), such that the reverse nucleotide sequence has a sequence order which is the reverse of the sequence order of the nucleotide sequence according to subparagraph (a); or(d) a nucleic acid capable of hybridizing to a nucleic acid according to any one of paragraphs (a)-(c), under conditions that permit formation of a nucleic acid duplex at a temperature from about 40.degree. C. and 48.degree. C. below the melting temperature of the nucleic acid duplex.

2. The isolated nucleic acid molecule according to claim 1, which has the nucleotide sequence according to any one of those sequences present in the Sequence Listing.

3. The isolated nucleic acid molecule according to claim 1, wherein said amino acid sequence comprises a polypeptide according to any one of the consensus sequences set forth in Tables 1-5, 2-6, 3-5 or 4-1.

4. The isolated nucleic acid molecule according to claim 1, wherein said amino acid sequence has a sequence according to any one of those sequences present in the Sequence Listing.

5. A vector construct comprising:a) a first nucleic acid having a regulatory sequence capable of causing transcription and/or translation in a plant; andb) a second nucleic acid having the sequence of the isolated nucleic acid molecule according to claim 1;wherein said first and second nucleic acids are operably linked andwherein said second nucleic acid is heterologous to any element in said vector construct.

6. The vector construct according to claim 5, wherein said first nucleic acid is native to said second nucleic acid.

7. The vector construct according to claim 5, wherein said first nucleic acid is heterologous to said second nucleic acid.

8. A host cell comprising an isolated nucleic acid molecule according to claim 1 wherein said nucleic acid molecule is flanked by exogenous sequence.

9. A host cell comprising a vector construct according to claim 5.

10. An isolated polypeptide comprising an amino acid sequence exhibiting at least 85% sequence identity to any of those sequences present in the Sequence Listing.

11. A method of introducing an isolated nucleic acid into a host cell comprising:a) providing an isolated nucleic acid molecule according to claim 1; andb) contacting said isolated nucleic acid with said host cell under conditions that permit insertion of said nucleic acid into said host cell.

12. A method of transforming a host cell that comprises contacting a host cell with a vector construct according to claim 5.

13. A method for detecting a nucleic acid in a sample which comprises:a) providing an isolated nucleic acid molecule according to claim 1;b) contacting said isolated nucleic acid molecule with a sample under conditions which permit a comparison of the sequence of said isolated nucleic acid molecule with the sequence of DNA in said sample; andc) analyzing the result of said comparison.

14. A plant, plant cell, plant material or seed of a plant which comprises a nucleic acid molecule according to claim 1 which is exogenous or heterologous to said plant or plant cell.

15. A plant, plant cell, plant material or seed of a plant which comprises a vector construct according to claim 5.

16. A plant that has been regenerated from a plant cell or seed according to claim 14.

17. A plant, plant cell, plant material or seed of a plant which comprises a nucleic acid molecule according to claim 1, wherein said plant has improved pH tolerance or phosphate use efficiency characteristics as compared to a wild-type plant cultivated under the same conditions.

18. A method for increasing pH tolerance or phosphate use efficiency in a plant comprising transforming a plant with a nucleic acid sequence according to claim 1.

19. A transgenic plant having a gene construct comprising a nucleic acid encoding a pH tolerance or phosphate use efficiency component operably linked to a plant promoter so that the pH tolerance or phosphate use efficiency component is ectopically overexpressed in the transgenic plant, and the transgenic plant exhibits:i) faster rate of growth,ii) greater fresh or dry weight at maturation,iii) greater fruit or seed yield,iv) higher tolerance to pH,v) higher tolerance to low phosphate concentration, orvi) higher tolerance to low nitrogen concentrationthan a progenitor plant which does not contain the polynucleotide construct, when the transgenic plant and the progenitor plant are cultivated under identical environmental conditions, wherein the pH or phosphate use efficiency component is any one of the polypeptides set forth in the Sequence Listing, or any one of the consensus sequences in claim 3.

20. A method for pH tolerance or phosphate use efficiency in a plant which comprises transforming a plant with a nucleic acid sequence that encodes a polypeptide that comprises at least one of the following:(a) an amino acid sequence that comprises the residues at positions 29-154 of the consensus sequence of Table 1-5,(b) an amino acid sequence that comprises the residues at positions 18-128 of the consensus sequence of Table 2-6,(c) an amino acid sequence that comprises the residues at positions 57-230 of the consensus sequence of Table 3-5,(d) an amino acid sequence that comprises the residues at positions 234-248 of the consensus sequence of Table 3-5, and(e) an amino acid sequence that comprises the residues at positions 10-276 of the consensus sequence of Table 4-1.

21. A plant, plant cell, plant material of a plant with improved pH tolerance or phosphate use efficiency characteristics as compared to a wild-type plant cultivated under the same conditions which comprises a nucleic acid sequence that encodes a polypeptide that comprises at least one of the following:(a) an amino acid sequence that comprises the residues at positions 29-154 of the consensus sequence of Table 1-5,(b) an amino acid sequence that comprises the residues at positions 18-128 of the consensus sequence of Table 2-6,(c) an amino acid sequence that comprises the residues at positions 57-230 of the consensus sequence of Table 3-5,(d) an amino acid sequence that comprises the residues at positions 234-248 of the consensus sequence of Table 3-5, and(e) an amino acid sequence that comprises the residues at positions 10-276 of the consensus sequence of Table 4-1.

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