Methods and compositions for providing tolerance to multiple herbicides

Abstract

Methods and compositions are provided related to improved plants that are tolerant to more than one herbicide. Particularly, the invention provides plants that are tolerant of glyphosate and are tolerant to at least one ALS inhibitor, and methods of use thereof. The glyphosate/ALS inhibitor-tolerant plants comprise a polynucleotide that encodes a polypeptide that confers tolerance to glyphosate and a polynucleotide that encodes an ALS inhibitor-tolerant polypeptide. In specific embodiments, a plant of the invention expresses a GAT polypeptide and an HRA polypeptide. Methods to control weeds, improve plant yield, and increase transformation efficiencies are provided.

1 . A plant comprising a heterologous polynucleotide comprising a first polynucleotide encoding a polypeptide that confers tolerance to glyphosate, a second polynucleotide encoding a polypeptide that confers tolerance to an Acetolactate Synthase (ALS) and a third polynucleotide that encodes a polypeptide that confers tolerance to a synthetic auxin. 
     
     
         2 . The plant of  claim 1 , wherein said polynucleotide encoding the polypeptide that confers tolerance to glyphosate encodes a glyphosate N-acetyltransferase. 
     
     
         3 . The plant of  claim 1 , wherein said polynucleotide encoding the polypeptide that confers tolerance to glyphosate encodes a modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) that is not inhibited by glyphosate. 
     
     
         4 . The plant of  claim 1 , wherein said synthetic auxin comprises a phenoxycarboxylic acid, a benzoic acid, a pyridine carboxylic acid, quinoline carboxylic acid, or benazolin-ethyl. 
     
     
         5 . The plant of  claim 4 , wherein said synthetic auxin comprises clomeprop, 2,4-D or mecoprop. 
     
     
         6 . The plant of  claim 4 , wherein said synthetic auxin comprises dicamba or chloramben. 
     
     
         7 . The plant of  claim 1 , wherein said polynucleotide that encodes said polypeptide that confers tolerance to said synthetic auxin comprises an aryloxyalkanoate dioxygenase. 
     
     
         8 . The plant of  claim 1 , wherein said plant is a dicot. 
     
     
         9 . The plant of  claim 1 , wherein said dicot is soybean, canola, sunflower, cotton, alfalfa, a sugar beet, or an  Arabidopsis.    
     
     
         10 . A transgenic seed produced from the plant of  claim 1 , wherein said seed comprises the heterologous polynucleotide comprising the first polynucleotide encoding the polypeptide that confers tolerance to glyphosate, the second polynucleotide encoding a polypeptide that confer tolerance to the Acetolactate Synthase (ALS) and the third polynucleotide that encodes a polypeptide that confers tolerance to the synthetic auxin. 
     
     
         11 . A method for controlling weeds in an area of cultivation comprising:
 a) planting crop seeds or crop plants in the area of cultivation, wherein the crop seeds or crop plants are tolerant to ALS herbicides, glyphosate and synthetic auxins; and,   b) applying to said crop seeds, said crop plants, said area of cultivation or a combination thereof, a sufficient amount of at least one herbicide combination to control the weeds without significantly affecting the crop.   
     
     
         12 . The method of  claim 11 , wherein the at least one herbicide combination comprises glyphosate, a synthetic auxin or an ALS inhibitor. 
     
     
         13 . The method of  claim 11 , wherein the herbicides of the at least one herbicide combination are applied sequentially. 
     
     
         14 . The method of  claim 11 , wherein the herbicides of the at least one herbicide combination are applied simultaneously. 
     
     
         15 . The method of  claim 11 , wherein the tolerance to glyphosate is by expression of a glyphosate N-acetyltransferase. 
     
     
         16 . The method of  claim 11 , wherein the tolerance to glyphosate is by expression of a modified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) that is not inhibited by glyphosate. 
     
     
         17 . The method of  claim 11 , wherein said synthetic auxin comprises a phenoxyearboxylic acid, a benzoic acid, a pyridine carboxylic acid, quinoline carboxylic acid, or benazolin-ethyl. 
     
     
         18 . The method of  claim 17 , wherein said synthetic auxin comprises 2,4-D or dicamba. 
     
     
         19 . The method of  claim 17 , wherein said benzoic acid comprises dicamba or chloramben. 
     
     
         20 . The method of  claim 11 , wherein said polynucleotide that encodes a polypeptide that confers tolerance to a synthetic auxin comprises an aryloxyalkanoate dioxygenase. 
     
     
         21 . The method of  claim 11 , wherein said plant is a dicot. 
     
     
         22 . The method of  claim 21 , wherein said dicot is soybean, canola, sunflower, cotton, alfalfa, a sugar beet, or an  Arabidopsis.    
     
     
         23 . A method for controlling weeds in an area of cultivation comprising:
 a) planting crop seeds or crop plants in the area of cultivation, wherein the crop seeds or crop plants express a glyphosate N-acetyltransferase and a polypeptide that confers tolerance to a synthetic auxin selected from the group consisting of 2,4-D and dicamba; and   b) applying to said crop seeds, said crop plants, said area of cultivation or a combination thereof, a sufficient amount of at least one herbicide combination to control the weeds without significantly affecting the crop.   
     
     
         24 . The method of  claim 23 , wherein the at least one herbicide combination comprises glyphosate and a synthetic auxin. 
     
     
         25 . The method of  claim 23 , wherein the herbicides of the at least one herbicide combination are applied sequentially. 
     
     
         26 . The method of  claim 23 , wherein the herbicides of the at least one herbicide combination are applied simultaneously. 
     
     
         27 . The method of  claim 23 , wherein said polypeptide that confers tolerance to a synthetic auxin comprises an aryloxyalkanoate dioxygenase. 
     
     
         28 . The method of  claim 23 , wherein said plant is a dicot. 
     
     
         29 . The method of  claim 23 , wherein said dicot is soybean, canola, sunflower, cotton, alfalfa, a sugar beet, or an  Arabidopsis.    
     
     
         30 . The method of  claim 23 , wherein said glyphosate N-acetyltransferase comprises an amino acid sequence having at least 80% or 85% or 90% sequence identity to the sequence set forth in SEQ ID NO: 56 or 69.