AXMI-010, a delta-endotoxin gene and methods for its use

Abstract

Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a delta-endotoxin 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 delta-endotoxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in SEQ ID NO:2, or the nucleotide sequence set forth in SEQ ID NO: 1, as well as variants thereof.

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 complement thereof;    b) a nucleic acid molecule comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:1, or a complement thereof;    c) a nucleic acid molecule comprising a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:1, or a complement thereof;    d) a nucleic acid molecule that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2;    e) a nucleic acid molecule that encodes a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2; and,    f) a nucleic acid molecule that encodes a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2.    
     
     
         2 . A vector comprising the nucleic acid molecule of  claim 1 .  
     
     
         3 . The vector of  claim 2 , further comprising a nucleic acid molecule encoding a heterologous polypeptide.  
     
     
         4 . A host cell that contains the vector of  claim 2 .  
     
     
         5 . The host cell of  claim 4  that is a bacterial host cell.  
     
     
         6 . The host cell of  claim 4  that is a plant cell.  
     
     
         7 . A transgenic plant comprising the host cell of  claim 6 .  
     
     
         8 . The transgenic plant of  claim 7 , wherein said plant is selected from the group consisting of maize, sorghum, wheat, cabbage, sunflower, tomato, crucifers, peppers, potato, cotton, rice, soybean, sugarbeet, sugarcane, tobacco, barley, and oilseed rape.  
     
     
         9 . Transformed seed of the plant of  claim 8 .  
     
     
         10 . 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;    b) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2;    c) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2;    d) a polypeptide that is encoded by the nucleotide sequence of SEQ ID NO:1;    e) a polypeptide that is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:1; and,    f) a polypeptide that is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:1.    
     
     
         11 . The polypeptide of  claim 10  further comprising heterologous amino acid sequences.  
     
     
         12 . A composition comprising the polypeptide of  claim 10 .  
     
     
         13 . The composition of  claim 12 , wherein said composition is selected from the group consisting of a powder, dust, pellet, granule, spray, emulsion, colloid, and solution.  
     
     
         14 . The composition of  claim 12 , wherein said composition is prepared by desiccation, lyophilization, homogenization, extraction, filtration, centrifugation, sedimentation, or concentration of a culture of  Bacillus thuringiensis  cells.  
     
     
         15 . The composition of  claim 12 , comprising from about 1% to about 99% by weight of said polypeptide.  
     
     
         16 . A method for controlling a lepidopteran or coleopteran pest population comprising contacting said population with a pesticidally effective amount of a polypeptide of  claim 10 .  
     
     
         17 . A method for killing a lepidopteran or coleopteran pest, comprising contacting said pest with, or feeding to said pest, a pesticidally effective amount of a polypeptide of  claim 10 .  
     
     
         18 . A method for producing a polypeptide with pesticidal activity, comprising culturing the host cell of  claim 4  under conditions in which a nucleic acid molecule comprising a nucleotide sequence 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;    c) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2;    d) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2;    e) a polypeptide that is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:1; and,    f) a polypeptide that is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:1.    
     
     
         19 . 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) the nucleotide sequence of SEQ ID NO:1;    b) a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:1;    c) a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:1;    d) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2;    e) a nucleotide sequence encoding a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2; and    f) a nucleotide sequence encoding a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2;    wherein said nucleotide sequence is operably linked to a promoter that drives expression of a coding sequence in a plant cell.    
     
     
         20 . A 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 of SEQ ID NO:1;    b) a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:1;    c) a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:1;    d) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2;    e) a nucleotide sequence encoding a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2;    f) a nucleotide sequence encoding a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2; and,    wherein 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, 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 comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:1;    c) a nucleic acid molecule comprising a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:1;    d) a nucleic acid molecule that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2;    e) a nucleic acid molecule that encodes a polypeptide having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:2; and,    f) a nucleic acid molecule that encodes a polypeptide having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:2.    
     
     
         22 . The method of  claim 21 , wherein said plant produces a pesticidal polypeptide having pesticidal activity against a lepidopteran or coleopteran pest.