AXMI477 toxin variants and methods for its use
Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a toxin polypeptide are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated toxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed, and antibodies specifically binding to those amino acid sequences. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in any of SEQ ID NO: 22 to 42, or the nucleotide sequence set forth in any of SEQ ID NO: 1 to 21, as well as variants and fragments thereof.
1 . A recombinant nucleic acid molecule comprising a nucleotide sequence encoding an amino acid sequence having pesticidal activity, wherein said nucleotide sequence is selected from the group consisting of:
a) the nucleotide sequence set forth in SEQ ID NO: 14;
b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 35;
c) a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35; and
d) a nucleotide sequence that encodes a polypeptide comprising SEQ ID NO: 35, wherein said polypeptide has modified amino acid at positions 178, 182, 194, 278, 283, 288, 497, and 501.
2 . The recombinant nucleic acid encoding the polypeptide of claim 1 , wherein said polypeptide confers resistance against Helicoverpa zea.
3 . The polypeptide according to claim 1 , wherein the modified amino acid at position 178 is Leucine, 182 is Phenylalanine, 194 is Asparagine, 278 is Methionine, 283 is Methionine, 288 is Leucine, 497 is Arginine, and 501 is Tryptophan.
4 . The recombinant nucleic acid molecule of claim 1 , wherein said nucleotide sequence is a synthetic sequence that has been designed for expression in a plant.
5 . The recombinant nucleic acid molecule of claim 1 , wherein said nucleotide sequence is operably linked to a promoter capable of directing expression of said nucleotide sequence in a plant cell.
6 . A vector comprising the recombinant nucleic acid molecule of claim 1 .
7 . The vector of claim 6 , further comprising a nucleic acid molecule encoding a heterologous polypeptide.
8 . A host cell that contains the recombinant nucleic acid of claim 1 .
9 . The host cell of claim 8 , wherein said host cell is a bacterial host cell.
10 . The host cell of claim 8 , wherein said host cell is a plant cell.
11 . A transgenic plant comprising the host cell of claim 10 .
12 . The transgenic plant of claim 11 , 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.
13 . A transgenic seed comprising the nucleic acid molecule of claim 1 .
14 . A recombinant polypeptide with pesticidal activity, selected from the group consisting of:
a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 35;
b) a polypeptide comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35; and
c) a polypeptide comprising SEQ ID NO: 35, wherein said polypeptide has modified amino acid at positions 178, 182, 194, 278, 283, 288, 497, and 501.
15 . The polypeptide of claim 14 , further comprising heterologous amino acid sequences.
16 . A composition comprising the polypeptide of claim 14 .
17 . The composition of claim 16 , wherein said composition is selected from the group consisting of a powder, dust, pellet, granule, spray, emulsion, colloid, and solution.
18 . The composition of claim 16 , wherein said composition is prepared by desiccation, lyophilization, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of bacterial cells.
19 . The composition of claim 16 , comprising from about 1% to about 99% by weight of said polypeptide.
20 . A method for controlling a Helicoverpa zea , lepidopteran, hemipteran, coleopteran, nematode, or dipteran pest population comprising contacting said population with a pesticidally-effective amount of the polypeptide of claim 14 .
21 . A method for killing a Helicoverpa zea , lepidopteran, hemipteran, coleopteran, nematode, or dipteran pest, comprising contacting said pest with, or feeding to said pest, a pesticidally-effective amount of the polypeptide of claim 14 .
22 . A method for producing a polypeptide with pesticidal activity, comprising culturing the host cell of claim 8 under conditions in which the nucleic acid molecule encoding the polypeptide is expressed.
23 . A plant or plant cell 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) the nucleotide sequence set forth in SEQ ID NO: 14;
b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 35;
c) a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35; and
d) a nucleotide sequence that encodes a polypeptide comprising SEQ ID NO: 35, wherein said polypeptide has modified amino acid at positions 178, 182, 194, 278, 283, 288, 497, and 501.
24 . A method for protecting a plant from a pest, comprising expressing in a plant or cell thereof a nucleotide sequence that encodes a pesticidal polypeptide, wherein said nucleotide sequence is selected from the group consisting of:
a) the nucleotide sequence set forth in SEQ ID NO: 14;
b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 35;
c) a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35; and
d) a nucleotide sequence that encodes a polypeptide comprising SEQ ID NO: 35, wherein said polypeptide has modified amino acid at positions 178, 182, 194, 278, 283, 288, 497, and 501.
25 . The method of claim 24 , wherein said plant produces a pesticidal polypeptide having pesticidal activity against a Helicoverpa zea , lepidopteran, hemipteran, coleopteran, nematode, or dipteran pest.
26 . A method for increasing yield in a plant comprising growing in a field a plant of or a seed thereof 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) the nucleotide sequence set forth in SEQ ID NO: 14;
b) a nucleotide sequence that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 35;
c) a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35; and
d) a nucleotide sequence that encodes a polypeptide comprising SEQ ID NO: 35, wherein said polypeptide has modified amino acid at positions 178, 182, 194, 278, 283, 288, 497, and 501.
27 . A commodity product comprising the nucleic acid molecule of claim 1 , or a protein encoded thereby, wherein said product is selected from the group consisting of whole or processed seeds or grain, animal feed, corn or soy meal, soybean meal, flakes, soy protein concentrate, soy protein isolates, texturized soy protein concentrate, cosmetics, hair care products, soy nut butter, natto, tempeh, hydrolyzed soy protein, whipped topping, shortening, lecithin, edible whole soybeans, soy yogurt, soy cheese, tofu, yuba, and cooked, polished, steamed, baked or parboiled grain.