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CN-122028797-A - Double-stranded RNA methods and compositions for pesticides

CN122028797ACN 122028797 ACN122028797 ACN 122028797ACN-122028797-A

Abstract

Disclosed herein are pesticidal compositions and methods for controlling plant pests. Pesticidal compositions and methods comprise recombinant RNA molecules for RNA interference-mediated inhibition of a target gene in plant pests. Transgenic plants expressing recombinant RNA molecules useful for controlling plant pests are further provided, as are plant cells and plant products obtained thereby.

Inventors

  • M. Montar
  • A. KUMAR

Assignees

  • 核糖核酸复兴股份有限责任公司

Dates

Publication Date
20260512
Application Date
20240815
Priority Date
20230816

Claims (20)

  1. 1. An insecticidal composition comprising an pesticidally effective amount of a recombinant RNA molecule comprising a nucleotide sequence of at least 17 or more contiguous nucleotides having a sequence of at least about 95% to about 100% complementarity to a portion of a target gene of an insect that infests plants, said target gene encoding an mRNA sequence selected from the group consisting of SEQ ID nos 1,4, 6 and 8.
  2. 2. The insecticidal composition of claim 1, wherein the recombinant RNA molecule comprises at least one RNA strand having a sequence from about 95% to about 100% identical or complementary to a sequence selected from the group consisting of SEQ ID NOs 3, 5, and 10.
  3. 3. The insecticidal composition of claim 1, wherein the recombinant RNA molecule is a double-stranded RNA (dsRNA), a microrna (miRNA), a small interfering RNA (siRNA), a hairpin RNA (hpRNA), or a piwi-interacting RNA (pi-RNA).
  4. 4. The insecticidal composition of claim 1, wherein the recombinant RNA molecule is a double-stranded RNA (dsRNA) comprising an RNA strand having a sequence selected from the group consisting of SEQ ID NOs 3, 5 and 10.
  5. 5. The insecticidal composition of claim 1, wherein the dsRNA is at least 17 base pairs in length.
  6. 6. The insecticidal composition of claim 1, wherein the insect is an adult or is in the larval or nymph stage.
  7. 7. The insecticidal composition of claim 1, wherein the plant-infesting insect is a coleopteran insect.
  8. 8. The insecticidal composition of claim 7, wherein the plant-infesting insect is an insect from the genus leptinotarsa.
  9. 9. The insecticidal composition according to claim 8 wherein the plant-infesting insect is a potato beetle (colorado potato beetle) and the target gene encodes an mRNA sequence comprising the nucleotide sequence of SEQ ID No: 1.
  10. 10. The insecticidal composition according to claim 1 wherein the plant-infesting insect is a potato beetle (colorado potato beetle) and the target gene encodes an mRNA sequence comprising the nucleotide sequence of SEQ ID No. 4 or 6.
  11. 11. The insecticidal composition according to claim 1 wherein the plant-infesting insect is a potato beetle (colorado potato beetle) and the target gene encodes an mRNA sequence comprising the nucleotide sequence of SEQ ID No: 8.
  12. 12. The insecticidal composition according to claim 1 wherein said plant-infesting insect is potato beetle (colorado potato beetle) and said recombinant RNA molecule comprises at least one RNA strand having a sequence from about 95% to about 100% identical or complementary to a sequence selected from the group consisting of SEQ ID nos. 3, 5 and 10.
  13. 13. The insecticidal composition according to claim 1 wherein the plant-infesting insect is a potato beetle (colorado potato beetle) and the recombinant RNA molecule is a dsRNA comprising an RNA strand having a sequence selected from the group consisting of SEQ ID nos 3, 5 and 10.
  14. 14. The insecticidal composition of claim 1, further comprising at least one component selected from the group consisting of a carrier, an excipient, a diluent, a surfactant, a silicone, a polynucleotide herbicidal molecule, a non-polynucleotide pesticide, a safener, an insect attractant, and an insect growth regulator.
  15. 15. The insecticidal composition of claim 1, further comprising at least one pesticide agent.
  16. 16. The insecticidal composition of claim 14, wherein the pesticide agent is selected from the group consisting of patatin, phytolectin, phytoecdysteroid, and insecticidal protein.
  17. 17. The insecticidal composition according to claim 1 in a form selected from the group consisting of solids, liquids, powders, suspensions, emulsions, sprays, encapsulates, microbeads, carrier particles, films, solid matrices, soil drenches, insect foods, insect baits, and seed treatments.
  18. 18. A plant or seed thereof treated with the insecticidal composition of claim 1, wherein the plant exhibits improved resistance to the insect.
  19. 19. A recombinant DNA construct comprising a heterologous promoter operably linked to DNA encoding an RNA transcript comprising a sequence having about 95% to about 100% identity or complementarity to a sequence selected from the group consisting of SEQ ID NOs 3, 5 and 10.
  20. 20. The recombinant DNA construct according to claim 19, wherein said heterologous promoter is functional for expression of RNA transcripts in bacteria or fungi.

Description

Double-stranded RNA methods and compositions for pesticides Cross Reference to Related Applications The present application claims priority from U.S. provisional patent application Ser. No. 63/520,023, filed 8/16 of 2023, and entitled "DOUBLE STRANDED RNA METHODS AND COMPOSITIONS FOR INSECTICIDE," which is incorporated herein by reference in its entirety. Incorporation of the sequence Listing The present application contains a sequence listing that has been submitted in XML format via patent center (PATENT CENTER) and is incorporated by reference herein in its entirety. The WIPO sequence table was created at 2024, 8, 13, with an XML copy named 066803-818366_Sequence listing.xml and a size of 41 kilobytes. Technical Field The present disclosure relates to the field of RNA interference (RNAi) -mediated gene silencing in insect pests. Background The control of insect pests on agriculturally important crops is important to maximize crop yield. Insect pests that damage plants belonging to the Solanaceae family (Solanaceae) include many coleopteran pests, especially potato (Solanum tuberosum), tomato (Solanumlycopersicum)), eggplant (Solanum melongena)), capsicum (cherokee rose (Solanum capsicum)), and solanum plants (e.g., solanum lycopersicum (Solanum aculeastrum), castanopsis cumingii (s. Bulbocastanum), sweet potato (s. Cardiophyllum), s. Douglasii, euonymus alatus (s. Dulcamara), s. Lanceolatum, s. Robustum, and s. Triquetrum). Although progress has been made in the last few decades in developing more effective methods and compositions for controlling insect infestation in plants using chemical pesticides, the resistance of insect pests to the compounds developed is a significant problem. Biological control methods such as biopesticides based on RNA interference (RNAi) offer an alternative and unique approach to pest integrated management. RNAi is a natural biological process found in most eukaryotes to combat viruses or to play a role in the regulation of messenger RNA (mRNA) stability and translation. In some insects, such as Colorado Potato Beetles (CPB) and other coleoptera species, RNAi pathways can be triggered upon insect ingestion of exogenous double-stranded RNA, potentially providing such a technique to control such pests. Thus, there is an unmet need to identify new targets in the insect genome and to develop RNAi-based pesticides to cause death, growth arrest or sterility of insect pests. Disclosure of Invention In some aspects, the present disclosure encompasses pesticidal compositions comprising a pesticidally effective amount of a recombinant RNA molecule comprising a nucleotide sequence of at least 17 or more contiguous nucleotides having a sequence of at least about 95% to about 100% complementarity to a portion of a target gene of an insect that infects a plant, said target gene encoding an mRNA sequence selected from the group consisting of SEQ ID nos. 1, 4, 6 and 8. In some aspects, the disclosed pesticidal compositions include a recombinant RNA molecule comprising at least one RNA strand having a sequence from about 95% to about 100% identical or complementary to a sequence selected from the group consisting of SEQ ID NOs 3, 5, and 10. In some aspects, the recombinant RNA molecule is a double-stranded RNA (dsRNA), a microrna (miRNA), a small interfering RNA (siRNA), a hairpin RNA (hpRNA), or a piwi-interacting RNA (pi-RNA). In some aspects, the disclosed pesticidal compositions comprise a recombinant RNA molecule that is a double-stranded RNA (dsRNA) comprising an RNA strand having a sequence selected from the group consisting of SEQ ID NOs 3,5, and 10. In some aspects, the dsRNA is at least 17 base pairs in length. In some aspects, the insect is an adult, or is in the larval or nymph stage. In some aspects, the plant-infesting insect is a coleopteran (Coleopteran) insect. In some aspects, the plant-invasive insect is an insect from the genus leptinotarsa (Leptinotarsa). In some aspects, the plant-infecting insect is a potato beetle (Leptinotarsa decemlineata) (Colorado potato beetle), and the target gene encodes an mRNA sequence comprising the nucleotide sequence of SEQ ID No. 1. In some aspects, the plant-infesting insect is a potato beetle (Colorado potato beetle), and the target gene encodes an mRNA sequence comprising the nucleotide sequence of SEQ ID No. 4 or 6. In some aspects, the plant-infesting insect is a potato beetle (Colorado potato beetle), and the target gene encodes an mRNA sequence comprising the nucleotide sequence of SEQ ID No. 8. In some aspects, the plant-infesting insect is a potato beetle (Colorado potato beetle), and the recombinant RNA molecule comprises at least one RNA strand having a sequence from about 95% to about 100% identical or complementary to a sequence selected from the group consisting of SEQ ID Nos. 3,5, and 10. In some aspects, the plant-invasive insect is a potato beetle (Colorado potato beetle), and the recombinant RNA molecule is a dsRNA compri