Endotoxins having nematocidal activity and methods of use thereof
AssigneePioneer Hi-Bred International, Inc.
InventorsUte Schellenberger, Jun-Zhi Wei, Genhai Zhu
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Abstract
The invention provides novel δ-endotoxin polypeptides, and variants and fragments thereof, obtained from Bacillus thuringiensis having pesticidal activity against nematodes. Particular embodiments of the invention provide isolated nucleic acids encoding pesticidal proteins, biopesticide compositions, expression cassettes, and transformed microorganisms and plants comprising a nucleic acid of the invention. These compositions find use in methods for controlling pests, especially plant parasitic nematodes.
Claims19
An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:
- (a) an amino acid sequence comprising SEQ ID NO:
- 1;
- (b) an amino acid sequence comprising at least 93% sequence identity to SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (c) an amino acid sequence comprising at least 178 consecutive amino acids of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (d) an amino acid sequence having at least 91% sequence identity to residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (e) an amino acid sequence comprising at least 89 consecutive amino acids from residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- and
- (f) an amino acid sequence encoded by a nucleotide sequence that hybridizes under stringent conditions to the complement of SEQ ID NO:
- 2, wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C., wherein said polypeptide has nematicidal activity.
An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of:
- (a) a nucleotide sequence comprising SEQ ID NO:
- 2;
- (b) a nucleotide sequence encoding an amino acid sequence comprising SEQ ID NO:
- 1;
- (c) a nucleotide sequence comprising at least 90% sequence identity to SEQ ID NO:
- 2, wherein said polynucleotide encodes a polypeptide having nematicidal activity;
- (d) a nucleotide sequence that hybridizes under stringent conditions to the complement of the nucleotide sequence of a) or b), wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C., wherein said polynucleotide encodes a polypeptide having nematicidal activity;
- (e) a nucleotide sequence encoding an amino acid sequence having at least 93% sequence identity to SEQ ID NO:
- 1, wherein said amino acid sequence has nematicidal activity;
- (f) a nucleotide sequence encoding an amino acid sequence comprising at least 178 consecutive amino acids of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (g) a nucleotide sequence encoding an amino acid sequence having at least 91% sequence identity to residues 23-684 of SEQ ID NO:
- 1, wherein said amino acid sequence has nematicidal activity;
- and
- (h) a nucleotide sequence encoding an amino acid sequence comprising at least 89 consecutive amino acids from residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity.
A plant comprising a heterologous polynucleotide operably linked to a promoter that drives expression in the plant, wherein said heterologous polynucleotide comprises a nucleotide sequence selected from the group consisting of:
- (a) a nucleotide sequence comprising SEQ ID NO:
- 2;
- (b) a nucleotide sequence encoding an amino acid sequence comprising SEQ ID NO:
- 1;
- (c) a nucleotide sequence comprising at least 90% sequence identity to SEQ ID NO:
- 2, wherein said heterologous polynucleotide encodes a polypeptide having nematicidal activity;
- (d) a nucleotide sequence that hybridizes under stringent conditions to the complement of the nucleotide sequence of a) or b), wherein said stringent conditions comprise hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1×SSC at 60° C. to 65° C., wherein said heterologous polynucleotide encodes a polypeptide having nematicidal activity;
- (e) a nucleotide sequence encoding an amino acid sequence having at least 93% sequence identity to SEQ ID NO:
- 1, wherein said amino acid sequence has nematicidal activity;
- (f) a nucleotide sequence encoding an amino acid sequence comprising at least 178 consecutive amino acids of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (g) a nucleotide sequence encoding an amino acid sequence having at least 91% sequence identity to residues 23-684 of SEQ ID NO:
- 1, wherein said amino acid sequence has nematicidal activity;
- and
- (h) a nucleotide sequence encoding an amino acid sequence comprising at least 89 consecutive amino acids from residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity.
A method of enhancing nematode resistance in a plant, said method comprising providing to said plant a polypeptide selected from the group consisting of:
- (a) the amino acid sequence set forth in SEQ ID NO:
- 1;
- (b) an amino acid sequence having at least 93% sequence identity to SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (c) an amino acid sequence comprising at least 178 consecutive amino acids of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (d) an amino acid sequence having at least 91% sequence identity to residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- and
- (e) an amino acid sequence comprising at least 89 consecutive amino acids from residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity.
A microorganism comprising at least one heterologous polynucleotide operably linked to a promoter that drives expression in the microorganism, wherein said heterologous polynucleotide comprises a nucleotide sequence selected from the group consisting of:
- (a) the nucleotide sequence set forth in SEQ ID NO:
- 2;
- (b) a nucleotide sequence encoding the amino acid sequence comprising SEQ ID NO:
- 1;
- (c) a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:
- 2, wherein said polynucleotide encodes a polypeptide having nematicidal activity;
- (d) a nucleotide sequence encoding an amino acid sequence having at least 93% sequence identity to SEQ ID NO:
- 1, wherein said polynucleotide encodes a polypeptide having nematicidal activity;
- (e) a nucleotide sequence encoding an amino acid sequence comprising at least 178 consecutive amino acids of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- (f) a nucleotide sequence encoding an amino acid sequence having at least 91% sequence identity to residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity;
- and
- (g) a nucleotide sequence encoding an amino acid sequence comprising at least 89 consecutive amino acids from residues 23-684 of SEQ ID NO:
- 1, wherein said polypeptide has nematicidal activity.
Dependencies are inferred from the claim text.