Methods and Compositions for Weed Control Using EPSPS Polynucleotides

Abstract

Provided are novel polynucleotide compositions for enhancing the herbicidal activity of glyphosate. Specifically provided are methods and compositions for modulating 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in plant species. The present compositions and methods are useful in controlling glyphosate resistant weeds.

We claim: 
     
         1 . A method of controlling growth, development or reproductive ability of a plant, comprising: topically treating the plant with a composition comprising a bioactive trigger polynucleotide and a transfer agent, wherein the bioactive trigger polynucleotide comprises a nucleotide sequence that is essentially identical or essentially complementary to SEQ ID NO: 3, 5, or 9-66, or a fragment thereof, whereby the growth, development or reproductive ability of the plant is reduced. 
     
     
         2 . The method of  claim 1 , wherein the bioactive trigger polynucleotide comprises a nucleotide sequence that is essentially identical or essentially complementary to SEQ ID NO 3 or SEQ ID NO 5. 
     
     
         3 . The method of  claim 1 , wherein the transfer agent is an organosilicone surfactant. 
     
     
         4 . The method of  claim 3 , wherein the organosilicone surfactant is BREAK-THRU® S 321, BREAK-THRU® S 200, BREAK-THRU® OE 441, BREAK-THRU® S 278, BREAK-THRU® S 243, SILWET L-77®, SILWET® HS 429, SILWET® HS 312, or BREAK-THRU® S 233. 
     
     
         5 . The method of  claim 1 , wherein the transfer agent comprises a cationic lipid reagent. 
     
     
         6 . The method of  claim 5 , wherein the cationic lipid reagent is N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate (DOTAP). 
     
     
         7 . The method of  claim 1 , wherein the transfer agent is a plant hormone and wherein the plant hormone is Brassinosteroid. 
     
     
         8 . The method of  claim 1 , wherein the bioactive trigger polynucleotide is selected from the group consisting of single-stranded DNA, single-stranded RNA, double-stranded RNA, double-stranded DNA, and double-stranded DNA/RNA hybrids. 
     
     
         9 . The method of  claim 8 , wherein the bioactive trigger polynucleotide is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 3 and 4. 
     
     
         10 . The method of  claim 8 , wherein the bioactive trigger polynucleotide is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 5 and 6. 
     
     
         11 . The method of  claim 1 , wherein the plant is selected from the group consisting of  Amaranthus palmeri, Amaranthus rudis, Amaranthus albus, Amaranthus chlorostachys, Amaranthus graecizans, Amaranthus hybridus, Amaranthus lividus, Amaranthus spinosus, Amaranthus thunbergii, Amaranthus viridis, Lolium multiflorum, Lolium rigidum, Ambrosia artemisiifolia, Ambrosia trifida, Euphorbia heterophylla, Kochia scoparia, Abutilon theophrasti, Sorghum halepense, Chenopodium album, Commelina diffusa, Convolvulus arvensis, Conyza canadensis, Digitaria sanguinalis , and  Xanthium strumarium.    
     
     
         12 . The method of  claim 1 , wherein the composition further comprises an EPSPS-inhibitor herbicide. 
     
     
         13 . The method of  claim 12 , wherein the composition further comprises one or more herbicides different from the EPSPS-inhibitor herbicide. 
     
     
         14 . The method of  claim 12 , wherein the composition further comprises an auxin-like herbicide. 
     
     
         15 . The method of  claim 14 , wherein the auxin-like herbicide is dicamba or 2,4-D. 
     
     
         16 . The method of  claim 1 , wherein the composition comprises a combination of two or more different bioactive trigger polynucleotides. 
     
     
         17 . A composition comprising: one or more bioactive trigger polynucleotides and a transfer agent, wherein one or more bioactive trigger polynucleotides comprises a nucleotide sequence that is essentially identical or essentially complementary to SEQ ID NO: 3, 5, or 9-66, or a fragment thereof. 
     
     
         18 . The composition of  claim 17 , wherein one or more bioactive trigger polynucleotides comprises a nucleotide sequence that is essentially identical or essentially complementary to SEQ ID NO 3 or SEQ ID NO 5. 
     
     
         19 . The composition of  claim 17 , wherein one or more bioactive trigger polynucleotides is selected from the group consisting of single-stranded DNA, single-stranded RNA, double-stranded RNA, double-stranded DNA, and double-stranded DNA/RNA hybrids. 
     
     
         20 . The composition of  claim 19 , wherein one or more bioactive trigger polynucleotides is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 3 and 4. 
     
     
         21 . The composition of  claim 19 , wherein one or more bioactive trigger polynucleotides is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 5 and 6. 
     
     
         22 . The composition of  claim 17 , wherein the transfer agent is selected from the group consisting of an organosilicone surfactant, a cationic lipid reagent and Brassinosteroid. 
     
     
         23 . The composition of  claim 22 , wherein the composition further comprises ammonium sulfate. 
     
     
         24 . The composition of  claim 22 , wherein the transfer agent is an organosilicone surfactant selected from the group consisting of: BREAK-THRU® S 321, BREAK-THRU® S 200, BREAK-THRU® OE 441, BREAK-THRU® S 278, BREAK-THRU® S 243, SILWET L-77®, SILWET® HS 429, SILWET® HS 312, and BREAK-THRU® S 233. 
     
     
         25 . The composition of  claim 22 , wherein the transfer agent is DOTAP. 
     
     
         26 . The composition of  claim 17 , further comprising an EPSPS-inhibitor herbicide. 
     
     
         27 . The composition of  claim 26 , wherein the EPSPS-inhibitor herbicide is glyphosate. 
     
     
         28 . The composition of  claim 26 , further comprising a non-EPSPS-inhibitor herbicide. 
     
     
         29 . The composition of  claim 28 , wherein the non-EPSPS-inhibitor herbicide is dicamba or 2,4-D. 
     
     
         30 . A bioactive trigger polynucleotide comprising: a nucleotide sequence that is essentially identical or essentially complementary to SEQ ID NO: 3, 5, or 9-35, or a fragment thereof. 
     
     
         31 . The bioactive trigger polynucleotide of  claim 30 , wherein the bioactive trigger polynucleotide comprises a nucleotide sequence that is essentially identical or essentially complementary to SEQ ID NO 3 or SEQ ID NO 5. 
     
     
         32 . The bioactive trigger polynucleotide of  claim 30 , wherein the bioactive trigger polynucleotide is single-stranded DNA, single-stranded RNA, double-stranded RNA, double-stranded DNA, or a double-stranded DNA/RNA hybrid. 
     
     
         33 . The bioactive trigger polynucleotide of  claim 32 , wherein the bioactive trigger polynucleotide is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 3 and 4. 
     
     
         34 . The bioactive trigger polynucleotide of  claim 32 , wherein the bioactive trigger polynucleotide is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 5 and 6. 
     
     
         35 . A plant cell comprising the bioactive trigger polynucleotide of any one of  claims 30 - 34 . 
     
     
         36 . A growing plant comprising the bioactive trigger polynucleotide of any one of  claims 30 - 34 . 
     
     
         37 . A seed comprising the bioactive trigger polynucleotide of any one of  claims 30 - 34 . 
     
     
         38 . A method of sensitizing a weed plant to an EPSPS-inhibitor herbicide comprising:
 treating the weed with a bioactive trigger polynucleotide that is essentially identical or essentially complementary to a nucleotide sequence selected from the group consisting of SEQ ID NO: 3, 5, and 9-66, or a fragment thereof, whereby the weed is more sensitive to an EPSPS-inhibitor herbicide relative to a weed not treated with the bioactive trigger polynucleotide.   
     
     
         39 . The method of  claim 38 , wherein the EPSPS-inhibitor herbicide is glyphosate. 
     
     
         40 . The method of  claim 38 , wherein the weed is resistant to one or more of glyphosate, dicamba and sulfonylurea. 
     
     
         41 . The method of  claim 40 , wherein the weed is growing in a field of herbicide-resistant crop plants. 
     
     
         42 . The method of  claim 40 , wherein the weed is selected from the group consisting of  Amaranthus palmeri, Amaranthus rudis, Amaranthus albus, Amaranthus chlorostachys, Amaranthus graecizans, Amaranthus hybridus, Amaranthus lividus, Amaranthus spinosus, Amaranthus thunbergii, Amaranthus viridis, Lolium multiflorum, Lolium rigidum, Ambrosia artemisiifolia, Ambrosia trifida, Euphorbia heterophylla, Kochia scoparia, Abutilon theophrasti, Sorghum halepense, Chenopodium album, Commelina diffusa, Convolvulus arvensis, Conyza canadensis, Digitaria sanguinalis , and  Xanthium strumarium.    
     
     
         43 . The method of  claim 38 , wherein the bioactive trigger polynucleotide is single-stranded DNA, single-stranded RNA, double-stranded RNA, double-stranded DNA, or a double-stranded DNA/RNA hybrid. 
     
     
         44 . The method of  claim 38 , wherein the bioactive trigger polynucleotide is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 3 and 4. 
     
     
         45 . The method of  claim 38 , wherein the bioactive trigger polynucleotide is double-stranded RNA and the double-stranded RNA comprises SEQ ID NOs: 5 and 6.