Compositions and methods for controlling Leptinotarsa

Patent metadata

Field	Value
Publication number	US-9850496-B2
Application number	US-201514603347-A
Country	US
Kind code	B2
Filing date	Jan 22, 2015
Priority date	Jul 19, 2013
Publication date	Dec 26, 2017
Grant date	Dec 26, 2017

Abstract

Disclosed herein are methods of controlling insect pests, in particular Leptinotarsa spp. which infest crop plants, and methods of providing plants resistant to such pests. Also disclosed are polynucleotides and recombinant DNA molecules and constructs useful in such methods, insecticidal compositions such as topical sprays containing insecticidal double-stranded RNAs, and solanaceous plants with improved resistance to infestation by Leptinotarsa spp. Further disclosed are methods of selecting target genes for RNAi-mediated silencing and control of Leptinotarsa spp.

First claim

What is claimed is:

1. A method for controlling a Leptinotarsa species infestation of a plant comprising:
(a) contacting said Leptinotarsa species with a double-stranded RNA (dsRNA) comprising a strand comprising a nucleotide sequence that is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730; or
(b) providing in the diet of said Leptinotarsa species a dsRNA comprising a strand comprising a nucleotide sequence that is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730; or
(c) causing mortality or stunting in larvae of said Leptinotarsa species by providing in the diet of said larvae at least one dsRNA comprising at least one silencing element comprising a strand comprising 22 contiguous nucleotides that are complementary to a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730; or
(d) topically applying to said plant a composition comprising at least one dsRNA comprising a strand comprising a nucleotide sequence that is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said at least one polynucleotide is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one polynucleotide; or
(e) expressing in said plant at least one dsRNA comprising at least one segment that is identical or complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said at least one polynucleotide is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one polynucleotide; or
(f) providing to said plant at least one dsRNA comprising at least one segment that is identical or complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730; or
(g) contacting said Leptinotarsa species with a dsRNA comprising at least one segment that is identical or complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said dsRNA is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one polynucleotide.

2. The method of claim 1 , wherein said double-stranded RNA is chemically synthesized or is produced by expression in a microorganism or by expression in a plant cell.

3. The method of claim 1 , wherein said double-stranded RNA comprises a strand comprising a sequence selected from the group consisting of SEQ ID NOs:989, 988, 1103 1105, and 1116-1119.

4. The method of claim 1 , wherein said method comprises topically applying to said plant a composition comprising at least one dsRNA comprising a strand comprising a nucleotide sequence that is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, and wherein said composition further comprises one or more components selected from the group consisting of a carrier agent, a surfactant, a cationic lipid, an organosilicone, an organosilicone surfactant, a polynucleotide herbicidal molecule, a non-polynucleotide herbicidal molecule, a non-polynucleotide pesticide, a safener, and an insect growth regulator.

5. The method of claim 1 , wherein said method comprises contacting said Leptinotarsa species with an effective amount of a solution comprising a double-stranded RNA, wherein at least one strand of the double-stranded RNA is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said Leptinotarsa species is Leptinotarsa decemlineata , and wherein RNA interference is induced and Leptinotarsa decemlineata mortality occurs.

6. The method of claim 5 , wherein said solution further comprises one or more components selected from the group consisting of an organosilicone surfactant or a cationic lipid.

7. The method of claim 1 , wherein said method comprises topically applying to said plant a composition comprising a double-stranded RNA comprising a strand comprising a nucleotide sequence that is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, and wherein said Leptinotarsa species is Leptinotarsa decemlineata.

8. The method of claim 1 , wherein said Leptinotarsa species is selected from the group consisting of: Leptinotarsa behrensi, Leptinotarsa collinsi, Leptinotarsa decemlineata (Colorado potato beetle), Leptinotarsa defecta, Leptinotarsa haldemani (Haldeman's green potato beetle), Leptinotarsa heydeni, Leptinotarsa juncta (false potato beetle), Leptinotarsa lineolata (burrobrush leaf beetle), Leptinotarsa peninsularis, Leptinotarsa rubiginosa, Leptinotarsa texana, Leptinotarsa tlascalana, Leptinotarsa tumamoca , and Leptinotarsa typographica.

9. A plant having improved resistance to a Leptinotarsa species infestation, provided by the method of claim 1 , or a fruit, seed, or propagatable part of said plant.

10. The plant of claim 9 , wherein said plant is selected from the group consisting of potato, tomato, and eggplant.

11. An insecticidal composition for controlling a Leptinotarsa species, comprising:
(a) at least one double-stranded RNA (dsRNA) consisting of two strands, wherein a first strand comprises at least 22 contiguous nucleotides that are complementary to a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said at least one dsRNA causes mortality or stunting of growth in said Leptinotarsa species when ingested or contacted by said Leptinotarsa species; or
(b) at least one dsRNA consisting of at least one silencing element that is complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said at least one dsRNA causes mortality or stunting of growth in said Leptinotarsa species when ingested or contacted by said Leptinotarsa species; or
(c) at least one dsRNA consisting of two strands, wherein a first strand comprises a sequence that is identical or complementary to at least 22 contiguous nucleotides of a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730, wherein said at least one dsRNA causes mortality or stunting of growth in said Leptinotarsa species when ingested or contacted by said Leptinotarsa species; or
(d) a dsRNA molecule that causes mortality or stunting of growth in a Leptinotarsa species when ingested or contacted by said Leptinotarsa species, wherein said dsRNA molecule consists of two strands, wherein a first strand comprises at least 22 contiguous nucleotides that are complementary to a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730; or
(e) an insecticidal double-stranded RNA molecule that causes mortality or stunting of growth in a Leptinotarsa species when ingested or contacted by said Leptinotarsa species, wherein at least one strand of said insecticidal double-stranded RNA molecule comprises 22 contiguous nucleotides that are complementary to a sequence corresponding to positions 227 to 758 of SEQ ID NO: 730; or
(f) at least one double-stranded RNA consisting of two strands, wherein the sequence of a first strand is selected from the group consisting of SEQ ID NO:989, 988, 1103 1105, and 1116-1119, wherein said at least one polynucleotide causes mortality or stunting of growth in said Leptinotarsa species when ingested or contacted by said Leptinotarsa species.

12.

Assignees

• Monsanto Technology Llc

Inventors

• Beattie Jodi Lynn
• Crawford Michael John
• Eads Brian Donovan
• Flagel Lex Evan
• Kapoor Mahak
• Taylor Christina Marie

Classifications

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C07K14/43563

  from insects · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• A01N57/16

  containing heterocyclic radicals · CPC title