Plant cinnamyl-alcohol dehydrogenase

This application claims priority benefit of U.S. Provisional Application No. 60/119,585 filed Feb. 10, 1999, now abandoned.

This invention is in the field of plant molecular biology. More specifically, this invention pertains to nucleic acid fragments encoding lignin biosynthetic enzymes in plants and seeds.

Plant cells and tissues can respond to mechanical, chemical or pathogen induced injury by producing various phenolic compounds including mono- or dimethoxylated lignin precursors derived from cinnamic acid via a complex series of biochemical reactions. These lignin precursors are eventually used by the plant to produce the phenolic macromolecule lignin which helps in wound repair by adding hydrophobicity, a physical barrier against pathogen infection and mechanical strength to the injured tissue (Vance, C. P., et al., 1980 , Annu Rev Phytopathol 18:259-288).

Cinnamyl-alcohol dehydrogenase (CAD) catalyzes the final step in the production of lignin monomers and has been implicated in plant defense response to fungal infection. Because of lignins importance in cell wall architecture and wound repair mechanisms there is considerable interest in the prospects for altering lignin quantity or quality by genetic engineering. For example, chemical treatments needed to remove lignin during the paper-pulping process are expensive and environmentally unfriendly. Plants with altered lignin quantity or quality could benefit this industry (Boudet, A., et al., 1996 , Mol Breeding 2:25-39; Campbell, M., et al., 1996 , Plant Physiol 110:3-13). Also, during soybean seed storage and processing lignin monomers are often oxidized, via polyphenol oxidase, to compounds that impart an undesirable yellow color to soybean seeds and seed products. It may be possible to reduce the amount of these phenolic compounds by suppressing CAD activity.

Cinnamyl-alcohol dehydrogenase genes described herein appear to be part of a multigene family. This application details the discovery of several different CAD genes from rice, soybean and wheat. Based on homology each of the genes (which range from 24% to 82% in similarity) have been placed into one of five different sub-groups that make up the CAD multigene family. There is a great deal of interest in identifying the genes that encode proteins involved in the production of lignin in plants. These genes may be used in plant cells to control lignin production. Accordingly, the availability of nucleic acid sequences encoding all or a portion of an enzyme involved in the production of lignin would facilitate studies to better understand lignin production in plants and provide genetic tools to enhance or otherwise alter lignin biosynthesis which in turn could provide mechanisms to control cell wall architecture, host defense and injury repair mechanisms and reduce the pool of phenolic compounds in plant cells.

The present invention relates to isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide of at least 200 amino acids that has at least 92% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of a corn cinnamyl-alcohol dehydrogenase polypeptide of SEQ ID NO:8, rice cinnamyl-alcohol dehydrogenase polypeptides of SEQ ID NOs:2 and 10, soybean cinnamyl-alcohol dehydrogenase polypeptides of SEQ ID NOs:4, 12, 20, 28, 30 and 34, and wheat cinnamyl-alcohol dehydrogenase polypeptides of SEQ ID NOs:6, 14 and 32. The present invention also relates to an isolated polynucleotide comprising the complement of the nucleotide sequences described above.

The present invention also relates to isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide of at least 103 amino acids that has at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of a corn cinnamyl-alcohol dehydrogenase polypeptide of SEQ ID NO:26, a rice cinnamyl-alcohol dehydrogenase polypeptide of SEQ ID NO:18 and wheat cinnamyl-alcohol dehydrogenase polypeptides of SEQ ID NOs:22 and 24. The present invention also relates to an isolated polynucleotide comprising the complement of the nucleotide sequences described above.

The present invention also relates to isolated polynucleotides comprising a nucleotide sequence encoding a polypeptide of at least 186 amino acids that has at least 80% identity based on the Clustal method of alignment when compared to a corn cinnamyl-alcohol dehydrogenase polypeptide of SEQ ID NO:16. The present invention also relates to an isolated polynucleotide comprising the complement of the nucleotide sequences described above.

It is preferred that the isolated polynucleotides of the claimed invention consists of a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 that codes for the polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48. The present invention also relates to an isolated polynucleotide comprising a nucleotide sequences of at least one of 60 (preferably at least one of 40, most preferably at least one of 30) contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47and the complement of such nucleotide sequences.

The present invention relates to a chimeric gene comprising an isolated polynucleotide of the present invention operably linked to suitable regulatory sequences.

The present invention relates to an isolated host cell comprising a chimeric gene of the present invention or an isolated polynucleotide of the present invention. The host cell may be eukaryotic, such as a yeast or a plant cell, or prokaryotic, such as a bacterial cell. The present invention also relates to a virus, preferably a baculovirus, comprising an isolated polynucleotide of the present invention or a chimeric gene of the present invention.

The present invention relates to a process for producing an isolated host cell comprising a chimeric gene of the present invention or an isolated polynucleotide of the present invention, the process comprising either transforming or transfecting an isolated compatible host cell with a chimeric gene or isolated polynucleotide of the present invention.

The present invention relates to a cinnamyl-alcohol dehydrogenase polypeptide of at least 200 amino acids comprising at least 92% homology based on the Clustal method of alignment compared to a polypeptide selected from the group consisting of SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 20, 28, 30, 32 and 34.

The present invention relates to a cinnamyl-alcohol dehydrogenase polypeptide of at least 103 amino acids comprising at least 80% homology based on the Clustal method of alignment compared to a polypeptide selected from the group consisting of SEQ ID NOs:18, 22, 24 and 26.

The present invention relates to a cinnamyl-alcohol dehydrogenase polypeptide of at least 186 amino acids comprising at least 80% homology based on the Clustal method of alignment compared to a polypeptide of SEQ ID NO:16.

The present invention relates to a method of selecting an isolated polynucleotide that affects the level of expression of a cinnamyl-alcohol dehydrogenase polypeptide in a host cell, preferably a plant cell, the method comprising the steps of: (a) constructing an isolated polynucleotide of the present invention or an isolated chimeric gene of the present invention; (b) introducing the isolated polynucleotide or the isolated chimeric gene into a host cell; (c) measuring the level a cinnamyl-alcohol dehydrogenase polypeptide in the host cell containing the isolated polynucleotide; and (d) comparing the level of a cinnamnyl-alcohol dehydrogenase polypeptide in the host cell containing the isolated polynucleotide with the level of a cinnamyl-alcohol dehydrogenase polypeptide in the host cell that does not contain the isolated polynucleotide.

The present invention relates to a method of obtaining a nucleic acid fragment encoding a substantial portion of a cinnamyl-alcohol dehydrogenase polypeptide gene, preferably a plant cinnamyl-alcohol dehydrogenase polypeptide gene, comprising the steps of: synthesizing an oligonucleotide primer comprising a nucleotide sequence of at least one of 60 (preferably at least one of 40, most preferably at least one of 30) contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, and the complement of such nucleotide sequences; and amplifying a nucleic acid fragment (preferably a cDNA inserted in a cloning vector) using the oligonucleotide primer. The amplified nucleic acid fragment preferably will encode a portion of a cinnamyl-alcohol dehydrogenase amino acid sequence.

The present invention also relates to a method of obtaining a nucleic acid fragment encoding all or a substantial portion of the amino acid sequence encoding a cinnamyl-alcohol dehydrogenase polypeptide comprising the steps of: probing a cDNA or genomic library with an isolated polynucleotide of the present invention; identifying a DNA clone that hybridizes with an isolated polynucleotide of the present invention; isolating the identified DNA clone; and sequencing the cDNA or genomic fragment that comprises the isolated DNA clone.

The present invention relates to a composition, such as a hybridization mixture, comprising an isolated polynucleotide of the present invention.

The present invention relates to an isolated polynucleotide of the present invention comprising at least one of 30 contiguous nucleotides derived from a nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47.

The present invention relates to an expression cassette comprising an isolated polynucleotide of the present invention operably linked to a promoter.

The present invention relates to a method for positive selection of a transformed cell comprising: (a) transforming a host cell with the chimeric gene of the present invention or an expression cassette of the present invention; and (b) growing the transformed host cell, preferably plant cell, such as a monocot or a dicot, under conditions which allow expression of the cinnamyl-alcohol dehydrogenase polynucleotide in an amount sufficient to complement a null mutant with altered cinnamyl-alcohol dehydrogenase activity and lignin production to provide a positive selection means.

The invention can be more fully understood from the following detailed description and the accompanying Sequence Listing which form a part of this application.

Table 1 lists the polypeptides that are described herein, the designation of the cDNA clones that comprise the nucleic acid fragments encoding polypeptides representing all or a substantial portion of these polypeptides, and the corresponding identifier (SEQ ID NO:) as used in the attached Sequence Listing. Table 1 also identifies the cDNA clones as individual ESTs (“EST”), the sequences of the entire cDNA inserts comprising the indicated cDNA clones (“FIS”), contigs assembled from two or more ESTs (“Contig”), contigs assembled from an FIS and one or more ESTs (“Contig*”), or sequences encoding the entire protein derived from an FIS, a contig, or an FIS and PCR (“CGS”). Nucleotide sequences, SEQ ID NOs:1, 5, 9, 11, 13, 19 and 33 and amino acid sequences SEQ ID NOs:2, 6, 10, 12, 14, 20 and 34 were determined by further sequence analysis of cDNA clones encoding the amino acid sequences set forth in SEQ ID NOs:36, 38, 40, 42, 44, 46 and 48. Nucleotide SEQ ID NOs:35, 37, 39, 41, 43, 45 and 47 and amino acid SEQ ID NOs:36, 38, 40, 42, 44, 46 and 48 were presented in a U.S. Provisional Application No. 60/119,585, filed Feb. 10, 1999.

The sequence descriptions and Sequence Listing attached hereto comply with the rules governing nucleotide and/or amino acid sequence disclosures in patent applications as set forth in 37 C.F.R. §1.821-1.825.

The Sequence Listing contains the one letter code for nucleotide sequence characters and the three letter codes for amino acids as defined in conformity with the IUPAC-IUBMB standards described in Nucleic Acids Res . 13:3021-3030 (1985) and in the Biochemical J . 219 (No. 2):345-373 (1984) which are herein incorporated by reference. The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. §1.822.

In the context of this disclosure, a number of terms shall be utilized. As used herein, a “polynucleotide” is a nucleotide sequence such as a nucleic acid fragment. A polynucleotide may be a polymer of RNA or DNA that is single- or double-stranded, that optionally contains synthetic, non-natural or altered nucleotide bases. A polynucleotide in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA, synthetic DNA, or mixtures thereof. An isolated polynucleotide of the present invention may include at least one of 60 contiguous nucleotides, preferably at least one of 40 contiguous nucleotides, most preferably one of at least 30 contiguous nucleotides derived from SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47 or the complement of such sequences.

As used herein, “contig” refers to a nucleotide sequence that is assembled from two or more constituent nucleotide sequences that share common or overlapping regions of sequence homology. For example, the nucleotide sequences of two or more nucleic acid fragments can be compared and aligned in order to identify common or overlapping sequences. Where common or overlapping sequences exist between two or more nucleic acid fragments, the sequences (and thus their corresponding nucleic acid fragments) can be assembled into a single contiguous nucleotide sequence.

As used herein, “substantially similar” refers to nucleic acid fragments wherein changes in one or more nucleotide bases results in substitution of one or more amino acids, but do not affect the functional properties of the polypeptide encoded by the nucleotide sequence. “Substantially similar” also refers to nucleic acid fragments wherein changes in one or more nucleotide bases does not affect the ability of the nucleic acid fragment to mediate alteration of gene expression by gene silencing through for example antisense or co-suppression technology. “Substantially similar” also refers to modifications of the nucleic acid fragments of the instant invention such as deletion or insertion of one or more nucleotides that do not substantially affect the functional properties of the resulting transcript vis-a-vis the ability to mediate gene silencing or alteration of the functional properties of the resulting protein molecule. It is therefore understood that the invention encompasses more than the specific exemplary nucleotide or amino acid sequences and includes functional equivalents thereof.

Substantially similar nucleic acid fragments may be selected by screening nucleic acid fragments representing subfragments or modifications of the nucleic acid fragments of the instant invention, wherein one or more nucleotides are substituted, deleted and/or inserted, for their ability to affect the level of the polypeptide encoded by the unmodified nucleic acid fragment in a plant or plant cell. For example, a substantially similar nucleic acid fragment representing at least one of 30 contiguous nucleotides derived from the instant nucleic acid fragment can be constructed and introduced into a plant or plant cell. The level of the polypeptide encoded by the unmodified nucleic acid fragment present in a plant or plant cell exposed to the substantially similar nucleic fragment can then be compared to the level of the polypeptide in a plant or plant cell that is not exposed to the substantially similar nucleic acid fragment.

For example, it is well known in the art that antisense suppression and co-suppression of gene expression may be accomplished using nucleic acid fragments representing less than the entire coding region of a gene, and by nucleic acid fragments that do not share 100% sequence identity with the gene to be suppressed. Moreover, alterations in a nucleic acid fragment which result in the production of a chemically equivalent amino acid at a given site, but do not effect the functional properties of the encoded polypeptide, are well known in the art. Thus, a codon for the amino acid alanine, a hydrophobic amino acid, may be substituted by a codon encoding another less hydrophobic residue, such as glycine, or a more hydrophobic residue, such as valine, leucine, or isoleucine. Similarly, changes which result in substitution of one negatively charged residue for another, such as aspartic acid for glutamic acid, or one positively charged residue for another, such as lysine for arginine, can also be expected to produce a functionally equivalent product. Nucleotide changes which result in alteration of the N-terminal and C-terminal portions of the polypeptide molecule would also not be expected to alter the activity of the polypeptide. Each of the proposed modifications is well within the routine skill in the art, as is determination of retention of biological activity of the encoded products. Consequently, an isolated polynucleotide comprising a nucleotide sequence of at least one of 60 (preferably at least one of 40, most preferably at least one of 30) contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47 and the complement of such nucleotide sequences may be used in methods of selecting an isolated polynucleotide that affects the expression of a polypeptide (cinnamyl-alcohol dehydrogenase) in a host cell. A method of selecting an isolated polynucleotide that affects the level of expression of a polypeptide in a host cell (eukaryotic, such as plant or yeast, prokaryotic such as bacterial, or viral) may comprise the steps of: constructing an isolated polynucleotide of the present invention or an isolated chimeric gene of the present invention; introducing the isolated polynucleotide or the isolated chimeric gene into a host cell; measuring the level a polypeptide in the host cell containing the isolated polynucleotide; and comparing the level of a polypeptide in the host cell containing the isolated polynucleotide with the level of a polypeptide in a host cell that does not contain the isolated polynucleotide.

Moreover, substantially similar nucleic acid fragments may also be characterized by their ability to hybridize. Estimates of such homology are provided by either DNA-DNA or DNA-RNA hybridization under conditions of stringency as is well understood by those skilled in the art (Hames and Higgins, Eds. (1985) Nucleic Acid Hybridisation, IRL Press, Oxford, U.K.). Stringency conditions can be adjusted to screen for moderately similar fragments, such as homologous sequences from distantly related organisms, to highly similar fragments, such as genes that duplicate functional enzymes from closely related organisms. Post-hybridization washes determine stringency conditions. One set of preferred conditions uses a series of washes starting with 6×SSC, 0.5% SDS at room temperature for 15 min, then repeated with 2×SSC, 0.5% SDS at 45° C. for 30 min, and then repeated twice with 0.2×SSC, 0.5% SDS at 50° C. for 30 min. A more preferred set of stringent conditions uses higher temperatures in which the washes are identical to those above except for the temperature of the final two 30 min washes in 0.2×SSC, 0.5% SDS was increased to 60° C. Another preferred set of highly stringent conditions uses two final washes in 0.1×SSC, 0.1% SDS at 65° C.

Substantially similar nucleic acid fragments of the instant invention may also be characterized by the percent identity of the amino acid sequences that they encode to the amino acid sequences disclosed herein, as determined by algorithms commonly employed by those skilled in this art. Suitable nucleic acid fragments (isolated polynucleotides of the present invention) encode polypeptides that are at least about 70% identical, preferably at least about 80% identical to the amino acid sequences reported herein. Preferred nucleic acid fragments encode amino acid sequences that are about 85% identical to the amino acid sequences reported herein. More preferred nucleic acid fragments encode amino acid sequences that are at least about 90% identical to the amino acid sequences reported herein. Most preferred are nucleic acid fragments that encode amino acid sequences that are at least about 95% identical to the amino acid sequences reported herein. Suitable nucleic acid fragments not only have the above homologies but typically encode a polypeptide having at least 50 amino acids, preferably at least 100 amino acids, more preferably at least 150 amino acids, still more preferably at least 200 amino acids, and most preferably at least 250 amino acids. Sequence alignments and percent identity calculations were performed using the Megalign program of the LASERGENE bioinformatics computing suite (DNASTAR Inc., Madison, Wis.). Multiple alignment of the sequences was performed using the Clustal method of alignment (Higgins and Sharp (1989) CABIOS . 5:151-153) with the default parameters (GAP PENALTY=10, GAP LENGTH PENALTY=10). Default parameters for pairwise alignments using the Clustal method were KTUPLE 1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5.

A “substantial portion” of an amino acid or nucleotide sequence comprises an amino acid or a nucleotide sequence that is sufficient to afford putative identification of the protein or gene that the amino acid or nucleotide sequence comprises. Amino acid and nucleotide sequences can be evaluated either manually by one skilled in the art, or by using computer-based sequence comparison and identification tools that employ algorithms such as BLAST (Basic Local Alignment Search Tool; Altschul et al. (1993) J. Mol. Biol . 215:403-410). In general, a sequence of ten or more contiguous amino acids or thirty or more contiguous nucleotides is necessary in order to putatively identify a polypeptide or nucleic acid sequence as homologous to a known protein or gene. Moreover, with respect to nucleotide sequences, gene-specific oligonucleotide probes comprising 30 or more contiguous nucleotides may be used in sequence-dependent methods of gene identification (e.g., Southern hybridization) and isolation (e.g., in situ hybridization of bacterial colonies or bacteriophage plaques). In addition, short oligonucleotides of 12 or more nucleotides may be used as amplification primers in PCR in order to obtain a particular nucleic acid fragment comprising the primers. Accordingly, a “substantial portion” of a nucleotide sequence comprises a nucleotide sequence that will afford specific identification and/or isolation of a nucleic acid fragment comprising the sequence. The instant specification teaches amino acid and nucleotide sequences encoding polypeptides that comprise one or more particular plant proteins. The skilled artisan, having the benefit of the sequences as reported herein, may now use all or a substantial portion of the disclosed sequences for purposes known to those skilled in this art. Accordingly, the instant invention comprises the complete sequences as reported in the accompanying Sequence Listing, as well as substantial portions of those sequences as defined above.

“Codon degeneracy” refers to divergence in the genetic code permitting variation of the nucleotide sequence without effecting the amino acid sequence of an encoded polypeptide. Accordingly, the instant invention relates to any nucleic acid fragment comprising a nucleotide sequence that encodes all or a substantial portion of the amino acid sequences set forth herein. The skilled artisan is well aware of the “codon-bias” exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid. Therefore, when synthesizing a nucleic acid fragment for improved expression in a host cell, it is desirable to design the nucleic acid fragment such that its frequency of codon usage approaches the frequency of preferred codon usage of the host cell.

“Synthetic nucleic acid fragments” can be assembled from oligonucleotide building blocks that are chemically synthesized using procedures known to those skilled in the art. These building blocks are ligated and annealed to form larger nucleic acid fragments which may then be enzymatically assembled to construct the entire desired nucleic acid fragment. “Chemically synthesized”, as related to nucleic acid fragment, means that the component nucleotides were assembled in vitro. Manual chemical synthesis of nucleic acid fragments may be accomplished using well established procedures, or automated chemical synthesis can be performed using one of a number of commercially available machines. Accordingly, the nucleic acid fragments can be tailored for optimal gene expression based on optimization of nucleotide sequence to reflect the codon bias of the host cell. The skilled artisan appreciates the likelihood of successful gene expression if codon usage is biased towards those codons favored by the host. Determination of preferred codons can be based on a survey of genes derived from the host cell where sequence information is available.

“Gene” refers to a nucleic acid fragment that expresses a specific protein, including regulatory sequences preceding (5′ non-coding sequences) and following (3′ non-coding sequences) the coding sequence. “Native gene” refers to a gene as found in nature with its own regulatory sequences. “Chimeric gene” refers any gene that is not a native gene, comprising regulatory and coding sequences that are not found together in nature. Accordingly, a chimeric gene may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different than that found in nature. “Endogenous gene” refers to a native gene in its natural location in the genome of an organism. A “foreign” gene refers to a gene not normally found in the host organism, but that is introduced into the host organism by gene transfer. Foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes. A “transgene” is a gene that has been introduced into the genome by a transformation procedure. “Coding sequence” refers to a nucleotide sequence that codes for a specific amino acid sequence. “Regulatory sequences” refer to nucleotide sequences located upstream (5′ non-coding sequences), within, or downstream (3′ non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include promoters, translation leader sequences, introns, and polyadenylation recognition sequences.

“Promoter” refers to a nucleotide sequence capable of controlling the expression of a coding sequence or functional RNA. In general, a coding sequence is located 3′ to a promoter sequence. The promoter sequence consists of proximal and more distal upstream elements, the latter elements often referred to as enhancers. Accordingly, an “enhancer” is a nucleotide sequence which can stimulate promoter activity and may be an innate element of the promoter or a heterologous element inserted to enhance the level or tissue-specificity of a promoter. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic nucleotide segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. Promoters which cause a nucleic acid fragment to be expressed in most cell types at most times are commonly referred to as “constitutive promoters”. New promoters of various types useful in plant cells are constantly being discovered; numerous examples may be found in the compilation by Okamuro and Goldberg (1989) Biochemistry of Plants 15:1-82. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, nucleic acid fragments of different lengths may have identical promoter activity.

The “translation leader sequence” refers to a nucleotide sequence located between the promoter sequence of a gene and the coding sequence. The translation leader sequence is present in the filly processed mRNA upstream of the translation start sequence. The translation leader sequence may affect processing of the primary transcript to mRNA, mRNA stability or translation efficiency. Examples of translation leader sequences have been described (Turner and Foster (1995) Mol. Biotechnol . 3:225-236).

The “3′ non-coding sequences” refer to nucleotide sequences located downstream of a coding sequence and include polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal is usually characterized by affecting the addition of polyadenylic acid tracts to the 3′ end of the mRNA precursor. The use of different 3′ non-coding sequences is exemplified by Ingelbrecht et al. (1989) Plant Cell 1:671-680.

“RNA transcript” refers to the product resulting from RNA polymerase-catalyzed transcription of a DNA sequence. When the RNA transcript is a perfect complementary copy of the DNA sequence, it is referred to as the primary transcript or it may be a RNA sequence derived from posttranscriptional processing of the primary transcript and is referred to as the mature RNA. “Messenger RNA (mRNA)” refers to the RNA that is without introns and that can be translated into polypeptide by the cell. “cDNA” refers to a double-stranded DNA that is complementary to and derived from mRNA. “Sense” RNA refers to an RNA transcript that includes the mRNA and so can be translated into a polypeptide by the cell. “Antisense RNA” refers to an RNA transcript that is complementary to all or part of a target primary transcript or mRNA and that blocks the expression of a target gene (see U.S. Pat. No. 5,107,065, incorporated herein by reference). The complementarity of an antisense RNA may be with any part of the specific nucleotide sequence, i.e., at the 5′ non-coding sequence, 3′ non-coding sequence, introns, or the coding sequence. “Functional RNA” refers to sense RNA, antisense RNA, ribozyme RNA, or other RNA that may not be translated but yet has an effect on cellular processes.

The term “operably linked” refers to the association of two or more nucleic acid fragments on a single nucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably linked with a coding sequence when it is capable of affecting the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory sequences in sense or antisense orientation.

The term “expression”, as used herein, refers to the transcription and stable accumulation of sense (mRNA) or antisense RNA derived from the nucleic acid fragment of the invention. Expression may also refer to translation of mRNA into a polypeptide. “Antisense inhibition” refers to the production of antisense RNA transcripts capable of suppressing the expression of the target protein. “Overexpression” refers to the production of a gene product in transgenic organisms that exceeds levels of production in normal or non-transformed organisms. “Co-suppression” refers to the production of sense RNA transcripts capable of suppressing the expression of identical or substantially similar foreign or endogenous genes (U.S. Pat. No. 5,231,020, incorporated herein by reference).

“Altered levels” refers to the production of gene product(s) in transgenic organisms in amounts or proportions that differ from that of normal or non-transformed organisms.

“Mature” protein refers to a post-translationally processed polypeptide; i.e., one from which any pre- or propeptides present in the primary translation product have been removed. “Precursor” protein refers to the primary product of translation of mRNA; i.e., with pre- and propeptides still present. Pre- and propeptides may be but are not limited to intracellular localization signals.

A “chloroplast transit peptide” is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the chloroplast or other plastid types present in the cell in which the protein is made. “Chloroplast transit sequence” refers to a nucleotide sequence that encodes a chioroplast transit peptide. A “signal peptide” is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the secretory system (Chrispeels (1991) Ann. Rev. Plant Phys. Plant Mol. Biol . 42:21-53). If the protein is to be directed to a vacuole, a vacuolar targeting signal (supra) can further be added, or if to the endoplasmic reticulum, an endoplasmic reticulum retention signal (supra) may be added. If the protein is to be directed to the nucleus, any signal peptide present should be removed and instead a nuclear localization signal included (Raikhel (1992) Plant Phys . 100: 1627-1632).

“Transformation” refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in genetically stable inheritance. Host organisms containing the transformed nucleic acid fragments are referred to as “transgenic” organisms. Examples of methods of plant transformation include Agrobacterium-mediated transformation (De Blaere et al. (1987) Meth. Enzymol . 143:277) and particle-accelerated or “gene gun” transformation technology (Klein et al. (1987) Nature ( London ) 327:70-73; U.S. Patent No. 4,945,050, incorporated herein by reference).

Standard recombinant DNA and molecular cloning techniques used herein are well known in the art and are described more fully in Sambrook et al. Molecular Cloning: A Laboratory Manual ; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, 1989 (hereinafter “Maniatis”).

Nucleic acid fragments encoding at least a portion of several lignin biosynthetic enzymes have been isolated and identified by comparison of random plant cDNA sequences to public databases containing nucleotide and protein sequences using the BLAST algorithms well known to those skilled in the art. The nucleic acid fragments of the instant invention may be used to isolate cDNAs and genes encoding homologous proteins from the same or other plant species. Isolation of homologous genes using sequence-dependent protocols is well known in the art. Examples of sequence-dependent protocols include, but are not limited to, methods of nucleic acid hybridization, and methods of DNA and RNA amplification as exemplified by various uses of nucleic acid amplification technologies (e.g., polymerase chain reaction, ligase chain reaction).

For example, genes encoding other cinnamyl-alcohol dehydrogenase proteins, either as cDNAs or genomic DNAs, could be isolated directly by using all or a portion of the instant nucleic acid fragments as DNA hybridization probes to screen libraries from any desired plant employing methodology well known to those skilled in the art. Specific oligonucleotide probes based upon the instant nucleic acid sequences can be designed and synthesized by methods known in the art (Maniatis). Moreover, the entire sequences can be used directly to synthesize DNA probes by methods known to the skilled artisan such as random primer DNA labeling, nick translation, or end-labeling techniques, or RNA probes using available in vitro transcription systems. In addition, specific primers can be designed and used to amplify a part or all of the instant sequences. The resulting amplification products can be labeled directly during amplification reactions or labeled after amplification reactions, and used as probes to isolate full length cDNA or genomic fragments under conditions of appropriate stringency.

In addition, two short segments of the instant nucleic acid fragments may be used in polymerase chain reaction protocols to amplify longer nucleic acid fragments encoding homologous genes from DNA or RNA. The polymerase chain reaction may also be performed on a library of cloned nucleic acid fragments wherein the sequence of one primer is derived from the instant nucleic acid fragments, and the sequence of the other primer takes advantage of the presence of the polyadenylic acid tracts to the 3′ end of the mRNA precursor encoding plant genes. Alternatively, the second primer sequence may be based upon sequences derived from the cloning vector. For example, the skilled artisan can follow the RACE protocol (Frohman et al. (1988) Proc. Natl. Acad. Sci. USA 85:8998-9002) to generate cDNAs by using PCR to amplify copies of the region between a single point in the transcript and the 3′ or 5′ end. Primers oriented in the 3′ and 5′ directions can be designed from the instant sequences. Using commercially available 3′ RACE or 5′ RACE systems (BRL), specific 3′ or 5′ cDNA fragments can be isolated (Ohara et al. (1989) Proc. Natl. Acad. Sci. USA 86:5673-5677; Loh et al. (1989) Science 243:217-220). Products generated by the 3′ and 5′ RACE procedures can be combined to generate full-length cDNAs (Frohman and Martin (1989) Techniques 1:165). Consequently, a polynucleotide comprising a nucleotide sequence of at least one of 60 (preferably one of at least 40, most preferably one of at least 30) contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, the complement of such nucleotide sequences may be used in such methods to obtain a nucleic acid fragment encoding a substantial portion of an amino acid sequence of a polypeptide. The present invention relates to a method of obtaining a nucleic acid fragment encoding a substantial portion of a polypeptide of a gene (such as cinnamyl-alcohol dehydrogenase) preferably a substantial portion of a plant polypeptide of a gene, comprising the steps of: synthesizing an oligonucteotide primer comprising a nucleotide sequence of at least one of 60 (preferably at least one of 40, most preferably at least one of 30) contiguous nucleotides derived from a nucleotide sequence selected from the group consisting of SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47 and the complement of such nucleotide sequences; and amplifying a nucleic acid fragment (preferably a cDNA inserted in a cloning vector) using the oligonucleotide primer. The amplified nucleic acid fragment preferably will encode a portion of a polypeptide (cinnamyl-alcohol dehydrogenase).

Availability of the instant nucleotide and deduced amino acid sequences facilitates immunological screening of cDNA expression libraries. Synthetic peptides representing portions of the instant amino acid sequences may be synthesized. These peptides can be used to immunize animals to produce polyclonal or monoclonal antibodies with specificity for peptides or proteins comprising the amino acid sequences. These antibodies can be then be used to screen cDNA expression libraries to isolate full-length cDNA clones of interest (Lemer (1984) Adv. Immunol . 36:1-34; Maniatis).

The nucleic acid fragments of the instant invention may be used to create transgenic plants in which the disclosed polypeptides are present at higher or lower levels than normal or in cell types or developmental stages in which they are not normally found. This would have the effect of altering the level of cinnamyl-alcohol dehydrogenase activity in those cells.

Overexpression of the proteins of the instant invention may be accomplished by first constructing a chimeric gene in which the coding region is operably linked to a promoter capable of directing expression of a gene in the desired tissues at the desired stage of development. The chimeric gene may comprise promoter sequences and translation leader sequences derived from the same genes. 3′ Non-coding sequences encoding transcription termination signals may also be provided. The instant chimeric gene may also comprise one or more introns in order to facilitate gene expression.

Plasmid vectors comprising the isolated polynucleotide (or chimeric gene) may be constructed. The choice of plasmid vector is dependent upon the method that will be used to transform host plants. The skilled artisan is well aware of the genetic elements that must be present on the plasmid vector in order to successfully transform, select and propagate host cells containing the chimeric gene. The skilled artisan will also recognize that different independent transformation events will result in different levels and patterns of expression (Jones et al. (1985) EMBO J . 4:2411-2418; De Almeida et al. (1989) Mol. Gen. Genetics 218:78-86), and thus that multiple events must be screened in order to obtain lines displaying the desired expression level and pattern. Such screening may be accomplished by Southern analysis of DNA, Northern analysis of mRNA expression, Western analysis of protein expression, or phenotypic analysis.

For some applications it may be useful to direct the instant polypeptides to different cellular compartments, or to facilitate its secretion from the cell. It is thus envisioned that the chimeric gene described above may be further supplemented by directing the coding sequence to encode the instant polypeptides with appropriate intracellular targeting sequences such as transit sequences (Keegstra (1989) Cell 56:247-253), signal sequences or sequences encoding endoplasmic reticulum localization (Chrispeels (1991) Ann. Rev. Plant Phys. Plant Mol. Biol . 42:21-53), or nuclear localization signals (Raikhel (1992) Plant Phys . 100:1627-1632) with or without removing targeting sequences that are already present. While the references cited give examples of each of these, the list is not exhaustive and more targeting signals of use may be discovered in the future.

It may also be desirable to reduce or eliminate expression of genes encoding the instant polypeptides in plants for some applications. In order to accomplish this, a chimeric gene designed for co-suppression of the instant polypeptide can be constructed by linking a gene or gene fragment encoding that polypeptide to plant promoter sequences. Alternatively, a chimeric gene designed to express antisense RNA for all or part of the instant nucleic acid fragment can be constructed by linking the gene or gene fragment in reverse orientation to plant promoter sequences. Either the co-suppression or antisense chimeric genes could be introduced into plants via transformation wherein expression of the corresponding endogenous genes are reduced or eliminated.

Molecular genetic solutions to the generation of plants with altered gene expression have a decided advantage over more traditional plant breeding approaches. Changes in plant phenotypes can be produced by specifically inhibiting expression of one or more genes by antisense inhibition or cosuppression (U.S. Pat. Nos. 5,190,931, 5,107,065 and 5,283,323). An antisense or cosuppression construct would act as a dominant negative regulator of gene activity. While conventional mutations can yield negative regulation of gene activity these effects are most likely recessive. The dominant negative regulation available with a transgenic approach may be advantageous from a breeding perspective. In addition, the ability to restrict the expression of specific phenotype to the reproductive tissues of the plant by the use of tissue specific promoters may confer agronomic advantages relative to conventional mutations which may have an effect in all tissues in which a mutant gene is ordinarily expressed.

The person skilled in the art will know that special considerations are associated with the use of antisense or cosuppression technologies in order to reduce expression of particular genes. For example, the proper level of expression of sense or antisense genes may require the use of different chimeric genes utilizing different regulatory elements known to the skilled artisan. Once transgenic plants are obtained b one of the methods described above, it will be necessary to screen individual transgenics for those that most effectively display the desired phenotype. Accordingly, the skilled artisan will develop methods for screening large numbers of transformants. The nature of these screens will generally be chosen on practical grounds. For example, one can screen by looking for changes in gene expression by using antibodies specific for the protein encoded by the gene being suppressed, or one could establish assays that specifically measure enzyme activity. A preferred method will be one which allows large numbers of samples to be processed rapidly, since it will be expected that a large number of transformants will be negative for the desired phenotype.

The instant polypeptides (or portions thereof) may be produced in heterologous host cells, particularly in the cells of microbial hosts, and can be used to prepare antibodies to the these proteins by methods well known to those skilled in the art. The antibodies are useful for detecting the polypeptides of the instant invention in situ in cells or in vitro in cell extracts. Preferred heterologous host cells for production of the instant polypeptides are microbial hosts. Microbial expression systems and expression vectors containing regulatory sequences that direct high level expression of foreign proteins are well known to those skilled in the art. Any of these could be used to construct a chimeric gene for production of the instant polypeptides. This chimeric gene could then be introduced into appropriate microorganisms via transformation to provide high level expression of the encoded lignin biosynthetic enzyme. An example of a vector for high level expression of the instant polypeptides in a bacterial host is provided (Example 6).

All or a substantial portion of the nucleic acid fragments of the instant invention may also be used as probes for genetically and physically mapping the genes that they are a part of, and as markers for traits linked to those genes. Such information may be useful in plant breeding in order to develop lines with desired phenotypes. For example, the instant nucleic acid fragments may be used as restriction fragment length polymorphism (RFLP) markers. Southern blots (Maniatis) of restriction-digested plant genomic DNA may be probed with the nucleic acid fragments of the instant invention. The resulting banding patterns may then be subjected to genetic analyses using computer programs such as MapMaker (Lander et al. (1987) Genomics 1:174-181) in order to construct a genetic map. In addition, the nucleic acid fragments of the instant invention may be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs of a set of individuals representing parent and progeny of a defined genetic cross. Segregation of the DNA polymorphisms is noted and used to calculate the position of the instant nucleic acid sequence in the genetic map previously obtained using this population (Botstein et al. (1980) Am. J. Hum. Genet . 32:314-331).

The production and use of plant gene-derived probes for use in genetic mapping is described in Bernatzky and Tanksley (1986) Plant Mol. Biol. Reporter 4:37-41. Numerous publications describe genetic mapping of specific cDNA clones using the methodology outlined above or variations thereof. For example, F2 intercross populations, backcross populations, randomly mated populations, near isogenic lines, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes derived from the instant nucleic acid sequences may also be used for physical mapping (i.e., placement of sequences on physical maps; see Hoheisel et al. In: Nonmammalian Genomic Analysis: A Practical Guide , Academic press 1996, pp. 319-346, and references cited therein).

In another embodiment, nucleic acid probes derived from the instant nucleic acid sequences may be used in direct fluorescence in situ hybridization (FISH) mapping (Trask (1991) Trends Genet . 7:149-154). Although current methods of FISH mapping favor use of large clones (several to several hundred KB; see Laan et al. (1995) Genome Res . 5:13-20), improvements in sensitivity may allow performance of FISH mapping using shorter probes.

A variety of nucleic acid amplification-based methods of genetic and physical mapping may be carried out using the instant nucleic acid sequences. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med . 11:95-96), polymorphism of PCR-amplified fragments (CAPS; Sheffield et al. (1993) Genomics 16:325-332), allele-specific ligation (Landegren et al. (1988) Science 241:1077-1080), nucleotide extension reactions (Sokolov (1990) Nucleic Acid Res . 18:3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet . 7:22-28) and Happy Mapping (Dear and Cook (1989) Nucleic Acid Res . 17:6795-6807). For these methods, the sequence of a nucleic acid fragment is used to design and produce primer pairs for use in the amplification reaction or in primer extension reactions. The design of such primers is well known to those skilled in the art. In methods employing PCR-based genetic map-ping, it may be necessary to identify DNA sequence differences between the parents of the mapping cross in the region corresponding to the instant nucleic acid sequence. This, however, is generally not necessary for mapping methods.

Loss of function mutant phenotypes may be identified for the instant cDNA clones either by targeted gene disruption protocols or by identifying specific mutants for these genes contained in a maize population carrying mutations in all possible genes (Ballinger and Benzer (1989) Proc. Natl. Acad Sci USA 86:9402-9406; Koes et al. (1995) Proc. Natl. Acad. Sci USA 92:8149-8153; Bensen et al. (1995) Plant Cell 7:75-84). The latter approach may be accomplished in two ways. First, short segments of the instant nucleic acid fragments may be used in polymerase chain reaction protocols in conjunction with a mutation tag sequence primer on DNAs prepared from a population of plants in which Mutator transposons or some other mutation-causing DNA element has been introduced (see Bensen, supra). The amplification of a specific DNA fragment with these primers indicates the insertion of the mutation tag element in or near the plant gene encoding the instant polypeptides. Alternatively, the instant nucleic acid fragment may be used as a hybridization probe against PCR amplification products generated from the mutation population using the mutation tag sequence primer in conjunction with an arbitrary genomic site primer, such as that for a restriction enzyme site-anchored synthetic adaptor. With either method, a plant containing a mutation in the endogenous gene encoding the instant polypeptides can be identified and obtained. This mutant plant can then be used to determine or confirm the natural function of the instant polypeptides disclosed herein.

The present invention is further defined in the following Examples, in which all parts and percentages are by weight and degrees are Celsius, unless otherwise stated. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Oryza sativa

1

gcacgaggtt ctaaccgccg gcgagaaacc tgcagagggc gagggcgagt gcggcggagc 60

aggggatgga ggagcatggc aaggcggctg tcggctgggc ggccagggac gactccggcg 120

tcctctcccc gtacaacttc tccaggaggg ctcagaagga cgatgacgtc accatcaagg 180

tgctctactg cggcatctgc cacactgacc tgcacatcgt caagaatgat tggggcaacg 240

ccatgtaccc cgtcgtcccc ggccacgaga tcgtcggcgt cgtcaccggc gtcggcgccg 300

gcgtgaccaa gttcaaggcc ggcgacacgg tgggcgtggg ctacttcgtc gcgtcgtgcc 360

gcggctgcga gtgctgcggg aatgggtacg agaactactg cgccaagatg gtaaccacct 420

gcaacggcgt cgaccacgac cacggcggcg gcgcggccac ccagggcggc ttctccgacg 480

ccatcgtcgt caacgagcac tacgtgctcc gcgtgccggc ggggctgccg ctggacagcg 540

cggcgccgct gctgtgcgcc ggcgtgacgg tgtacagccc gatggtgatc cacggcctga 600

acgcgccggg gaagcacgtc ggcgtcgtcg gcctcggcgg gctcggccac gtcgccgtca 660

agttcgccaa ggcgttcggg atgcgcgtca ccgtcatcag cacgtcgccg gggaagcggc 720

aggaggcact ggagcacctc ggcgccgacg agttcctcgt cagccgcgac gcgggccaga 780

tggcggcggc ggcggccacc atggacggga tcctcaacac ggtgtcggcc tggcacccga 840

tcgcgccgct gttctcgctg atgaagccga tggcgcagat ggtgttcgtc ggcgggccga 900

ccaggccgct ggagctgccg gcgtacgcca tcgtgcccgg agggaagggc atcaccggca 960

actgcgtcgg cggcatcagg gactgccagg ccatgctcga cttcgccggc gagcacggca 1020

tcactgccga ggtggaggtc atcaagatgg actacgtcaa cacggcgatg gagcggctgg 1080

agaagaacga tgtccgctac cgcttcgtca tcgacgttgc cggcagcagc ctcgccggct 1140

ccggcgacgc caagatataa agctcgccgg cttgcttgct tgcttgtgtc gacggcgccg 1200

gcgccgccta gcttgcttgt tgtgtgtcag tggccttgaa tgtttggtga tgtgcaaact 1260

actacgaaat ggagtaagaa ataagtggag catatgtctc catcacttgg ttattaatta 1320

aaaaaaaaaa aaaaaaaaa 1339

2

364

Oryza sativa

2

Met Glu Glu His Gly Lys Ala Ala Val Gly Trp Ala Ala Arg Asp Asp

1 5 10 15

Ser Gly Val Leu Ser Pro Tyr Asn Phe Ser Arg Arg Ala Gln Lys Asp

20 25 30

Asp Asp Val Thr Ile Lys Val Leu Tyr Cys Gly Ile Cys His Thr Asp

35 40 45

Leu His Ile Val Lys Asn Asp Trp Gly Asn Ala Met Tyr Pro Val Val

50 55 60

Pro Gly His Glu Ile Val Gly Val Val Thr Gly Val Gly Ala Gly Val

65 70 75 80

Thr Lys Phe Lys Ala Gly Asp Thr Val Gly Val Gly Tyr Phe Val Ala

85 90 95

Ser Cys Arg Gly Cys Glu Cys Cys Gly Asn Gly Tyr Glu Asn Tyr Cys

100 105 110

Ala Lys Met Val Thr Thr Cys Asn Gly Val Asp His Asp His Gly Gly

115 120 125

Gly Ala Ala Thr Gln Gly Gly Phe Ser Asp Ala Ile Val Val Asn Glu

130 135 140

His Tyr Val Leu Arg Val Pro Ala Gly Leu Pro Leu Asp Ser Ala Ala

145 150 155 160

Pro Leu Leu Cys Ala Gly Val Thr Val Tyr Ser Pro Met Val Ile His

165 170 175

Gly Leu Asn Ala Pro Gly Lys His Val Gly Val Val Gly Leu Gly Gly

180 185 190

Leu Gly His Val Ala Val Lys Phe Ala Lys Ala Phe Gly Met Arg Val

195 200 205

Thr Val Ile Ser Thr Ser Pro Gly Lys Arg Gln Glu Ala Leu Glu His

210 215 220

Leu Gly Ala Asp Glu Phe Leu Val Ser Arg Asp Ala Gly Gln Met Ala

225 230 235 240

Ala Ala Ala Ala Thr Met Asp Gly Ile Leu Asn Thr Val Ser Ala Trp

245 250 255

His Pro Ile Ala Pro Leu Phe Ser Leu Met Lys Pro Met Ala Gln Met

260 265 270

Val Phe Val Gly Gly Pro Thr Arg Pro Leu Glu Leu Pro Ala Tyr Ala

275 280 285

Ile Val Pro Gly Gly Lys Gly Ile Thr Gly Asn Cys Val Gly Gly Ile

290 295 300

Arg Asp Cys Gln Ala Met Leu Asp Phe Ala Gly Glu His Gly Ile Thr

305 310 315 320

Ala Glu Val Glu Val Ile Lys Met Asp Tyr Val Asn Thr Ala Met Glu

325 330 335

Arg Leu Glu Lys Asn Asp Val Arg Tyr Arg Phe Val Ile Asp Val Ala

340 345 350

Gly Ser Ser Leu Ala Gly Ser Gly Asp Ala Lys Ile

355 360

3

1463

Glycine max

3

attacccaag ytcgaaatka acctcactaa agggaacaaa agctggagct ccaccgcggt 60

ggcggccgct ctagaactag tggatccccc gggctgcagg aattcggcac gagcagcaaa 120

gggaaaagca aacagagtgt ctaatcgaag atagaagaag agatggctaa atctccagaa 180

actgaacttc ccgtcgaagc tttcggttgg gctgcttcag acacttctgg cacccttgca 240

ccttttcatt tctccagaag agaaaatggc gttgatgacg tcactctcaa gattctcttc 300

tgcggggttt gccactcgga tctccacact ctcaaaaacg actggggttt caccacttac 360

cctgtcgttc ctgggcatga aattgttggt gttgtgacaa aagttggcaa caatgtgaaa 420

aacttcaaag tgggtgataa agttggagtt ggtgtgatag tggagtcttg caaggaatgt 480

gaaagttgcc aacaagatct ggagagttac tgtccccgcc ctgtttttac ctataactct 540

ccttattatg atgggacacg aaccaaacgt ggttattcta acatcatggt tgtgcaccag 600

cgttatgtgc tccgatttcc tgagaacttg cctcttgatg ctggtgctcc tctgctgtgt 660

gctgggataa ctgtgtatag cccgatgaaa tattatggga tgacagagcc aggcaaacat 720

ttgggagtgg cagggcttgg tgggttaggt catgttgcaa tcaaacttgc taaggcgttt 780

gggctgaagt tactgtcatc cagtagctct cctaacaagc aagctgaggc cattgaccga 840

ctcggggctg attctttcct tgtttcatca gaccctgcaa aaatgaaggt ggctttggga 900

accatggact atatcataga cacaatttct gctgttcatt ccctgattcc actgcttggt 960

ctgctgaagc tgaatgggaa gctagttact gtagggttgc ctaacaagcc ccttgagctt 1020

cctatctttc cattagttgc aggacggaaa cttatagggg ggagcaactt tggagggatc 1080

aaagaaactc aggaaatgct tgatttctgc gcaaagcaca acataactgc agatattgag 1140

ctgattaaga tggatcagat caacacagcc atggaaaggc ttagcaaagc tgatgtgaaa 1200

tatcgctttg tgattgatgt ggcgaactct ttcgcaagtt tgtagaagcc accatgattc 1260

tccaactctt tgcatgtgtg gcctttagat gtctcagtat tttgaaagta gattggtggt 1320

ataagtagtc tataagatgt tgaactttaa agatgagaac atctgctaat taataatact 1380

ttgggttttg aatttatatt atgttccttt aaaaaaaaaa aaaaaaaaaa aaactcgagg 1440

gggggcccgg tacccaattc gcc 1463

4

360

Glycine max

4

Met Ala Lys Ser Pro Glu Thr Glu Leu Pro Val Glu Ala Phe Gly Trp

1 5 10 15

Ala Ala Ser Asp Thr Ser Gly Thr Leu Ala Pro Phe His Phe Ser Arg

20 25 30

Arg Glu Asn Gly Val Asp Asp Val Thr Leu Lys Ile Leu Phe Cys Gly

35 40 45

Val Cys His Ser Asp Leu His Thr Leu Lys Asn Asp Trp Gly Phe Thr

50 55 60

Thr Tyr Pro Val Val Pro Gly His Glu Ile Val Gly Val Val Thr Lys

65 70 75 80

Val Gly Asn Asn Val Lys Asn Phe Lys Val Gly Asp Lys Val Gly Val

85 90 95

Gly Val Ile Val Glu Ser Cys Lys Glu Cys Glu Ser Cys Gln Gln Asp

100 105 110

Leu Glu Ser Tyr Cys Pro Arg Pro Val Phe Thr Tyr Asn Ser Pro Tyr

115 120 125

Tyr Asp Gly Thr Arg Thr Lys Arg Gly Tyr Ser Asn Ile Met Val Val

130 135 140

His Gln Arg Tyr Val Leu Arg Phe Pro Glu Asn Leu Pro Leu Asp Ala

145 150 155 160

Gly Ala Pro Leu Leu Cys Ala Gly Ile Thr Val Tyr Ser Pro Met Lys

165 170 175

Tyr Tyr Gly Met Thr Glu Pro Gly Lys His Leu Gly Val Ala Gly Leu

180 185 190

Gly Gly Leu Gly His Val Ala Ile Lys Leu Ala Lys Ala Phe Gly Leu

195 200 205

Lys Leu Leu Ser Ser Ser Ser Ser Pro Asn Lys Gln Ala Glu Ala Ile

210 215 220

Asp Arg Leu Gly Ala Asp Ser Phe Leu Val Ser Ser Asp Pro Ala Lys

225 230 235 240

Met Lys Val Ala Leu Gly Thr Met Asp Tyr Ile Ile Asp Thr Ile Ser

245 250 255

Ala Val His Ser Leu Ile Pro Leu Leu Gly Leu Leu Lys Leu Asn Gly

260 265 270

Lys Leu Val Thr Val Gly Leu Pro Asn Lys Pro Leu Glu Leu Pro Ile

275 280 285

Phe Pro Leu Val Ala Gly Arg Lys Leu Ile Gly Gly Ser Asn Phe Gly

290 295 300

Gly Ile Lys Glu Thr Gln Glu Met Leu Asp Phe Cys Ala Lys His Asn

305 310 315 320

Ile Thr Ala Asp Ile Glu Leu Ile Lys Met Asp Gln Ile Asn Thr Ala

325 330 335

Met Glu Arg Leu Ser Lys Ala Asp Val Lys Tyr Arg Phe Val Ile Asp

340 345 350

Val Ala Asn Ser Phe Ala Ser Leu

355 360

5

1414

Triticum aestivum

5

gcacgagtcg tggttagtga ttacgaccag caagtcacaa gtgagggcgg gagaggttga 60

ggttgaggaa gagcagagga atccattcca ggagtgttta atggcgggag gcaaggaagc 120

gcacgggtgg gcagccaggg atgtctctgg tcacctctcc ccttaccact tctcacggag 180

ggttcagaga gacgacgacg tcaccatcaa ggtgctcttc tgcgggcttt gccacactga 240

cctccacgtc atcaagaacg agtttggcaa cgccaagtac cccgtcgttc ccgggcacga 300

gatcgtcggc gtcgtcaccg acgtcggctc cggcgtcaca agcttcaagc ccggcgacac 360

ggtgggcgtg ggctacttcg tcgactcctg ccgcagctgc gacagctgca gcaaggggta 420

cgagagctac tgcccgcagc tcgtggagac gtccaacggc gtgagcctgg acgacgatga 480

cggcggcgcc accaccaagg gcggcttctc cgacgccctc gtcgtccacc agcgctacgt 540

ggtgcgggtc ccggccagcc tgccgcccgc cggggccgcg ccgctgctgt gcgccggcgt 600

caccgtgttc agccccatgg tgcagtacgg cctgaacgcg ccggggaagc acctgggcgt 660

cgtcggcctc ggcggcctcg gccacctggc cgtccgcttc ggcaaggcgt tcgggatgaa 720

ggtcaccgtc atcagcacgt cgctgggcaa gcgggacgag gccctcggcc gcctcggtgc 780

cgacgcgttc ctggtcagcc gcgaccccga gcagatgagg gcggcggcgg gcaccttgga 840

cggcgtcatc gacacggtgt cggccgacca ccctgtcgtg ccgctgctgg acctgctcaa 900

gccgatgggc cagatggtcg tcgtcggcct gcccaccaag ccgctccagg tgcctgcctt 960

cagcctcgtc gccggcggga agcgcgtggc cgggagtgcc ggcggcggcg tcggggagtg 1020

ccaggccatg ctcgactttg ccggcgagca cgggatcacc gcggatgtgg aggtcgtcgg 1080

gatggactac gtcaataccg ccatccagcg cctagagagg aacgatgtca ggtaccgctt 1140

cgttgtcgac gtcgcgagca gcaagattgg aggctaggca tcaccattcc tagtgttctg 1200

tcgatcgacg tgtgatttgc ttcttcctcg agcgtgtctt attgttctgg ttggagcacg 1260

tacgcggcca tcacacgcag gcgtggacaa taaacaaggt agagtttcgg gttgtgtcgt 1320

taaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1380

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 1414

6

358

Triticum aestivum

6

Met Ala Gly Gly Lys Glu Ala His Gly Trp Ala Ala Arg Asp Val Ser

1 5 10 15

Gly His Leu Ser Pro Tyr His Phe Ser Arg Arg Val Gln Arg Asp Asp

20 25 30

Asp Val Thr Ile Lys Val Leu Phe Cys Gly Leu Cys His Thr Asp Leu

35 40 45

His Val Ile Lys Asn Glu Phe Gly Asn Ala Lys Tyr Pro Val Val Pro

50 55 60

Gly His Glu Ile Val Gly Val Val Thr Asp Val Gly Ser Gly Val Thr

65 70 75 80

Ser Phe Lys Pro Gly Asp Thr Val Gly Val Gly Tyr Phe Val Asp Ser

85 90 95

Cys Arg Ser Cys Asp Ser Cys Ser Lys Gly Tyr Glu Ser Tyr Cys Pro

100 105 110

Gln Leu Val Glu Thr Ser Asn Gly Val Ser Leu Asp Asp Asp Asp Gly

115 120 125

Gly Ala Thr Thr Lys Gly Gly Phe Ser Asp Ala Leu Val Val His Gln

130 135 140

Arg Tyr Val Val Arg Val Pro Ala Ser Leu Pro Pro Ala Gly Ala Ala

145 150 155 160

Pro Leu Leu Cys Ala Gly Val Thr Val Phe Ser Pro Met Val Gln Tyr

165 170 175

Gly Leu Asn Ala Pro Gly Lys His Leu Gly Val Val Gly Leu Gly Gly

180 185 190

Leu Gly His Leu Ala Val Arg Phe Gly Lys Ala Phe Gly Met Lys Val

195 200 205

Thr Val Ile Ser Thr Ser Leu Gly Lys Arg Asp Glu Ala Leu Gly Arg

210 215 220

Leu Gly Ala Asp Ala Phe Leu Val Ser Arg Asp Pro Glu Gln Met Arg

225 230 235 240

Ala Ala Ala Gly Thr Leu Asp Gly Val Ile Asp Thr Val Ser Ala Asp

245 250 255

His Pro Val Val Pro Leu Leu Asp Leu Leu Lys Pro Met Gly Gln Met

260 265 270

Val Val Val Gly Leu Pro Thr Lys Pro Leu Gln Val Pro Ala Phe Ser

275 280 285

Leu Val Ala Gly Gly Lys Arg Val Ala Gly Ser Ala Gly Gly Gly Val

290 295 300

Gly Glu Cys Gln Ala Met Leu Asp Phe Ala Gly Glu His Gly Ile Thr

305 310 315 320

Ala Asp Val Glu Val Val Gly Met Asp Tyr Val Asn Thr Ala Ile Gln

325 330 335

Arg Leu Glu Arg Asn Asp Val Arg Tyr Arg Phe Val Val Asp Val Ala

340 345 350

Ser Ser Lys Ile Gly Gly

355

7

cagaagtaga atcattcgat tccccaaagc gaaatccgca gcctcgcctc tctcgccatt 60

ttccaggcgc gcgctcggcg cggccgccgc cagcgataca gatccttgca cggacacaag 120

ggctccgcgc tcccaccgga tcggatcccg tcccgatggc tcccactccc actcacacgg 180

cggcggcggt ggcgctcgcc gcgcacgact cgtccggcca cctcgcgccg ctcaccatca 240

cccgcaggag caccggggac gacgatgtan cgataaagat cctgtactgt gggatgtgtc 300

actctgacct tcacagcatc aagaacgagt ggcacaacgc cacgtacccc atggtccctg 360

gtcacgagat cgccggagtg gtgacggang tgggcaagaa cgtgaccagc ttcaaggccg 420

gggaccgcgt cggcgtgggc tgcatggtga actcgtgcca gtcgtgcggc agctgcgacg 480

aggacttcga gaacagctgc cgggccgtca tcttcaccta caactccgtc gaccgcgacg 540

gcaccgtcac ctacggcggc tactccagca gcgtggtggt gcacgagcgc ttcgtggtcc 600

ggttccccga cgcgatgccg ctggatcggg gggcgccgct gctgtgcgcg ggcatcacag 660

tgtacagccc catgaagtac cacggcctga acgtgccggg gaagcacgtc ggcgtgatgg 720

ggctgggagg cctgggccac gtcgccgtca agttcgccaa ggcgttcggg gccaaggtca 780

cggtcatcag cacgtcgcct gggaagaagc aggaggcgct cgaccggctt ggcgccgacg 840

ccttcgtcgt cagcaaggac gccgaagaaa tgaaggcggc tgcgagcacc atggacggca 900

tcatcaacac ggtgtcggcg aacctcacct tggccccgta catgggcctg ctcaagccca 960

acggcaagat gatcatggtc ggcctgccca ccaagcctct cgagatcccg ccctttgatc 1020

tcatcattgg gaacaagacc ctggccggga gctgcatcgg cggcatgagg gacacgcagg 1080

agatgatcaa cgtggcggcc aagcacggcg tgacggcgga catagaactg gtggccgcgg 1140

actacgtgaa cacggccatg gagcgcctgg ccaaggctga cgtcaggtac cgcttcgtca 1200

tcgacatcgg caacacctca agaatcagag tgtcagacta aaacccggcg acactctact 1260

gctgctgctg ttgtccttcg ctttatacat gaacgctcct ttttcntgtc atgataatat 1320

taactaagca actnactant ccgaggatct ccanctgtat gaattggtct tatgtgttcc 1380

taaacagata natcacattt taaaaaaaa 1409

8

Met Ala Pro Thr Pro Thr His Thr Ala Ala Ala Val Ala Leu Ala Ala

1 5 10 15

His Asp Ser Ser Gly His Leu Ala Pro Leu Thr Ile Thr Arg Arg Ser

20 25 30

Thr Gly Asp Asp Asp Val Xaa Ile Lys Ile Leu Tyr Cys Gly Met Cys

35 40 45

His Ser Asp Leu His Ser Ile Lys Asn Glu Trp His Asn Ala Thr Tyr

50 55 60

Pro Met Val Pro Gly His Glu Ile Ala Gly Val Val Thr Xaa Val Gly

65 70 75 80

Lys Asn Val Thr Ser Phe Lys Ala Gly Asp Arg Val Gly Val Gly Cys

85 90 95

Met Val Asn Ser Cys Gln Ser Cys Gly Ser Cys Asp Glu Asp Phe Glu

100 105 110

Asn Ser Cys Arg Ala Val Ile Phe Thr Tyr Asn Ser Val Asp Arg Asp

115 120 125

Gly Thr Val Thr Tyr Gly Gly Tyr Ser Ser Ser Val Val Val His Glu

130 135 140

Arg Phe Val Val Arg Phe Pro Asp Ala Met Pro Leu Asp Arg Gly Ala

145 150 155 160

Pro Leu Leu Cys Ala Gly Ile Thr Val Tyr Ser Pro Met Lys Tyr His

165 170 175

Gly Leu Asn Val Pro Gly Lys His Val Gly Val Met Gly Leu Gly Gly

180 185 190

Leu Gly His Val Ala Val Lys Phe Ala Lys Ala Phe Gly Ala Lys Val

195 200 205

Thr Val Ile Ser Thr Ser Pro Gly Lys Lys Gln Glu Ala Leu Asp Arg

210 215 220

Leu Gly Ala Asp Ala Phe Val Val Ser Lys Asp Ala Glu Glu Met Lys

225 230 235 240

Ala Ala Ala Ser Thr Met Asp Gly Ile Ile Asn Thr Val Ser Ala Asn

245 250 255

Leu Thr Leu Ala Pro Tyr Met Gly Leu Leu Lys Pro Asn Gly Lys Met

260 265 270

Ile Met Val Gly Leu Pro Thr Lys Pro Leu Glu Ile Pro Pro Phe Asp

275 280 285

Leu Ile Ile Gly Asn Lys Thr Leu Ala Gly Ser Cys Ile Gly Gly Met

290 295 300

Arg Asp Thr Gln Glu Met Ile Asn Val Ala Ala Lys His Gly Val Thr

305 310 315 320

Ala Asp Ile Glu Leu Val Ala Ala Asp Tyr Val Asn Thr Ala Met Glu

325 330 335

Arg Leu Ala Lys Ala Asp Val Arg Tyr Arg Phe Val Ile Asp Ile Gly

340 345 350

Asn Thr Ser Arg Ile Arg Val Ser Asp

355 360

9

1480

Oryza sativa

9

gcacgagctt acatgtaagc tcgtgccgaa ttcggcacga gcttacactc tcatcctctc 60

ctctcccact ccacacccag gcacccagct cgagcgcgac tcggcaaatc catggctccc 120

acggcggcgt cggcggcggc gggggaggag cagcaggcgg tggggctggc ggcgcgcgac 180

tcctccggcc acctctcccc tttcgccatc tcccggagga gcactggaga tgacgatgtg 240

gcgataaaga ttctgttctg tggaatctgc cactctgacc tgcactgcat caagaatgag 300

tggaagcact ccatctaccc acttgttcct gggcacgaga tcgccggtgt ggtgacggag 360

gtggggaaga acgtcaccag gttcaaggcc ggcgacaggg tcggcgtggg gtgcatggtg 420

aactcgtgcc ggtcatgcga gagctgtaac aatggcttcg agaaccactg cccggagggc 480

gtcttcacct acaactccgt cgacaaggac ggcaccgtca cctacggcgg ttactcgagc 540

atggtggtgg tgcacgagcg cttcgtggtc atgttcccgg aggcgatgcc gctcgacgtc 600

ggcgccccgc tgctgtgcgc cggcatcacc gtgtacaccc ccatgaagta ccacggcctg 660

aacgcgccgg ggaagcacgt cggcgtgctc ggcctgggcg ggctcggcca cgtcgcggtg 720

aagttcgcca gggcgttcgg gttgaaggtg acggtgatca gctcgtcgcc ggggaagaag 780

cgggaggcgc tggagcggct cggcgccgac gcgttcgtcg tgagcagcag cgccgaggag 840

atggaggccg cgaggtccac catggacggc gtcatcaaca cggtctccgc caacaccccc 900

atggcgccgt acctggcgtt gctgaagccc aacgggaaga tgatcctcgt cggcctcccg 960

gagaatcccc tggaggtccc tcccttctct ctcgtccacg ggaacaggac tctggcaggg 1020

agcaacatcg gcggcatggc ggacacgcag gagatgatcg agctggcggc gaagcacggc 1080

gtgacggcgg acatcgaggt gatcggcgcc gacgacgtga acacggccat ggagcgcctc 1140

gccaaggcgg atgtcaggta tcgcttcgtc atcgacgtcg ggaacaccct ccacgccgcc 1200

gccgccgagt gatggcatcg cctgcaaact attcgtgtgg tctgtaatct gtatgaaatt 1260

gttcatggta cgtacacgat atatagattg tggcgtgtca tggcgtatag gaatgaaccc 1320

gatcgaggaa gctattgttg gtagtctgat gctagtcagc tcagtgtttg actgtttgca 1380

tttacggatt gaaattgagc tttgtgtggc ctaagtttta ttttctaaag atttcaagtg 1440

aatttcgttt tattcgaatg aaaaaaaaaa aaaaaaaaaa 1480

10

366

Oryza sativa

10

Met Ala Pro Thr Ala Ala Ser Ala Ala Ala Gly Glu Glu Gln Gln Ala

1 5 10 15

Val Gly Leu Ala Ala Arg Asp Ser Ser Gly His Leu Ser Pro Phe Ala

20 25 30

Ile Ser Arg Arg Ser Thr Gly Asp Asp Asp Val Ala Ile Lys Ile Leu

35 40 45

Phe Cys Gly Ile Cys His Ser Asp Leu His Cys Ile Lys Asn Glu Trp

50 55 60

Lys His Ser Ile Tyr Pro Leu Val Pro Gly His Glu Ile Ala Gly Val

65 70 75 80

Val Thr Glu Val Gly Lys Asn Val Thr Arg Phe Lys Ala Gly Asp Arg

85 90 95

Val Gly Val Gly Cys Met Val Asn Ser Cys Arg Ser Cys Glu Ser Cys

100 105 110

Asn Asn Gly Phe Glu Asn His Cys Pro Glu Gly Val Phe Thr Tyr Asn

115 120 125

Ser Val Asp Lys Asp Gly Thr Val Thr Tyr Gly Gly Tyr Ser Ser Met

130 135 140

Val Val Val His Glu Arg Phe Val Val Met Phe Pro Glu Ala Met Pro

145 150 155 160

Leu Asp Val Gly Ala Pro Leu Leu Cys Ala Gly Ile Thr Val Tyr Thr

165 170 175

Pro Met Lys Tyr His Gly Leu Asn Ala Pro Gly Lys His Val Gly Val

180 185 190

Leu Gly Leu Gly Gly Leu Gly His Val Ala Val Lys Phe Ala Arg Ala

195 200 205

Phe Gly Leu Lys Val Thr Val Ile Ser Ser Ser Pro Gly Lys Lys Arg

210 215 220

Glu Ala Leu Glu Arg Leu Gly Ala Asp Ala Phe Val Val Ser Ser Ser

225 230 235 240

Ala Glu Glu Met Glu Ala Ala Arg Ser Thr Met Asp Gly Val Ile Asn

245 250 255

Thr Val Ser Ala Asn Thr Pro Met Ala Pro Tyr Leu Ala Leu Leu Lys

260 265 270

Pro Asn Gly Lys Met Ile Leu Val Gly Leu Pro Glu Asn Pro Leu Glu

275 280 285

Val Pro Pro Phe Ser Leu Val His Gly Asn Arg Thr Leu Ala Gly Ser

290 295 300

Asn Ile Gly Gly Met Ala Asp Thr Gln Glu Met Ile Glu Leu Ala Ala

305 310 315 320

Lys His Gly Val Thr Ala Asp Ile Glu Val Ile Gly Ala Asp Asp Val

325 330 335

Asn Thr Ala Met Glu Arg Leu Ala Lys Ala Asp Val Arg Tyr Arg Phe

340 345 350

Val Ile Asp Val Gly Asn Thr Leu His Ala Ala Ala Ala Glu

355 360 365

11

1389

Glycine max

11

gcacgagcag agatggagca tccaaagaag gtttttggtt gggctgctcg ggactcatct 60

gggcttctct ccccattcaa cttctccaga agagaaacgg gggagaaaga tttggtattc 120

aaagtgcaat actgtggaat atgccactca gatctacata tgttaaagaa cgagtggggc 180

aacacaacct atcctctggt tcctgggcac gagattgctg gtgtggtaac agaagttgga 240

agcaaggtgc aaaagttcaa agttggagac cgggtaggtg tggggtgcat gattggatcc 300

tgtcgttcat gtgaaagctg tgatgaaaat cttgagaatt actgccccaa aatgattctc 360

acatacggtg tcaagtattt tgatggcacc ataacacatg gaggctactc tgacttgatg 420

gttgcggatg aacactttgt ggttagaatt ccggataacc taccacttga tgctgctgct 480

cctctcctct gtgctgggat cactgtgtat agccctttga gatactatgg acttgacaag 540

cctggtctga atcttggtgt ggttggtctt ggtggacttg gtcacatggc tgtcaagttt 600

gccaaagctc ttggagctaa tgtcacagtg attagcacat ctcctaacaa aaagaaggaa 660

gccattgaga acataggagc tgactcattt gtggttagcc gtgaacaaga tcagatgcag 720

gctgtaatgg gaaccatgga tggtatcatt gacacagttt ctgctgtcca tcctcttgtg 780

cctttgattg gcctgttgaa gcctcatggt aaactagtca tggttggtgc accagagaag 840

cctttggagt tgccagtatt ttctttactt atggggagga aaatggttgg tggcagtagc 900

atcggaggga tgaaagagac gcaagaaatg attgattttg ctgctaagca tggagtgaag 960

cctgatattg aggttattcc catagattat gtgaacactg caatagagcg cctcgccaaa 1020

gcagatgtga aatatcgatt tgtaattgac attggaaaca cgttgaagcc aagctcttga 1080

gtaaataaag aaatatgcat gcgtgattca gacttcagat actgaacgta catggggcaa 1140

gagataaaga ttgagaattg ataataattt ctttaagcat tatatgtgtt ttccgtaatg 1200

agtttttgtt tcaaggattt agctacgcta ttccttgata aataagcaat taagtgtggg 1260

gtgtgcaatt gcttaagctt ggttataatc cctttgggac gaaaattgtt ttgtactata 1320

tatacatttt aaagagtgaa attgtgtgag attatgaaca aaaaaaaaaa aaaaaaaaaa 1380

aaaaaaaaa 1389

12

355

Glycine max

12

Met Glu His Pro Lys Lys Val Phe Gly Trp Ala Ala Arg Asp Ser Ser

1 5 10 15

Gly Leu Leu Ser Pro Phe Asn Phe Ser Arg Arg Glu Thr Gly Glu Lys

20 25 30

Asp Leu Val Phe Lys Val Gln Tyr Cys Gly Ile Cys His Ser Asp Leu

35 40 45

His Met Leu Lys Asn Glu Trp Gly Asn Thr Thr Tyr Pro Leu Val Pro

50 55 60

Gly His Glu Ile Ala Gly Val Val Thr Glu Val Gly Ser Lys Val Gln

65 70 75 80

Lys Phe Lys Val Gly Asp Arg Val Gly Val Gly Cys Met Ile Gly Ser

85 90 95

Cys Arg Ser Cys Glu Ser Cys Asp Glu Asn Leu Glu Asn Tyr Cys Pro

100 105 110

Lys Met Ile Leu Thr Tyr Gly Val Lys Tyr Phe Asp Gly Thr Ile Thr

115 120 125

His Gly Gly Tyr Ser Asp Leu Met Val Ala Asp Glu His Phe Val Val

130 135 140

Arg Ile Pro Asp Asn Leu Pro Leu Asp Ala Ala Ala Pro Leu Leu Cys

145 150 155 160

Ala Gly Ile Thr Val Tyr Ser Pro Leu Arg Tyr Tyr Gly Leu Asp Lys

165 170 175

Pro Gly Leu Asn Leu Gly Val Val Gly Leu Gly Gly Leu Gly His Met

180 185 190

Ala Val Lys Phe Ala Lys Ala Leu Gly Ala Asn Val Thr Val Ile Ser

195 200 205

Thr Ser Pro Asn Lys Lys Lys Glu Ala Ile Glu Asn Ile Gly Ala Asp

210 215 220

Ser Phe Val Val Ser Arg Glu Gln Asp Gln Met Gln Ala Val Met Gly

225 230 235 240

Thr Met Asp Gly Ile Ile Asp Thr Val Ser Ala Val His Pro Leu Val

245 250 255

Pro Leu Ile Gly Leu Leu Lys Pro His Gly Lys Leu Val Met Val Gly

260 265 270

Ala Pro Glu Lys Pro Leu Glu Leu Pro Val Phe Ser Leu Leu Met Gly

275 280 285

Arg Lys Met Val Gly Gly Ser Ser Ile Gly Gly Met Lys Glu Thr Gln

290 295 300

Glu Met Ile Asp Phe Ala Ala Lys His Gly Val Lys Pro Asp Ile Glu

305 310 315 320

Val Ile Pro Ile Asp Tyr Val Asn Thr Ala Ile Glu Arg Leu Ala Lys

325 330 335

Ala Asp Val Lys Tyr Arg Phe Val Ile Asp Ile Gly Asn Thr Leu Lys

340 345 350

Pro Ser Ser

355

13

1418

Triticum aestivum

13

gcacgagcga acgtctaatc gcctcacaca caggaaaggc aaaaaagaaa ccaaaacctc 60

cggggagtta ctctagagat ttccggcgga acctcgcgtc aggatggcac ccacggcgac 120

ggcggcggag cagagccagc agcacacgaa gaaggcggtg ggtctggcgg cgctcgacgc 180

ctccggccac ctctccccgc tcgccatcac ccgaaggagc accggagatg acgatgtcgt 240

gataaagatt ctgtactgcg ggatctgcca ctctgacctg cacagcatca agaacgactg 300

gaagaacgcc aagtacccca tgattcccgg gcacgagatc gccggcgagg tcaccgaggt 360

cggcaagaac gtgaccaagt tcaaggcagg cgaccgtgtc ggcgtcgggt gcatggtgaa 420

ctcgtgccag tcctgcgaga gctgcgacaa gggcttcgag aatcactgcc cgggcatgat 480

tttcacctac aactcggtcg accgtgacgg cacccgcacc cacggtggct actccagcat 540

ggtagtggtg cacgagcgct tcgtggtccg gtttccagac gccatgccgc tggacaaggg 600

cgcgcccctg ctgtgcgccg gcatcaccgt gtacagcccc atgaagtacc acgggctgaa 660

cgcccctggg atgcacctcg gcgtgcttgg actgggcggg ctgggccacg tcgccgtcaa 720

gttcggcaag gccttcggga tgaaggtgac cgtgatcagc tcgtcgccgg ggaagaagca 780

ggaggccctc gagaggctag gcgccgacgc gttcgttgtc agcaaggacg ccgacgagat 840

gaaggctact atgagcacca tggatggcat cataaacacg gtgtctgcaa acgtgcccat 900

ggcccctctc ttcgggctac tcaagcccaa cggcaagatg atcatggtcg gcctcccgga 960

gaagcctatc gaggtccctc cctttgctct ggttgccagg aacaagaccc tggcggggag 1020

ctgcatcggc ggcatgaggg acacccagga gatgctggac ctcgcggcga agcacggcgt 1080

gacggcagac atcgaggtga tcggcgcgga gtacgtgaac acggccatgg agcgccttgc 1140

caaggccgac gtcaggtatc gattcgtcat cgacatcgcc aacaccctcg acaaggccgc 1200

cgccgccaca accgagtgac cgccgacgta actctcagca ctactcacgg tccacgttgg 1260

tctctggttc tctgctatta tcacatactt tatctccagt agtaacaata cacagtctaa 1320

aattattgtc agtgatgtat tttttagaca aacttgtcag tgatgtatgc atggtgcgat 1380

gttgttcgac gatgggtgaa aaaaaaaaaa aaaaaaaa 1418

14

371

Triticum aestivum

14

Met Ala Pro Thr Ala Thr Ala Ala Glu Gln Ser Gln Gln His Thr Lys

1 5 10 15

Lys Ala Val Gly Leu Ala Ala Leu Asp Ala Ser Gly His Leu Ser Pro

20 25 30

Leu Ala Ile Thr Arg Arg Ser Thr Gly Asp Asp Asp Val Val Ile Lys

35 40 45

Ile Leu Tyr Cys Gly Ile Cys His Ser Asp Leu His Ser Ile Lys Asn

50 55 60

Asp Trp Lys Asn Ala Lys Tyr Pro Met Ile Pro Gly His Glu Ile Ala

65 70 75 80

Gly Glu Val Thr Glu Val Gly Lys Asn Val Thr Lys Phe Lys Ala Gly

85 90 95

Asp Arg Val Gly Val Gly Cys Met Val Asn Ser Cys Gln Ser Cys Glu

100 105 110

Ser Cys Asp Lys Gly Phe Glu Asn His Cys Pro Gly Met Ile Phe Thr

115 120 125

Tyr Asn Ser Val Asp Arg Asp Gly Thr Arg Thr His Gly Gly Tyr Ser

130 135 140

Ser Met Val Val Val His Glu Arg Phe Val Val Arg Phe Pro Asp Ala

145 150 155 160

Met Pro Leu Asp Lys Gly Ala Pro Leu Leu Cys Ala Gly Ile Thr Val

165 170 175

Tyr Ser Pro Met Lys Tyr His Gly Leu Asn Ala Pro Gly Met His Leu

180 185 190

Gly Val Leu Gly Leu Gly Gly Leu Gly His Val Ala Val Lys Phe Gly

195 200 205

Lys Ala Phe Gly Met Lys Val Thr Val Ile Ser Ser Ser Pro Gly Lys

210 215 220

Lys Gln Glu Ala Leu Glu Arg Leu Gly Ala Asp Ala Phe Val Val Ser

225 230 235 240

Lys Asp Ala Asp Glu Met Lys Ala Thr Met Ser Thr Met Asp Gly Ile

245 250 255

Ile Asn Thr Val Ser Ala Asn Val Pro Met Ala Pro Leu Phe Gly Leu

260 265 270

Leu Lys Pro Asn Gly Lys Met Ile Met Val Gly Leu Pro Glu Lys Pro

275 280 285

Ile Glu Val Pro Pro Phe Ala Leu Val Ala Arg Asn Lys Thr Leu Ala

290 295 300

Gly Ser Cys Ile Gly Gly Met Arg Asp Thr Gln Glu Met Leu Asp Leu

305 310 315 320

Ala Ala Lys His Gly Val Thr Ala Asp Ile Glu Val Ile Gly Ala Glu

325 330 335

Tyr Val Asn Thr Ala Met Glu Arg Leu Ala Lys Ala Asp Val Arg Tyr

340 345 350

Arg Phe Val Ile Asp Ile Ala Asn Thr Leu Asp Lys Ala Ala Ala Ala

355 360 365

Thr Thr Glu

370

15

aaacaatcta cggttgccag gtctgcgacg tcagtgccga cgaactcatg gctcgcgaga 60

ggcccgtaaa aacatgatag ccagatctgc gacgtcacgg ctgtcgccag tgatcgccat 120

tcgccaccct ctccgttcca ttccaatccg ttcagcggtc agcggaacat gatacggtct 180

gcgcccttcc aagcaatcac atcttcttcc ccgttataaa tcgctcactc gctatcgtcg 240

gctacctgta tcctggcgac acccaacagc ctgcttgccg cgcttgcatt gaggggagtt 300

cggggagcgg ccggtgcgga aggaatcgac acagagttca gttcacatgg ctgctgaatc 360

agagaacggc aactgcaaag cttgggcagc gagagatcct tctggagttc tctcaccaca 420

cagttttaac cgcagagcag tgcaaagcgg cgatgtttcg ctgaagatca tatactgtgg 480

agtctgttat gcagacgttg cttggacgcg gaataagctc aatgattcaa aatatccttt 540

ggttcctggg catgagatag ctggagttgt aactcaggtt ggtgcagatg tcaaaggctt 600

taaagtgggt gaccatgtag gtgttggaac atatgtgaac tcttgccgtg attgtgagaa 660

ctgcaatatc tctctagaca accattgccc aaaagcagtt tatactttca atggcattga 720

tacagatgga actgtcacaa aggggggtta ctccactcac atttgtagtc caggaaaggt 780

attgcttcaa aatacccgat ggctaccttt ggcgaangna gcacctcttc tgtgttgctg 840

gaataccgtg tatactncaa tgatgcgaca caacatgaac caacctggaa agtcacttgg 900

ggtcat 906

16

Met Ala Ala Glu Ser Glu Asn Gly Asn Cys Lys Ala Trp Ala Ala Arg

1 5 10 15

Asp Pro Ser Gly Val Leu Ser Pro His Ser Phe Asn Arg Arg Ala Val

20 25 30

Gln Ser Gly Asp Val Ser Leu Lys Ile Ile Tyr Cys Gly Val Cys Tyr

35 40 45

Ala Asp Val Ala Trp Thr Arg Asn Lys Leu Asn Asp Ser Lys Tyr Pro

50 55 60

Leu Val Pro Gly His Glu Ile Ala Gly Val Val Thr Gln Val Gly Ala

65 70 75 80

Asp Val Lys Gly Phe Lys Val Gly Asp His Val Gly Val Gly Thr Tyr

85 90 95

Val Asn Ser Cys Arg Asp Cys Glu Asn Cys Asn Ile Ser Leu Asp Asn

100 105 110

His Cys Pro Lys Ala Val Tyr Thr Phe Asn Gly Ile Asp Thr Asp Gly

115 120 125

Thr Val Thr Lys Gly Gly Tyr Ser Thr His Ile Cys Ser Pro Gly Lys

130 135 140

Val Leu Leu Gln Asn Thr Arg Trp Leu Pro Leu Ala Xaa Xaa Ala Pro

145 150 155 160

Leu Leu Cys Cys Trp Asn Thr Val Tyr Thr Xaa Met Met Arg His Asn

165 170 175

Met Asn Gln Pro Gly Lys Ser Leu Gly Val

180 185

17

gcgccaagac gagtcgcggc tcaacagcgg agaggcggcg atctcccatc tggcgagcag 60

agcaggggag gggaagggat cttgttagag tggcatggct gctgaatgtg gaagtggcaa 120

ctgtgatgct tgggcagcga gagatccttc agggatcctc tccccgtaca agttcaaccg 180

cagggctgta caaagtgacg atgtttcctt gaggatcaca cactgtggtg tttgttatgc 240

tgatgttgca tggacaagga atatactcaa caattcgatg taccctttag tccctgggca 300

tgagatagca ggagttgtaa ctgaggttgg tgcagacgtc aagagcttca aagtgggtga 360

ccatgtaggt gttggcacat acgttaattc aagcccggga ctgtgagaac tgcaataact 420

ctctagagaa ctnactgctc acaacaa 447

18

111

Oryza sativa

18

Met Ala Ala Glu Cys Gly Ser Gly Asn Cys Asp Ala Trp Ala Ala Arg

1 5 10 15

Asp Pro Ser Gly Ile Leu Ser Pro Tyr Lys Phe Asn Arg Arg Ala Val

20 25 30

Gln Ser Asp Asp Val Ser Leu Arg Ile Thr His Cys Gly Val Cys Tyr

35 40 45

Ala Asp Val Ala Trp Thr Arg Asn Ile Leu Asn Asn Ser Met Tyr Pro

50 55 60

Leu Val Pro Gly His Glu Ile Ala Gly Val Val Thr Glu Val Gly Ala

65 70 75 80

Asp Val Lys Ser Phe Lys Val Gly Asp His Val Gly Val Gly Thr Tyr

85 90 95

Val Asn Ser Ser Pro Asp Cys Glu Asn Cys Asn Asn Ser Leu Glu

100 105 110

19

1310

Glycine max

19

gcacgagcac aagtgtgaga aaaattatga gttccaaagg tgttggtgag gattgcctgg 60

gatgggcagc aagagatgca tccggagttc tatcacctta caaattcagt cgcaggactc 120

ttgggaatga agatgttcat attaaaatta cgcactgtgg tgtttgcttc gctgatgtag 180

tttggacaag gaataaacat ggtgactcaa agtatcctgt cgtgcctggt catgagattg 240

ctgggattgt gacaaaggtt ggcgccaatg tccaccattt taaggttggc gaccatgttg 300

gagtggggac ttatatcaac tcatgtaggg attgtgagta ttgtaatgat ggacaagaag 360

ttcattgtac caagggatct gtttacactt ttaatggtgt tgattttgat ggtacaatta 420

caaaaggagg atactccagt tacatagtag tccatgagag gtactgcttc atgataccaa 480

aaagctatcc attggcttcc gcagctcctt tgctttgtgc tggaattact gtttattcac 540

cgatggtccg ccacaagatg aatcaacctg gtaaatctct aggagtgatt ggtcttggtg 600

gcctcggtca tatggcagtg aaatttggaa aggcatttgg tttgagtgta acggttttca 660

gcactagtat atccaagaaa gaggaggcac tgagcctgct tggtgcagac aaatttgttg 720

tttcatctaa tcaagaggaa atgacggcgt tggctaaatc gttggacttt ataatcgaca 780

cagcatctgg tgatcactcg tttgatcctt acatgtcact gctgaagaca tatggtgttt 840

ttgtcctagt tggtttccct agtcaagtca aatttatccc tgcaagcctt aatataggat 900

caaagactgt tgccggaagt gttacaggtg gtacaaaaga tatacaggag atgattggct 960

tctgtgctgc aaacgagatt cacccaaata tagaggtgat tccaatcgag tatgccaatg 1020

aagctcttga gaggctcata aatagggacg tcaagtaccg gtttgtaata gatgttgaga 1080

attccctgaa agaaaaatga gttgttgcct cccaaattgg acattattgg acttcacctt 1140

gttcgataaa taatgatggt cggagttcgt aatttactta catagagttg atttgaattt 1200

tctttaatta ttttgtgaac taatattatg tgattataag aatattttgg atttttaaat 1260

aataataatt ttgggaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1310

20

357

Glycine max

20

Met Ser Ser Lys Gly Val Gly Glu Asp Cys Leu Gly Trp Ala Ala Arg

1 5 10 15

Asp Ala Ser Gly Val Leu Ser Pro Tyr Lys Phe Ser Arg Arg Thr Leu

20 25 30

Gly Asn Glu Asp Val His Ile Lys Ile Thr His Cys Gly Val Cys Phe

35 40 45

Ala Asp Val Val Trp Thr Arg Asn Lys His Gly Asp Ser Lys Tyr Pro

50 55 60

Val Val Pro Gly His Glu Ile Ala Gly Ile Val Thr Lys Val Gly Ala

65 70 75 80

Asn Val His His Phe Lys Val Gly Asp His Val Gly Val Gly Thr Tyr

85 90 95

Ile Asn Ser Cys Arg Asp Cys Glu Tyr Cys Asn Asp Gly Gln Glu Val

100 105 110

His Cys Thr Lys Gly Ser Val Tyr Thr Phe Asn Gly Val Asp Phe Asp

115 120 125

Gly Thr Ile Thr Lys Gly Gly Tyr Ser Ser Tyr Ile Val Val His Glu

130 135 140

Arg Tyr Cys Phe Met Ile Pro Lys Ser Tyr Pro Leu Ala Ser Ala Ala

145 150 155 160

Pro Leu Leu Cys Ala Gly Ile Thr Val Tyr Ser Pro Met Val Arg His

165 170 175

Lys Met Asn Gln Pro Gly Lys Ser Leu Gly Val Ile Gly Leu Gly Gly

180 185 190

Leu Gly His Met Ala Val Lys Phe Gly Lys Ala Phe Gly Leu Ser Val

195 200 205

Thr Val Phe Ser Thr Ser Ile Ser Lys Lys Glu Glu Ala Leu Ser Leu

210 215 220

Leu Gly Ala Asp Lys Phe Val Val Ser Ser Asn Gln Glu Glu Met Thr

225 230 235 240

Ala Leu Ala Lys Ser Leu Asp Phe Ile Ile Asp Thr Ala Ser Gly Asp

245 250 255

His Ser Phe Asp Pro Tyr Met Ser Leu Leu Lys Thr Tyr Gly Val Phe

260 265 270

Val Leu Val Gly Phe Pro Ser Gln Val Lys Phe Ile Pro Ala Ser Leu

275 280 285

Asn Ile Gly Ser Lys Thr Val Ala Gly Ser Val Thr Gly Gly Thr Lys

290 295 300

Asp Ile Gln Glu Met Ile Gly Phe Cys Ala Ala Asn Glu Ile His Pro

305 310 315 320

Asn Ile Glu Val Ile Pro Ile Glu Tyr Ala Asn Glu Ala Leu Glu Arg

325 330 335

Leu Ile Asn Arg Asp Val Lys Tyr Arg Phe Val Ile Asp Val Glu Asn

340 345 350

Ser Leu Lys Glu Lys

355

21

gtttcccgtt tgataccgca ccgctgtagc tggagtacat agtagctgac gtcaatgccc 60

tacacacgta gtgtctgacg ttcgatccaa ctcgtcagct cgcctattta tccccaacct 120

cgtggcctgg cagtgtcatc cggatatttt tgagcaggaa atcgaagtgc caggaccgaa 180

gaagcagagc aagcagtcat caatggcgaa aggcacgcct gcgaccggtt gggccgcgaa 240

ggacccgtcc ggtgtcctct ccccctacag cttctcacga agggctctga aggacgacga 300

tgtcaccatc aaggtgctct tctgcggatt ctgccacacc gacctccaca tcgccaagaa 360

cgagtgggga aacgccctct acccgtcgtc cccggggcat gagattgttg gcgtcctcac 420

cgacgctggg cccggcgtca aagaattcaa ggccggggac acngtgggcg tgggctactt 480

cgtcgantcc tgccgctcct gcgtnaactg cggcancggg cacaagaaca ctgcccaagc 540

tcgtgctcac tccaacgccg tggactacga cggngccaca accaaggang gtctcnaagt 600

ctcntcntca caagg 615

22

Met Ala Lys Gly Thr Pro Ala Thr Gly Trp Ala Ala Lys Asp Pro Ser

1 5 10 15

Gly Val Leu Ser Pro Tyr Ser Phe Ser Arg Arg Ala Leu Lys Asp Asp

20 25 30

Asp Val Thr Ile Lys Val Leu Phe Cys Gly Phe Cys His Thr Asp Leu

35 40 45

His Ile Ala Lys Asn Glu Trp Gly Asn Ala Leu Tyr Pro Ser Ser Pro

50 55 60

Gly His Glu Ile Val Gly Val Leu Thr Asp Ala Gly Pro Gly Val Lys

65 70 75 80

Glu Phe Lys Ala Gly Asp Thr Val Gly Val Gly Tyr Phe Val Xaa Ser

85 90 95

Cys Arg Ser Cys Val Asn Cys

100

23

453

Triticum aestivum

23

ctcgtgccga attcggcacg aggtaccctt tagtccctgg gcatgagatt gctggagttg 60

taaccgaggt tggttcagac gtcaagggct tcaaactggg cgaccatgtg gctgttggga 120

catatgtcaa ctcatgccgt gactgtgaca actgcaatag cttcctcgat aaccactgct 180

caaagtttgt tttcactttc aatggtgttg atactgacgg tactgtcaca aagggaggat 240

attccagtca cattgttgtt catgaacggt attgctataa aatacctgat ggctacccac 300

tggaaaaggc cgcaccttta gtttgtgctg ggatcactgt gtacagtccg atgatgcgac 360

ataacatgaa ccagccagga aagtcactcg gtgtcattgg tctcggtggg ttgggtcaca 420

tggcagtgaa atttgggaaa gcctttggac tga 453

24

150

Triticum aestivum

24

Arg Ala Glu Phe Gly Thr Arg Tyr Pro Leu Val Pro Gly His Glu Ile

1 5 10 15

Ala Gly Val Val Thr Glu Val Gly Ser Asp Val Lys Gly Phe Lys Leu

20 25 30

Gly Asp His Val Ala Val Gly Thr Tyr Val Asn Ser Cys Arg Asp Cys

35 40 45

Asp Asn Cys Asn Ser Phe Leu Asp Asn His Cys Ser Lys Phe Val Phe

50 55 60

Thr Phe Asn Gly Val Asp Thr Asp Gly Thr Val Thr Lys Gly Gly Tyr

65 70 75 80

Ser Ser His Ile Val Val His Glu Arg Tyr Cys Tyr Lys Ile Pro Asp

85 90 95

Gly Tyr Pro Leu Glu Lys Ala Ala Pro Leu Val Cys Ala Gly Ile Thr

100 105 110

Val Tyr Ser Pro Met Met Arg His Asn Met Asn Gln Pro Gly Lys Ser

115 120 125

Leu Gly Val Ile Gly Leu Gly Gly Leu Gly His Met Ala Val Lys Phe

130 135 140

Gly Lys Ala Phe Gly Leu

145 150

25

458

Zea mays

25

gacctctcaa atcattcaat tccccagtac tgccactcgt ggtgtattca agcaaaccac 60

gatcgaaaca cctgcacttg ccaacaatga agtgctgatc aagatcctcg ctagcggtgt 120

ctgtcacaca gactgtggct tcatgggtca agatggcctt gtccttggtc atgaatccgt 180

gggccgtgtt gttcagatcg gatccgatgt caaaaagatt caagtgggtg atgtggttgg 240

tagttcctat ttgaaggaat cctgtttgag ctgtccctct tgtaattcgg gtgtagatac 300

caattgtgat aatagaaaga tgttccctga aggaaactca aatggattcg cctctcatca 360

aatcgctgac agtcgctttg tctacaagat tcctgatcat ttagaaccca agcatgctgg 420

tcccttgatg tgtgctggta tcactgtctt taatgctc 458

26

152

Zea mays

26

Thr Ser Gln Ile Ile Gln Phe Pro Ser Thr Ala Thr Arg Gly Val Phe

1 5 10 15

Lys Gln Thr Thr Ile Glu Thr Pro Ala Leu Ala Asn Asn Glu Val Leu

20 25 30

Ile Lys Ile Leu Ala Ser Gly Val Cys His Thr Asp Cys Gly Phe Met

35 40 45

Gly Gln Asp Gly Leu Val Leu Gly His Glu Ser Val Gly Arg Val Val

50 55 60

Gln Ile Gly Ser Asp Val Lys Lys Ile Gln Val Gly Asp Val Val Gly

65 70 75 80

Ser Ser Tyr Leu Lys Glu Ser Cys Leu Ser Cys Pro Ser Cys Asn Ser

85 90 95

Gly Val Asp Thr Asn Cys Asp Asn Arg Lys Met Phe Pro Glu Gly Asn

100 105 110

Ser Asn Gly Phe Ala Ser His Gln Ile Ala Asp Ser Arg Phe Val Tyr

115 120 125

Lys Ile Pro Asp His Leu Glu Pro Lys His Ala Gly Pro Leu Met Cys

130 135 140

Ala Gly Ile Thr Val Phe Asn Ala

145 150

27

catcactcta ccacctagca tctctctaac tttgagaaag tgtgtgttgt gtgttctctt 60

ctctcactaa aaaaggtttc attaattcta tattatggct canaccacag ngaatcanac 120

agtgaccgta tctgggtggg cagctcatga ttcttcgggc aagattaccc cctacacttt 180

caaaagaagg ggagaatggt gtgaacgatg tgacaataaa gatactgtat tgtgggatct 240

gccacacgga cctccactat gctaaaaatg aatggggcat tactatgtat cctgtagtac 300

ccggacacga gatcattggg gtggtaacta aggtggggag agacgtgaag gggttcaagg 360

aaggagacag agtgggtgtg gggtgtttgg cggcttcatg tttggaatgt gagcactgca 420

agaccgatca ggaaaattac tgtgaaaagt tgcaatttgt ctacaacgga gtattctggg 480

atggcagcat cacctatggg ggctactctc aaatttttgt cgcagattac agatatgtgg 540

tacacattcc ggaaaaccta gcaatggatg cagcggctcc tctgctatgc gcaggaataa 600

cagtgttcaa tcctttgaaa gaccacgatt tggtagcatc accgggaaag aagattgggg 660

tggttggtct gggaggcctt gggcacattg ccgttaaatt tggcaaggct ttcggccacc 720

atgttaccgt catcagcact tctccctcca aggaggctga agctaagcag cgtttgggtg 780

ctgatgattt cattgtcagc tccaacccta aacaattaca ggctgcaagg agaagcatag 840

atttcatatt ggacactgtt tcggcagagc actctctgtt gcccatcttg gaattgctca 900

aagtcaacgg gaccttattt ctcgtgggtg caccaagaca aagcccttac aattgccagc 960

tttcccattg atatttggca agagatcagt gaaaggtggg atcatagggg gaataaaaga 1020

gac 1023

28

Met Ile Leu Arg Ala Arg Leu Pro Pro Thr Leu Ser Lys Glu Gly Glu

1 5 10 15

Asn Gly Val Asn Asp Val Thr Ile Lys Ile Leu Tyr Cys Gly Ile Cys

20 25 30

His Thr Asp Leu His Tyr Ala Lys Asn Glu Trp Gly Ile Thr Met Tyr

35 40 45

Pro Val Val Pro Gly His Glu Ile Ile Gly Val Val Thr Lys Val Gly

50 55 60

Arg Asp Val Lys Gly Phe Lys Glu Gly Asp Arg Val Gly Val Gly Cys

65 70 75 80

Leu Ala Ala Ser Cys Leu Glu Cys Glu His Cys Lys Thr Asp Gln Glu

85 90 95

Asn Tyr Cys Glu Lys Leu Gln Phe Val Tyr Asn Gly Val Phe Trp Asp

100 105 110

Gly Ser Ile Thr Tyr Gly Gly Tyr Ser Gln Ile Phe Val Ala Asp Tyr

115 120 125

Arg Tyr Val Val His Ile Pro Glu Asn Leu Ala Met Asp Ala Ala Ala

130 135 140

Pro Leu Leu Cys Ala Gly Ile Thr Val Phe Asn Pro Leu Lys Asp His

145 150 155 160

Asp Leu Val Ala Ser Pro Gly Lys Lys Ile Gly Val Val Gly Leu Gly

165 170 175

Gly Leu Gly His Ile Ala Val Lys Phe Gly Lys Ala Phe Gly His His

180 185 190

Val Thr Val Ile Ser Thr Ser Pro Ser Lys Glu Ala Glu Ala Lys Gln

195 200 205

Arg Leu Gly Ala Asp Asp Phe Ile Val Ser Ser Asn Pro Lys Gln Leu

210 215 220

Gln Ala Ala Arg Arg Ser Ile Asp Phe Ile Leu Asp Thr Val Ser Ala

225 230 235 240

Glu His Ser Leu Leu Pro Ile Leu Glu Leu Leu Lys Val Asn Gly Thr

245 250 255

Leu Phe Leu Val Gly Ala Pro Xaa Thr Lys Pro Leu Gln Leu Pro Ala

260 265 270

Phe Pro Leu Ile Phe Gly Lys Arg Ser Val Lys Gly Gly Ile Ile Gly

275 280 285

Gly Ile Lys Glu

290

29

aancctttta gcaagttaac aacacttttg tcttcgttcc attatctagt tctcaattct 60

cttcaccctt tgcctctgaa agaaatgggt agtcttgaag cggaaagaac aacagtagga 120

ttggcagcaa gagacccttc agggattctc tccccataca cctacaatct aagaaacaca 180

ggccctgatg atgtttacat taaagttcac tactgtggaa tttgccattc tgatctccac 240

cagattaaaa atgatcttgg catgtccaac tatcccatgg tcccaggaca tgaagtggtt 300

ggtgaggtac tggaggtggg atcaaacgtt agcaggttca gagtgggaga gttggtggga 360

gttggactcc tcgtaggttg ctgcaaaaac tgccagccat gccaacaaga cattgagaat 420

tactgcagca agaagatctg gtcttacaat gatgtttatg tcgatggaaa acccacacag 480

ggtggctttg ctgaaaccat gatcgttgag caaaagtttg tggtgaagat tccagagggt 540

ttggcgccag agcaagtggc accattactg tgtgcgggtg tgacagtgta cagtccactg 600

gtacactttg ggctgaaaga gagtgggcta agaggtggaa tattggggct tggaggagtg 660

ggacacatgg gggtgaagat agcanaggca ctgggtcatc atgtgactgt gataagctct 720

tctgacaaga agaaacaaga ggcacttgaa caccttggag ctgatcaata tctcgttagc 780

tctgatgcca ctgccatgca ggaagctgct gattcacttg actacatcat tgacactgtg 840

cctgttggtc accctcttga gccttatctc tccttgctca aacttgatgg caagttgatc 900

ttgatggggg tcatcaacac ccctcttcaa tttgttagcc ccatggtcat gctagggagg 960

aagtcaataa cgggaagctt cattgggagc atgaaggaga cagaggagat gttggagttc 1020

tggaaagaga agggtctgag ttcgatgatt gaaatggtaa acatggatta cattaacaaa 1080

gcctttgaaa ggttggagaa gaatgacgtg agatacaggt ttgttgtgga tgtcaaaggc 1140

agcaaacttg tcgatcagta aaagaaagaa a 1171

30

Met Gly Ser Leu Glu Ala Glu Arg Thr Thr Val Gly Leu Ala Ala Arg

1 5 10 15

Asp Pro Ser Gly Ile Leu Ser Pro Tyr Thr Tyr Asn Leu Arg Asn Thr

20 25 30

Gly Pro Asp Asp Val Tyr Ile Lys Val His Tyr Cys Gly Ile Cys His

35 40 45

Ser Asp Leu His Gln Ile Lys Asn Asp Leu Gly Met Ser Asn Tyr Pro

50 55 60

Met Val Pro Gly His Glu Val Val Gly Glu Val Leu Glu Val Gly Ser

65 70 75 80

Asn Val Ser Arg Phe Arg Val Gly Glu Leu Val Gly Val Gly Leu Leu

85 90 95

Val Gly Cys Cys Lys Asn Cys Gln Pro Cys Gln Gln Asp Ile Glu Asn

100 105 110

Tyr Cys Ser Lys Lys Ile Trp Ser Tyr Asn Asp Val Tyr Val Asp Gly

115 120 125

Lys Pro Thr Gln Gly Gly Phe Ala Glu Thr Met Ile Val Glu Gln Lys

130 135 140

Phe Val Val Lys Ile Pro Glu Gly Leu Ala Pro Glu Gln Val Ala Pro

145 150 155 160

Leu Leu Cys Ala Gly Val Thr Val Tyr Ser Pro Leu Val His Phe Gly

165 170 175

Leu Lys Glu Ser Gly Leu Arg Gly Gly Ile Leu Gly Leu Gly Gly Val

180 185 190

Gly His Met Gly Val Lys Ile Ala Xaa Ala Leu Gly His His Val Thr

195 200 205

Val Ile Ser Ser Ser Asp Lys Lys Lys Gln Glu Ala Leu Glu His Leu

210 215 220

Gly Ala Asp Gln Tyr Leu Val Ser Ser Asp Ala Thr Ala Met Gln Glu

225 230 235 240

Ala Ala Asp Ser Leu Asp Tyr Ile Ile Asp Thr Val Pro Val Gly His

245 250 255

Pro Leu Glu Pro Tyr Leu Ser Leu Leu Lys Leu Asp Gly Lys Leu Ile

260 265 270

Leu Met Gly Val Ile Asn Thr Pro Leu Gln Phe Val Ser Pro Met Val

275 280 285

Met Leu Gly Arg Lys Ser Ile Thr Gly Ser Phe Ile Gly Ser Met Lys

290 295 300

Glu Thr Glu Glu Met Leu Glu Phe Trp Lys Glu Lys Gly Leu Ser Ser

305 310 315 320

Met Ile Glu Met Val Asn Met Asp Tyr Ile Asn Lys Ala Phe Glu Arg

325 330 335

Leu Glu Lys Asn Asp Val Arg Tyr Arg Phe Val Val Asp Val Lys Gly

340 345 350

Ser Lys Leu Val Asp Gln

355

31

1308

Triticum aestivum

31

gcaaaggcaa gtgcaagtcc tcccaaatcc acgcatccct cgcccgcgtc tcctcccgtt 60

gccgcagcag caagagaatg ggcagcgtcg acgcctccga gacgacggtc accgggtggg 120

ccgccaggga cgccaccggc cacctctccc cctacagata caccctcagg aaaacaggac 180

ctgaagacgt ggtgttgaag gttaagtact gcggcatctg ccacactgac gtccaccagg 240

tcaagaacga cctcggtgct tcgaagtacc ccatggtccc agggcatgag gtggttggcg 300

aggtggtgga ggtcgggccg gaggtgagca agttccgggc cggcgacgtg gtgggcgtgg 360

gggtgatcgt ggggtgctgc cgcgactgcc ggccgtgcaa ggccaacgtc gagcagtact 420

gcaacaagaa gatctggtcg tacaacgacg tctacaccga cggcaagccg acgcagggcg 480

gcttcgcctc cgccatggtc gtcgatcaga agttcgtggt gaagatcccc gccgggctgg 540

cgccggagca ggcggcgccg ctgctgtgcg cgggggtgac ggtgtacagc ccgctgaagc 600

acttcgggct catgaccccg ggcctccgcg gcgggatact gggcctgggc ggcgtgggcc 660

acatgggcgt gaaggtggcc aagtccatgg gccaccacgt gacggtgatc agctcgtcca 720

acaagaagcg ggccgaggcc atggacgacc tgggcgccga cgcgtacctc gtcagctccg 780

acaccgacca gatggccgcc gccgccgact cgctggacta catcatcgac accgtgcccg 840

ccaagcaccc gctggagccc tacctggcgc tgctcaagat ggacggcaag ctggtgctga 900

tgggcgtgat cgcggagccg ctcagcttcg tgtcccccat ggtgatgctg gggaggaaga 960

ccatcacggg gagcttcatc gggagcatgg acgagacgga ggaggtgctc cagttctgcg 1020

tcgacaaagg gctcacctcg cagatcgagg tcgtcaagat ggactacgtc aaccaggcgt 1080

tcgagaggct cgagcgcaac gacgtccgct accgcttcgt cgtcgacgtc ggcgggagca 1140

acatcgagga cgccgcctga tcgatgacgg ccgggaaaaa gagagagcta gaccgagcgg 1200

cgagtgcaat gacagtatca tgtgtgcccg tgccatttgc ctgtacttga tgcagtacat 1260

ttcgtgacct gttccgtttt catcaaattc gcgcgttcgt ttcggtgg 1308

32

360

Triticum aestivum

32

Met Gly Ser Val Asp Ala Ser Glu Thr Thr Val Thr Gly Trp Ala Ala

1 5 10 15

Arg Asp Ala Thr Gly His Leu Ser Pro Tyr Arg Tyr Thr Leu Arg Lys

20 25 30

Thr Gly Pro Glu Asp Val Val Leu Lys Val Lys Tyr Cys Gly Ile Cys

35 40 45

His Thr Asp Val His Gln Val Lys Asn Asp Leu Gly Ala Ser Lys Tyr

50 55 60

Pro Met Val Pro Gly His Glu Val Val Gly Glu Val Val Glu Val Gly

65 70 75 80

Pro Glu Val Ser Lys Phe Arg Ala Gly Asp Val Val Gly Val Gly Val

85 90 95

Ile Val Gly Cys Cys Arg Asp Cys Arg Pro Cys Lys Ala Asn Val Glu

100 105 110

Gln Tyr Cys Asn Lys Lys Ile Trp Ser Tyr Asn Asp Val Tyr Thr Asp

115 120 125

Gly Lys Pro Thr Gln Gly Gly Phe Ala Ser Ala Met Val Val Asp Gln

130 135 140

Lys Phe Val Val Lys Ile Pro Ala Gly Leu Ala Pro Glu Gln Ala Ala

145 150 155 160

Pro Leu Leu Cys Ala Gly Val Thr Val Tyr Ser Pro Leu Lys His Phe

165 170 175

Gly Leu Met Thr Pro Gly Leu Arg Gly Gly Ile Leu Gly Leu Gly Gly

180 185 190

Val Gly His Met Gly Val Lys Val Ala Lys Ser Met Gly His His Val

195 200 205

Thr Val Ile Ser Ser Ser Asn Lys Lys Arg Ala Glu Ala Met Asp Asp

210 215 220

Leu Gly Ala Asp Ala Tyr Leu Val Ser Ser Asp Thr Asp Gln Met Ala

225 230 235 240

Ala Ala Ala Asp Ser Leu Asp Tyr Ile Ile Asp Thr Val Pro Ala Lys

245 250 255

His Pro Leu Glu Pro Tyr Leu Ala Leu Leu Lys Met Asp Gly Lys Leu

260 265 270

Val Leu Met Gly Val Ile Ala Glu Pro Leu Ser Phe Val Ser Pro Met

275 280 285

Val Met Leu Gly Arg Lys Thr Ile Thr Gly Ser Phe Ile Gly Ser Met

290 295 300

Asp Glu Thr Glu Glu Val Leu Gln Phe Cys Val Asp Lys Gly Leu Thr

305 310 315 320

Ser Gln Ile Glu Val Val Lys Met Asp Tyr Val Asn Gln Ala Phe Glu

325 330 335

Arg Leu Glu Arg Asn Asp Val Arg Tyr Arg Phe Val Val Asp Val Gly

340 345 350

Gly Ser Asn Ile Glu Asp Ala Ala

355 360

33

1347

Glycine max

33

gcacgagatt tatacttgag aaatcccttt cacttcttcc tctcttcttc ttctatttcc 60

tcataccatt actcccatca tctatctgaa ctctcacact tctccgttca gaaatctcaa 120

catataatca tattacttac ccctccaaac ctacccctcc atcaccccca taataaaacc 180

aaaatgggat atcccgagac ctttacagga ttccaaattg aagatcctgc aaaatggacc 240

gagtttcata agaaccaata tacacctaag ccattcggcg actttgatgt cgatatcaag 300

attttggcct gtggtgtttg cgcgagtgat ttgcatactg ttagtggagg gtggggtgaa 360

cagaagtttc cgctttgtgt ggggcatgaa attgtaggag aagccctccg cgtcggccca 420

aaagtaactc tcatcaagcc cggtcaacgt gttggtgttg gtgctcaatc atactcctgt 480

ctccaatgtc gtcagtgcaa aaatgacaac gagacatact gtcttcacca actcgatacc 540

tatggagccg tctggcccga aacgggtatc gtttcccaag gtggctatgc tagccatgtg 600

agaactcatg agcattgggt tttccccatt ccagatggat tgaagaccga agaggcggct 660

cctatgttgt gtgctggttt gacggcttat agtcctttgg ttagaaatgg ctgtggacct 720

ggaaagaagg tgggtattgt gggattggga ggaattggtc attttggtgt tatgtttgct 780

aaggctttgg gcgctgagac ttgggctatt tcgagaaccc acgccaaaga agccgatgca 840

aagaaactcg gcgctgatgg ctttctcgcc acagccgaca aggactggaa caaagagcac 900

atcatgccct tcgacttgat agtcaacaca gcttcttcct cctcaggatt caatttctcc 960

gagtacctcg agctcctcga cgtacacggc cactgggtat ccgtcggact ccccgaaggc 1020

gaaggtcaaa cgatcgataa ctttcaacta ttgaaaaacg gtgttctcat cggaggaagt 1080

catctaggta gcagaaaaga ggttctggcc atgttggatt tagccgtgga gaagggtatc 1140

aatagttggg ttgagacgat tgatattagt gaggagggat tgaagacggc cttgacgagg 1200

ttgcataata atgatgttag gtatagattt acgatggtgg gttatgagaa gtgttttggt 1260

acttagaatg ggttattact cgtttgtaag attttatgta ggcgagatag taagctttac 1320

atctttacca aaaaaaaaaa aaaaaaa 1347

34

360

Glycine max

34

Met Gly Tyr Pro Glu Thr Phe Thr Gly Phe Gln Ile Glu Asp Pro Ala

1 5 10 15

Lys Trp Thr Glu Phe His Lys Asn Gln Tyr Thr Pro Lys Pro Phe Gly

20 25 30

Asp Phe Asp Val Asp Ile Lys Ile Leu Ala Cys Gly Val Cys Ala Ser

35 40 45

Asp Leu His Thr Val Ser Gly Gly Trp Gly Glu Gln Lys Phe Pro Leu

50 55 60

Cys Val Gly His Glu Ile Val Gly Glu Ala Leu Arg Val Gly Pro Lys

65 70 75 80

Val Thr Leu Ile Lys Pro Gly Gln Arg Val Gly Val Gly Ala Gln Ser

85 90 95

Tyr Ser Cys Leu Gln Cys Arg Gln Cys Lys Asn Asp Asn Glu Thr Tyr

100 105 110

Cys Leu His Gln Leu Asp Thr Tyr Gly Ala Val Trp Pro Glu Thr Gly

115 120 125

Ile Val Ser Gln Gly Gly Tyr Ala Ser His Val Arg Thr His Glu His

130 135 140

Trp Val Phe Pro Ile Pro Asp Gly Leu Lys Thr Glu Glu Ala Ala Pro

145 150 155 160

Met Leu Cys Ala Gly Leu Thr Ala Tyr Ser Pro Leu Val Arg Asn Gly

165 170 175

Cys Gly Pro Gly Lys Lys Val Gly Ile Val Gly Leu Gly Gly Ile Gly

180 185 190

His Phe Gly Val Met Phe Ala Lys Ala Leu Gly Ala Glu Thr Trp Ala

195 200 205

Ile Ser Arg Thr His Ala Lys Glu Ala Asp Ala Lys Lys Leu Gly Ala

210 215 220

Asp Gly Phe Leu Ala Thr Ala Asp Lys Asp Trp Asn Lys Glu His Ile

225 230 235 240

Met Pro Phe Asp Leu Ile Val Asn Thr Ala Ser Ser Ser Ser Gly Phe

245 250 255

Asn Phe Ser Glu Tyr Leu Glu Leu Leu Asp Val His Gly His Trp Val

260 265 270

Ser Val Gly Leu Pro Glu Gly Glu Gly Gln Thr Ile Asp Asn Phe Gln

275 280 285

Leu Leu Lys Asn Gly Val Leu Ile Gly Gly Ser His Leu Gly Ser Arg

290 295 300

Lys Glu Val Leu Ala Met Leu Asp Leu Ala Val Glu Lys Gly Ile Asn

305 310 315 320

Ser Trp Val Glu Thr Ile Asp Ile Ser Glu Glu Gly Leu Lys Thr Ala

325 330 335

Leu Thr Arg Leu His Asn Asn Asp Val Arg Tyr Arg Phe Thr Met Val

340 345 350

Gly Tyr Glu Lys Cys Phe Gly Thr

355 360

35

gttctaaccg ccggcgagaa acctgcagan ggcgagggcg agtgcggcgg acaggggatg 60

gaggagcatg gcaaggcggc tgtcggctgg gcggccaggg acgactccgg cgtcctctcc 120

ccgtacaact tctcnaggag ggctcagaag gacgatgacg tcaccatcaa ggtgctctac 180

tgcggcatct gccacactga cctgcacatc gtcaagaatg attggggcaa cgccatgtac 240

cccgtcgtcc ccggccacga gatcgtcggc gtcgtcaccg gcgtcggcgc cggcgtgacc 300

aagttcaagg ccggcgacac ggtgggcgtg ggctacttcg tcgcgtcgtg ccgcggctgc 360

gagtgctgcg ggaatgggta cgagaactac tgcgccaaga tggtaaccac ctgcaacggc 420

gtcgaccacg accacggcgg cggcgcggcc acccagggcg gcttctccga cgccatcgtc 480

gtcaacgagc actacgtgct ccgcgtgccg gcggggctgc cgctggacag cgcggcgccg 540

ctgctgtgcg ccggcgtgac ggtgtacagc ccgatggtga tccacggcct gaacccccgg 600

ggaacactcg gcgtctcggc tcggcggctc ggcactccct caattcgcca agcttcggga 660

tcgctcacct ctcacactcc cggggaagcg caggaggact ggacactcgg gccacgattc 720

tctcagcgcg agcggcaaat gcgnggngcg gcacaaggcg gatctaa 767

36

125

Oryza sativa

36

Met Glu Glu His Gly Lys Ala Ala Val Gly Trp Ala Ala Arg Asp Asp

1 5 10 15

Ser Gly Val Leu Ser Pro Tyr Asn Phe Ser Arg Arg Ala Gln Lys Asp

20 25 30

Asp Asp Val Thr Ile Lys Val Leu Tyr Cys Gly Ile Cys His Thr Asp

35 40 45

Leu His Ile Val Lys Asn Asp Trp Gly Asn Ala Met Tyr Pro Val Val

50 55 60

Pro Gly His Glu Ile Val Gly Val Val Thr Gly Val Gly Ala Gly Val

65 70 75 80

Thr Lys Phe Lys Ala Gly Asp Thr Val Gly Val Gly Tyr Phe Val Ala

85 90 95

Ser Cys Arg Gly Cys Glu Cys Cys Gly Asn Gly Tyr Glu Asn Tyr Cys

100 105 110

Ala Lys Met Val Thr Thr Cys Asn Gly Val Asp His Asp

115 120 125

37

tcgtggttag tgattacgac cagcaagtca caagtgaggg cgggagaggt tgaggttgag 60

gaagagcaga ggaatccatt ccaggagtgt ttaatggcgg gaggcaagga agcgcacggg 120

tgggcagcca gggatgtctc tggtcacctc tccccttacc acttctcacg gagggttcag 180

agagacgacg acgtcaccat caaggtgctc ttctgcgggc tttgccacac tgacctccac 240

gtcatcaaga acgagtttgg caacgccaag taccccgtcg ttcccgggca cgagatcgtc 300

ggcgtcgtca ccgacgtcgg ctccggcgtc acaagcttca agcccggcga cacggtgggc 360

gtgggctact tcgtccactc ctgccgcagc tgcgacagct gcagcaaggg gtacgagact 420

actgcccgca ctccgtggag acttccaacg cgttaacctg gacgacgatg acggnggccc 480

acaacaaagg cggttctccg accctctcgt caacaaccta ctttgtccgn cccggcaacc 540

tccgcccccg gggcccnccc tgtntcccgg ntaactt 577

38

114

Triticum aestivum

38

Met Ala Gly Gly Lys Glu Ala His Gly Trp Ala Ala Arg Asp Val Ser

1 5 10 15

Gly His Leu Ser Pro Tyr His Phe Ser Arg Arg Val Gln Arg Asp Asp

20 25 30

Asp Val Thr Ile Lys Val Leu Phe Cys Gly Leu Cys His Thr Asp Leu

35 40 45

His Val Ile Lys Asn Glu Phe Gly Asn Ala Lys Tyr Pro Val Val Pro

50 55 60

Gly His Glu Ile Val Gly Val Val Thr Asp Val Gly Ser Gly Val Thr

65 70 75 80

Ser Phe Lys Pro Gly Asp Thr Val Gly Val Gly Tyr Phe Val His Ser

85 90 95

Cys Arg Ser Cys Asp Ser Cys Ser Lys Gly Tyr Glu Thr Thr Ala Arg

100 105 110

Thr Pro

39

cttacatgta agctcgtgcc gaattcggca cgagcttaca ctctcatcct ctcctctccc 60

actccacacc caggcaccca gctcgagcgc gactcggcaa atccatggct cccacggcgg 120

cgtcggcggc ggcgggggag gagcagcagg cggtggggct ggcggcgcgc gactcctccg 180

gccacctctc ccctttcgcc atctcccgga ggagcactgg agatgacgat gtggcgataa 240

agattctgtt ctgtggaatc tgccactctg acctgcactg catcaagaat gagtggaagc 300

actccatcta cccacttgtt cctgggcacg agatcgccgg tgtggtgacg gaagtgggga 360

agaacgtcac caggtttaag gccggtgaca gggtcggcgt ggggtgcatg gtgaactcgt 420

gccggtcatc cgagagctgt aacaatggct tcgagaacca ctgcccggga gggcgtcttc 480

aacctacaac tccgtcgaca aggacgggaa cgtcacctan ngngggttac tcnagcatgg 540

tggtggtgca cgagcgcttc tgggcaagtt cccgggnggg 580

40

Met Ala Pro Thr Ala Ala Ser Ala Ala Ala Gly Glu Glu Gln Gln Ala

1 5 10 15

Val Gly Leu Ala Ala Arg Asp Ser Ser Gly His Leu Ser Pro Phe Ala

20 25 30

Ile Ser Arg Arg Ser Thr Gly Asp Asp Asp Val Ala Ile Lys Ile Leu

35 40 45

Phe Cys Gly Ile Cys His Ser Asp Leu His Cys Ile Lys Asn Glu Trp

50 55 60

Lys His Ser Ile Tyr Pro Leu Val Pro Gly His Glu Ile Ala Gly Val

65 70 75 80

Val Thr Glu Val Gly Lys Asn Val Thr Arg Phe Lys Ala Gly Asp Arg

85 90 95

Val Gly Val Gly Cys Met Val Asn Ser Cys Arg Ser Ser Glu Ser Cys

100 105 110

Asn Asn Gly Phe Glu Asn His Cys Pro Gly Gly Arg Leu Gln Pro Thr

115 120 125

Thr Pro Ser Thr Arg Thr Gly Thr Ser Pro Xaa Xaa Gly Tyr Ser Ser

130 135 140

Met Val Val Val His Glu Arg Phe

145 150

41

cagagatgga gcatccaaag aaggtttttg gttggnctgc tcgggactca tctggncttc 60

tctccccatt caacttctcc agaagagaaa cgggggagaa agatttggta ttcaaagtgc 120

aatactgtgg aatatgccac tcagatctac atatgttaaa gaacgagtgg ggcaacacaa 180

cctatcctct ggttcctggg cacgagattg ctggtgtggt aacagaagtt ggaagcaagg 240

tgcaaaagtt caaagttgga gaccgggtag gtgtggggtg catgattgga tcctgngttc 300

atgtgaaagc tgtgatgaaa tcttgagaat tactgcccca aaatgttctc acatacggtg 360

tcaagtattt tgtggaccat aacacatgga gntactctga cttgtggttn ggtgacactt 420

tgtgtngatt cggtaccnac cacttatgct gtctctctct ctgtctggat nctgtgnnag 480

ccttagncnt gacttacact gctnanttgt gtgtgcttgt gacttgnact gcgnagttcc 540

aannttgant atgncatata ccttctacaa agagga 576

42

Glu Met Glu His Pro Lys Lys Val Phe Gly Trp Xaa Ala Arg Asp Ser

1 5 10 15

Ser Gly Leu Leu Ser Pro Phe Asn Phe Ser Arg Arg Glu Thr Gly Glu

20 25 30

Lys Asp Leu Val Phe Lys Val Gln Tyr Cys Gly Ile Cys His Ser Asp

35 40 45

Leu His Met Leu Lys Asn Glu Trp Gly Asn Thr Thr Tyr Pro Leu Val

50 55 60

Pro Gly His Glu Ile Ala Gly Val Val Thr Glu Val Gly Ser Lys Val

65 70 75 80

Gln Lys Phe Lys Val Gly Asp Arg Val Gly Val Gly Cys Met Ile Gly

85 90 95

Ser

43

cgaacgtcta atcgcctcac acacaggaaa ggcaaaaaag aaaccaaaac ctccggggag 60

ttactctaga gatttccggc ggaacctcgc gtcaggatgg cacccacggc gacggcggcg 120

gagcagagcc agcagcacac gaagaaggcg gtgggtctgg cggcgctcga cgcctccggc 180

cacctctccc cgctcgccat cacccgaagg agcaccggag atgacgatgt cgtgataaag 240

attctgtact gcgggatctg ccactctgac ctgcacagca tcaagaacga ctggaagaac 300

gccaagtacc ccatgattcc cgggcacgag atcgccggcg aggtcaccga ggtcggcaag 360

aacgtgacca agttcaaggc angcgaccgt gtcggcgtcn ggtgcatggt gaactcctgc 420

caatcctgcg aaaactgcga caagggcttc gagaatcact gcccnggcat gatttcaact 480

acaactccgt cgacgtgacg ga 502

44

Met Ala Pro Thr Ala Thr Ala Ala Glu Gln Ser Gln Gln His Thr Lys

1 5 10 15

Lys Ala Val Gly Leu Ala Ala Leu Asp Ala Ser Gly His Leu Ser Pro

20 25 30

Leu Ala Ile Thr Arg Arg Ser Thr Gly Asp Asp Asp Val Val Ile Lys

35 40 45

Ile Leu Tyr Cys Gly Ile Cys His Ser Asp Leu His Ser Ile Lys Asn

50 55 60

Asp Trp Lys Asn Ala Lys Tyr Pro Met Ile Pro Gly His Glu Ile Ala

65 70 75 80

Gly Glu Val Thr Glu Val Gly Lys Asn Val Thr Lys Phe Lys Ala Xaa

85 90 95

Asp Arg Val Gly Val Xaa Cys Met Val Asn Ser Cys Gln Ser Cys Glu

100 105 110

Asn Cys Asp Lys Gly Phe Glu Asn His Cys Pro Gly Met Ile Ser Thr

115 120 125

45

cacaagtgtg agaaaaatta tgagttccaa aggtgttggt gaggattgcc tgggatgggc 60

agcaagagat gcatccggag ttctatcacc ttacaaattc agtcgcagga ctcttgggaa 120

tgaagatgtt catattaaaa ttacgcactg tggtgtttgc ttcgctgatg tagtttggac 180

aaggaataaa catggtgact caaagtatcc tgtcgtgcct ggtcacgaga ttgctgggat 240

tgtgacaaag gttggctcca atgtccaccg ttttaaggtt ggtgaccatg ttggagtggg 300

gacttatgtc aactcatgta gggattgtga acattgtaat gacagagaag aagttcattg 360

taccaaggga tctgttttca cttttaatgg tgttgatttt gatggtacaa ttacaaaagg 420

aggatactcc agttacatag tagtccatga gaggtactgc ttcacgatac caaaaagcta 480

tccattggct tcagcagctc ctttgctttg tgctggaatt acagtttatt caccgatngt 540

gcgccacaaa gatgaatcaa cctggtaaat cccntgggag tgattggncc tggtggcccc 600

ccgtcaaatg gcantgaaat ttggaaaagg catttggttt gagtgtgaca cntttcacca 660

ccatcatatc caacaaagag ggcggnnccn gaaccccctt nggtggncac naaattnttg 720

tttcncccca atcncaaagg naat 744

46

172

Glycine max

46

Met Ser Ser Lys Gly Val Gly Glu Asp Cys Leu Gly Trp Ala Ala Arg

1 5 10 15

Asp Ala Ser Gly Val Leu Ser Pro Tyr Lys Phe Ser Arg Arg Thr Leu

20 25 30

Gly Asn Glu Asp Val His Ile Lys Ile Thr His Cys Gly Val Cys Phe

35 40 45

Ala Asp Val Val Trp Thr Arg Asn Lys His Gly Asp Ser Lys Tyr Pro

50 55 60

Val Val Pro Gly His Glu Ile Ala Gly Ile Val Thr Lys Val Gly Ser

65 70 75 80

Asn Val His Arg Phe Lys Val Gly Asp His Val Gly Val Gly Thr Tyr

85 90 95

Val Asn Ser Cys Arg Asp Cys Glu His Cys Asn Asp Arg Glu Glu Val

100 105 110

His Cys Thr Lys Gly Ser Val Phe Thr Phe Asn Gly Val Asp Phe Asp

115 120 125

Gly Thr Ile Thr Lys Gly Gly Tyr Ser Ser Tyr Ile Val Val His Glu

130 135 140

Arg Tyr Cys Phe Thr Ile Pro Lys Ser Tyr Pro Leu Ala Ser Ala Ala

145 150 155 160

Pro Leu Leu Cys Ala Gly Ile Thr Val Tyr Ser Pro

47

ctcgtgccga attcggcacg aggaacgacc tcggtgcttc gaagtacccc atggtcccag 60

ggcatgaggt ggttggcgag gtggtggagg tcgggccgga ggtgagcaag ttccgggccg 120

gcgacgtggt gggcgtgggg gtgatcgtgg ggtgctgccg cgactgccgg ccgtgcaagg 180

ccaacgtcga gcagtactgc aacaagaaga tctggtcgta caacgacgtc tacaccgacg 240

gcaagccgac gcagggcggc ttcgcctccg ccatggtcgt cgatcagaag ttcgtggtga 300

agatccccgc cgggctggcg ccggagcagg cggcgccgct gctgtgcgcg ggggtgacgg 360

tgtacagccc gctgaagcac ttcgggctca tgaccccggg cctccgcggc gggatactgg 420

gcctgggcgg cgttgggcca catgggcgtg aangtggcaa atccatggcc acaacgtgac 480

gtgannactc gtccaacaag aacggccnag gcatggacac tgggngccga gcgtactcgt 540

nactccgaca ccgacaaang gccccgccgc cgancnctgg atanncanca cacgtgccgc 600

caacaccgct ggacctactg cctctaagtg acgnaactgt ctatgctgat ncgacc 656

48

Arg Ala Glu Phe Gly Thr Arg Asn Asp Leu Gly Ala Ser Lys Tyr Pro

1 5 10 15

Met Val Pro Gly His Glu Val Val Gly Glu Val Val Glu Val Gly Pro

20 25 30

Glu Val Ser Lys Phe Arg Ala Gly Asp Val Val Gly Val Gly Val Ile

35 40 45

Val Gly Cys Cys Arg Asp Cys Arg Pro Cys Lys Ala Asn Val Glu Gln

50 55 60

Tyr Cys Asn Lys Lys Ile Trp Ser Tyr Asn Asp Val Tyr Thr Asp Gly

65 70 75 80

Lys Pro Thr Gln Gly Gly Phe Ala Ser Ala Met Val Val Asp Gln Lys

85 90 95

Phe Val Val Lys Ile Pro Ala Gly Leu Ala Pro Glu Gln Ala Ala Pro

100 105 110

Leu Leu Cys Ala Gly Val Thr Val Tyr Ser Pro Leu Lys His Phe Gly

115 120 125

Leu Met Thr Pro Gly Leu Arg Gly Gly Ile Leu Gly Leu Gly Gly Val

130 135 140

Gly Pro His Gly Arg Glu Xaa Gly Lys Ser Met

145 150 155