Plant derived insecticidal proteins and methods for their use

Abstract

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions are insecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, Hemipteran, fungi and nematode pest populations and for producing compositions with insecticidal activity.

1 . A recombinant insecticidal polypeptide comprising an amino acid sequence selected from:
 a) an amino acid sequence having at least 70% identity to the amino acid sequence of SEQ ID NO: 2;   b) an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 56; and   c) an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 144.   
     
     
         2 . The recombinant insecticidal polypeptide of  claim 1 , wherein the polypeptide comprises a MAC/Perforin (MACPF) Pfam domain (PF01823). 
     
     
         3 . The recombinant insecticidal polypeptide of  claim 1 , wherein the insecticidal polypeptide is joined to a heterologous signal sequence or a transit sequence. 
     
     
         4 . The recombinant insecticidal polypeptide of  claim 1 , wherein the insecticidal activity is against a Coleopteran pest. 
     
     
         5 . The recombinant insecticidal polypeptide of  claim 4 , wherein the insecticidal activity is against a  Diabrotica  species. 
     
     
         6 . The recombinant insecticidal polypeptide of  claim 5 , wherein the insecticidal activity is against at least  Diabrotica virgifera, Diabrotica undecimpunctata howardi  or  Diabrotica barberi.    
     
     
         7 . A recombinant polynucleotide encoding the insecticidal polypeptide of  claim 1 . 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . The recombinant polynucleotide of  claim 7 , wherein the polynucleotide is a non-genomic polynucleotide. 
     
     
         11 . The recombinant polynucleotide of  claim 10 , wherein the polynucleotide is a cDNA. 
     
     
         12 . The recombinant polynucleotide of  claim 11 , wherein the polynucleotide is a synthetic polynucleotide. 
     
     
         13 . The recombinant polynucleotide of  claim 11 , wherein the polynucleotide has codons optimized for expression in an agriculturally important crop. 
     
     
         14 . A DNA construct comprising the polynucleotide of  claim 7  operably linked to a heterologous regulatory element. 
     
     
         15 . (canceled) 
     
     
         16 . (canceled) 
     
     
         17 . A DNA construct comprising a polynucleotide encoding a plant derived perform operably linked to a heterologous regulatory element. 
     
     
         18 . The DNA construct of  claim 17 , wherein the plant derived perform is from a fern or a primitive plant species. 
     
     
         19 . The DNA construct of  claim 18 , wherein the plant derived perform comprises a MAC/Perforin (MACPF) Pfam domain (PF01823). 
     
     
         20 . A transgenic plant or plant cell comprising the DNA construct of  claim 14 . 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . A transgenic plant or plant cell comprising the DNA construct of  claim 17 . 
     
     
         24 . A composition comprising the recombinant insecticidal polypeptide of  claim 1 . 
     
     
         25 . (canceled) 
     
     
         26 . A fusion protein comprising the recombinant insecticidal polypeptide of  claim 1 . 
     
     
         27 . A method for controlling a Coleopteran pest, comprising contacting the insect pest population with the insecticidal polypeptide of  claim 1 . 
     
     
         28 . A method of inhibiting growth or killing a Coleopteran pest or pest population, comprising contacting the insect pest with a composition comprising the insecticidal polypeptide of  claim 1 . 
     
     
         29 . A method of inhibiting growth or killing an insect pest or pest population comprising expressing in a transgenic plant a polynucleotide encoding a heterologous plant derived perforin polypeptide. 
     
     
         30 . The method of  claim 29 , wherein the plant derived perform polypeptide is the recombinant polypeptide of  claim 1 . 
     
     
         31 . The method of  claim 29 , wherein the plant derived perforin polypeptide comprises a MAC/Perforin (MACPF) Pfam domain (PF01823). 
     
     
         32 . The method of  claim 30 , wherein the insect pest is a Coleopteran. 
     
     
         33 . The method of  claim 32 , wherein the Coleopteran is a  Diabrotica  species. 
     
     
         34 . The method of  claim 33 , wherein the  Diabrotica  species is  Diabrotica virgifera, Diabrotica undecimpunctata howardi  or  Diabrotica barberi.    
     
     
         35 . The method of  claim 27 , wherein the pest species or population is resistant to at least one Cry insecticidal polypeptide. 
     
     
         36 . (canceled)