Compositions and methods for controlling Leptinotarsa
First Claim
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1. A method for controlling a Leptinotarsa species infestation of a plant comprising:
- (a) contacting said Leptinotarsa species with an inhibitory polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(b) providing in the diet of said Leptinotarsa species an inhibitory polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(c) causing mortality or stunting in larvae of said Leptinotarsa species by providing in the diet of said larvae at least one inhibitory polynucleotide comprising at least one silencing element comprising 21 contiguous nucleotides that are complementary to SEQ ID NO;
730, or an RNA transcribed therefrom;
or(d) topically applying to said plant a composition comprising at least one polynucleotide comprising an inhibitory nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(e) expressing in said plant at least one inhibitory polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730;
or(f) providing to said plant at least one inhibitory polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730 or an RNA transcribed therefrom;
or(g) contacting said Leptinotarsa species with an inhibitory polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom.
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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.
444 Citations
20 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 an inhibitory polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(b) providing in the diet of said Leptinotarsa species an inhibitory polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(c) causing mortality or stunting in larvae of said Leptinotarsa species by providing in the diet of said larvae at least one inhibitory polynucleotide comprising at least one silencing element comprising 21 contiguous nucleotides that are complementary to SEQ ID NO;
730, or an RNA transcribed therefrom;
or(d) topically applying to said plant a composition comprising at least one polynucleotide comprising an inhibitory nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(e) expressing in said plant at least one inhibitory polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730;
or(f) providing to said plant at least one inhibitory polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730 or an RNA transcribed therefrom;
or(g) contacting said Leptinotarsa species with an inhibitory polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A insecticidal composition for controlling a Leptinotarsa species, comprising:
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(a) at least one double-stranded RNA (dsRNA) comprising at least 21 contiguous nucleotides that are complementary to a SEQ ID NO;
730, or an RNA transcribed therefrom, 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 comprising at least one silencing element that is complementary to at least 21 contiguous nucleotides 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 comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom, 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) an RNA molecule that causes mortality or stunting of growth in a Leptinotarsa species when ingested or contacted by said Leptinotarsa species, wherein said RNA molecule comprises at least 21 contiguous nucleotides that are complementary to SEQ ID NO;
730, or an RNA transcribed therefrom;
or(e) an insecticidal dsRNA 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 dsRNA molecule comprises 21 contiguous nucleotides that are complementary to SEQ ID NO;
730, or an RNA transcribed therefrom;
or(f) at least one dsRNA comprising a sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1104, 1105, and 1110, wherein said at least one dsRNA causes mortality or stunting of growth in said Leptinotarsa species when ingested or contacted by said Leptinotarsa species. - View Dependent Claims (13, 14, 15)
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16. A recombinant DNA construct comprising a heterologous promoter operably linked to:
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(a) a DNA encoding an inhibitory nucleotide sequence that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(b) a DNA encoding 21 or more contiguous inhibitory nucleotide sequence having 100% identity to a fragment of equivalent length of a DNA having a sequence of SEQ ID NO;
730, or the DNA complement thereof;
or(c) a DNA encoding at least one silencing element that is complementary to at least 21 contiguous nucleotides of SEQ ID NO;
730, or an RNA transcribed therefrom;
or(d) a DNA encoding at least one silencing element comprising at least 21 contiguous nucleotides that are complementary to SEQ ID NO;
730, or an RNA transcribed therefrom;
or(e) a DNA encoding a RNA comprising at least 21 contiguous nucleotides that are complementary to a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1104, 1105, and 1110, or the complement thereof;
or(f) a DNA encoding a RNA comprising at least one double-stranded RNA region, at least one strand of which comprises at least 21 contiguous nucleotides that are complementary to a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1104, 1105, and 1110, or the complement thereat;
or(g) a DNA encoding RNA comprising a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 988, 1104, 1105, and 1110, or the complement thereof. - View Dependent Claims (17, 18, 19, 20)
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Specification