Polynucleotides encoding Δ-endotoxins toxic to lepidoptera and coleoptera, and method of use
AssigneeMonsanto Technology LLC
InventorsAnne-Marie Light Mettus, James A. Baum
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Abstract
Disclosed are novel polynucleotides encoding Lepidopteran- and Coleopteran-active δ-endotoxin polypeptides and compositions comprising the polypeptides. Also disclosed are vectors, transformed host cells, and transgenic plants that comprise the novel nucleic acid segments encoding the δ-endotoxin polypeptides. Also disclosed are methods of identifying related polynucleotides encoding δ-endotoxin polypeptides, methods of making and using transgenic cells comprising the novel sequences of the invention, as well as methods for controlling an insect population, such as the Western Corn Rootworm and Colorado potato beetle, and for conferring to a plant population resistance to the target insect species.
Claims55
A method of preparing a δ-endotoxin polypeptide having insecticidal activity against a coleopteran or lepidopteran insect, comprising:
- (a) culturing Bacillus thuringiensis, NRRL B-21885 or NRRL B-21886 cells under conditions effective to produce the δ-endotoxin polypeptide of SEQ ID NO:2;
- and
- (b) obtaining from said cells the polypeptide so produced.
A Bacillus thuringiensis cell having the NRRL accession number NRRL B-21886.
A Bacillus thuringiensis cell having the NRRL accession number NRRL B-21885.
An isolated polynucleotide encoding a Bacillus thuringiensis δ-endotoxin polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle.
A pluripotent plant cell population comprising a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2, and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle.
A plant callus or embryo comprising a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2, and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle.
A transgenic plant having incorporated into its genome a selected polynucleotide, said polynucleotide encoding a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle.
A method of preparing an insect resistant plant, wherein the method comprises:
- (a) contacting a recipient plant cell with a transgene comprising a promoter active in said plant cell operably linked to a nucleic acid that encodes a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle, wherein said contacting occurs under conditions permitting the uptake of said transgene by said cell;
- (b) selecting a recipient cell in which the transgene is chromosomally incorporated to produce a selected cell;
- (c) regenerating a transgenic plant from said selected cell;
- and
- (d) identifying a fertile transgenic plant;
- wherein said plant has enhanced insect resistance relative to the corresponding non-transformed plant.
An insect resistant, crossed fertile transgenic plant prepared by a method comprising:
- (a) obtaining a fertile transgenic plant comprising a chromosomally incorporated transgene comprising a promoter active in said plant operably linked to a nucleic acid that encodes a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle;
- (b) crossing said fertile transgenic plant with a second plant lacking said transgene to obtain a third plant comprising said transgene;
- and
- (c) backcrossing said third plant to obtain a backcrossed fertile plant comprising the transgene.
An insect resistant hybrid, transgenic plant prepared by a method comprising:
- crossing a first and a second inbred plant, wherein one or both of said first and said second inbred plants comprises a chromosomally incorporated transgene comprising a promoter active in said plant operably linked to a nucleic acid that encodes a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle, to produce an insect resistant hybrid comprising the transgene.
A method of producing transgenic seed, wherein the method comprises the steps of:
- (a) obtaining a fertile transgenic plant comprising a chromosomally integrated transgene comprising a promoter active in said plant operably linked to a nucleic acid that encodes a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle;
- and
- (b) growing said plant under appropriate conditions to produce said transgenic seed.
A method of producing progeny of any generation of an insect resistance-enhanced fertile transgenic plant, wherein the method comprises:
- (a) collecting transgenic seed from a transgenic plant, said transgenic seed comprising a chromosomally integrated transgene comprising a promoter active in said plant operably linked to a nucleic acid that encodes a polypeptide comprising an amino acid sequence at least 99% identical to SEQ ID NO:2 and exhibiting insecticidal activity against the Western corn root worm or the Colorado potato beetle;
- (b) planting the collected transgenic seed;
- and
- (c) growing progeny transgenic plants from said seed.
Dependencies are inferred from the claim text.