PEST CONTROL SYSTEM

• US 20180195137A1
• Filed: 06/08/2016
• Published: 07/12/2018
• Est. Priority Date: 06/11/2015
• Status: Abandoned Application

• Current Assignee: Swansea University
• Original Assignee: Swansea University
• Inventors: Dyson, Paul, Whitten, Miranda
• Application Number:
  US15/580,042
• Publication Number:
  US 20180195137A1
• Time in Patent Office: Days
• CPC Class Codes: A01N 25/006 insecticidal
• CPC Class Codes: A01N 37/46 N-acyl derivatives
• CPC Class Codes: A01N 63/14 Insects
• CPC Class Codes: A01N 63/60 Isolated nucleic acids
• CPC Class Codes: C07K 16/1036 Retroviridae, e.g. leukemia viruses
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 15/113 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 Class Codes: C12N 15/746 for lactic acid bacteria (Streptococcus; Lactococcus; Lactobacillus; Pediococcus; Enterococcus; Leuconostoc; Propionibacterium; Bifidobacterium; Sporolactobacillus)
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12R 2001/01 Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Abstract

The invention relates to a genetically transformed or transfected bacterial cell of a gut symbiont of an insect belonging to the Order Thysanoptera wherein said cell is transformed to express double-stranded RNA (dsRNA) active against at least one selected insect gene; a vector for transforming or transfecting said bacterial cell; an insect including said transformed bacterial cell and a method of pest control employing the use of said bacterial cell and/or said insect.

Claims

1. A genetically transformed or transfected bacterial cell wherein said bacteria is a gut symbiont of an insect belonging to the Order Thysanoptera characterised in that said bacterial cell is transformed or transfected with nucleic acid to express dsRNA against at least a part of tubulin gene or at least a part of elongation factor gene of the insect.

2. The genetically transformed or transfected bacterial cell according to claim 1 wherein said dsRNA is against at least a part of tubulin alpha-1 chain gene or at least a part of elongation factor 1-alpha gene of the insect.

3. The genetically transformed or transfected bacterial cell according to claim 1, wherein said dsRNA comprises a strand of RNA that shares 50% complementarity to at least one of said genes.

4. The genetically transformed or transfected bacterial cell according to claim 3, wherein said dsRNA comprises a strand of RNA that shares at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% complementarity to at least one of said genes.

5. The genetically transformed or transfected bacterial cell according to claim 3, wherein said dsRNA active against said tubulin gene or said elongation factor gene of the insect is complementary to at least a part of the sequence of SEQ ID NO:

17 or 18.

6. The genetically transformed or transfected bacterial cell according to claim 3, wherein dsRNA comprises a strand of RNA that shares at least 75% 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% complementarity to ata part of the sequence of SEQ ID NO:

17 or 18.

7. The genetically transformed or transfected bacterial cell of claim 1, wherein said insect belongs to the Thripidae family.

8. The genetically transformed or transfected bacterial cell according to claim 7 wherein said insect belongs to the genus Frankliniella.

9. The genetically transformed or transfected bacterial cell according to claim 8 wherein said insect belongs to the species Frankliniella occidentalis.

10. The genetically transformed or transfected bacterial cell of claim 1, wherein said bacteria is a gut symbiont related to bacteria of the genus Pantoea;

or an Erwinia species gut symbiont.

11. The genetically transformed or transfected bacterial cell according to claim 10 wherein said bacteria is BFo2 or BFo1.

23. A method for modulating the expression of a target gene of an insect belonging to the Order Thysanoptera comprising:

contaminating a composition to be ingested by the insect with a bacterial cell according to claim 1;

whereupon ingestion of said bacterial cell by said insect results in said bacterial cell colonising the gut of said insect wherein it synthesises dsRNA against at least a part of tubulin gene or at least a part of elongation factor gene to modulate said insect gene expression.

24. The method according to claim 23 wherein said dsRNA is against at least a part of tubulin alpha-1 chain gene or at least a part of elongation factor 1-alpha of the insect.

25. The method according to claim 23, wherein said bacterial cell is transformed or genetically modified such that recombinant DNA is stably integrated into the host cell genome.

26. The method of claim 25 wherein said nucleic acid or recombinant DNA is stably integrated in the RNaseIII gene.

27. The method according to claim 23, wherein modulating the expression of a target gene causes insect death, prevents transmission of a pathogenic organism, or both.

28. The method according to claim 27, wherein said insect death occurs at the larval stage.

30. The method according to claim 27, wherein said pathogenic organism is a tospovirus.

31. The method according to claim 23, wherein said composition comprises a food source for the insect, a plant, faeces or frass.

12. An expression vector for transforming or transfecting a bacterial cell that is a gut symbiont of an insect belonging to the Order Thysanoptera wherein said vector comprises a nucleic acid sequence that expresses dsRNA against at least a part of tubulin gene or at least a part of elongation factor gene of the insect.

13. The expression vector according to claim 12 wherein said dsRNA is against at least a part of tubulin alpha-1 chain gene or at least a part of elongation factor 1-alpha gene of the insect.

14. The expression vector according to claim 12, wherein said vector comprises at least one constitutive promoter.

15. The expression vector according to claim 14 wherein said constitutive promoter is Ptac.

16. The expression vector according to claim 14, wherein said vector comprises a pair of said promoters.

17. The expression vector according to claim 16 wherein said pair of promoters are configured to drive transcription in a convergent manner.

18. The expression vector according to claim 12, wherein said vector is selected from the group comprising Pex A, pTub3, pElong1, pRN1, and pRN2.

19. An insect belonging to the Order Thysanoptera characterised in that said insect comprises a genetically transformed or transfected bacterial cell wherein said bacteria is a gut symbiont of said insect and is transformed to express dsRNA against at least a part of tubulin gene or at least a part of elongation factor gene of the insect.

20. The insect according to claim 19 wherein said dsRNA is against at least a part of tubulin alpha-1 chain gene or at least a part of elongation factor 1-alpha of the insect.

21. (canceled)

22. (canceled)

29. (canceled)

32. (canceled)