Glyphosate-resistant plants

Abstract

This invention relates to glyphosate-resistant transgenic plants and methods of making the same. In a preferred embodiment, a DNA fragment which comprises an EPSPS 5' regulatory sequence and a glyphosate-resistant EPSPS coding sequence is introduced into regenerable plant cells. The encoded EPSPS has a chloroplast transit peptide. The DNA fragment does not contain a non-EPSPS enhancer. Cells are selected for stable transformation, and the selected cells can be used to regenerate glyphosate-resistant transgenic plants. The DNA fragment used for transformation preferably comprises a modified plant genomic sequence, such as SEQ ID NO: 2, SEQ ID NO:4 or SEQ ID NO: 6. In one embodiment, two DNA fragments of this invention are stably transformed into a plant to confer glyphosate-resistance.

Description

TECHNICAL FIELD

This invention relates to glyphosate-resistant transgenic plants and methods of making the same.

BACKGROUND

Glyphosate is a widely used component in herbicides. Glyphosate inhibits 5-enolpyruvyl-3-phosphoshikimic acid synthase (EPSP synthase, or EPSPS), which is involved in the synthesis of aromatic amino acids in plant cells. Inhibition of EPSPS effectively disrupts protein synthesis and thereby kills the affected plant cells. Because glyphosate is non-selective, it kills both weeds and crop plants. Accordingly, there is a need to produce transgenic crop plants that are resistant to glyphosate.

Recombinant DNA technology has been used to create mutant EPSP synthases that are glyphosate-resistant. These mutant EPSP synthases can be transformed into plants and confer glyphosate-resistance upon the transformed plants. Examples of mutant EPSP synthases and glyphosate-resistant transgenic plants are illustrated in U.S. Pat. Nos. 6,040,497 and 5,554,798, 5,310,667 and WO 00/66748.

Current plant transformation technology employs chimeric expression vectors. These vectors include regulatory sequences, such as enhancers or promoters, that are heterologous to the EPSPS genes. For instance, WO 00/66748 fuses enhancers from CaMV 35S, FMV 35S, rice actin 1, rice GOS2, maize polyubiquitin, or barley plastocyanin genes to a glyphosate-resistant EPSPS coding sequence in order to enhance the expression of the glyphosate-resistant EPSPS in transformed plant cells.

No one has used a complete expression cassette of the EPSP synthase gene isolated from the genome of a donor plant and mutated to give glyphosate resistance. In one embodiment of the present invention, the expression cassette of the EPSP synthase gene consists of a native EPSPS 5' regulator sequence, a coding sequence (with or without introns) encoding a glyphosate-resistant EPSPS which includes a native transit peptide, and a native EPSPS 3' regulatory sequence (such as an EPSPS transcriptional terminator). The fact that such an expression cassette is sufficient to provide glyphosate resistance is surprising. Moreover, the use of the native EPSPS 5' and/or 3' regulatory sequences simplifies the process of constructing expression vectors suitable for plant transformation.

Suitable sources of EPSP synthase genes include dicotyledonous plants, such as Arabidopsis thaliana, and mono-cotyledonous plants, such as Zea mays, Arabidopsis thaliana has two EPSP synthase genes (epm1 and epm2). The present invention includes use of one or both of mutated epm1 and epm2 to confer resistance to glyphosate. Mutated EPSP synthase genes from Zea mays or other plants can also be used for transforming plant cells to make glyphosate-resistant plants.

SUMMARY OF THE INVENTION

In accordance with one aspect of the present invention, a DNA fragment which comprises an EPSPS 5' regulatory sequence and a glyphosate-resistant EPSPS coding sequence (including a chloroplast transit peptide coding sequence) is introduced into regenerable plant cells. The DNA fragment does not contain a non-EPSPS enhancer. Cells are selected for stable transformation. The selected cells are then used to regenerate glyphosate-resistant, transgenic plants.

In one embodiment, the DNA fragment used for transformation comprises a modified plant genomic sequence. The unmodified plant genomic sequence comprises at least part of an EPSPS gene, and includes an EPSPS 5' regulatory sequence and a glyphosate-sensitive EPSPS coding sequence (including a chloroplast transit peptide coding sequence). The glyphosate-sensitive EPSPS coding sequence is modified to make the encoded EPSPS glyphosate-resistant. The DNA fragment comprising the modified plant genomic sequence is stably transformed into plant cells, from which glyphosate-resistant plants are regenerated.

In a preferred embodiment, the DNA fragment used for transformation comprises SEQ ID NO: 2. In another preferred embodiment, the DNA fragment used for transformation comprises SEQ ID NO: 4. In yet another preferred embodiment, the DNA fragment comprises SEQ ID NO: 6. In a further preferred embodiment, any two sequences selected from SEQ ID. NO: 2, SEQ ID NO: 4, and SEQ ID NO: 6 are used to transform plant cells. In one embodiment, the transgenic plant comprises transformed SEQ ID. NO: 2 and SEQ ID NO: 4.

Other features, objects, and advantages of the present invention are apparent in the detailed description that follows. It should be understood, however, that the detailed description, while indicating preferred embodiments of the invention, are given by way of illustration only, not limitation. Various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from the detailed description.

DETAILED DESCRIPTION

This invention relates to methods of making glyphosate-resistant plants. In accordance with one aspect of the invention, a DNA fragment is introduced into regenerable, glyphosate-sensitive recipient plant cells. The DNA fragment comprises an EPSPS 5' regulatory sequence, and a coding sequence encoding a glyphosate-resistant EPSPS. The EPSPS 5' regulatory sequence is operably linked to the EPSPS coding sequence. The glyphosate-resistant EPSPS includes a chloroplast transit peptide. The DNA fragment does not contain a non-EPSPS enhancer. The recipient plant cells are selected for glyphosate-resistance and stable transformation. The cells thus selected can be used to regenerate glyphosate-resistant plants. As used herein, a "DNA fragment" may be either linear or circular. Preferably, the DNA fragment used for transformation is a linear DNA fragment. A "coding sequence" encoding an EPSPS refers to a nucleic acid sequence transcription and translation of which produce a functional EPSPS. The boundaries of the coding sequence are generally determined by a translation start codon at its 5' end and a translation stop co don at its 3, end. A coding sequence of EPSPS may be a cDNA, or a plant genomic sequence which consists of all of the exons and introns of an EPSPS gene. An EPSPS gene refers to the plant genomic sequence which includes the EPSPS 5' regulatory sequence, the EPSPS coding sequence (including the sequence encoding the chloroplast transit peptide), and the EPSPS 3' regulatory sequence (such as an EPSPS transcriptional terminator). A "plant genomic sequence" refers to a nucleotide sequence found in the genome of the plant.

A chloroplast transit peptide functions post-translationally to direct a polypeptide to chloroplast. Either endogenous or heterologous chloroplast peptides can be used in the present invention. As used herein, "heterologous" means derived from a different source, and "endogenous" means derived from the same source. In a preferred embodiment, the endogenous transit peptide encoded by a native EPSPS gene is used.

As used herein, an EPSPS 5' regulatory sequence refers to a nucleotide sequence located upstream (5') to the start codon of the EPSPS coding sequence in an EPSPS gene in a plant or plant cell which has not been subject to genetic engineering. The 5' regulatory sequence generally includes an EPSPS promoter which directs the transcription of the EPSPS gene. Preferably, the EPSPS 5' regulatory sequence comprises one or more EPSPS enhancers operably linked to the promoter. In one embodiment, the 5' regulatory sequence comprises at least 200 bp. Preferably, the 5' regulatory sequence comprises at least 400, 600, 800, 1000, 1,200 or 1,800 bp.

An EPSPS 3' regulatory sequence refers to a nucleotide sequence located downstream (3') to the stop codon of the EPSPS coding sequence in an EPSPS gene in a plant or plant cell which has not been subject to genetic engineering. The 3, regulatory sequence generally includes a transcription terminator which controls the termination of the transcription of the EPSPS gene.

"Operably linked" refers to a juxtaposition of genetic elements, wherein the elements are in a relationship permitting them to operate in the expected manner. For instance, a 5' regulatory sequence is operably linked to a coding sequence if the 5' regulatory sequence functions to initiate transcription of the coding sequence.

Preferably, the DNA fragment used for transformation does not include a non-EPSPS enhancer. As used in the present invention, a "non-EPSPS enhancer" refers to an enhancer which is not used by an EPSPS gene in a plant or plant cell which has not been subject to genetic engineering. Non-EPSPS enhancers include, but are not limited to, enhancers that are associated with CaMV 35S, FMV 35S, rice actin 1, rice GOS2, maize polyubiquitin, or barley plastocyanin genes.

As used herein, a "glyphosate-resistant" cell or plant refers to a cell or plant that can survive or continue to grow in the presence of certain concentrations of glyphosate that typically kill or inhibit the growth of other cells or plants. Growth includes, for instance, photosynthesis, increased rooting, increased height, increased mass, or development of new leaves. In one embodiment, a glyphosate-resistant cell can grow and divide on a culture medium containing 50 mg/l or more glyphosate. Preferably, a glyphosate-resistant cell can grow and divide on a culture medium containing 100 mg/l or more glyphosate, such as 200 mg/l, 300 mg/l or 400 mg/l glyphosate. More preferably, a glyphosate-resistant cell can grow and divide on a culture medium containing 500 mg/l or more glyphosate, such as 600 mg/l. For purposes of the present invention, the term "glyphosate" includes any herbicidally effective form of N-phosphonomethylglycine (including any salt thereof) and other forms which result in the production of the glyphosate anion in plants.

Regenerable glyphosate-resistant plant cells may be used to regenerate glyphosate-resistant plants. In one embodiment, the glyphosate-resistant plant thus regenerated can survive or continue to grow after being sprayed with glyphosate at a rate of 25 g/ha (grams per hectare) or more. Preferably, the glyphosate-resistant plant thus regenerated can survive or continue to grow after being sprayed with glyphosate at a rate of 50 g/ha or more, such as 100 g/ha, 200 g/ha, 400 g/ha, or 800 g/ha. More preferably, the glyphosate-resistant plant thus regenerated can survive or continue to grow after being sprayed with glyphosate at a rate of 1000 g/ha or more, such as 2000 g/ha and 3000 g/ha. The spray may preferably be carried out at or after the growth stage of v2, such as v3, v4, v5 or later stages. In another embodiment, the regenerated glyphosate-resistant plant can tolerate the spray of glyphosate at between 0.1 M and 0.4 M.

As used herein, a "glyphosate-resistant" EPSPS refers to an EPSPS the expression of which in a plant cell confers glyphosate resistance upon the plant cell. An EPSPS is "glyphosate-sensitive" if it does not confer glyphosate-resistance when being expressed in plant cells.

A variety of EPSPS mutations have been known to be glyphosate-resistant and capable of conferring glyphosate resistance upon transformed plants. For instance, EPSPS of Zea mays (GenBank Accession No. X63374) can be mutated at amino acid residues 102 (substitution of He for Thr) and 106 (substitution of Ser for Pro). EPSPS encoded by epm1 gene of Arabidopsis thaliana can be mutated at amino acid residues 179 (substitution of He for Thr) and 183 (substitution of Ser for Pro). EPSPS encoded by epm2 gene of Arabidopsis thaliana can be mutated at amino acid residues 177 (substitution of He for Thr) and 182 (substitution of Ser for Pro). These mutated EPSPSs are glyphosate-resistant and capable of conferring glyphosate resistance upon transformed plants. Other mutated or modified EPSPSs, such as those described in U.S. Pat. Nos. 5,310,667, 5,866,775, 6,225,114, and 6,248,876, or natural EPSPS variants showing glyphosate-resistance, can be used in the present invention. In addition, bacteria-derived, glyphosate-resistant EPSPSs, after fusion with a chloroplast transit peptide, can also be used.

The DNA fragment comprising the EPSPS 5' regulatory sequence and the glyphosate-resistant EPSPS coding sequence can be stably transformed into a regenerable plant cell. As used herein, stable transformation refers to integration of the DNA fragment into the genome of the transformed plant cell.

In one embodiment, the EPSPS 5' regulatory sequence in the DNA fragment used for transformation comprises an EPSPS enhancer and an EPSPS promoter. In another embodiment, the DNA fragment used for transformation further comprises an EPSPS 3' regulatory sequence, such as an EPSPS transcriptional terminator, which is operably linked to the coding sequence encoding the glyphosate-resistant EPSPS.

In yet another embodiment, the DNA fragment used for transformation comprises a modified plant genomic sequence that encodes a glyphosate-resistant EPSPS. Without modification, the plant genomic sequence encodes a glyphosate-sensitive EPSPS. Modifications that are capable of converting a glyphosate-sensitive EPSPS to a glyphosate-resistant EPSPS are known in the art.

In a preferred embodiment, the DNA fragment used for transformation is modified from a plant genomic sequence. Before modification, the plant genomic sequence comprises an EPSPS regulatory sequence, a coding sequence encoding a glyphosate-sensitive EPSPS which includes a chloroplast transit peptide, and preferably an EPSPS 3, regulatory sequence, such as an EPSPS transcriptional terminator. The genomic sequence may be obtained by fragmenting the genome of a plant of interest, or isolated from bacterial artificial chromosome clones. Other methods for obtaining genomic sequences can also be used, such as PCR or DNA synthesis.

The EPSPS-coding sequence in this plant genomic sequence is then subject to nucleotide modification(s) to render the encoded EPSPS glyphosate resistant. Suitable modifications for this purpose, such as nucleotide substitutions, are well known in the art. The DNA fragment comprising the genomic sequence thus modified can be stably transformed into glyphosate-sensitive recipient plant cells. These transformed plant cells are selected for glyphosate resistance and then used to regenerate glyphosate-resistant plants.

The recipient plant cells are regenerable. They can be derived from immature embryos or meristematic tissues which contain cells that have not yet terminally differentiated. Juvenile leaf basal regions, immature tassels and gametic cells can be used to provide regenerable recipient cells for Zea mays. The preferred source of recipient cells for soybean includes the immature cotyledon.

In another preferred embodiment, two or more DNA fragments can be stably transformed into a recipient plant cell. Each of these DNA fragments includes an EPSPS 5' regulatory sequence, a coding sequence encoding a glyphosate-resistant EPSPS which contains a chloroplast transit peptide, and preferably an EPSPS 3' regulatory sequence (such as an EPSPS transcriptional terminator). These DNA fragments can be modified plant genomic sequences. They can be derived from the same or different plant species. They can be derived from the same EPSPS gene, or from different EPSPS genes of the same plant species, such as emp1 and emp2 of Arabidopsis thaliana.

Transformation of plant cells can be carried out using various methods. These methods include, but are not limited to, Agrobacterium tumefaciens mediated DNA transfer, PEG or liposome mediated DNA transfer, electroporation, micro-injection, microprojectile or particle bombardment, receptor-mediated DNA transfer, and viral or other vector mediated DNA transfer. Preferably, transformation is carried out using aerosol beam injection as described in U.S. patent application Ser. No. 09/450,226, which is incorporated herein by reference.

Selection for stably transformed plant cells can be performed using methods as appreciated by one of ordinary skill in the art. For instance, the transformed cells can be grown and selected on media containing glyphosate. Preferably, the introduced DNA fragment is stably transformed and integrated into a chromosome of the transformed plant cell. A variety of assays can be used to confirm stable transformation. Suitable assays include molecular biological assays, such as Southern and Northern Blotting and PCR, or biochemical assays, such as ELISA and Western Blot. In addition, plant part assays, such as leaf and root assays, or analysis of the phenotype of the whole regenerated plant, can be used to confirm stable transformation.

Plants can be regenerated from the selected, stably transformed cells. Progeny can be recovered from the regenerated plants and tested for glyphosate resistance. Seeds or other parts of the regenerated transgenic plants can also be obtained. In one embodiment, glyphosate-resistant plants are made by crossing.

Both monocotyledonous and dicotyledonous plants can be transformed using the methods of the present invention. The glyphosate-resistant EPSPS coding sequence can be derived from either monocotyledonous or dicotyledonous plants. The representative monocotyledonous and dicotyledonous plants used in the present invention include, but are not limited to, Oryza sativa, Zea mays, Hordeum vulgare, Triticum aestivum, Avena sativa, turf grasses including species of the genera Poa, Festuca, Lolium, Zoysia, and Cynodon among others, Glycine max, Gossypium hirsutii, Lycopersicum esculentum, Solanum tuberosum, Phaseolus species, Beta vulgaris, and Brassica species.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature used herein and the laboratory procedures in cell culture and molecular genetics described herein are those well known and commonly employed in the art. Standard techniques can be used for recombinant nucleic acid methods, polynucleotide synthesis, plant cell culture, cell culture, tissue culture, and plant transformation and regeneration. Generally, enzymatic reactions and purification and/or isolation steps are performed according to the manufacturers' specifications. The techniques and procedures are generally performed according to conventional methodology disclosed, for example, in Molecular Cloning A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989), and Current Protocols in Molecular Biology (John Wiley & Sons, Baltimore, Md., 1989).

It should be understood that the above-described embodiments and the following examples are given by way of illustration, not limitation. Various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art from the present description.

EXAMPLE 1

Preparation and Mutation of Arabidopsis Genomic Fragments Containing EPSPS Genes

Two bacterial artificial chromosome (BAC) clones, F27K7 and F4L23, were obtained from the Arabidopsis Biological Resource Center, DNA Stock Center, at the Ohio State University. F27K7 and F4L23 contain the EPSPS genes found on chromosome 1 and 2 of Arabidopsis thaliana, respectively. The F27K7 clone was digested using Sac II and Bam HI restriction enzymes to produce a 4.7 kb fragment, the sequence of which is shown as SEQ ID NO: 1. The 4.7 kb fragment comprises the complete EPSPS gene (epm1) found on chromosome which includes an EPSPS 5' regulatory sequence (the sequence before nucleotide residue 1290), an EPSPS coding sequence (from nucleotide residue 1290 to nucleotide residue 3729), and an EPSPS 3, regulatory sequence (the sequence after nucleotide residue 3729). The EPSPS coding sequence also encodes a chloroplast transit peptide (from nucleotide residue 1290 to nucleotide residue 1612). The sequence encoding this chloroplast transit peptide can be predicted using the computer program PSORT maintained on the public accessible GenomeNet at Kyoto University, Japan.

The 4.7 kb fragment was cloned into a pbluescript II vector (Stratagene), and two nucleotide substitutions were introduced into the EPSPS coding sequence using QuikChange.RTM. Site-Directed Mutagenesis Kit (Stratagene) according to the instructions of the manufacturer. The two nucleotide substitutions are a cytosine to thymine substitution at nucleotide 2007 and a cytosine to thymine substitution at nucleotide 2018. The mutated sequence is shown as SEQ ID NO: 2. The mutated sequence encodes a glyphosate-resistant EPSPS which has, as compared to the EPSPS encoded by SEQ ID NO: 1, a Thr to Ile mutation at amino acid 179 and a Pro to Ser mutation at amino acid 183. The amino acid sequence of the glyphosate-resistant EPSPS is shown as SEQ ID NO: 7. The pbluescript II vector containing SEQ ID NO: 2 is referred to as epm1 vector.

The BAC F4L23 clone was digested using Eco RI restriction enzyme to produce a 5.2 kb fragment, the sequence of which is shown as SEQ ID NO: 3. The 5.2 kb fragment comprises the complete EPSPS gene (epm2) from chromosome 2, which includes an EPSPS 5 'regulatory sequence (the sequence before nucleotide 1515), and EPSPS coding sequence (from nucleotide 1515 to nucleotide 3872), and an EPSPS 3' regulatory sequence (the sequence after nucleotide 3872). The EPSPS coding sequence also encodes a chloroplast transit peptide (from nucleotide 1515 to nucleotide 1665). The sequence encoding this chloroplast transit peptide can be predicted using the computer program PSORT maintained on the public accessible GenomeNet at Kyoto University, Japan.

The 5.2 kb fragment was cloned into a pbluescript II vector, and then subject to site-directed mutagenesis using QuikChange.RTM. Site-Directed Mutagenesis Kit (Stratagene). SEQ ID NO: 4 shows the mutated sequence which has two nucleotide substitutions in the EPSPS coding sequence as compared to SEQ ID NO: 3. The two substitutions are a cytosine to thymine substitution at nucleotide 2134 and a cytosine to thymine substitution at nucleotide 2145. The mutated sequence encodes a glyphosate-resistant EPSPS which has, as compared to the EPSPS encoded by SEQ ID NO: 3, a Thr to Ile mutation at amino acid 178 and a Pro to Ser mutation at amino acid 182. The amino acid sequence of the .Iadd.(putative) .Iaddend.glyphosate-resistant EPSPS is shown as SEQ ID: 8. The pbluescript II vector containing SEQ ID NO: 4 is referred to as epm2 vector.

EXAMPLE 2

Transformation of Soybean

The Bam HI/Sac II fragment (SEQ ID NO: 2) of epm1 vector and the Eco RI fragment (SEQ ID NO: 4) of epm2 vector were used to transform soybean embryogenic callus using an aerosol beam injector as described in U.S. patent application Ser. No. 09/450,226, which is incorporated herein by reference. These fragments comprised mutant epm1 and mutant epm2 which encode .Iadd.(putative) .Iaddend.glyphosate-resistant EPSPSs. These fragments were used either alone or, preferably, together.

The transformed tissue .[.was.]. .Iadd.is .Iaddend.selected for glyphosate resistance using the method described below. First, the beamed embryogenic callus .[.was.]. .Iadd.is .Iaddend.maintained for one month on B1-30 3Co5My0.01PA medium. Table 1 shows the composition of B1-30 3Co5My0.01PA medium.

TABLE-US-00001 TABLE 1 Ingredients in 1 liter B1-30 3Co5My0.10PA Medium (pH 5.8) MS Salts* 4.43 g NaEDTA 37.3 mg 2,4 dichlorophenoxyacetic acid 30 mg Phytagar 8 g Coconut water 30 ml Myo-inositol 5 g Phytic acid 10 mg *Sigma Plant Culture catalogue, reference M5519

The tissue .[.was.]. .Iadd.is .Iaddend.then transferred to the same medium but now containing 300 mg/l glyphosate. After a number of passages (up to 5 passages, each passage may last for about a month) on this latter medium, resistant clonal material may be identified. After an optional further few passages on B1-30 3Co5My0.01PA medium but containing 500 mg/l glyphosate, the growing tissue .[.was.]. .Iadd.is .Iaddend.transferred to a regeneration media as described in U.S. patent Ser. No. 09/450,226. Regenerated plants .[.were.]. .Iadd.are .Iaddend.transferred to pots in a greenhouse. These plants and their progenies .[.were.]. .Iadd.are .Iaddend.sprayed with glyphosate at commercial rates, and complete resistance to glyphosate .[.was.]. .Iadd.is expected to be .Iaddend.observed. Progenies .[.segregated.]. .Iadd.are expected to segregate .Iaddend.3:1 for glyphosate resistance as would be expected for Mendelian inheritance of a transgene.

Preferably, both mutant epm1 (such as SEQ ID NO: 2) and mutant epm2 (such as SEQ ID NO: 4) are stably transformed into a plant cell, from which glyphosate-resistant plants can be regenerated.

EXAMPLE 3

Preparation and Mutation of Corn Genomic Fragments Containing EPSPS Gene

A corn (B 73) BAC library was screened with a probe containing a sequence of a corn EPSPS gene published in Genbank accession number X63374 by Incyte Genomics Inc. Four BAC clones were identified. Southern blot analysis indicated that all four clones contained the same EPSPS gene. One BAC clone was further characterized by nucleotide sequencing which resulted in identification of a 6.0 kb genomic fragment flanked by unique Cla I and Eco RV sites. The sequence of the 6.0 kb fragment was shown as SEQ ID NO: 5. The 6.0 kb fragment includes an EPSPS 5' regulatory sequence (the sequence before nucleotide 1868), an EPSPS coding sequence (from nucleotide 1868 to nucleotide 5146), and an EPSPS 3, regulatory sequence (the sequence after nucleotide 5146). The EPSPS coding sequence also encodes a chloroplast transit peptide (from nucleotide 1868 to nucleotide 2041). The sequence encoding this chloroplast transit peptide can be predicted using the computer program PS ORT maintained on the public accessible GenomeNet at Kyoto University, Japan.

The 6.0 kb fragment was cloned into the Cla I and Eco RV sites of a pbluescript vector, and then subject to site directed mutagenesis using QuikChange Site-Directed Mutagenesis Kit (Stratagene). Two mutations were introduced into the EPSPS coding sequence: the first mutation being a cytosine to thymine substitution at nucleotide 2886 and the second mutation being a cytosine to thymine substitution at nucleotide 2897. The mutated sequence is shown as SEQ ID NO: 6. The mutations changed the encoded amino acid residue Thr to Ile at position 164 and Pro to Ser at position 168. This mutated EPSPS amino acid sequence is shown as SEQ ID NO: 9. The mutated EPSPS is glyphosate-resistant. The pBluescript vector comprising SEQ ID NO: 6 is referred to as HCEM.

EXAMPLE 4

Transformation of Corn

The Cla I and Eco RV fragment (SEQ ID NO: 6) of HCEM was introduced into cultured immature corn embryos using an aerosol beam injector according to U.S. patent application Ser. No. 09/450,226. The Cla I-Eco RV fragment comprised the glyphosate-resistant EPSPS coding sequence.

Selection was carried out as follows: the beamed embryos were allowed to remain on DN62A0S20G medium for 5 days. Table 2 shows the composition of DN62A0S20G medium.

TABLE-US-00002 TABLE 2 Ingredients in 1 liter Culture Medium (pH 5.8) DN62A0S20G DN62A0S20GLC N6 Salts* 3.98 g 3.98 g N6 Vitamins 1 ml 1 ml Asparagine 800 mg 800 mg Myoinositol 100 g 100 g Proline 1400 mg 1400 mg Casamino acids 100 mg 100 mg 2,4 dichlorophenoxyacetic acid 1 mg 1 mg Glucose 20 g 20 g Silver nitrate 10 mg 10 mg Cefotaxime 0 mg 50 mg *Sigma Plant Culture catalogue, reference C1416

The beamed embryos were then transferred to DN62A100RR a medium containing 100 mg/l glyphosate. Table 3 lists the composition of DN62A100RR and other media. After two 14-day passages on DN62A100RR, actively growing tissue .[.was.]. .Iadd.is .Iaddend.transferred to DN62A300RR medium which contains 300 mg/l (Table 3). After two 14-day passages on this medium, tissue was finally transferred to DN62540RR medium which contains 540 mg/l glyphosate (Table 3). Stable transformation allowed continued growth on 540 mg/l glyphosate. Regeneration .[.was.]. .Iadd.is .Iaddend.carried out as described in U.S. patent application Ser. No. 09/450,226.

TABLE-US-00003 TABLE 3 Ingredients in 1 liter Culture Medium (pH 5.8) DN62A100R R DN62ALC180R R DN62A300R R DN62540RR N6 Salts* 3.98 g 3.98 g 3.98 g 3.98 g N6 Vitamins 1 ml 1 ml 1 ml 1 ml Asparagine 800 mg 800 mg 800 mg 800 mg Myoinositol 100 mg 100 mg 100 mg 100 mg Proline 1400 mg 1400 mg 1400 mg 1400 mg Casamino acids 100 mg 100 mg 100 mg 100 mg 2,4 1 mg 1 mg 1 mg 1 mg dichlorophenoxyacetic acid Sucrose 20 g 20 g 20 g 20 g Silver nitrate 10 mg 10 mg 10 mg Glyphosate 100 mg 180 mg 300 mg 540 mg Cefotaxime 0 mg 50 mg 0 mg 0 mg *Sigma Plant Culture catalogue, reference C1416

Transformation can also be accomplished using Agrobacterium-mediated DNA delivery. In this case, the transformation and regeneration .[.were.]. .Iadd.are .Iaddend.performed according to the methods as described in U.S. patent application Ser. No. 09/203,679, which is herein incorporated by reference. Briefly, after culturing on DN62A0S20GLC (Table 2) for five days, co-cultivated embryos .[.were.]. .Iadd.are .Iaddend.transferred to DN62ALC180RR medium which contains 180 mg/l glyphosate (Table 3). After two 14-day passages on this medium, actively growing tissue .[.was.]. .Iadd.is .Iaddend.transferred to DN62540RR medium containing 540 mg/l glyphosate (Table 3). Stable transformation .[.allowed.]. .Iadd.will allow .Iaddend.continued growth on 540 mg/l glyphosate. Regeneration .[.was.]. .Iadd.is .Iaddend.carried out as described in U.S. Ser. No. 09/203,679.

Resistance to glyphosate in regenerants .[.was.]. .Iadd.is .Iaddend.confirmed by spraying them with glyphosate at commercial rates. Seed from the regenerants .[.segregated.]. .Iadd.is expected to segregate .Iaddend.3:1 for resistance as would be expected with Mendelian inheritance of a transgene. Seeds from backcrossed individuals .[.segregated.]. .Iadd.are expected to segregate .Iaddend.1:1. Corn transformation may also be accomplished by other means including, for example, particle bombardment or electroporation of competent cells.

SEQUENCE LISTINGS

1

914706DNAArabidopsis thaliana 1 ccgcggtggc ggcttggact caaccggaaa caaaagtgtc gaaagtagcc g gtgatgatg 60 aagaatcacg gcgcgttatg agaaaaatac agagctctgg gagattgtct c gctcctgtc 120 gtgttaccgc cacgtggccg gaggagaagc tccttatgaa tgtaggaagt t acgatagta 180 accttcccgt cggcgagttt gatcttgccg aggcgtggag actggtgatc g tgaccgtga 240 gcttggatgg gatgattagg acattccata actatggact gccacgtaca t tttagagat 300 cgataatcta tttttgtttt aaataggaga aaacaaaaat tgattttttg t ttggttttt 360 gttttggtgt ctaaatatat gtagattttc tagtttctag tccttcgctt c caagttcct 420 cgatggctca atgccaattt agtcagataa gattatgctg caactgctag a gcgactctc 480 tcctacttca ttttagtcca tactagttca ttttatctcc aagaaaatga c tccgttttg 540 cttcaaagct caaagatgat tcttttttta atgggccctc ttgaaaaatg g gtaaaatct 600 tggtcttttt acagatagca aacatataca aattatggag aaaacaggat t atattgatg 660 cagcttcgtt gtagacagac ttgtagtcgt tttcttcatt ttcccattcc t ctgctaaat 720 tattaaatcc aacaaaaaca aattttcttc tgttgcttga taaaatctat g tggaatatc 780 tatatctaca cactactgaa gaccaagaag taaacattag tttgcctgat c ttcacctac 840 ctcaaacgag aagtagaagt ttttatggtg acttttgtat ttagagaaac a atgggattc 900 cagtttaagt tggctctttt taactttgct aacatatgcc tgaaagtgga a gacaagaac 960 ttggtttaag aaacctcaag cgattgcgat tttgggtctt aagcttgaaa a aagtgttgt 1020 atggaacaaa caaactaaca tatcggaaca agcttggctt tggttttaaa g ctgatagat 1080 aatggtcgaa ccataaccgg tatggcccaa gatgttcatg ttttttaaaa c tcaccaaag 1140 ctatatcact aacccacaca ttcttgcaga aggttttaga atcacaaagc a taactcacc 1200 tacccctaaa ccaactccaa tttctctcct cctctattaa atctttctca a tcatctttc 1260 tttgagtctt ttgccttgga atcctgatca tggcgtcttc tctcacttcc a aatccattc 1320 tcggatgcac caaacccgct tcttcttctt ttcttccgtc ggagctccgt c gtctctctt 1380 ctcccgccgt tcagatatct ctccattcac aaaccaggaa gaacttccgt g agttctctg 1440 attcttttca aaatttttag atttgaagcc tgtatctagc ttaaagaaga a agatgtgtg 1500 atttgaatct ctagggcagt cgtggggatt gaagaagagt gatctgatgc t aaatggttc 1560 tgagattcgt cctgtgaagg ttagggcttc tgtttccacg gcggagaaag c ttcggagat 1620 tgtgcttcaa cccattagag aaatctcggg tctcattaag cttcctggct c caagtctct 1680 ctctaatcga attctgcttc tcgctgctct atctgaggta tatataaatg t atcacttca 1740 tttcttcctt ctctgtactc cgaatttaga ttattaaaga tataaacttt a ccattttgc 1800 tgtgcttata tagggaacta ctgtagtgga caacttgttg aacagtgatg a catcaatta 1860 catgcttgat gcgttgaaga tattgggact taatgtggaa actcacagtg a aaacaatcg 1920 tgctgtagtt gaaggatgtg gcggggtatt tccagcttcc attgattcca a gagtgatat 1980 cgaactttac ctcggcaatg caggaacagc aatgcgtcca cttaccgccg c agttactgc 2040 tgcaggtggc aacgcaaggt atattgaagg agtaaatgct gaatagtttt g atttcttaa 2100 gaatcgatct tgttttgatg cttttcaatc ggtttatttc agttatgtcc t tgatggggt 2160 gcctcggatg agagagagac ctatagggga tttggttgtt ggtcttaagc a gcttggtgc 2220 tgatgttgaa tgtactcttg gcactaactg ccctcctgtt cgtgtcaacg c taatggtgg 2280 ccttcctggt ggaaaggtga gatcttgcaa atggcatgtg aatttataac t ttataaaca 2340 cttgcagcaa tttgtgttca tcatagcctt acttgacaag atttcatttt t tttgtttgt 2400 tgtcaatgta ttgttcctga aaacgaattg ttttttttta gtagggatta g ttttctctc 2460 ttgattaccc ttttccttgt atggtttctt tattgacgca tcgaacattt t ttgcatttg 2520 caggtgaagc tttctggatc tattagtagt cagtacttga ccgctctgct c atggcagct 2580 cccttagctc ttggagacgt cgaaattgaa attgtcgata aattgatttc t gttccgtat 2640 gttgaaatga cattgaagtt gatggaacgt tttggggtaa gtgctgagca t agtgaaagc 2700 tgggatcgtt tctttgttaa gggtgggcaa aaatacaagt aagagttatt a ttctcttcc 2760 ttttctgaaa tcacatacct tagattgaca aaataatgac taatatggga a atgattcag 2820 gtcgccgggt aatgcttacg tagaaggtga tgcttctagt gctagttatt t cctggctgg 2880 tgctgccatt accggtgaaa ctgtcactgt tgaaggttgt ggaacgacca g tttgcaggt 2940 aatatttgta cactgaatca tcaaagaggc tgttaagttt atagtgaaat t cgtttaggt 3000 caaagtttca tctttttaag gctttgacaa gttgtatgta acatattcgc a agaatctaa 3060 gttcaatttt tgtgatgaat ctctagggag atgtgaaatt tgccgaggtt c ttgagaaaa 3120 tgggatgtaa agtgtcctgg acagagaaca gtgtgactgt gacagggccg t ctagagatg 3180 cttttggaat gagacacttg cgggctattg atgtcaacat gaacaaaatg c ctgatgtag 3240 caatgactct tgccgtcgtt gctctctttg ccgatggtcc aaccaccatt a gagatggta 3300 agtaaaaagc tctctcttat aatataaggt ttctcaagat tcatggtcac t taattctat 3360 ttggtcaata tagtggctag ctggagagta aaggagacgg aaaggatgat t gccatttgc 3420 acagagctta gaaaagtaaa attcttcttc tctctctctc tttctgttta c agtgctcat 3480 tctaagaaat tttgcggtat ttgtgtccag ctgggagcta cagtggaaga a ggttcagat 3540 tattgtgtga ttactccgcc gaaaaaggtg aaaccggcag agattgatac a tatgatgat 3600 catagaatgg caatggcatt ctctcttgca gcttgtgctg atgttccaat c accatcaat 3660 gaccccggtt gcaccaggaa aaccttcccc gactacttcc aagtccttga a agaatcaca 3720 aagcattaaa caaaaaaact ctaaaatctc cactgttttt tcttctgatc c aagcttatc 3780 tgtttccatt tttcttgtct ctgtaacatt attagaaagc aagagtagtg t ttgtttgtg 3840 tgtacctgaa ctgagtgaga tttgagatgc aatcattgaa tcggctttgg t atatcattt 3900 tactctgttt ttcagggtgt ttgttcaggt tctctctagt tatcatccac t ccaaacagg 3960 tcccatgatg tctaacgttt tggttctaag aatgaacaga acaaacaata c actgcgata 4020 accggtgctt ggaagttgtg ttaattgaag aaacaatggc aatagctgca t acttatagt 4080 tgcaggagtg aaaaatgaga taagaggaat gcaaatatgc aattgcaggt t ctatttttt 4140 ttttgctgcc aatgttatta ccaaaagggc tacaagtgag tattctccaa g cttggatga 4200 ggttattcag ggtaataggg tatcaagtta gtaataagag tcagagatac c atgaaagga 4260 ttccaagttg tagtaagaac aactcaaatt caaagtgaag ttttgtgagt t gtgtaattg 4320 tgttggagtt ttgcacaaat gagaagactc ttatagaaac agaggggttg a agaagaagc 4380 gatatttgcc catctcactt gaaaacacta accggagata aaccaaatta a ttggaacta 4440 ttctcagtta tggtttggtc ctcgtcgttt tgggttagtg ttgttggtag g tggagaatt 4500 ttgcatttgc atttgcacaa cgaagaagaa gaccacaaga gccatttgca a ttaggcata 4560 atatatgtcc taactcacca accccctcaa aattgccacc aacttcaaat t tctctcctt 4620 taaacctttc tcaatcatct ttcttctgcc ttggaatcct gatcatggcg t cgtcttctc 4680 tcacttcgaa atccattctc ggatcc 4706 24706DNAArabidopsis thalianamisc_feature (2007)..(2007)substituting thymine for cytosine at position 2007 as compared to SEQ ID NO 1 2 ccgcggtggc ggcttggact caaccggaaa caaaagtgtc gaaagtagcc g gtgatgatg 60 aagaatcacg gcgcgttatg agaaaaatac agagctctgg gagattgtct c gctcctgtc 120 gtgttaccgc cacgtggccg gaggagaagc tccttatgaa tgtaggaagt t acgatagta 180 accttcccgt cggcgagttt gatcttgccg aggcgtggag actggtgatc g tgaccgtga 240 gcttggatgg gatgattagg acattccata actatggact gccacgtaca t tttagagat 300 cgataatcta tttttgtttt aaataggaga aaacaaaaat tgattttttg t ttggttttt 360 gttttggtgt ctaaatatat gtagattttc tagtttctag tccttcgctt c caagttcct 420 cgatggctca atgccaattt agtcagataa gattatgctg caactgctag a gcgactctc 480 tcctacttca ttttagtcca tactagttca ttttatctcc aagaaaatga c tccgttttg 540 cttcaaagct caaagatgat tcttttttta atgggccctc ttgaaaaatg g gtaaaatct 600 tggtcttttt acagatagca aacatataca aattatggag aaaacaggat t atattgatg 660 cagcttcgtt gtagacagac ttgtagtcgt tttcttcatt ttcccattcc t ctgctaaat 720 tattaaatcc aacaaaaaca aattttcttc tgttgcttga taaaatctat g tggaatatc 780 tatatctaca cactactgaa gaccaagaag taaacattag tttgcctgat c ttcacctac 840 ctcaaacgag aagtagaagt ttttatggtg acttttgtat ttagagaaac a atgggattc 900 cagtttaagt tggctctttt taactttgct aacatatgcc tgaaagtgga a gacaagaac 960 ttggtttaag aaacctcaag cgattgcgat tttgggtctt aagcttgaaa a aagtgttgt 1020 atggaacaaa caaactaaca tatcggaaca agcttggctt tggttttaaa g ctgatagat 1080 aatggtcgaa ccataaccgg tatggcccaa gatgttcatg ttttttaaaa c tcaccaaag 1140 ctatatcact aacccacaca ttcttgcaga aggttttaga atcacaaagc a taactcacc 1200 tacccctaaa ccaactccaa tttctctcct cctctattaa atctttctca a tcatctttc 1260 tttgagtctt ttgccttgga atcctgatca tggcgtcttc tctcacttcc a aatccattc 1320 tcggatgcac caaacccgct tcttcttctt ttcttccgtc ggagctccgt c gtctctctt 1380 ctcccgccgt tcagatatct ctccattcac aaaccaggaa gaacttccgt g agttctctg 1440 attcttttca aaatttttag atttgaagcc tgtatctagc ttaaagaaga a agatgtgtg 1500 atttgaatct ctagggcagt cgtggggatt gaagaagagt gatctgatgc t aaatggttc 1560 tgagattcgt cctgtgaagg ttagggcttc tgtttccacg gcggagaaag c ttcggagat 1620 tgtgcttcaa cccattagag aaatctcggg tctcattaag cttcctggct c caagtctct 1680 ctctaatcga attctgcttc tcgctgctct atctgaggta tatataaatg t atcacttca 1740 tttcttcctt ctctgtactc cgaatttaga ttattaaaga tataaacttt a ccattttgc 1800 tgtgcttata tagggaacta ctgtagtgga caacttgttg aacagtgatg a catcaatta 1860 catgcttgat gcgttgaaga tattgggact taatgtggaa actcacagtg a aaacaatcg 1920 tgctgtagtt gaaggatgtg gcggggtatt tccagcttcc attgattcca a gagtgatat 1980 cgaactttac ctcggcaatg caggaatagc aatgcgttca cttaccgccg c agttactgc 2040 tgcaggtggc aacgcaaggt atattgaagg agtaaatgct gaatagtttt g atttcttaa 2100 gaatcgatct tgttttgatg cttttcaatc ggtttatttc agttatgtcc t tgatggggt 2160 gcctcggatg agagagagac ctatagggga tttggttgtt ggtcttaagc a gcttggtgc 2220 tgatgttgaa tgtactcttg gcactaactg ccctcctgtt cgtgtcaacg c taatggtgg 2280 ccttcctggt ggaaaggtga gatcttgcaa atggcatgtg aatttataac t ttataaaca 2340 cttgcagcaa tttgtgttca tcatagcctt acttgacaag atttcatttt t tttgtttgt 2400 tgtcaatgta ttgttcctga aaacgaattg ttttttttta gtagggatta g ttttctctc 2460 ttgattaccc ttttccttgt atggtttctt tattgacgca tcgaacattt t ttgcatttg 2520 caggtgaagc tttctggatc tattagtagt cagtacttga ccgctctgct c atggcagct 2580 cccttagctc ttggagacgt cgaaattgaa attgtcgata aattgatttc t gttccgtat 2640 gttgaaatga cattgaagtt gatggaacgt tttggggtaa gtgctgagca t agtgaaagc 2700 tgggatcgtt tctttgttaa gggtgggcaa aaatacaagt aagagttatt a ttctcttcc 2760 ttttctgaaa tcacatacct tagattgaca aaataatgac taatatggga a atgattcag 2820 gtcgccgggt aatgcttacg tagaaggtga tgcttctagt gctagttatt t cctggctgg 2880 tgctgccatt accggtgaaa ctgtcactgt tgaaggttgt ggaacgacca g tttgcaggt 2940 aatatttgta cactgaatca tcaaagaggc tgttaagttt atagtgaaat t cgtttaggt 3000 caaagtttca tctttttaag gctttgacaa gttgtatgta acatattcgc a agaatctaa 3060 gttcaatttt tgtgatgaat ctctagggag atgtgaaatt tgccgaggtt c ttgagaaaa 3120 tgggatgtaa agtgtcctgg acagagaaca gtgtgactgt gacagggccg t ctagagatg 3180 cttttggaat gagacacttg cgggctattg atgtcaacat gaacaaaatg c ctgatgtag 3240 caatgactct tgccgtcgtt gctctctttg ccgatggtcc aaccaccatt a gagatggta 3300 agtaaaaagc tctctcttat aatataaggt ttctcaagat tcatggtcac t taattctat 3360 ttggtcaata tagtggctag ctggagagta aaggagacgg aaaggatgat t gccatttgc 3420 acagagctta gaaaagtaaa attcttcttc tctctctctc tttctgttta c agtgctcat 3480 tctaagaaat tttgcggtat ttgtgtccag ctgggagcta cagtggaaga a ggttcagat 3540 tattgtgtga ttactccgcc gaaaaaggtg aaaccggcag agattgatac a tatgatgat 3600 catagaatgg caatggcatt ctctcttgca gcttgtgctg atgttccaat c accatcaat 3660 gaccccggtt gcaccaggaa aaccttcccc gactacttcc aagtccttga a agaatcaca 3720 aagcattaaa caaaaaaact ctaaaatctc cactgttttt tcttctgatc c aagcttatc 3780 tgtttccatt tttcttgtct ctgtaacatt attagaaagc aagagtagtg t ttgtttgtg 3840 tgtacctgaa ctgagtgaga tttgagatgc aatcattgaa tcggctttgg t atatcattt 3900 tactctgttt ttcagggtgt ttgttcaggt tctctctagt tatcatccac t ccaaacagg 3960 tcccatgatg tctaacgttt tggttctaag aatgaacaga acaaacaata c actgcgata 4020 accggtgctt ggaagttgtg ttaattgaag aaacaatggc aatagctgca t acttatagt 4080 tgcaggagtg aaaaatgaga taagaggaat gcaaatatgc aattgcaggt t ctatttttt 4140 ttttgctgcc aatgttatta ccaaaagggc tacaagtgag tattctccaa g cttggatga 4200 ggttattcag ggtaataggg tatcaagtta gtaataagag tcagagatac c atgaaagga 4260 ttccaagttg tagtaagaac aactcaaatt caaagtgaag ttttgtgagt t gtgtaattg 4320 tgttggagtt ttgcacaaat gagaagactc ttatagaaac agaggggttg a agaagaagc 4380 gatatttgcc catctcactt gaaaacacta accggagata aaccaaatta a ttggaacta 4440 ttctcagtta tggtttggtc ctcgtcgttt tgggttagtg ttgttggtag g tggagaatt 4500 ttgcatttgc atttgcacaa cgaagaagaa gaccacaaga gccatttgca a ttaggcata 4560 atatatgtcc taactcacca accccctcaa aattgccacc aacttcaaat t tctctcctt 4620 taaacctttc tcaatcatct ttcttctgcc ttggaatcct gatcatggcg t cgtcttctc 4680 tcacttcgaa atccattctc ggatcc 4706 35193DNAArabidopsis thaliana 3 ctgtctagaa tctttccatt tttgcttaca aatatggcac aaccggaaat t cccttactt 60 atttatggaa aaacaaaagt gtcatgtgtt atatatattg gtcccttttt a tgtttgtag 120 ggctatcact accaatccac aatggaagat tcacaaccgt gtacatcacg t aacctagat 180 catgccgaaa actccacagc aagcattcct cttaaactca tcttcgaaat a ctatcgagg 240 ctttcgacga aatttatcat cagattccga tccgtgtcga tgctgtgacg c tctatcatc 300 gatagtaaag atttcgtcga cgcctttctg aatcctcttt tttcaatgaa g actcttttt 360 ttagtccgct ttatggaata atacttatgt tttaacgtaa tgaatcttta t atctatttg 420 tatttttcag ttacactttt gtttggacgt tttgttttca aagaacttct c aagttttct 480 tttccttttt tcttcttgta gtttgccttg ctcgttggtg aatacactac a ttaattgag 540 ggcacttgcg attatgatgt tctcccgcgt gaaagagcat tgacctctct t ttattcaaa 600 acatcacata acagagagtc tgtgaggttt gggacatcgc attctcaaat c tcaatataa 660 gtggttttgg tcgaatgcta gaaaagggtg acttgtatgt tgtgtcgccg c tagtgttcg 720 tacatacagt ttatgatcta acgtatctat aacatattat taatagatat t attttgttt 780 taattttgca cattgtttat aaattaagtg caaaaatttt tagctccaag a gatcgcctt 840 tgctaatcaa taaaattgaa cgcatttaat aaaattgtaa gagcaaactc g cactgatac 900 gtagaataaa tttgttgctt ttgccttcac gacaccatta cctatagtta a tactccaaa 960 agaaatagca gattcaacat acaacgtgca agaccaaaaa acaaatgact c gtaatctcc 1020 agagaatcat aattcataac atgggagatt gtccacaaaa aacataaatt c cctttcatg 1080 tctttttgtt agaaaaccca tttcttaagg cccaacaaaa aacataatcc c ctttcatgt 1140 ctttttgtta gaaaccccat ttatctttct tgaggcccaa tttgaaaacc c acattttct 1200 ttcacctaac ccaccaaagc ctttgcacat gttgacgtga acaccaaact a acacgtgtc 1260 atactgccag tggttatgat aaatgctcat accataccag agtcatagag t ttttggttg 1320 gtgaaagatt tgacggatgc cttcttctca tttctcacca actccctcca a acccaacaa 1380 aagtgtttat attagcaaag ccgccaaagt gtaaacgaaa gtttataaat t tcatttctg 1440 tgatcttacg taattggagg aagatcaaaa ttttcaatcc ccattcttcg a ttgcttcaa 1500 ttgaagtttc tccgatggcg caagttagca gaatctgcaa tggtgtgcag a acccatctc 1560 ttatctccaa tctctcgaaa tccagtcaac gcaaatctcc cttatcggtt t ctctgaaga 1620 cgcagcagca tccacgagct tatccgattt cgtcgtcgtg gggattgaag a agagtggga 1680 tgacgttaat tggctctgag cttcgtcctc ttaaggtcat gtcttctgtt t ccacggcgg 1740 agaaagcgtc ggagattgta cttcaaccca ttagagaaat ctccggtctt a ttaagcttc 1800 ctggctccaa gtctctatca aatcggatcc tgcttctcgc tgctctgtct g aggtatata 1860 tcacttcgtt tcgtccttct ctgtaatctg aacttagatt ataaagattg a tactttacc 1920 attttgctgt ggttttatag ggaacaactg tagtggacaa cttgttgaat a gcgatgaca 1980 tcaattacat gcttgatgcg ttgaagagat tgggacttaa tgtggaaact g acagtgaaa 2040 ataatcgtgc tgtagttgaa ggatgtggcg ggatattccc agcttccata g attcaaaga 2100 gtgatatcga actttacctc ggtaatgcag gaacagcaat gcgtccactt a ccgctgcgg 2160 tcactgctgc aggtggaaac gcaaggtaga ttgaaggagt tgatgcttct t ggtatttga 2220 tgtttaagga atggagcttt tgttgatgct ttatgatcca tttattccag t tatgtgctt 2280 gatggggtgc ctcgtatgag agaaagacct ataggggatt tggttgttgg t cttaagcag 2340 cttggtgctg atgttgaatg tactcttgga actaactgcc ctcctgttcg t gtcaacgct 2400 aatggtggcc ttcccggtgg aaaggttaga tcttgcaaat ggcatgtgaa t atgtaatct 2460 cgttccttac tctatgaaca cttgcagaaa tgtgtgttca tcatagcctt a gcttgacaa 2520 gatttcagtt tttaatctac tctcaacgga tggatcctaa aatagaatcg g atttggtga 2580 ttggttttcg ttctcgatta ccgttttcgt tgtatgattt cttgattaac a attaggaga 2640 catgttatgc atttgcaggt gaagctttct ggatcaatta gtagtcagta c ttgactgct 2700 ctgctcatgt ctgctccctt agctcttgga gacgtcgaga ttgagattgt c gataaatta 2760 atttctgttc catatgttga aatgacattg aagttgatgg aacgtttcgg g gttagtgtc 2820 gagcatagtg atagctggga tcgtttcttt gtcaagggcg ggcaaaaata c aagtaggag 2880 ttattctttt cttccttttc tgaaatcaca tcccttagct tgacaatata a tgactaaaa 2940 ggtgaatgat tcaggtctcc gggtaatgcg tatgtagaag gtgatgcttc t agtgctagt 3000 tatttcttgg ctggtgctgc cattaccggt gaaactgtca cagtcgaagg t tgtggaact 3060 accagcttgc aggtaatatt tgtacactga atcatcgacg aggctgttaa g tttatagtg 3120 aaattcgtct aggtcaaagt ttcatctttt gacaagttgt atataacata t tcgcaagaa 3180 tctaagctca atttttgtga tgaatctcta gggagatgta aaattcgccg a ggtccttga 3240 gaaaatggga tgtaaagtgt cctggacaga gaacagtgtg actgtgacag g accacctag 3300 agatgctttt ggaatgagac acttgcgggc tattgatgtc aacatgaaca a aatgcctga 3360 tgtagccatg acccttgccg tcgttgctct ctttgctgac ggtccaacca c cattagaga 3420 tggtaagtaa aaagctctct cttataatta aggtttctca atattcatga t cacttaatt 3480 ctgtttggtt aatatagtgg ctagctggag agtaaaggag acagaaagga t gattgccat 3540 ttgcacagag cttagaaaag taagagattc ttatctctct ctttctgtct c ttgacagtg 3600 ctcattctaa gtaattagct cataaatttt gtgtgtttgt gttcagctgg g agctacagt 3660 ggaagaaggt tcagattatt gtgtgataac tccgcccaaa aaggtgaaaa c ggcagagat 3720 tgatacatat gatgatcata gaatggcaat ggcattctct cttgcagctt g tgctgatgt 3780 tccaatcacc atcaacgatc ctggttgcac caggaaaacc ttccccgact a cttccaagt 3840 acttgaaaga atcacaaagc actaaacaat aaactctgtt ttttcttctg a tccaagctt 3900 atctgtttcc atttttcttg tctctgtaat attattagaa accgagagtg t ttgtttgcg 3960 tgtaactgaa ctgagcgagt tttgagatgc aatcatttga gttcgattga g agaaatgaa 4020 tgtgtagaga tttcctttta tcttgatgga aagaaattga gttttccttc t tctcttttt

4080 tttccaattc ctaggtcgtc gactcgaata tataaagaca gcagccacga t cgtctcttt 4140 tgatcactta ttagagacaa taatgttgga aagacatggt tcctctagtt t ggtattgaa 4200 aagacatcgt tcttgtttgg aattgctgcc acacgatgta gtagagctca t cctcgagag 4260 acacagttga tcggtcacga gaacaatatt atagatgaag ctcaaaggag g agaatagtg 4320 ttgttggggt cgtcatcgta attagcaaac taactgtgag ttcccgtcaa a gaccattta 4380 tggcccacgt aaaacgacgc cgtttaaatc tgagtcaaag cccatttgtg g cccacgtcc 4440 taacacagtc gtttctctcc gactagtaaa ctaaaatccc ggaaattctc a tccgcatga 4500 gctccggtga aaaatggaga ccaagagaaa gtaagcagga gcctctcgtc t ctgcaatct 4560 gagacatcga aagaccgaaa atccttcaac aggtaacatt tcaatttcgc c ttcgcctag 4620 aaagaagctc gtgtttgttt ttgggtttta gctaagaatt ttagggaaaa g cttgaaaca 4680 aatttggctc tcttatcaat tgcatttgtt ttggagttat gattctgtgt g gaatcgaat 4740 caaaattatc aatctgaaag tgacaataat cccttgtttg tcttttgtgt t ttttatttg 4800 agttcggttt acatggtttc gaacttttca attgattttt gggtttcggt t tgcattgga 4860 attaataagg ttttgagaag agaaaagaaa aaaaggcacg cacgcgaggc g tttttagag 4920 aggggcgagt gtggttcaaa ataggcgttt tggtgggtta gaacccacag a aattggatt 4980 cacgcgccaa acgcaagatg ggcgagagtg ggtatgaaat ggtaagatcg g tgagaatgg 5040 ggtatgacat ggagggctct gattggctaa taaactcaaa atagtagaca t atagctcct 5100 cccttcctcc tctcataata atagtagtta ttattactta gtcttatatg c gaagaaaca 5160 atgaatgaaa aaaccttact tgggtcggaa ttc 5193 45193DNAArabidopsis thalianamisc_feature (2134)..(2134)substituting thymine for cytosine at position 2134 as compared to SEQ ID NO 3 4 ctgtctagaa tctttccatt tttgcttaca aatatggcac aaccggaaat t cccttactt 60 atttatggaa aaacaaaagt gtcatgtgtt atatatattg gtcccttttt a tgtttgtag 120 ggctatcact accaatccac aatggaagat tcacaaccgt gtacatcacg t aacctagat 180 catgccgaaa actccacagc aagcattcct cttaaactca tcttcgaaat a ctatcgagg 240 ctttcgacga aatttatcat cagattccga tccgtgtcga tgctgtgacg c tctatcatc 300 gatagtaaag atttcgtcga cgcctttctg aatcctcttt tttcaatgaa g actcttttt 360 ttagtccgct ttatggaata atacttatgt tttaacgtaa tgaatcttta t atctatttg 420 tatttttcag ttacactttt gtttggacgt tttgttttca aagaacttct c aagttttct 480 tttccttttt tcttcttgta gtttgccttg ctcgttggtg aatacactac a ttaattgag 540 ggcacttgcg attatgatgt tctcccgcgt gaaagagcat tgacctctct t ttattcaaa 600 acatcacata acagagagtc tgtgaggttt gggacatcgc attctcaaat c tcaatataa 660 gtggttttgg tcgaatgcta gaaaagggtg acttgtatgt tgtgtcgccg c tagtgttcg 720 tacatacagt ttatgatcta acgtatctat aacatattat taatagatat t attttgttt 780 taattttgca cattgtttat aaattaagtg caaaaatttt tagctccaag a gatcgcctt 840 tgctaatcaa taaaattgaa cgcatttaat aaaattgtaa gagcaaactc g cactgatac 900 gtagaataaa tttgttgctt ttgccttcac gacaccatta cctatagtta a tactccaaa 960 agaaatagca gattcaacat acaacgtgca agaccaaaaa acaaatgact c gtaatctcc 1020 agagaatcat aattcataac atgggagatt gtccacaaaa aacataaatt c cctttcatg 1080 tctttttgtt agaaaaccca tttcttaagg cccaacaaaa aacataatcc c ctttcatgt 1140 ctttttgtta gaaaccccat ttatctttct tgaggcccaa tttgaaaacc c acattttct 1200 ttcacctaac ccaccaaagc ctttgcacat gttgacgtga acaccaaact a acacgtgtc 1260 atactgccag tggttatgat aaatgctcat accataccag agtcatagag t ttttggttg 1320 gtgaaagatt tgacggatgc cttcttctca tttctcacca actccctcca a acccaacaa 1380 aagtgtttat attagcaaag ccgccaaagt gtaaacgaaa gtttataaat t tcatttctg 1440 tgatcttacg taattggagg aagatcaaaa ttttcaatcc ccattcttcg a ttgcttcaa 1500 ttgaagtttc tccgatggcg caagttagca gaatctgcaa tggtgtgcag a acccatctc 1560 ttatctccaa tctctcgaaa tccagtcaac gcaaatctcc cttatcggtt t ctctgaaga 1620 cgcagcagca tccacgagct tatccgattt cgtcgtcgtg gggattgaag a agagtggga 1680 tgacgttaat tggctctgag cttcgtcctc ttaaggtcat gtcttctgtt t ccacggcgg 1740 agaaagcgtc ggagattgta cttcaaccca ttagagaaat ctccggtctt a ttaagcttc 1800 ctggctccaa gtctctatca aatcggatcc tgcttctcgc tgctctgtct g aggtatata 1860 tcacttcgtt tcgtccttct ctgtaatctg aacttagatt ataaagattg a tactttacc 1920 attttgctgt ggttttatag ggaacaactg tagtggacaa cttgttgaat a gcgatgaca 1980 tcaattacat gcttgatgcg ttgaagagat tgggacttaa tgtggaaact g acagtgaaa 2040 ataatcgtgc tgtagttgaa ggatgtggcg ggatattccc agcttccata g attcaaaga 2100 gtgatatcga actttacctc ggtaatgcag gaatagcaat gcgttcactt a ccgctgcgg 2160 tcactgctgc aggtggaaac gcaaggtaga ttgaaggagt tgatgcttct t ggtatttga 2220 tgtttaagga atggagcttt tgttgatgct ttatgatcca tttattccag t tatgtgctt 2280 gatggggtgc ctcgtatgag agaaagacct ataggggatt tggttgttgg t cttaagcag 2340 cttggtgctg atgttgaatg tactcttgga actaactgcc ctcctgttcg t gtcaacgct 2400 aatggtggcc ttcccggtgg aaaggttaga tcttgcaaat ggcatgtgaa t atgtaatct 2460 cgttccttac tctatgaaca cttgcagaaa tgtgtgttca tcatagcctt a gcttgacaa 2520 gatttcagtt tttaatctac tctcaacgga tggatcctaa aatagaatcg g atttggtga 2580 ttggttttcg ttctcgatta ccgttttcgt tgtatgattt cttgattaac a attaggaga 2640 catgttatgc atttgcaggt gaagctttct ggatcaatta gtagtcagta c ttgactgct 2700 ctgctcatgt ctgctccctt agctcttgga gacgtcgaga ttgagattgt c gataaatta 2760 atttctgttc catatgttga aatgacattg aagttgatgg aacgtttcgg g gttagtgtc 2820 gagcatagtg atagctggga tcgtttcttt gtcaagggcg ggcaaaaata c aagtaggag 2880 ttattctttt cttccttttc tgaaatcaca tcccttagct tgacaatata a tgactaaaa 2940 ggtgaatgat tcaggtctcc gggtaatgcg tatgtagaag gtgatgcttc t agtgctagt 3000 tatttcttgg ctggtgctgc cattaccggt gaaactgtca cagtcgaagg t tgtggaact 3060 accagcttgc aggtaatatt tgtacactga atcatcgacg aggctgttaa g tttatagtg 3120 aaattcgtct aggtcaaagt ttcatctttt gacaagttgt atataacata t tcgcaagaa 3180 tctaagctca atttttgtga tgaatctcta gggagatgta aaattcgccg a ggtccttga 3240 gaaaatggga tgtaaagtgt cctggacaga gaacagtgtg actgtgacag g accacctag 3300 agatgctttt ggaatgagac acttgcgggc tattgatgtc aacatgaaca a aatgcctga 3360 tgtagccatg acccttgccg tcgttgctct ctttgctgac ggtccaacca c cattagaga 3420 tggtaagtaa aaagctctct cttataatta aggtttctca atattcatga t cacttaatt 3480 ctgtttggtt aatatagtgg ctagctggag agtaaaggag acagaaagga t gattgccat 3540 ttgcacagag cttagaaaag taagagattc ttatctctct ctttctgtct c ttgacagtg 3600 ctcattctaa gtaattagct cataaatttt gtgtgtttgt gttcagctgg g agctacagt 3660 ggaagaaggt tcagattatt gtgtgataac tccgcccaaa aaggtgaaaa c ggcagagat 3720 tgatacatat gatgatcata gaatggcaat ggcattctct cttgcagctt g tgctgatgt 3780 tccaatcacc atcaacgatc ctggttgcac caggaaaacc ttccccgact a cttccaagt 3840 acttgaaaga atcacaaagc actaaacaat aaactctgtt ttttcttctg a tccaagctt 3900 atctgtttcc atttttcttg tctctgtaat attattagaa accgagagtg t ttgtttgcg 3960 tgtaactgaa ctgagcgagt tttgagatgc aatcatttga gttcgattga g agaaatgaa 4020 tgtgtagaga tttcctttta tcttgatgga aagaaattga gttttccttc t tctcttttt 4080 tttccaattc ctaggtcgtc gactcgaata tataaagaca gcagccacga t cgtctcttt 4140 tgatcactta ttagagacaa taatgttgga aagacatggt tcctctagtt t ggtattgaa 4200 aagacatcgt tcttgtttgg aattgctgcc acacgatgta gtagagctca t cctcgagag 4260 acacagttga tcggtcacga gaacaatatt atagatgaag ctcaaaggag g agaatagtg 4320 ttgttggggt cgtcatcgta attagcaaac taactgtgag ttcccgtcaa a gaccattta 4380 tggcccacgt aaaacgacgc cgtttaaatc tgagtcaaag cccatttgtg g cccacgtcc 4440 taacacagtc gtttctctcc gactagtaaa ctaaaatccc ggaaattctc a tccgcatga 4500 gctccggtga aaaatggaga ccaagagaaa gtaagcagga gcctctcgtc t ctgcaatct 4560 gagacatcga aagaccgaaa atccttcaac aggtaacatt tcaatttcgc c ttcgcctag 4620 aaagaagctc gtgtttgttt ttgggtttta gctaagaatt ttagggaaaa g cttgaaaca 4680 aatttggctc tcttatcaat tgcatttgtt ttggagttat gattctgtgt g gaatcgaat 4740 caaaattatc aatctgaaag tgacaataat cccttgtttg tcttttgtgt t ttttatttg 4800 agttcggttt acatggtttc gaacttttca attgattttt gggtttcggt t tgcattgga 4860 attaataagg ttttgagaag agaaaagaaa aaaaggcacg cacgcgaggc g tttttagag 4920 aggggcgagt gtggttcaaa ataggcgttt tggtgggtta gaacccacag a aattggatt 4980 cacgcgccaa acgcaagatg ggcgagagtg ggtatgaaat ggtaagatcg g tgagaatgg 5040 ggtatgacat ggagggctct gattggctaa taaactcaaa atagtagaca t atagctcct 5100 cccttcctcc tctcataata atagtagtta ttattactta gtcttatatg c gaagaaaca 5160 atgaatgaaa aaaccttact tgggtcggaa ttc 5193 56010DNAZea mays 5 atcgattgca accttcaaat tctcttcgat cttccttcca aattcatcta a tatttcatc 60 ctcggcaagg aaatcttcta acgtgtaaac ttcactggga tccaactcga a aggatcaaa 120 ctctccctct ggttcgtttg acattgtgga tggagtgact aacctgctaa c accctgcaa 180 caatttatac aggagcatat cctcatgcac acgcaaaact gatgttgtcc a caagacacg 240 cacaggacac gcacaggaca cgcaaacagt ttcagactca tgcacacgca c atcagtttc 300 agactcaggc acacgcacat caaatcacct tcgcttgtcg atgagtcgca g ccgcatcgt 360 acaatggcga ttttaccgac gataaggcat gggagcacga gccgtcgccg t cgccttgcg 420 agacgacggg agcgatctct cccttcattt aatctcttcc acgtcaggtt a ttttgctga 480 gatggcagta tacagacggc aaagttaatg ccgttgtaca tgcccttaga c tcttccgtc 540 accaactcac ttagattttt acaacggaac ataaggttcg cttgcagact t acatataag 600 gtatagttgc ataataatcg ccttatgctg tacattgcga cacccgtaaa t attcgatga 660 aatattagta cacaatatta aataagaacg aacaatacat atattatcat t gatcttagt 720 atctcctttt gctcctcgta gaacaattct gtgtaaatta tgcgtaaaat t cgaggacca 780 aaacattggc tagaaaaata cctaaaatca gttttgcaat tgtttctgat t ttcctcata 840 ttttcttgct tataaagttt tccaaaagta ccattttgga tgaaaaaacg g aaaacaacg 900 ctggtctact tgtaaatttg gtagtgacat ttgggaccgt ctagacacga c ctaaaaata 960 gtagtctaaa acatagtctg acacgatgcc ttaaaaatag acgacaaagc a caacacgat 1020 tagatgtgtc gtgttttgac cgacacgaca caaagtaagg cacgatttaa a acccaataa 1080 ataatatttt aatggttatt ttatgttcca ataattttca tctcttcaaa a aaatgttat 1140 agaaatcatt gatacttagt tgaatatcct aacacaatat atatatatat a ttaatatat 1200 atatatatca attttaagtc actttgctag acatagtaat atattttaaa t attttctct 1260 ttcttgtata tttttaaaat acacatcagt ttttatatgt gtcgtgcttg a accgacacg 1320 atataatcat cggttcgccg tacttctaga tcatgatgtt cctaggtttt a atattaaga 1380 gacggtctat attaactcaa aactatttcg tgaaaggcta actcgaaaaa a aaatgaatg 1440 taatcacggc ccgtcctgga ttcgagattc taacgtttca ttcgtgtcca g tgtgcacac 1500 ttgtggaaaa ggaagacgaa gaaaaaaacc aacaactaac tccggcccgc c ggatgcgcc 1560 cacctacttc cccctcgccc ctctcatggt ctctctcgcg cccagatctg c tactagacg 1620 gcaccgctgc agcgcgtcgt gtcgcggggg ttggtggcag gcagcgagag c ttgccgttc 1680 ctctctctca gttgtcaggt cctaggctca cctcaccggc tcccagcccg c ttctatttc 1740 ttcctccccg accccgtgca ggtggcagtc cagtccacgc caccaaccgc g aggcgaacc 1800 aaaccaaccc actctcccca accccgcgcg cccaggccgc ccgccctacc a accatcggc 1860 gtcggcaatg gcggccatgg cgaccaaggc cgccgcgggc accgtgtcgc t ggacctcgc 1920 cgcgccgtcg cgccgccacc accgcccgag ctcggcgcgc ccgcccgccc g ccccgccgt 1980 ccgcgggctg cgggcgcctg ggcgccgcgt gatcgccgcg ccgccggcgg c ggcagcggc 2040 ggcggcggtg caggcgggtg ccgaggagat cgtgctgcag cccatcaagg a gatctccgg 2100 caccgtcaag ctgccggggt ccaagtcgct ttccaaccgg atcctcctgc t cgccgccct 2160 gtccgaggtg agcgattttg gtgcttgctg cgctgccctg tctcactgct a cctaaatgt 2220 tttgcctgtc gaataccatg gattctcggt gtaatccatc tcacgatcag a tgcaccgca 2280 tgtcgcatgc ctagctctct ctaatttgtc tagtagtttg tatacggatt a atattgata 2340 aatcggtacc gcaaaagcta ggtgtaaata aacactagaa aattggatgt t cccctatcg 2400 gcctgtactc ggctactcgt tcttgtgatg gcatgctgtc tcttcttggt g tttggtgaa 2460 caaccttatg aaatttgggc gcaaagaact cgccctcaag ggttgatctt a tgccatcgt 2520 catgataaac agtggagcac ggacgatcct ttacgttgtt tttaacaaac t ttgtcagaa 2580 aactagcatc attaacttct taatgacgat ttcacaacaa aaaaaggtaa c ctcgctact 2640 aacataacaa aatacttgtt gcttattaat tatatgtttt ttaatctttg a tcaggggac 2700 aacagtggtt gataacctgt tgaacagtga ggatgtccac tacatgctcg g ggccttgag 2760 gactcttggt ctctctgtcg aagcggacaa agctgccaaa agagctgtag t tgttggctg 2820 tggtggaaag ttcccagttg aggattctaa agaggaagtg cagctcttct t ggggaatgc 2880 tggaactgca atgcggccat tgacagcagc tgttactgct gctggtggaa a tgcaacgta 2940 tgtttcctct ctttctctct acaatacttg ctggagttag tatgaaaccc a tgggtatgt 3000 ctagtggctt atggtgtatt ggtttttgaa cttcagttac gtgcttgatg g agtaccaag 3060 aatgagggag agacccattg gcgacttggt tgtcggattg aagcagcttg g tgcagatgt 3120 tgattgtttc cttggcactg actgcccacc tgttcgtgtc aatggaatcg g agggctacc 3180 tggtggcaag gttagctact aagggccaca tgttacattc ttctgtaaat g gtacaacta 3240 ttgtcgagct tttgcatttg taaggaaagc attgattgat ctgaatttga t gctacacca 3300 caaaatatcc tacaaatggt catccctaac tagcaaacaa tgaagtaata c ttggcatgt 3360 gtttatcaaa ttaatttcca tcttctgggg cattgcctgt tttctagtct a atagcattt 3420 gtttttagca ttaattagct cttacaattg ttatgttcta caggtcaagc t gtctggctc 3480 catcagcagt cagtacttga gtgccttgct gatggctgct cctttggctc t tggggatgt 3540 ggagattgaa atcattgata aattaatctc cattccctac gtcgaaatga c attgagatt 3600 gatggagcgt tttggtgtga aagcagagca ttctgatagc tgggacagat t ctacattaa 3660 gggaggtcaa aaatacaagt aagctctgta atgtatttca ctactttgat g ccaatgttt 3720 cagttttcag ttttccaaac agtcgcatca atatttgaat agatgcactg t agaaaaaaa 3780 atcattgcag ggaaaaacta gtactgagta ttttgactgt aaattatttt a ccagtcgga 3840 atatagtcag tctattggag tcaagagcgt gaaccgaaat agccagttaa t tatcccatt 3900 atacagagga caaccatgta tactattgaa acttggttta taagagaatc t aggtagctg 3960 gactcgtagc tgcttggcat ggataccttc ttatctttag gaaaagacac t tgatttttt 4020 ttttctgtgg ccctctatga tgtgtgaacc tgcttctcta ttgctttaga a ggatatatc 4080 tatgtcgtta tgcaacatgc ttcccttagc catttgtact gaaatcagtt t cataagttc 4140 gttagtggtt ccctaaacga aaccttgttt ttctttgcaa tcaacaggtc c cctaaaaat 4200 gcctatgttg aaggtgatgc ctcaagcgca agctatttct tggctggtgc t gcaattact 4260 ggagggactg tgactgtgga aggttgtggc accaccagtt tgcaggtaaa g atttcttgg 4320 ctggtgctac aataactgct tttgtctttt tggtttcagc attgttctca g agtcactaa 4380 ataacattat catctgcaaa tgtcaaatag acatacttag gtgaattcat g taaccgttt 4440 ccttacaaat ttgctgaaac ctcagggtga tgtgaagttt gctgaggtac t ggagatgat 4500 gggagcgaag gttacatgga ccgagactag cgtaactgtt actggcccac c gcgggagcc 4560 atttgggagg aaacacctca aggcgattga tgtcaacatg aacaagatgc c tgatgtcgc 4620 catgactctt gctgtggttg ccctctttgc cgatggcccg acagccatca g agacggtaa 4680 aacattctca gccctacaac catgcctctt ctacatcact acttgacaag a ctaaaaact 4740 attggctcgt tggcagtggc ttcctggaga gtaaaggaga ccgagaggat g gttgcgatc 4800 cggacggagc taaccaaggt aaggctacat acttcacatg tctcacgtcg t ctttccata 4860 gctcgctgcc tcttagcggc ttgcctgcgg tcgctccatc ctcggttgct g tctgtgttt 4920 tccacagctg ggagcatctg ttgaggaagg gccggactac tgcatcatca c gccgccgga 4980 gaagctgaac gtgacggcga tcgacacgta cgacgaccac aggatggcca t ggccttctc 5040 ccttgccgcc tgtgccgagg tccccgtgac catccgggac cctgggtgca c ccggaagac 5100 cttccccgac tacttcgatg tgctgagcac tttcgtcaag aattaataaa g cgtgcgata 5160 ctaccacgca gcttgattga agtgataggc ttgtgctgag gaaatacatt t cttttgttc 5220 tgttttttct ctttcacggg attaagtttt gagtctgtaa cgttagttgt t tgtagcaag 5280 tttctatttc ggatcttaag tttgtgcact gtaagccaaa tttcatttca a gagtggttc 5340 gttggaataa taagaataat aaattacgtt tcagtggctg tcaagcctgc t gctacgttt 5400 taggagatgg cattagacat tcatcatcaa caacaataaa accttttagc c tcaaacaat 5460 aatagtgaag ttatttttta gtcctaaaca agttgcatta ggatatagtt a aaacacaaa 5520 agaagctaaa gttagggttt agacatgtgg atattgtttt ccatgtatag t atgttcttt 5580 ctttgagtct catttaacta cctctacaca taccaacttt agtttttttt c tacctcttc 5640 atgttactat ggtgccttct tatcccactg agcattggta tatttagagg t ttttgttga 5700 acatgcctaa atcatctcaa tcaacgatgg acaatctttt cttcgattga g ctgaggtac 5760 gtcatctaca ggataggacc ttgagaatat gtgtccgtca atagctaacc c tctactaat 5820 tttttcaatc aagcaaccta ttggcttgac tttaattcgt accggcttct a ctacttcta 5880 cagtattttg tctctataaa ttgcagctac aacagtcaga acggctggct t taaaatcaa 5940 atggcctaag gatcattgaa aggcatctta gcaatgtcta aaattattac c ttctctaga 6000 cgttgatatc 6010 66010DNAZea maysmisc_feature (2886)..(2886)substituting thymine for cytosine at position 2886 as compared to SEQ ID NO 5 6 atcgattgca accttcaaat tctcttcgat cttccttcca aattcatcta a tatttcatc 60 ctcggcaagg aaatcttcta acgtgtaaac ttcactggga tccaactcga a aggatcaaa 120 ctctccctct ggttcgtttg acattgtgga tggagtgact aacctgctaa c accctgcaa 180 caatttatac aggagcatat cctcatgcac acgcaaaact gatgttgtcc a caagacacg 240 cacaggacac gcacaggaca cgcaaacagt ttcagactca tgcacacgca c atcagtttc 300 agactcaggc acacgcacat caaatcacct tcgcttgtcg atgagtcgca g ccgcatcgt 360 acaatggcga ttttaccgac gataaggcat gggagcacga gccgtcgccg t cgccttgcg 420 agacgacggg agcgatctct cccttcattt aatctcttcc acgtcaggtt a ttttgctga 480 gatggcagta tacagacggc aaagttaatg ccgttgtaca tgcccttaga c tcttccgtc 540 accaactcac ttagattttt acaacggaac ataaggttcg cttgcagact t acatataag 600 gtatagttgc ataataatcg ccttatgctg tacattgcga cacccgtaaa t attcgatga 660 aatattagta cacaatatta aataagaacg aacaatacat atattatcat t gatcttagt 720 atctcctttt gctcctcgta gaacaattct gtgtaaatta tgcgtaaaat t cgaggacca 780 aaacattggc tagaaaaata cctaaaatca gttttgcaat tgtttctgat t ttcctcata 840 ttttcttgct tataaagttt tccaaaagta ccattttgga tgaaaaaacg g aaaacaacg 900 ctggtctact tgtaaatttg gtagtgacat ttgggaccgt ctagacacga c ctaaaaata 960 gtagtctaaa acatagtctg acacgatgcc ttaaaaatag acgacaaagc a caacacgat

1020 tagatgtgtc gtgttttgac cgacacgaca caaagtaagg cacgatttaa a acccaataa 1080 ataatatttt aatggttatt ttatgttcca ataattttca tctcttcaaa a aaatgttat 1140 agaaatcatt gatacttagt tgaatatcct aacacaatat atatatatat a ttaatatat 1200 atatatatca attttaagtc actttgctag acatagtaat atattttaaa t attttctct 1260 ttcttgtata tttttaaaat acacatcagt ttttatatgt gtcgtgcttg a accgacacg 1320 atataatcat cggttcgccg tacttctaga tcatgatgtt cctaggtttt a atattaaga 1380 gacggtctat attaactcaa aactatttcg tgaaaggcta actcgaaaaa a aaatgaatg 1440 taatcacggc ccgtcctgga ttcgagattc taacgtttca ttcgtgtcca g tgtgcacac 1500 ttgtggaaaa ggaagacgaa gaaaaaaacc aacaactaac tccggcccgc c ggatgcgcc 1560 cacctacttc cccctcgccc ctctcatggt ctctctcgcg cccagatctg c tactagacg 1620 gcaccgctgc agcgcgtcgt gtcgcggggg ttggtggcag gcagcgagag c ttgccgttc 1680 ctctctctca gttgtcaggt cctaggctca cctcaccggc tcccagcccg c ttctatttc 1740 ttcctccccg accccgtgca ggtggcagtc cagtccacgc caccaaccgc g aggcgaacc 1800 aaaccaaccc actctcccca accccgcgcg cccaggccgc ccgccctacc a accatcggc 1860 gtcggcaatg gcggccatgg cgaccaaggc cgccgcgggc accgtgtcgc t ggacctcgc 1920 cgcgccgtcg cgccgccacc accgcccgag ctcggcgcgc ccgcccgccc g ccccgccgt 1980 ccgcgggctg cgggcgcctg ggcgccgcgt gatcgccgcg ccgccggcgg c ggcagcggc 2040 ggcggcggtg caggcgggtg ccgaggagat cgtgctgcag cccatcaagg a gatctccgg 2100 caccgtcaag ctgccggggt ccaagtcgct ttccaaccgg atcctcctgc t cgccgccct 2160 gtccgaggtg agcgattttg gtgcttgctg cgctgccctg tctcactgct a cctaaatgt 2220 tttgcctgtc gaataccatg gattctcggt gtaatccatc tcacgatcag a tgcaccgca 2280 tgtcgcatgc ctagctctct ctaatttgtc tagtagtttg tatacggatt a atattgata 2340 aatcggtacc gcaaaagcta ggtgtaaata aacactagaa aattggatgt t cccctatcg 2400 gcctgtactc ggctactcgt tcttgtgatg gcatgctgtc tcttcttggt g tttggtgaa 2460 caaccttatg aaatttgggc gcaaagaact cgccctcaag ggttgatctt a tgccatcgt 2520 catgataaac agtggagcac ggacgatcct ttacgttgtt tttaacaaac t ttgtcagaa 2580 aactagcatc attaacttct taatgacgat ttcacaacaa aaaaaggtaa c ctcgctact 2640 aacataacaa aatacttgtt gcttattaat tatatgtttt ttaatctttg a tcaggggac 2700 aacagtggtt gataacctgt tgaacagtga ggatgtccac tacatgctcg g ggccttgag 2760 gactcttggt ctctctgtcg aagcggacaa agctgccaaa agagctgtag t tgttggctg 2820 tggtggaaag ttcccagttg aggattctaa agaggaagtg cagctcttct t ggggaatgc 2880 tggaattgca atgcggtcat tgacagcagc tgttactgct gctggtggaa a tgcaacgta 2940 tgtttcctct ctttctctct acaatacttg ctggagttag tatgaaaccc a tgggtatgt 3000 ctagtggctt atggtgtatt ggtttttgaa cttcagttac gtgcttgatg g agtaccaag 3060 aatgagggag agacccattg gcgacttggt tgtcggattg aagcagcttg g tgcagatgt 3120 tgattgtttc cttggcactg actgcccacc tgttcgtgtc aatggaatcg g agggctacc 3180 tggtggcaag gttagctact aagggccaca tgttacattc ttctgtaaat g gtacaacta 3240 ttgtcgagct tttgcatttg taaggaaagc attgattgat ctgaatttga t gctacacca 3300 caaaatatcc tacaaatggt catccctaac tagcaaacaa tgaagtaata c ttggcatgt 3360 gtttatcaaa ttaatttcca tcttctgggg cattgcctgt tttctagtct a atagcattt 3420 gtttttagca ttaattagct cttacaattg ttatgttcta caggtcaagc t gtctggctc 3480 catcagcagt cagtacttga gtgccttgct gatggctgct cctttggctc t tggggatgt 3540 ggagattgaa atcattgata aattaatctc cattccctac gtcgaaatga c attgagatt 3600 gatggagcgt tttggtgtga aagcagagca ttctgatagc tgggacagat t ctacattaa 3660 gggaggtcaa aaatacaagt aagctctgta atgtatttca ctactttgat g ccaatgttt 3720 cagttttcag ttttccaaac agtcgcatca atatttgaat agatgcactg t agaaaaaaa 3780 atcattgcag ggaaaaacta gtactgagta ttttgactgt aaattatttt a ccagtcgga 3840 atatagtcag tctattggag tcaagagcgt gaaccgaaat agccagttaa t tatcccatt 3900 atacagagga caaccatgta tactattgaa acttggttta taagagaatc t aggtagctg 3960 gactcgtagc tgcttggcat ggataccttc ttatctttag gaaaagacac t tgatttttt 4020 ttttctgtgg ccctctatga tgtgtgaacc tgcttctcta ttgctttaga a ggatatatc 4080 tatgtcgtta tgcaacatgc ttcccttagc catttgtact gaaatcagtt t cataagttc 4140 gttagtggtt ccctaaacga aaccttgttt ttctttgcaa tcaacaggtc c cctaaaaat 4200 gcctatgttg aaggtgatgc ctcaagcgca agctatttct tggctggtgc t gcaattact 4260 ggagggactg tgactgtgga aggttgtggc accaccagtt tgcaggtaaa g atttcttgg 4320 ctggtgctac aataactgct tttgtctttt tggtttcagc attgttctca g agtcactaa 4380 ataacattat catctgcaaa tgtcaaatag acatacttag gtgaattcat g taaccgttt 4440 ccttacaaat ttgctgaaac ctcagggtga tgtgaagttt gctgaggtac t ggagatgat 4500 gggagcgaag gttacatgga ccgagactag cgtaactgtt actggcccac c gcgggagcc 4560 atttgggagg aaacacctca aggcgattga tgtcaacatg aacaagatgc c tgatgtcgc 4620 catgactctt gctgtggttg ccctctttgc cgatggcccg acagccatca g agacggtaa 4680 aacattctca gccctacaac catgcctctt ctacatcact acttgacaag a ctaaaaact 4740 attggctcgt tggcagtggc ttcctggaga gtaaaggaga ccgagaggat g gttgcgatc 4800 cggacggagc taaccaaggt aaggctacat acttcacatg tctcacgtcg t ctttccata 4860 gctcgctgcc tcttagcggc ttgcctgcgg tcgctccatc ctcggttgct g tctgtgttt 4920 tccacagctg ggagcatctg ttgaggaagg gccggactac tgcatcatca c gccgccgga 4980 gaagctgaac gtgacggcga tcgacacgta cgacgaccac aggatggcca t ggccttctc 5040 ccttgccgcc tgtgccgagg tccccgtgac catccgggac cctgggtgca c ccggaagac 5100 cttccccgac tacttcgatg tgctgagcac tttcgtcaag aattaataaa g cgtgcgata 5160 ctaccacgca gcttgattga agtgataggc ttgtgctgag gaaatacatt t cttttgttc 5220 tgttttttct ctttcacggg attaagtttt gagtctgtaa cgttagttgt t tgtagcaag 5280 tttctatttc ggatcttaag tttgtgcact gtaagccaaa tttcatttca a gagtggttc 5340 gttggaataa taagaataat aaattacgtt tcagtggctg tcaagcctgc t gctacgttt 5400 taggagatgg cattagacat tcatcatcaa caacaataaa accttttagc c tcaaacaat 5460 aatagtgaag ttatttttta gtcctaaaca agttgcatta ggatatagtt a aaacacaaa 5520 agaagctaaa gttagggttt agacatgtgg atattgtttt ccatgtatag t atgttcttt 5580 ctttgagtct catttaacta cctctacaca taccaacttt agtttttttt c tacctcttc 5640 atgttactat ggtgccttct tatcccactg agcattggta tatttagagg t ttttgttga 5700 acatgcctaa atcatctcaa tcaacgatgg acaatctttt cttcgattga g ctgaggtac 5760 gtcatctaca ggataggacc ttgagaatat gtgtccgtca atagctaacc c tctactaat 5820 tttttcaatc aagcaaccta ttggcttgac tttaattcgt accggcttct a ctacttcta 5880 cagtattttg tctctataaa ttgcagctac aacagtcaga acggctggct t taaaatcaa 5940 atggcctaag gatcattgaa aggcatctta gcaatgtcta aaattattac c ttctctaga 6000 cgttgatatc 6010 7521PRTArabidopsis thaliana 7 Met Ala Ser Ser Leu Thr Ser Lys Ser Ile L eu Gly Cys Thr Lys Pro 1 5 10 15 Ala Ser Ser Ser Phe Leu Pro Ser Glu Leu A rg Arg Leu Ser Ser Pro 20 25 30 Ala Val Gln Ile Ser Leu His Ser Gln Thr A rg Lys Asn Phe Arg Gln 35 40 45 Ser Trp Gly Leu Lys Lys Ser Asp Leu Met L eu Asn Gly Ser Glu Ile 50 55 60 Arg Pro Val Lys Val Arg Ala Ser Val Ser T hr Ala Glu Lys Ala Ser 65 70 75 80 Glu Ile Val Leu Gln Pro Ile Arg Glu Ile S er Gly Leu Ile Lys Leu 85 90 95 Pro Gly Ser Lys Ser Leu Ser Asn Arg Ile L eu Leu Leu Ala Ala Leu 100 105 110 Ser Glu Gly Thr Thr Val Val Asp Asn Leu L eu Asn Ser Asp Asp Ile 115 120 125 Asn Tyr Met Leu Asp Ala Leu Lys Ile Leu G ly Leu Asn Val Glu Thr 130 135 140 His Ser Glu Asn Asn Arg Ala Val Val Glu G ly Cys Gly Gly Val Phe 145 150 155 160 Pro Ala Ser Ile Asp Ser Lys Ser Asp Ile G lu Leu Tyr Leu Gly Asn 165 170 175 Ala Gly Ile Ala Met Arg Ser Leu Thr Ala A la Val Thr Ala Ala Gly 180 185 190 Gly Asn Ala Ser Tyr Val Leu Asp Gly Val P ro Arg Met Arg Glu Arg 195 200 205 Pro Ile Gly Asp Leu Val Val Gly Leu Lys G ln Leu Gly Ala Asp Val 210 215 220 Glu Cys Thr Leu Gly Thr Asn Cys Pro Pro V al Arg Val Asn Ala Asn 225 230 235 240 Gly Gly Leu Pro Gly Gly Lys Val Lys Leu S er Gly Ser Ile Ser Ser 245 250 255 Gln Tyr Leu Thr Ala Leu Leu Met Ala Ala P ro Leu Ala Leu Gly Asp 260 265 270 Val Glu Ile Glu Ile Val Asp Lys Leu Ile S er Val Pro Tyr Val Glu 275 280 285 Met Thr Leu Lys Leu Met Glu Arg Phe Gly V al Ser Ala Glu His Ser 290 295 300 Glu Ser Trp Asp Arg Phe Phe Val Lys Gly G ly Gln Lys Tyr Lys Ser 305 310 315 320 Pro Gly Asn Ala Tyr Val Glu Gly Asp Ala S er Ser Ala Ser Tyr Phe 325 330 335 Leu Ala Gly Ala Ala Ile Thr Gly Glu Thr V al Thr Val Glu Gly Cys 340 345 350 Gly Thr Thr Ser Leu Gln Gly Asp Val Lys P he Ala Glu Val Leu Glu 355 360 365 Lys Met Gly Cys Lys Val Ser Trp Thr Glu A sn Ser Val Thr Val Thr 370 375 380 Gly Pro Ser Arg Asp Ala Phe Gly Met Arg H is Leu Arg Ala Ile Asp 385 390 395 400 Val Asn Met Asn Lys Met Pro Asp Val Ala M et Thr Leu Ala Val Val 405 410 415 Ala Leu Phe Ala Asp Gly Pro Thr Thr Ile A rg Asp Val Ala Ser Trp 420 425 430 Arg Val Lys Glu Thr Glu Arg Met Ile Ala I le Cys Thr Glu Leu Arg 435 440 445 Lys Leu Gly Ala Thr Val Glu Glu Gly Ser A sp Tyr Cys Val Ile Thr 450 455 460 Pro Pro Lys Lys Val Lys Pro Ala Glu Ile A sp Thr Tyr Asp Asp His 465 470 475 480 Arg Met Ala Met Ala Phe Ser Leu Ala Ala C ys Ala Asp Val Pro Ile 485 490 495 Thr Ile Asn Asp Pro Gly Cys Thr Arg Lys T hr Phe Pro Asp Tyr Phe 500 505 510 Gln Val Leu Glu Arg Ile Thr Lys His 515 520 8520PRTArabidopsis thaliana 8 Met Ala Gln Val Ser Arg Ile Cys Asn Gly V al Gln Asn Pro Ser Leu 1 5 10 15 Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg L ys Ser Pro Leu Ser Val 20 25 30 Ser Leu Lys Thr Gln Gln His Pro Arg Ala T yr Pro Ile Ser Ser Ser 35 40 45 Trp Gly Leu Lys Lys Ser Gly Met Thr Leu I le Gly Ser Glu Leu Arg 50 55 60 Pro Leu Lys Val Met Ser Ser Val Ser Thr A la Glu Lys Ala Ser Glu 65 70 75 80 Ile Val Leu Gln Pro Ile Arg Glu Ile Ser G ly Leu Ile Lys Leu Pro 85 90 95 Gly Ser Lys Ser Leu Ser Asn Arg Ile Leu L eu Leu Ala Ala Leu Ser 100 105 110 Glu Gly Thr Thr Val Val Asp Asn Leu Leu A sn Ser Asp Asp Ile Asn 115 120 125 Tyr Met Leu Asp Ala Leu Lys Arg Leu Gly L eu Asn Val Glu Thr Asp 130 135 140 Ser Glu Asn Asn Arg Ala Val Val Glu Gly C ys Gly Gly Ile Phe Pro 145 150 155 160 Ala Ser Ile Asp Ser Lys Ser Asp Ile Glu L eu Tyr Leu Gly Asn Ala 165 170 175 Gly Ile Ala Met Arg Ser Leu Thr Ala Ala V al Thr Ala Ala Gly Gly 180 185 190 Asn Ala Ser Tyr Val Leu Asp Gly Val Pro A rg Met Arg Glu Arg Pro 195 200 205 Ile Gly Asp Leu Val Val Gly Leu Lys Gln L eu Gly Ala Asp Val Glu 210 215 220 Cys Thr Leu Gly Thr Asn Cys Pro Pro Val A rg Val Asn Ala Asn Gly 225 230 235 240 Gly Leu Pro Gly Gly Lys Val Lys Leu Ser G ly Ser Ile Ser Ser Gln 245 250 255 Tyr Leu Thr Ala Leu Leu Met Ser Ala Pro L eu Ala Leu Gly Asp Val 260 265 270 Glu Ile Glu Ile Val Asp Lys Leu Ile Ser V al Pro Tyr Val Glu Met 275 280 285 Thr Leu Lys Leu Met Glu Arg Phe Gly Val S er Val Glu His Ser Asp 290 295 300 Ser Trp Asp Arg Phe Phe Val Lys Gly Gly G ln Lys Tyr Lys Ser Pro 305 310 315 320 Gly Asn Ala Tyr Val Glu Gly Asp Ala Ser S er Ala Ser Tyr Phe Leu 325 330 335 Ala Gly Ala Ala Ile Thr Gly Glu Thr Val T hr Val Glu Gly Cys Gly 340 345 350 Thr Thr Ser Leu Gln Gly Asp Val Lys Phe A la Glu Val Leu Glu Lys 355 360 365 Met Gly Cys Lys Val Ser Trp Thr Glu Asn S er Val Thr Val Thr Gly 370 375 380 Pro Pro Arg Asp Ala Phe Gly Met Arg His L eu Arg Ala Ile Asp Val 385 390 395 400 Asn Met Asn Lys Met Pro Asp Val Ala Met T hr Leu Ala Val Val Ala 405 410 415 Leu Phe Ala Asp Gly Pro Thr Thr Ile Arg A sp Val Ala Ser Trp Arg 420 425 430 Val Lys Glu Thr Glu Arg Met Ile Ala Ile C ys Thr Glu Leu Arg Lys 435 440 445 Leu Gly Ala Thr Val Glu Glu Gly Ser Asp T yr Cys Val Ile Thr Pro 450 455 460 Pro Lys Lys Val Lys Thr Ala Glu Ile Asp T hr Tyr Asp Asp His Arg 465 470 475 480 Met Ala Met Ala Phe Ser Leu Ala Ala Cys A la Asp Val Pro Ile Thr 485 490 495 Ile Asn Asp Pro Gly Cys Thr Arg Lys Thr P he Pro Asp Tyr Phe Gln 500 505 510 Val Leu Glu Arg Ile Thr Lys His 515 520 9506PRTZea mays 9 Met Ala Ala Met Ala Thr Lys Ala Ala Ala G ly Thr Val Ser Leu Asp 1 5 10 15 Leu Ala Ala Pro Ser Arg Arg His His Arg P ro Ser Ser Ala Arg Pro 20 25 30 Pro Ala Arg Pro Ala Val Arg Gly Leu Arg A la Pro Gly Arg Arg Val 35 40 45 Ile Ala Ala Pro Pro Ala Ala Ala Ala Ala A la Ala Val Gln Ala Gly 50 55 60 Ala Glu Glu Ile Val Leu Gln Pro Ile Lys G lu Ile Ser Gly Thr Val 65 70 75 80 Lys Leu Pro Gly Ser Lys Ser Leu Ser Asn A rg Ile Leu Leu Leu Ala 85 90 95 Ala Leu Ser Glu Gly Thr Thr Val Val Asp A sn Leu Leu Asn Ser Glu 100 105 110 Asp Val His Tyr Met Leu Gly Ala Leu Arg T hr Leu Gly Leu Ser Val 115 120 125 Glu Ala Asp Lys Ala Ala Lys Arg Ala Val V al Val Gly Cys Gly Gly 130 135 140 Lys Phe Pro Val Glu Asp Ser Lys Glu Glu V al Gln Leu Phe Leu Gly 145 150 155 160 Asn Ala Gly Ile Ala Met Arg Ser Leu Thr A la Ala Val Thr Ala Ala 165 170 175 Gly Gly Asn Ala Thr Tyr Val Leu Asp Gly V al Pro Arg Met Arg Glu 180 185 190 Arg Pro Ile Gly Asp Leu Val Val Gly Leu L ys Gln Leu Gly Ala Asp 195 200 205 Val Asp Cys Phe Leu Gly Thr Asp Cys Pro P ro Val Arg Val Asn Gly 210 215 220 Ile Gly Gly Leu Pro Gly Gly Lys Val Lys L eu Ser Gly Ser Ile Ser 225 230 235 240 Ser Gln Tyr Leu Ser Ala Leu Leu Met Ala A la Pro Leu Ala Leu Gly 245 250 255 Asp Val Glu Ile Glu Ile Ile Asp Lys Leu I le Ser Ile Pro Tyr Val 260 265 270 Glu Met Thr Leu Arg Leu Met Glu Arg Phe G ly Val Lys Ala Glu His 275 280 285 Ser Asp Ser Trp Asp Arg Phe Tyr Ile Lys G ly Gly Gln Lys Tyr Lys 290 295 300 Ser Pro Lys Asn Ala Tyr Val Glu Gly Asp A la Ser Ser Ala Ser Tyr 305 310 315 320 Phe Leu Ala Gly Ala Ala Ile Thr Gly Gly T hr Val Thr Val Glu Gly 325 330 335 Cys Gly Thr Thr Ser Leu Gln Gly Asp Val L ys Phe Ala Glu Val Leu 340 345 350 Glu Met Met Gly Ala Lys Val Thr Trp Thr G lu Thr Ser Val Thr Val 355 360 365 Thr Gly Pro Pro Arg Glu Pro Phe Gly Arg L ys His Leu Lys Ala Ile 370 375 380 Asp Val Asn Met Asn Lys Met Pro Asp Val A la Met Thr Leu Ala Val 385 390 395 400 Val Ala Leu Phe Ala Asp Gly Pro Thr Ala I le Arg Asp Val Ala Ser 405 410 415 Trp Arg Val Lys Glu Thr Glu Arg Met Val A la Ile Arg Thr Glu Leu 420 425 430 Thr Lys Leu Gly Ala Ser Val Glu Glu Gly P ro Asp Tyr Cys Ile Ile 435

440 445 Thr Pro Pro Glu Lys Leu Asn Val Thr Ala I le Asp Thr Tyr Asp Asp 450 455 460 His Arg Met Ala Met Ala Phe Ser Leu Ala A la Cys Ala Glu Val Pro 465 470 475 480 Val Thr Ile Arg Asp Pro Gly Cys Thr Arg L ys Thr Phe Pro Asp Tyr 485 490 495 Phe Asp Val Leu Ser Thr Phe Val Lys Asn 500 505

Claims

What is claimed is:

1. A method of making a glyphosate-resistant plant cell, comprising: (a) introducing a first DNA fragment into a plurality of regenerable plant cells, the first DNA fragment comprising a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4 and SEQ ID NO: 6; and (b) selecting from said regenerable plant cells a glyphosate-resistant plant cell which is stably transformed with the first DNA fragment.

2. The method according to claim 1, further comprising introducing a second DNA fragment with the first DNA fragment into said regenerable plant cells, wherein the second DNA fragment comprises a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 4 and SEQ ID NO: 6, and the sequence of the first DNA fragment is different from the sequence of the second DNA fragment.

3. The method according to claim 2, wherein the first DNA fragment comprises SEQ ID NO: 2, and the second DNA fragment comprises SEQ ID NO: 4.

4. A glyphosate-resistant plant cell made according to the method of claim 3, which is stably transformed with said first DNA fragment and said second DNA fragment.

5. A plant regenerated from the glyphosate-resistant plant cell of claim 4.

6. A regenerable, glyphosate-resistant plant cell comprising an introduced, chromosomally integrated DNA sequence which comprises a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4 and SEQ ID NO: 6.

7. The glyphosate-resistant plant cell according to claim 6, comprising another introduced, chromosomally integrated DNA sequence, wherein said another introduced, integrated DNA sequence comprises a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4 and SEQ ID NO: 6.

8. The glyphosate-resistant plant cell according to claim 6, wherein the introduced, chromosomally integrated DNA sequence comprises SEQ ID NO: 2, and wherein the plant cell comprises another introduced, chromosomally integrated DNA sequence which comprises SEQ ID NO: 4.

9. A plant regenerated from the glyphosate-resistant plant cell of claim 6.

10. A plant regenerated from the glyphosate-resistant plant cell of claim 8.

11. An isolated polynucleotide comprising the sequence depicted in SEQ ID NO: 2.

12. An isolated polynucleotide comprising the sequence depicted in SEQ ID NO: 4.

13. An isolated polynucleotide comprising the sequence depicted in SEQ ID NO: 6.