Compositions and methods for controlling plant pests
View Patent ↗Novel insecticidal proteins isolated from Bacillus thuringiensis that are active against lepidopteran insect pests are disclosed. The DNA encoding the insecticidal proteins can be used to transform various prokaryotic and eukaryotic organisms to express the insecticidal proteins. These recombinant organisms can be used to control lepidopteran insects in various environments.
1. A chimeric gene comprising a heterologous promoter operably linked to a nucleic acid molecule comprising a nucleotide sequence that encodes a protein toxic to black cutworm ( Agrotis ipsilon ), wherein the nucleotide sequence (a) comprises SEQ ID NO:10; or (b) encodes a protein comprising an amino acid sequence that has at least 99% sequence identity with SEQ ID NO:18; or (c) is a synthetic sequence of (a) or (b) that has codons optimized for expression in a transgenic organism.
2. The chimeric gene of claim 1 , wherein the nucleotide sequence comprises SEQ ID NO:10.
3. The chimeric gene of claim 1 , wherein the nucleotide sequence encodes a protein comprising SEQ ID NO:18.
4. A recombinant Cry9C protein that is toxic to black cutworm ( Agrotis ipsilon ), wherein the recombinant protein comprises an amino acid sequence that has at least 99% sequence identity to SEQ ID NO: 18 and wherein the amino acid sequence comprises one or more amino acid substitutions in an antigenic region corresponding to amino acid positions 342-354 of SEQ ID NO: 18 that allows said recombinant protein to be antigenically distinguished from other Cry9C proteins in a protein detection assay.
5. The Cry9C protein of claim 4 , wherein the amino acids within said antigenic region that allow said insecticidal protein to be antigenically distinguished from other Cry9C proteins are at positions that correspond to amino acids 350, 351 and 354 of SEQ ID NO:18.
6. The Cry9C protein of claim 5 , wherein the amino acid at a position corresponding to amino acid position 350 is an isoleucine (I), the amino acid at a position corresponding to amino acid position 351 is glutamine (Q) and the amino acid at a position corresponding to amino acid position 354 is serine (S).
7. The Cry9C protein of claim 6 , wherein the protein comprises SEQ ID NO:18.
8. An insecticidal composition comprising the recombinant protein of claim 4 and an agriculturally acceptable carrier.
9. A recombinant vector comprising the chimeric gene of claim 1 .
10. A transgenic plant comprising the chimeric gene of claim 1 .
11. Transgenic seed of the transgenic plant of claim 10 , wherein said seed comprises the chimeric gene.
12. A harvested product derived from the transgenic plant of claim 10 , wherein the harvested product comprises the chimeric gene or the protein.
13. A processed product derived from the harvested product of claim 12 , wherein the processed product is selected from the group consisting of flour, meal, oil, and starch, wherein the processed product comprises the chimeric gene or protein.
14. An extract from the transgenic plant of claim 10 , wherein the extract comprises the chimeric gene or the protein.
15. A method of producing an insect-resistant transgenic plant, comprising: introducing into a plant the chimeric gene of claim 1 , wherein the insecticidal protein is expressed in the plant, thereby conferring to the plant resistance to black cutworm ( Agrotis ipsilon ), and producing an insect-resistant transgenic plant.
16. A method of controlling black cutworm ( Agrotis ipsilon ) insects, comprising delivering to the insects an effective amount of the protein of claim 4 .
17. A synthetic nucleic acid molecule comprising a nucleotide sequence that encodes a protein that is active against black cutworm ( Agrotis ipsilon ), wherein the nucleotide sequence (a) comprises SEQ ID NO:10; or (b) encodes an amino acid sequence that has at least 99% sequence identity with SEQ ID NO:18.
18. The synthetic nucleic acid molecule of claim 17 , wherein the nucleotide sequence comprises SEQ ID NO:10.
19. The synthetic nucleic acid molecule of claim 17 , wherein the nucleotide sequence encodes SEQ ID NO: 18.