초록 ▼

The present invention relates to the production of a non-transgenic plant resistant or tolerant to a herbicide of the phosphonomethylglycine family, e.g., glyphosate. The present invention also relates to the use of a recombinagenic oligonucleobase to make a desired mutation in the chromosomal or ep

The present invention relates to the production of a non-transgenic plant resistant or tolerant to a herbicide of the phosphonomethylglycine family, e.g., glyphosate. The present invention also relates to the use of a recombinagenic oligonucleobase to make a desired mutation in the chromosomal or episomal sequences of a plant in the gene encoding for 5-enol pyruvylshikimate-3-phosphate synthase (EPSPS). The mutated protein, which substantially maintains the catalytic activity of the wild-type protein, allows for increased resistance or tolerance of the plant to a herbicide of the phosphonomethylglycine family, and allows for the substantially normal growth or development of the plant, its organs, tissues or cells as compared to the wild-type plant irrespective of the presence or absence of the herbicide. The present invention also relates to a non-transgenic plant cell in which the EPSPS gene has been mutated, a non-transgenic plant regenerated therefrom, as well as a plant resulting from a cross using a regenerated non-transgenic plant having a mutated EPSPS gene.

대표청구항 ▼

1. A non-transgenic herbicide resistant plant which comprises a plant that expresses a mutant EPSPS gene product instead of a wild-type EPSPS gene product instead of a wild-type EPSPS gene product wherein the wild-type EPSPS gene product has been mutated at one or more amino acid positions, said pos

1. A non-transgenic herbicide resistant plant which comprises a plant that expresses a mutant EPSPS gene product instead of a wild-type EPSPS gene product instead of a wild-type EPSPS gene product wherein the wild-type EPSPS gene product has been mutated at one or more amino acid positions, said positions selected from the group consisting of Leu173, Ala179, Met180, Arg181, Ser98, Ser255 and Leu198 in SEQ ID NO:2 or at an analogous amino acid residue in an EPSPS paralog and said plant has substantially normal growth as compared to a plant expressing the wild-type EPSPS gene product; wherein the plant is selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eucalyptus, apple, lettuce, peas, lentils and grape. 2. A non-transgenic herbicide resistant plant which comprises a plant that expresses a mutant EPSPS gene product instead of a wild-type EPSPS gene product instead of a wild-type EPSPS gene product wherein the wild-type EPSPS gene product has been mutated at one or more amino acid positions, said positions selected from the group consisting of Leu173, Ala179, Met180, Arg181, Ser98, Ser255 and Leu198 in SEQ ID NO:2 or at an analogous amino acid residue in an EPSPS paralog and said plant has substantially the same catalytic activity as compared to a plant expressing the wild-type EPSPS gene product; wherein the plant is selected from the group consisting of corn, wheat, rice, barley, soybean, cotton, sugarbeet, oilseed rape, canola, flax, sunflower, potato, tobacco, tomato, alfalfa, poplar, pine, eucalyptus, apple, lettuce, peas, lentils and grape. 3. The plant according to claim 1 in which the herbicide is a member of the phosphonomethylglycine family. 4. The plant according to claim 3 in which the member of the phosphonomethylglycine family is glyphosate. 5. The plant according to claim 1 in which the positions in the Zea mays paralog are selected from the group consisting of Leu97, Ala103, Met104, Arg105, Ser23, Ser179 and Leu194. 6. The plant according to claim 1 in which the positions in the Brassica napus paralog are selected from the group consisting of Leu169, Ala175, Met176, Arg177, Ser94, Ser251 and Leu194. 7. The plant according to claim 1 in which the positions in the Petunia hybrida are selected from the group consisting of Leu169, Ala175, Met176, Arg177, Ser94, Ser251 and Leu194. 8. The plant according to claim 1 in which the mutated gene results in one or more of the following amino acid substitutions in the EPSPS enzyme in comparison with the wild-type sequence: (i)Leu173 - Phe(ii)Ala179 - Gly(iii)Met180 - Cys(iv)Arg181 - Leu or Ser(v)Ser98 - Asp(vi)Ser255 - Ala(vii)Leu198 - Lys. 9. The plant according to claim 5 in which the mutated gene results in one or more of the following amino acid substitutions in the EPSPS enzyme in comparison with the wild-type sequence: (i)Leu97 - Phe(ii)Ala103 - Gly(iii)Met104 - Cys(iv)Arg105 - Leu or Ser(v)Ser23 - Asp(vi)Ser179 - Ala(vii)Leu122 - Lys. 10. The plant according to claim 6 in which the mutated gene results in one or more of the following amino acid substitutions in the EPSPS enzyme in comparison with the wild-type sequence: (i)Leu169 - Phe(ii)Ala175 - Gly(iii)Met176 - Cys(iv)Arg177 - Leu or Ser(v)Ser94 - Asp(vi)Ser251 - Ala(vii)Leu194 - Lys. 11. The plant according to claim 7 in which the mutated gene results in one or more of the following amino acid substitutions in the EPSPS enzyme in comparison with the wild-type sequence: (i)Leu169 - Phe(ii)Ala175 - Gly(iii)Met176 - Cys(iv)Arg177 - Leu or Ser(v)Ser94 - Asp(vi)Ser251 - Ala(vii)Leu194 - Lys. 12. The plant according to claim 1 in which the plant expresses a mutant EPSPS gene product instead of a wild-type EPSPS gene product wherein the wild-type EPSPS gene product has been mutated at one or more amino acid positions, said positions selected from the group consisting of Ser98, Ser255 and Leu198 in Arabidopsis or at an analogous amino acid residue in an EPSPS paralog. 13. The plant according to claim 1 in which the mutated gene results in a Ala to Gly amino acid substitution at the position corresponding to Ala179 of the Arabidopsis EPSPS enzyme. 14. The plant according to claim 1 in which the mutated gene results in a Met to Cys amino acid substitution at the position corresponding to Ala180 of the Arabidopsis EPSPS enzyme. 15. The plant according to claim 1 in which the mutated gene results in a Arg to Leu or Ser amino acid substitution at the position corresponding to Arg181 of the Arabidopsis EPSPS enzyme. 16. The plant according to claim 1 in which the mutated gene results in a Ser to Asp amino acid substitution at the position corresponding to Ser98 of the Arabidopsis EPSPS enzyme. 17. The plant according to claim 1 in which the mutated gene results in a Ser to Ala amino acid substitution at the position corresponding to Ser255 of the Arabidopsis EPSPS enzyme. 18. The plant according to claim 1 in which the mutated gene results in a Leu to Lys amino acid substitution at the position corresponding to Leu198 of the Arabidopsis EPSPS enzyme. 19. The plant according to claim 1 in which the plant expresses an EPSPS gene product that is mutated in at least two amino acid positions, wherein said at least two amino acid positions include Thr178 and Pro182 in Arabidopsis EPSPS protein or at an analogous amino acid residue in an EPSPS paralog; and wherein the mutated gene results in a Thr to Val or Leu amino acid substitution at the position corresponding to Thr178 of the Arabidopsis EPSPS enzyme and a Pro to Ser amino acid substitution at the position corresponding to Pro182 of the Arabidopsis EPSPS enzyme. 20. The plant according to claim 19 in which the amino acid positions are Thr102 and Pro106 in the Zea mays paralog. 21. The plant according to claim 19 in which the amino acid positions are Thr174 and Pro178 in a Brassica sp paralog. 22. The plant according to claim 19 in which the amino acid positions are Thr174 and Pro178 in the Petunia hybrida paralog. 23. A non-transgenic herbicide resistant plant which comprises a plant that expresses a mutant EPSPS gene product instead of a wild-type EPSPS gene product instead of a wild-type EPSPS gene product wherein the wild-type EPSPS gene product has been mutated at one or more amino acid positions, said positions selected from the group consisting of Leu173, Ala179, Met180, Arg181, Ser98, Ser255 and Leu198 in Arabidopsis or at an analogous amino acid residue in an EPSPS paralog and said plant has substantially normal growth as compared to a plant expressing the wild-type EPSPS gene product; wherein said non-transgenic herbicide resistant plant is selected from the group consisting of corn, wheat, and sugarbeet and wherein said herbicide is glyphosate. 24. The non-transgenic herbicide resistant plant of claim 23, wherein said plant is corn. 25. The non-transgenic herbicide resistant plant of claim 23, wherein said plant is wheat. 26. The non-transgenic herbicide resistant plant of claim 23, wherein said plant is sugarbeet. 27. The plant according to claim 23 in which the plant expresses an EPSPS gene product that is mutated in at least two amino acid positions, wherein said at least two amino acid positions include Thr178 and Pro182 in Arabidopsis EPSPS protein or at an analogous amino acid residue in an EPSPS paralog; and wherein the mutated gene results in a Thr to Val or Leu amino acid substitution at the position corresponding to Thr178 of the Arabidopsis EPSPS enzyme and a Pro to Ser amino acid substitution at the position corresponding to Pro182 of the Arabidopsis EPSPS enzyme. 28. The non-transgenic herbicide resistant plant of claim 27, wherein said plant is corn. 29. The non-transgenic herbicide resistant plant of claim 27, wherein said plant is wheat. 30. The non-transgenic herbicide resistant plant of claim 27, wherein said plant is sugarbeet. 31. A non-transgenic herbicide resistant plant which comprises a plant that expresses a mutant EPSPS gene product instead of a wild-type EPSPS gene product instead of a wild-type EPSPS gene product wherein the wild-type EPSPS gene product has been mutated at one or more amino acid positions, said positions selected from the group consisting of Leu173, Ala179, Met180, Arg181, Ser98, Ser255 and Leu198 in Arabidopsis or at an analogous amino acid residue in an EPSPS paralog and said plant has substantially the same catalytic activity as compared to a plant expressing the wild-type EPSPS gene product; wherein said non-transgenic herbicide resistant plant is selected from the group consisting of corn, wheat, and sugarbeet and wherein said herbicide is glyphosate. 32. The non-transgenic herbicide resistant plant of claim 31, wherein said plant is corn. 33. The non-transgenic herbicide resistant plant of claim 31, wherein said plant is wheat. 34. The non-transgenic herbicide resistant plant of claim 31, wherein said plant is sugarbeet.