Herbicide resistance genes

Abstract

The subject invention provides novel plants that are not only resistant to 2,4-D, but also to pyridyloxyacetate herbicides. Heretofore, there was no expectation or suggestion that a plant with both of these advantageous properties could be produced by the introduction of a single gene. The subject invention also includes plants that produce one or more enzymes of the subject invention “stacked” together with one or more other herbicide resistance genes. The subject invention enables novel combinations of herbicides to be used in new ways. Furthermore, the subject invention provides novel methods of preventing the development of, and controlling, strains of weeds that are resistant to one or more herbicides such as glyphosate. The preferred enzyme and gene for use according to the subject invention are referred to herein as AAD-12 (AryloxyAlkanoate Dioxygenase). This highly novel discovery is the basis of significant herbicide tolerant crop trait and selectable marker opportunities.

1 . A polynucleotide comprising a heterologous regulatory element operably linked to a coding sequence, said coding sequence encoding an AAD-12 polypeptide that exhibits aryloxyalkanoate dioxygenase activity wherein said activity enzymatically degrades a phenoxy auxin herbicide and a pyridyloxy auxin herbicide, further wherein said AAD-12 polypeptide comprises:
 i) an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 2; and   ii) an AAD-12 motif having the general formula of:
 HX 109 D(X) 111-134 T(X) 136-261 H(X) 263-272 R, wherein
 X 109  represents a single amino acid at position 109, relative to the sequence of SEQ ID NO: 2; 
 (X) 111-134  represents a sequence of 24 amino acids; 
 (X) 136-261  represents a sequence of 126 amino acids; and 
 (X) 263-272  represents a sequence of 10 amino acids. 
 
   
     
     
         2 . The polynucleotide of  claim 1  wherein the AAD-12 polypeptide comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 2. 
     
     
         3 . The polynucleotide of  claim 1  wherein said heterologous regulatory element is a promoter that functions in plants. 
     
     
         4 . The polynucleotide of  claim 3  wherein said promoter is functional in a dicotyledonous plant. 
     
     
         5 . The polynucleotide of  claim 3  wherein said promoter is functional in a monocotyledonous plant. 
     
     
         6 . The polynucleotide of  claim 1  wherein said polynucleotide comprises the plant optimized codons of SEQ ID NO: 3. 
     
     
         7 . An expression cassette comprising
 a polynucleotide that encodes an AAD-12 polypeptide that exhibits aryloxyalkanoate dioxygenase activity wherein said activity enzymatically degrades a phenoxy auxin herbicide and a pyridyloxy auxin herbicide, further wherein said AAD-12 polypeptide comprises:
 i) an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 2; and 
 ii) an AAD-12 motif having the general formula of:
 HX 109 D(X) 111-134 T(X) 136-261 H(X) 263-272 R, wherein
 X 109  represents a single amino acid at position 109, relative to the sequence of SEQ ID NO: 2; 
 (X) 111-134  represents a sequence of 24 amino acids; 
 (X) 136-261  represents a sequence of 126 amino acids; and 
 
 (X) 263-272  represents a sequence of 10 amino acids; and 
 
   a heterologous promoter operably linked to said polynucleotide.   
     
     
         8 . The expression cassette of  claim 7  wherein the AAD-12 polypeptide comprises an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 2. 
     
     
         9 . The expression cassette of  claim 7  wherein said promoter is functional in a plant cell. 
     
     
         10 . The expression cassette of  claim 7  wherein said promote is a plant promoter. 
     
     
         11 . The expression cassette of  claim 10  wherein said promoter is native to a soybean plant. 
     
     
         12 . The expression cassette of  claim 10  wherein said promoter is native to a maize plant. 
     
     
         13 . The expression cassette of  claim 7  wherein said promoter is selected from the group consisting of cassava vein mosaic virus promoter, CaMV 35S promoter, Figwort Mosaic Virus promoter, rice actin promoter, phaseolin promoter,  Arabidopsis thaliana  Ubiquitin 10 promoter, maize ubiquitin promoter,  Arabidopsis thaliana  Act2 promoter,  Arabidopsis thaliana  Ubiquitin 11 promoter, and  Arabidopsis thaliana  Ubiquitin 3 promoter. 
     
     
         14 . The expression cassette of  claim 7  further comprising a first and second T-DNA border sequences, wherein said promoter and polynucleotide are located between said first and second T-DNA border sequences. 
     
     
         15 . The expression cassette of  claim 14  further comprising a selectable marker gene. 
     
     
         16 . The expression cassette of  claim 7  further comprising nucleic acid sequences from an  Agrobacterium  broad spectrum vector. 
     
     
         17 . The expression cassette of  claim 14  wherein the  Agrobacterium  broad spectrum vector is pRK2 or a derivative thereof. 
     
     
         18 . The expression cassette of  claim 7  wherein said aryloxyalkanoate dioxygenase activity degrades 2,4-dichlorophenoxyacetic acid, and MCPA. 
     
     
         19 . The expression cassette of  claim 7  wherein said aryloxyalkanoate dioxygenase activity degrades triclopyr or fluroxypyr.