Compositions and methods for controlling leptinotarsa
First Claim
Patent Images
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 sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
or(b) providing in the diet of said Leptinotarsa species a dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
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 a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
or(d) topically applying to said plant a composition comprising at least one dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, wherein said at least one dsRNA is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one dsRNA;
or(e) expressing in said plant at least one dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, wherein said at least one dsRNA is ingested by a Leptinotarsa species feeding on said plant;
or(f) providing to said plant at least one dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, wherein said at least one dsRNA is ingested by a Leptinotarsa species feeding on said plant;
or(g) contacting said Leptinotarsa species with a dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105.
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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.
387 Citations
18 Claims
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1. A method for controlling a Leptinotarsa species infestation of a plant comprising:
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(a) contacting said Leptinotarsa species with a double-stranded RNA (dsRNA) comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
or(b) providing in the diet of said Leptinotarsa species a dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
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 a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
or(d) topically applying to said plant a composition comprising at least one dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, wherein said at least one dsRNA is provided in a manner that permits a Leptinotarsa species feeding on said plant to ingest said at least one dsRNA;
or(e) expressing in said plant at least one dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, wherein said at least one dsRNA is ingested by a Leptinotarsa species feeding on said plant;
or(f) providing to said plant at least one dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, wherein said at least one dsRNA is ingested by a Leptinotarsa species feeding on said plant;
or(g) contacting said Leptinotarsa species with a dsRNA comprising a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. An insecticidal composition for controlling a Leptinotarsa species;
- comprising;
(a) at least one dsRNA consisting of two strands, wherein a first strand comprises a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104; and
1105, 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 comprises a strand comprising a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, 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 having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or complementary to a sequence having at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, 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 is at least 98% identical to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
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 consists of a sequence comprising at least 98% identity to a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105;
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 NOs;
989;
988, 1103, 1104, and 1105, wherein said at least one double-stranded RNA causes mortality or stunting of growth in said Leptinotarsa species when ingested or contacted by said Leptinotarsa species. - View Dependent Claims (12, 13)
- comprising;
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14. A recombinant DNA construct comprising a heterologous promoter operably linked to:
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(a) DNA encoding an RNA comprising at least 98% identity to a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or the complement thereof;
or(b) DNA encoding an RNA comprising at least one double-stranded RNA region, at least one strand of which comprises at least 98% identity to the nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or the complement thereof;
or(c) DNA encoding an RNA comprising the nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1103, 1104, and 1105, or the complement thereof. - View Dependent Claims (15, 16, 17, 18)
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Specification