Epsps mutants

Abstract

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.

1 . An herbicide resistant plant that expresses a mutant EPSPS gene product wherein the EPSPS gene is mutated at a position to change one or more amino acid positions in the gene product, said amino acid positions selected from the group consisting of Leu 160 , Thr 179 , Pro 183 , Val 114 , Asp 164 , Asn 193 , and X 194 , in an Arabidopsis EPSPS protein (AF360224) or at an analogous amino acid position in an EPSPS homolog, wherein X is not Leu. 
     
     
         2 . The plant according to  claim 1  wherein the plant is Zea mays and the amino acid positions are selected from the group consisting of Leu 84 , Thr 102 , Pro 106 , Val 117 , Asp 164 , Asn 193 , and X 374 . 
     
     
         3 . The plant according to  claim 1  wherein the plant is Brassica napus and the amino acid positions are selected from the group consisting of Leu 155 , Thr 174 , Pro 178 , Val 114 , Asp 164 , Asn 193 , and X 189 . 
     
     
         4 . The plant according to  claim 1  wherein the plant is Petunia hybrida and the amino acid positions are selected from the group consisting of Leu 155 , Thr 174 , Pro 178 , Val 114 , Asp 164 , Asn 193 ,and X 189 . 
     
     
         5 . The plant according to  claim 1  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, grape, and turf grasses. 
     
     
         6 . 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 gene product in comparison with the wild-type sequence: Leu 160 Ser, Thr 179 Ile or Ala, Pro 183 Ala or Thr or Leu or Cys or Gly, Val 114 Ala, Asp 164 Ala, Asn 193 Ala, and X 194 Leu, wherein the amino acid to the left of the subscript number is the native amino acid, the amino acid to the right of the subscript number is the mutant amino acid and X is any amino acid except Leu. 
     
     
         7 . The plant according to  claim 2  in which the mutated gene results in one or more of the following amino acid substitutions in the Zea mays EPSPS gene product in comparison with the wild-type sequence: Leu 84 Ser, Thr 102 Ile or Ala, Pro 106 Ala or Thr or Leu or Cys or Gly, Val 114 Ala, Asp 164 Ala, Asn 193 Ala, and X 374 Leu, wherein the amino acid to the left of the subscript number is the native amino acid and the amino acid to the right of the subscript number is the mutant amino acid. 
     
     
         8 . The plant according to  claim 3  in which the mutated gene results in one or more of the following amino acid substitutions in the Brassica napus EPSPS gene product in comparison with the wild-type sequence: Leu 155 Ser, Thr 174 Ile or Ala, Pro 178 Ala or Thr or Leu or Cys or Gly, Val 114 Ala, Asp 164 Ala, Asn 193 Ala, and X 374 Leu, wherein the amino acid to the left of the subscript number is the native amino acid and the amino acid to the right of the subscript number is the mutant amino acid. 
     
     
         9 . The plant according to  claim 4  in which the mutated gene results in one or more of the following amino acid substitutions in the Petunia hybrida EPSPS gene product in comparison with the wild-type sequence: Leu 155 Ser, Thr 174 Ile or Ala, Pro 178 Ala or Thr or Leu or Cys or Gly, Val 114 Ala, Asp 164 Ala, Asn 193 Ala, and X 374 Leu, wherein the amino acid to the left of the subscript number is the native amino acid and the amino acid to the right of the subscript number is the mutant amino acid.