Patent
The invention provides nucleic acids, and variants and fragments thereof, obtained from strains of Bacillus thuringiensis encoding polypeptides having pesticidal activity against insect pests, including Lepidoptera. Particular embodiments of the invention provide isolated nucleic acids encoding pesticidal proteins, pesticidal compositions, DNA constructs, and transformed microorganisms and plants comprising a nucleic acid of the embodiments. These compositions find use in methods for controlling pests, especially plant pests.
1. An isolated nucleic acid molecule selected from the group consisting of:
(a) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, or a full-length complement thereof;
(b) a nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2; and
(c) a nucleotide sequence encoding a protein comprising an amino acid sequence characterized by at least 95% sequence identity to the amino acid sequence of (b) wherein the amino acid sequence has pesticidal activity against a lepidopteran pest.
2. The isolated nucleic acid molecule of claim 1 , wherein said nucleotide sequence is a synthetic sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein the nucleotide encodes a polypeptide having pesticidal activity against a lepidopteran pest.
3. A DNA construct comprising the nucleic acid molecule of claim 1 .
4. The DNA construct of claim 3 , further comprising a nucleic acid molecule encoding a heterologous polypeptide.
5. A host cell that contains the DNA construct of claim 3 .
6. The host cell of claim 5 that is a bacterial cell.
7. The host cell of claim 5 that is a plant cell.
8. A transgenic plant comprising the host cell of claim 7 .
9. The transgenic plant of claim 8 , wherein said plant is selected from the group consisting of maize, sorghum, wheat, cabbage, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugar beet, sugarcane, tobacco, barley, and oilseed rape.
10. Transformed seed of the plant of claim 9 , wherein the seed comprises the DNA construct.
11. An isolated polypeptide with pesticidal activity, selected from the group consisting of:
(a) a polypeptide comprising the amino acid sequence of SEQ ID NO:2; and
(b) a polypeptide that is encoded by the nucleotide sequence of SEQ ID NO:1.
12. The polypeptide of claim 11 further comprising heterologous amino acid sequences.
13. A composition comprising the polypeptide of claim 11 and at least one formulating agent.
14. The composition of claim 13 , wherein said composition is selected from the group consisting of a powder, dust, pellet, granule, spray, emulsion, colloid, and solution.
15. The composition of claim 13 , wherein said composition is prepared by desiccation, lyophilization, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of Bacillus thuringiensis cells.
16. The composition of claim 13 , comprising from about 1% to about 99% by weight of said polypeptide.
17. A method for controlling a lepidopteran pest population comprising contacting said population with a pesticidally-effective amount of a polypeptide of claim 11 .
18. A method for killing a lepidopteran pest comprising contacting said pest with, or feeding to said pest, a pesticidally-effective amount of a polypeptide of claim 11 .
19. A method for producing a polypeptide with pesticidal activity, comprising culturing the host cell of claim 5 under conditions in which a nucleic acid molecule encoding the polypeptide is expressed, said polypeptide being selected from the group consisting of:
(a) a polypeptide comprising the amino acid sequence of SEQ ID NO:2;
(b) a polypeptide that is encoded by the nucleotide sequence of SEQ ID NO:1; and
(c) a polypeptide sequence having at least 95% sequence identity to a polypeptide sequence of (a) or (b) wherein the amino acid sequence has pesticidal activity against a lepidopteran pest.
20. A plant having stably incorporated into its genome a DNA construct comprising a nucleotide sequence that encodes a protein having pesticidal activity, wherein said nucleotide sequence is selected from the group consisting of:
(a) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1;
(b) a nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2; and
(c) a nucleotide sequence encoding a protein comprising an amino acid sequence characterized by at least 95% sequence identity to an amino acid sequence of (b) wherein and the amino acid sequence has pesticidal activity against a lepidopteran pest and said nucleotide sequence is operably linked to a promoter that drives expression of a coding sequence in a plant cell.
21. A method for protecting a plant from a pest, comprising introducing into said plant or cell thereof at least one expression vector comprising a nucleotide sequence that encodes a pesticidal polypeptide expressed in a plant, wherein said nucleotide sequence is selected from the group consisting of:
(a) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1;
(b) a nucleic acid molecule which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2; and
(c) a nucleotide sequence encoding a protein comprising an amino acid sequence characterized by at least 95% sequence identity to an amino acid sequence of (b) wherein the amino acid sequence has pesticidal activity against a lepidopteran pest.
22. The method of claim 21 , wherein said plant produces a said pesticidal polypeptide having pesticidal activity against a lepidopteran pest.