Compositions and methods for controlling Leptinotarsa

Patent metadata

Field	Value
Publication number	US-9856495-B2
Application number	US-201514608951-A
Country	US
Kind code	B2
Filing date	Jan 29, 2015
Priority date	Jul 19, 2013
Publication date	Jan 2, 2018
Grant date	Jan 2, 2018

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) polynucleotide comprising a strand comprising a nucleotide sequence that is at least 98% identical to SEQ ID NO:989; or
(b) providing in the diet of said Leptinotarsa species a dsRNA polynucleotide comprising a strand comprising a nucleotide sequence that is at least 98% identical to SEQ ID NO:989; or
(c) causing mortality or stunting in larvae of said Leptinotarsa species by providing in the diet of said larvae at least one dsRNA polynucleotide comprising at least one silencing element comprising a strand comprising a nucleotide sequence that is at least 98% identical to SEQ ID NO:989; or
(d) topically applying to said plant a composition comprising at least one dsRNA polynucleotide comprising a strand comprising a nucleotide sequence that is at least 98% identical to SEQ ID NO:989, wherein said at least one dsRNA polynucleotide is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one dsRNA polynucleotide; or
(e) expressing in said plant at least one dsRNA polynucleotide comprising a strand comprising a sequence that is at least 98% identical to SEQ ID NO: 989, wherein said at least one dsRNA polynucleotide is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one dsRNA polynucleotide; or
(f) providing to said plant at least one dsRNA polynucleotide comprising a strand comprising a sequence that is at least 98% identical to SEQ ID NO: 989, wherein said at least one dsRNA polynucleotide is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one dsRNA 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 100% identical to SEQ ID NO:989.

4. The method of claim 1 , wherein said method comprises topically applying to said plant a composition comprising at least one dsRNA polynucleotide comprising a strand comprising a nucleotide sequence that is at least 98% identical to SEQ ID NO:989; 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 nonpolynucleotide 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 polynucleotide, wherein said Leptinotarsa species is Leptinotarsa decemlineata , wherein RNA interference is induced and Leptinotarsa decemlineata mortality occurs, and wherein at least one strand of said double-stranded RNA polynucleotide comprises the sequence of SEQ ID NO:989.

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

7. The method of claim 1 , wherein said method comprises topically applying to said plant a composition comprising at least one dsRNA polynucleotide in a manner such that an effective amount of said dsRNA polynucleotide is ingested by Leptinotarsa species feeding on said plant, wherein said Leptinotarsa species is Leptinotarsa decemlineata ; and wherein said dsRNA polynucleotide comprises a strand having a sequence 100% identical to SEQ ID NO:989.

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, a seed, or a 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 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, and wherein one strand comprises a nucleotide sequence that is at least 98% identical to SEQ ID NO:989.

12. The insecticidal composition of claim 11 , wherein said insecticidal composition is in the form of at least one selected from the group consisting of a solid, liquid, powder, suspension, emulsion, spray, encapsulation, microbeads, carrier particulates, film, matrix, seed treatment, soil drench, implantable formulation, and in-furrow formulation.

13. The insecticidal composition of claim 11 , further comprising at least one component 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 nonpolynucleotide pesticide, a safener, and an insect growth regulator.

14. The insecticidal composition of claim 11 , wherein said insecticidal composition comprises an insecticidal dsRNA molecule that causes mortality or stunting of growth in a Leptinotarsa species when ingested or contacted by said Leptinotarsa species, wherein said insecticidal dsRNA molecule comprises at least one segment that is at least 98% identical to SEQ ID NO:989.

15. A recombinant DNA construct comprising a heterologous promoter operably linked to: DNA encoding a RNA comprising at least one double-stranded RNA region, at least one strand of which comprises a nucleotide sequence that is at least 98% identical to SEQ ID NO:989.

16. A plant chromosome or a plastid or a recombinant plant virus vector or a recombinant baculovirus vector comprising the recombinant DNA construct of claim 15 .

17. A transgenic solanaceous plant cell having in its genome the recombinant DNA construct of claim 15 .

18. The transgenic solanaceous plant cell of claim 17 , wherein said transgenic solanaceous plant cell further has in its genome DNA encoding at least one pesticidal agent selected from the group consisting of a patatin, a plant lectin, a phytoecdysteroid, a Bacillus thuringiensis insecticidal protein, a Xenorhabdus insecticidal protein, a Photorhabdus insecticidal protein, a Bacillus laterosporous insecticidal protein, and a Bacillus sphaericus insecticidal protein.

19. A transgenic solanaceous plant comprising the transgenic solanaceous plant cell of claim 17 , or a fruit, a seed, or a propagatable part of said transgenic solanaceous plant.

Assignees

• Monsanto Technology Llc

Inventors

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

Classifications

• C12N15/8218

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

• C07K14/43563

  from insects · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N15/8286 Primary

  for insect resistance · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title