IPC분류정보
국가/구분 |
United States(US) Patent
등록
|
국제특허분류(IPC7판) |
|
출원번호 |
US-0160725
(2007-01-10)
|
등록번호 |
US-8268622
(2012-09-18)
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국제출원번호 |
PCT/US2007/000591
(2007-01-10)
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§371/§102 date |
20081117
(20081117)
|
국제공개번호 |
WO2007/084294
(2007-07-26)
|
발명자
/ 주소 |
- Gocal, Greg F. W.
- Knuth, Mark E.
- Beetham, Peter R.
|
출원인 / 주소 |
|
대리인 / 주소 |
|
인용정보 |
피인용 횟수 :
5 인용 특허 :
28 |
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. Additionally the present invention relates to mutant E. coli cells that contain mutated EPSPS genes.
대표청구항
▼
1. A method for producing a non-transgenic, herbicide resistant or tolerant plant comprising: introducing into plant cells a recombinagenic oligonucleobase with a targeted mutation in the EPSPS gene to produce plant cells with a mutant EPSPS gene that expresses a EPSPS protein that is mutated at ami
1. A method for producing a non-transgenic, herbicide resistant or tolerant plant comprising: introducing into plant cells a recombinagenic oligonucleobase with a targeted mutation in the EPSPS gene to produce plant cells with a mutant EPSPS gene that expresses a EPSPS protein that is mutated at amino acid positions Thr179 and Pro183in an Arabidopsis EPSPS protein (SEQ ID NO:19) or at an analogous amino acid residue in an EPSPS homolog wherein Thr179 is changed to Ile and Pro183 is changed to Thr or Ala;selecting a plant cell exhibiting improved tolerance to glyphosate as compared to a corresponding wild-type plant cell; andregenerating a nort-transgenic herbicide resistant or tolerant plant having as mutated EPSPS gene from said selected plant cell. 2. A method for producing a non-transgenic, herbicide resistant or tolerant plant comprising: introducing into plant cells a recombinagenic oligonucleobase with a targeted mutation in the EPSPS gene to produce plant cells with a mutant EPSPS gene that expresses a mutant EPSPS protein that is mutated at amino acid positions Thr179 and Pro183 in an Arabidopsis EPSPS protein (SEQ ID NO:19) or at an analogous amino acid residue in an EPSPS homolog wherein Thr179 is changed to Ile and Pro183 is changed to Thr or Ala;identifying a plant cell having mutant EPSPS protein that exhibits substantially the same catalytic activity as a wild type EPSPS protein, and which exhibits that activity even in the presence of glyphosate; and(c) regenerating a non-transgenic herbicide resistant or tolerant plant having a mutated EPSPS gene from said plant cell. 3. The method according to claim 1 or 2 in which the recombinagenic, oligonucleobase is a mixed duplex nucleotide or a SSMOV. 4. The method according to claim 3 in which the mixed duplex nucleotide contains a first homologous region which has a sequence identical to the sequence of at least 6 base pairs of the first fragment of the target EPSPS gene and a second homologous region which has a sequence identical to the sequence of at least 6 based pairs of a second fragment of the target EPSPS gene, and an intervening region which contains at least one nucleobase heterologous to the target EPSPS gene, which intervening region connects the first and second homologous region. 5. The method according to claim 1 or 2 in which the recombinagenic oligonucleobase is introduced by electroporation. 6. The method according to claim 1 in which the amino acid positions are Thr102 and Pro106, in the Zea mays homolog. 7. The method according to claim 1 in which the amino acid. positions are Thr174and Pro 178 in a Brassica sp homolog. 8. The method according to claim 1 in which the amino acid positions are Thr174and Pro178 in the Petunia hybrida homolog. 9. The plant according to claim 1 or 2 in which the plant cells are 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, grape, turf grasses, and Brassica sp. 10. The method according to claim 2 in which the amino acid positions are Thr102and Pro106 in the Zea mays homolog. 11. The method according to claim 2 in which the amino acid positions are Thr174 and Pro178 in a Brassica sp homolog. 12. The method according to claim 2 in which the amino acid positions are Thr174 and Pro178 in the Petunia hybrida homolog. 13. A method for producing a non-transgenic, herbicide resistant or tolerant plant comprising: introducing into plant cells a recombinagenic oligonucleobase with a targeted mutation in the EPSPS gene to produce plant cells with a mutant EPSPS gene that expresses an EPSPS protein that is mutated at amino acid positions Thr179 and Pro183 in an Arabidopsis EPSPS protein (SEQ ID NO:19) or at an analogous amino acid residue in an EPSPS homolog wherein the Thr179 is changed to Ile and Pro183 is changed to Thr or Ala;selecting a plant cell exhibiting improved tolerance to glyphosate as compared to a corresponding wild-type plant cell; andregenerating a non-transgenic herbicide resistant or tolerant plant having a mutated EPSPS gene from said selected plant cell;wherein said plant is selected from the group consisting of corn, wheat, and sugarbeet. 14. The method of claim 13, wherein said plant is corn. 15. The method of claim 13, wherein said plant is wheat. 16. The method of claim 13, wherein said plant is sugarbeet.
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