Pesticides
The present invention relates to the development of fusion proteins based on peptide toxins as pesticides. The invention also relates to transgenic plants encoding said peptides and their utility in pest management.
1. A fusion protein comprising: (i) an omega atracotoxin family member 1a protein from Hadronyche versuta (ω-ACTX-Hv1a) protein toxin comprising the amino acid sequence: SPTCIPSGQPCPYNENCCSQSCTFKENENGNTVKRCD (SEQ ID NO: 1), or a variant of ω-ACTX-Hv1a protein toxin having at least 95% identity to SEQ ID NO: 1, operably linked to (ii) a protein capable of mediating translocation of the fusion protein from the invertebrate gut, wherein the protein capable of mediating translocation is a snowdrop lectin (GNA) comprising the sequence of SEQ ID NO: 3 and wherein the fusion protein comprises the amino acid sequence of any of SEQ ID NOs: 4, 5, 6, 7, 8 or 9.
2. The fusion protein of claim 1 wherein the fusion protein further comprises an affinity tag to aid purification.
3. The fusion protein of claim 1 wherein the protein has pesticide activity against invertebrate pests selected from the group consisting of insects, arthropods, nematodes, and molluscs.
4. A method of preparing a fusion protein comprising culturing a host cell comprising a nucleic acid encoding the fusion protein of claim 1 under conditions suitable for expression of the fusion protein.
5. The method of claim 4 further comprising the step of recovering the fusion protein.
6. A pesticide composition comprising the fusion protein of claim 1 in an effective pesticidal amount, together with one or more suitable carriers, diluents, adjuvants, preservatives, dispersants, solvents, and emulsifying agents.
7. A method for the preparation of a pesticide composition which comprises mixing an effective pesticidal amount of the fusion protein of claim 1 with one or more suitable carriers, diluents, adjuvants, preservatives, dispersants, solvents, or emulsifying agents.
8. A method of preventing or treating a pest infection of a plant comprising applying a quantity of a composition comprising the fusion protein according to claim 1 to the plant or its locus of growth.
9. A method of preventing or treating a mollusc or nematode pest infection of a plant comprising applying a quantity of a composition comprising the fusion protein according to claim 1 to the plant or its locus of growth; or introducing to the plant a nucleic acid sequence encoding said fusion protein.
10. The method of claim 9 wherein the pest is a slug or snail.
11. A molluscicide bait composition comprising the fusion protein of claim 1 .
12. A transgenic plant or progeny thereof comprising the fusion protein of claim 1 .
13. A method of preventing or treating a pest infection of a plant comprising applying a quantity of the pesticide composition according to claim 6 , to the plant or its locus of growth.
14. A method of preventing or treating a mollusc or nematode pest infection of a plant comprising applying a quantity of the pesticide composition according to claim 6 , to the plant or its locus of growth; or introducing to the plant a nucleic acid sequence encoding said fusion protein by transformation with Agrobacterium , particle bombardment, electroporation or viral transformation.
15. A molluscicide bait composition comprising the pesticide composition according to claim 6 .
16. The fusion protein of claim 1 wherein the fusion protein comprises a w-ACTX-Hv1a protein toxin having the sequence of SEQ ID NO: 1.
17. The fusion protein of claim 1 , wherein the fusion protein comprises a variant of ω-ACTX-Hv1 a protein toxin having at least 95% identity to SEQ ID NO: 1, and wherein amino acid K34 of the ω-ACTX-Hv1a protein toxin of SEQ ID NO: 1 is modified.
18. The fusion protein of claim 17 wherein the variant of ω-ACTX-Hv1a protein toxin has a K34Q substitution.