Glyphosate resistance enhancement

Abstract

The present invention relates to a new method for improving glyphosate resistance of a plant. The method encompasses providing one or more specific mutations in a specific nucleotide sequence in said plant. In comparison to a plant not manipulated according to the method, the plant obtained by the method displays (improved) glyphosate resistance. Also provided are a (transgenic) plant, including a seed thereof, and a plant product that can be obtained by the method according to the invention.

1 . A nucleotide sequence encoding an EPSPS enzyme that confers glyphosate resistance to a plant, characterized in that the nucleotide sequence comprises at least one mutation that corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 44 in SEQ ID NO: 5. 
     
     
         2 .- 29 . (canceled) 
     
     
         30 . The nucleotide sequence of  claim 1 , wherein the EPSPS enzyme is derived from a plant EPSPS enzyme. 
     
     
         31 . The nucleotide sequence of  claim 1 , wherein the amino acid that is changed is Asparagine. 
     
     
         32 . The nucleotide sequence of  claim 1 , wherein the amino acid is changed into an Aspartic Acid. 
     
     
         33 . The nucleotide sequence of  claim 1 , characterized in that the nucleotide sequence comprises at least one further mutation that on its own renders said nucleotide sequence to encode an EPSPS enzyme that confers glyphosate resistance to a plant. 
     
     
         34 . The nucleotide sequence of  claim 33 , wherein the at least one further mutation corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 101, 106, or 179 in SEQ ID NO: 5. 
     
     
         35 . The nucleotide sequence of  claim 34 , wherein the amino acid that is changed corresponding to the position 101 is Glycine, the amino acid that is changed corresponding to the position 106 is Proline, and/or the amino acid that is changed corresponding to the position 179 is Serine. 
     
     
         36 . The nucleotide sequence of  claim 34 , wherein the amino acid corresponding to the position 101 is changed into an Alanine, the amino acid corresponding to the position 106 is changed into a Leucine, and/or the amino acid corresponding to the position 179 is changed into an Asparagine. 
     
     
         37 . An EPSPS enzyme that confers glyphosate resistance to a plant, encoded by the nucleotide sequence of  claim 1 . 
     
     
         38 . The EPSPS enzyme of  claim 37 , wherein the EPSPS enzyme is derived from a plant EPSPS enzyme. 
     
     
         39 . The EPSPS enzyme of  claim 37 , wherein the EPSPS enzyme is derived from a bacterial EPSPS enzyme. 
     
     
         40 . A vector comprising a nucleotide sequence according to  claim 1 . 
     
     
         41 . A host comprising the vector of  claim 40 . 
     
     
         42 . A plant or part thereof comprising the nucleotide sequence of  claim 1 . 
     
     
         43 . The plant or part thereof of  claim 42 , wherein the amino acid that is changed is Asparagine. 
     
     
         44 . The plant or part thereof of  claim 42 , wherein the amino acid is changed into an Aspartic Acid. 
     
     
         45 . The plant or part thereof of  claim 42 , wherein the nucleotide sequence comprises at least one further mutation that on its own renders said nucleotide sequence to encode an EPSPS enzyme that confers glyphosate resistance to a plant. 
     
     
         46 . The plant or part thereof of  claim 45 , wherein the at least one further mutation corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 101, 106, or 179 in SEQ ID NO: 5. 
     
     
         47 . The plant or part thereof of  claim 46 , wherein the amino acid that is changed corresponding to the position 101 is Glycine, the amino acid that is changed corresponding to the position 106 is Proline, and/or the amino acid that is changed corresponding to the position 179 is Serine. 
     
     
         48 . The plant or part thereof of  claim 46 , wherein the amino acid corresponding to the position 101 is changed into an Alanine, the amino acid corresponding to the position 106 is changed into a Leucine, and/or the amino acid corresponding to the position 179 is changed into an Asparagine. 
     
     
         49 . A seed derived from the plant or part thereof of  claim 42 . 
     
     
         50 . A method for creating a plant with resistance to glyphosate, the method comprising:
 a) providing a plant or plant cell that comprises a nucleotide sequence encoding an EPSPS enzyme; and   b) introducing at least one mutation into the nucleotide sequence wherein the mutation corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 44 in SEQ ID NO: 5.   
     
     
         51 . The method of  claim 50 , wherein the at least one mutation is introduced into the nucleotide sequence by introducing at least one oligonucleotide capable of hybridizing to the nucleotide sequence into the plant or plant cell of step a), wherein the oligonucleotide comprises at least one mismatch with respect to the nucleotide sequence of step a), and wherein the mismatch is positioned in the oligonucleotide so that it causes the mutation of step b). 
     
     
         52 . The method of  claim 50 , wherein the amino acid that is changed is Asparagine. 
     
     
         53 . The method of  claim 50 , wherein the amino acid is changed into an Aspartic Acid. 
     
     
         54 . A method for enhancing glyphosate resistance of a plant, the method comprising:
 a) providing a plant or plant cell that comprises a nucleotide sequence encoding an EPSPS enzyme that confers glyphosate resistance to the plant or plant cell; and   b) introducing at least one mutation into the nucleotide sequence wherein the mutation corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 44 in SEQ ID NO: 5.   
     
     
         55 . The method of  claim 54 , wherein the nucleotide sequence of step a) comprises at least one mutation that corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 101, 106, or 179 in SEQ ID NO: 5. 
     
     
         56 . The method of  claim 54 , wherein the at least one mutation is introduced into the nucleotide sequence in step b) by introducing at least one oligonucleotide capable of hybridizing to the nucleotide sequence into the plant or plant cell of step a), wherein the oligonucleotide comprises at least one mismatch with respect to the nucleotide sequence of step a), and wherein the mismatch is positioned in the oligonucleotide so that it causes the mutation of step b). 
     
     
         57 . The method of  claim 54 , wherein the amino acid that is changed is Asparagine. 
     
     
         58 . The method of  claim 54 , wherein the amino acid is changed into an Aspartic Acid. 
     
     
         59 . The method of  claim 55 , wherein the amino acid that is changed corresponding to the position 101 is Glycine, the amino acid that is changed corresponding to the position 106 is Proline, and/or the amino acid that is changed corresponding to the position 179 is Serine. 
     
     
         60 . The method of  claim 55 , wherein the amino acid corresponding to the position 101 is changed into an Alanine, the amino acid corresponding to the position 106 is changed into a Leucine, and/or the amino acid corresponding to the position 179 is changed into an Asparagine. 
     
     
         61 . A plant product obtained by processing a plant, or a part thereof, created by the method of  claim 50 , wherein the processing comprises cooking, grinding, drying, milling, baking, cutting, sieving, flaking, peeling, soaking, washing, heating, cooling, crushing, or wetting. 
     
     
         62 . The plant product of  claim 61 , wherein the plant product is a starch-based product or a plant-oil-based product. 
     
     
         63 . A plant product obtained by processing a plant, or a part thereof, created by the method of  claim 54 , wherein the processing comprises cooking, grinding, drying, milling, baking, cutting, sieving, flaking, peeling, soaking, washing, heating, cooling, crushing, or wetting. 
     
     
         64 . The plant product of  claim 63 , wherein the plant product is a starch-based product or a plant-oil-based product. 
     
     
         65 . A method for creating a plant with resistance to glyphosate, the method comprising:
 a) providing a plant or plant cell;   b) introducing a nucleotide sequence according to  claim 1 ; and   c) expressing the EPSPS enzyme encoded by the nucleotide sequence of step b) in the plant or plant cell.   
     
     
         66 . The method of  claim 65 , wherein the amino acid that is changed is Asparagine. 
     
     
         67 . The method of  claim 65 , wherein the amino acid is changed into an Aspartic Acid. 
     
     
         68 . The method of  claim 65 , wherein the nucleotide sequence encoding an EPSPS enzyme comprises at least one further mutation that on its own renders said nucleotide sequence to encode an EPSPS enzyme that confers glyphosate resistance to a plant. 
     
     
         69 . The method of  claim 68 , wherein the at least one further mutation corresponds to a change of an amino acid in the encoded EPSPS enzyme at a position corresponding to position 101, 106, or 179 in SEQ ID NO: 5. 
     
     
         70 . The method of  claim 69 , wherein the amino acid that is changed corresponding to the position 101 is Glycine, the amino acid that is changed corresponding to the position 106 is Proline, and/or the amino acid that is changed corresponding to the position 179 is Serine. 
     
     
         71 . The method of  claim 69 , wherein the amino acid corresponding to the position 101 is changed into an Alanine, the amino acid corresponding to the position 106 is changed into a Leucine, and/or the amino acid corresponding to the position 179 is changed into an Asparagine. 
     
     
         72 . The nucleotide sequence of  claim 1 , wherein the nucleotide sequence is a cDNA sequence. 
     
     
         73 . The vector of  claim 40 , wherein the nucleotide sequence is located downstream of a heterologous transcription regulatory sequence and is operably linked to the transcription regulatory sequence. 
     
     
         74 . The vector of  claim 73 , wherein the transcription regulatory sequence comprises a promoter. 
     
     
         75 . The vector of  claim 74 , wherein the promoter is a constitutive 35S promoter. 
     
     
         76 . The EPSPS enzyme of  claim 37 , wherein the EPSPS enzyme is fused to a peptide tag. 
     
     
         77 . The EPSPS enzyme of  claim 76 , wherein the peptide tag is a 6×-Histidine tag. 
     
     
         78 . The EPSPS enzyme of  claim 37 , wherein the amino acid that is changed is Asparagine. 
     
     
         79 . The EPSPS enzyme of  claim 37 , wherein the amino acid is changed into an Aspartic Acid. 
     
     
         80 . The EPSPS enzyme of  claim 37 , characterized in that it comprises at least one further amino acid change that on its own renders an EPSPS enzyme into an EPSPS enzyme that confers glyphosate resistance to a plant. 
     
     
         81 . The EPSPS enzyme of  claim 80 , wherein the at least one further amino acid change is at a position corresponding to position 101, 106, or 179 in SEQ ID NO: 5. 
     
     
         82 . The EPSPS enzyme of  claim 81 , wherein the amino acid that is changed corresponding to the position 101 is Glycine, the amino acid that is changed corresponding to the position 106 is Proline, and/or the amino acid that is changed corresponding to the position 179 is Serine. 
     
     
         83 . The EPSPS enzyme of  claim 81 , wherein the amino acid corresponding to the position 101 is changed into an Alanine, the amino acid corresponding to the position 106 is changed into a Leucine, and/or the amino acid corresponding to the position 179 is changed into an Asparagine. 
     
     
         84 . A seed derived from the plant or part thereof of  claim 43 . 
     
     
         85 . A seed derived from the plant or part thereof of  claim 44 . 
     
     
         86 . A seed derived from the plant or part thereof of  claim 45 . 
     
     
         87 . A seed derived from the plant or part thereof of  claim 46 . 
     
     
         88 . A seed derived from the plant or part thereof of  claim 47 . 
     
     
         89 . A seed derived from the plant or part thereof of  claim 48 . 
     
     
         90 . A plant product obtained by processing the plant, or a part thereof, of  claim 42 , wherein the processing comprises cooking, grinding, drying, milling, baking, cutting, sieving, flaking, peeling, soaking, washing, heating, cooling, crushing, or wetting. 
     
     
         91 . The plant product of  claim 90 , wherein the plant product is a starch-based product or a plant-oil-based product. 
     
     
         92 . The plant or part thereof of  claim 42 , wherein the plant is a recombinant or transgenic plant. 
     
     
         93 . The plant or part thereof of  claim 42 , wherein the nucleotide sequence is a cDNA sequence. 
     
     
         94 . A seed derived from the plant or part thereof of  claim 93 .