Control of coleopteran pests using RNA molecules
Disclosed are double stranded RNA molecules that are toxic to coleopteran insects. In particular, interfering RNA molecules capable of interfering with pest target genes and that are toxic to the target pest are provided. Further, methods of making and using the interfering RNA, for example in transgenic plants or as the active ingredient in a composition, to confer protection from insect damage are disclosed.
1. An interfering RNA molecule comprising a double-stranded RNA (dsRNA), the dsRNA consisting of annealed complementary strands, one strand of which consists of the nucleotide sequence of SEQ ID NO:390, and the other strand of which consists of the complement of SEQ ID NO:390, wherein the dsRNA has insecticidal activity on a coleopteran plant pest.
2. The interfering RNA molecule of claim 1 , wherein said coleopteran plant pest is a Diabrotica insect.
3. The interfering RNA molecule of claim 1 , wherein the annealed strands are substantially complementary.
4. The interfering RNA molecule of claim 1 , wherein the annealed strands are fully complementary.
5. The interfering RNA molecule of claim 2 , wherein the Diabrotica insect is selected from the group consisting of D. barberi, D. virgifera, D. undecimpunctata, D. balteata, D undecimpunctata, D significate , and D. speciosa.
6. A nucleic acid molecule encoding the interfering RNA molecule of claim 1 .
7. A recombinant vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes the interfering RNA molecule of claim 1 .
8. A transgenic plant, or part thereof, comprising the nucleic acid molecule of claim 6 , wherein the transgenic plant has enhanced resistance to a Diabrotica insect as compared to a control plant.
9. The transgenic plant, or part thereof, of claim 8 , wherein the transgenic plant comprises at least a second insecticidal agent for controlling Diabrotica insects.
10. The transgenic plant, or part thereof, of claim 9 , wherein the second insecticidal agent is a Bacillus thuringiensis insecticidal protein, a Bacillus cereus insecticidal protein, a Xenorhabdus spp. insecticidal protein, a Photorhabdus spp. insecticidal protein, a Brevibacillus laterosporous insecticidal protein, a Lysinibacillus sphearicus insecticidal protein, a Chromobacterium spp. insecticidal protein, a Yersinia entomophaga insecticidal protein, a Paenibacillus popiliae insecticidal protein, a Clostridium spp. insecticidal protein, a patatin, a protease, a protease inhibitor, a urease, an alpha-amylase inhibitor, a pore-forming protein, a lectin, an engineered antibody or antibody fragment, or a chitinase.
11. The transgenic plant, or part thereof, of claim 8 , wherein the transgenic plant, or part thereof, is a maize plant or part thereof.
12. Transgenic seed of the transgenic plant of claim 11 .
13. A commodity product produced from the transgenic plant, or part thereof, of claim 8 , wherein the commodity product comprises a detectable amount of said nucleic acid molecule.
14. The commodity product of claim 13 , selected from the group consisting of whole or processed seeds, beans, grains, kernels, hulls, meals, grits, flours, sugars, sugars, starches, protein concentrates, protein isolates, waxes, oils, extracts, juices, concentrates, liquids, syrups, feed, silage fiber, paper or other food or product produced from plants.
15. A method of controlling a Diabrotica insect comprising contacting the Diabrotica insect with a nucleic acid molecule that is or is capable of producing the interfering RNA molecule of claim 1 for inhibiting expression of a target gene in the Diabrotica insect thereby controlling the Diabrotica insect.
16. The method of claim 15 , wherein the Diabrotica insect is selected from the group consisting of D. barberi, D. virgifera, D. undecimpunctata, D. balteata, D. undecimpunctata, D. significata , and D. speciosa.
17. The method of claim 16 , wherein contacting comprises:
a) planting a transgenic seed capable of producing a transgenic plant that expresses the nucleic acid molecule, wherein the Diabrotica insect feeds on the transgenic plant, or part thereof, or
b) applying a composition comprising the nucleic acid molecule to a seed or plant, or part thereof, wherein the Diabrotica insect feeds on the seed, the plant, or a part thereof.
18. The method of claim 17 , wherein the transgenic seed and transgenic plant is a corn seed and a corn plant.
19. A method of controlling a Diabrotica insect comprising contacting the Diabrotica insect with a nucleic acid molecule that is or is capable of producing the interfering RNA molecule of claim 1 for inhibiting expression of a target gene in the Diabrotica insect, and contacting the Diabrotica insect with at least a second insecticidal agent for controlling Diabrotica.
20. The method of claim 19 , wherein said second insecticidal agent comprises a B. thuringiensis insecticidal protein, a Bacillus cereus insecticidal protein, a Xenorhabdus spp. insecticidal protein, a Photorhabdus spp. Insecticidal protein, a Brevibacillus laterosporous insecticidal protein, a Lysinibacillus sphearicus insecticidal protein, a Chromobacterium spp. insecticidal protein, a Yersinia entomophaga insecticidal protein, a Paenibacillus popiliae insecticidal protein, a Clostridium spp. insecticidal protein, a patatin, a protease, a protease inhibitor, a urease, an alpha-amylase inhibitor, a pore-forming protein, a lectin, an engineered antibody or antibody fragment, or a chitinase.
21. The interfering RNA molecule of claim 1 , wherein the interfering RNA molecule provides at least 87% mortality at 0.01 μg and at least 93% mortality at 0.1 λg after 7 days in western corn rootworm.