Novel bacillus thuringiensis insecticidal proteins

Abstract

The invention pertains to novel insecticidal compounds derived from Bacillus thuringiensis strains. New proteins designated Cry2Ae, Cry2Af, and Cry2Ag, and variants thereof are provided, as well as DNA sequences encoding these proteins or their variants. Further provided are recombinant hosts expressing such proteins, particularly plant cells and plants.

1 - 56 . (canceled) 
     
     
         57 . A method for controlling the insect  Anticarsia gemmatalis  comprising the steps of (1) selecting a habitat of  Anticarsia gemmatalis  to be protected; and (2) applying an insecticidal amount of protein insecticidal to  Anticarsia gemmatalis  to said habitat, wherein said insecticidal protein is:
 i) a protein insecticidal to  Anticarsia gemmatalis  comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO. 2,   ii) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence from the amino acid at position 1 to the amino acid at position 625 in SEQ ID No. 2,   iii) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence from the amino acid at position 50 to the amino acid at position 625 in SEQ ID No. 2,   iv) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence of SEQ ID No. 2,   v) a protein insecticidal to  Anticarsia gemmatalis  having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID NO: 2 without changing the insecticidal activity of the protein,   vi) a protein insecticidal to  Anticarsia gemmatalis  having less than 5 amino acids added, replaced or deleted in the sequence of SEQ ID NO: 2 without changing the insecticidal activity of the protein,   vii) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence of SEQ ID No. 8, or   viii) a protein insecticidal to  Anticarsia gemmatalis  encoded by the DNA of SEQ ID No 7 or 9, and   wherein said insecticidal amount is an amount of protein sufficient to limit damage on a plant by  Anticarsia gemmatalis  insects feeding on such plant, by killing said insects, wherein said insecticidal protein is the active ingredient in an insecticidal composition together with suitable carriers, diluents, emulsifiers and/or dispersants, and wherein said composition is formulated as a wettable powder, pellets, granules or dust or as a liquid formulation.   
     
     
         58 . The method of  claim 57 , wherein said plant is soybean, rice, corn or cotton. 
     
     
         59 . The method of  claim 57 , wherein said plant expresses another insecticidal protein, wherein said another insecticidal protein is a Cry1F protein or a hybrid derived from a Cry1F protein, a Cry1A protein or a toxic fragment thereof, a Cry1Ac protein or a hybrid derived from a Cry1Ac protein, or a VIP3Aa protein or a toxic fragment thereof. 
     
     
         60 . The method of  claim 57 , wherein said insecticidal protein has at least 98% sequence identity to the amino acid sequence of SEQ ID NO. 2. 
     
     
         61 . The method of  claim 57 , wherein said insecticidal protein is an insecticidal protein having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID No. 2 without changing the insecticidal activity of the protein. 
     
     
         62 . A method for controlling the insect  Anticarsia gemmatalis , comprising the steps of (1) selecting an area to be protected which is a habitat for  Anticarsia gemmatalis ; and (2) planting plants expressing an insecticidal amount of a protein insecticidal to  Anticarsia gemmatalis  in a said area, wherein said plants are infested by  Anticarsia gemmatalis , and wherein said insecticidal protein is:
 i) a protein insecticidal to  Anticarsia gemmatalis  comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO. 2,   ii) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence from the amino acid at position 1 to the amino acid at position 625 in SEQ ID No. 2,   iii) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence from the amino acid at position 50 to the amino acid at position 625 in SEQ ID No. 2,   iv) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence of SEQ ID No. 2,   v) a protein insecticidal to  Anticarsia gemmatalis  having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID NO: 2 without changing the insecticidal activity of the protein,   vi) a protein insecticidal to  Anticarsia gemmatalis  having less than 5 amino acids added, replaced or deleted in the sequence of SEQ ID NO: 2 without changing the insecticidal activity of the protein,   vii) a protein insecticidal to  Anticarsia gemmatalis  comprising the amino acid sequence of SEQ ID No. 8, or   viii) a protein insecticidal to  Anticarsia gemmatalis  encoded by the DNA of SEQ ID No 7 or 9; and   wherein said insecticidal amount is an amount of protein sufficient to limit damage on a plant by  Anticarsia gemmatalis  insects feeding on such plant, by killing said insects.   
     
     
         63 . The method of  claim 62 , wherein said plant is soybean, rice, corn or cotton. 
     
     
         64 . The method of  claim 62 , wherein said plant expresses another insecticidal protein, wherein said another insecticidal protein is a Cry1F protein or a hybrid derived from a Cry1F protein, a Cry1A protein or a toxic fragment thereof, a Cry1Ac protein or a hybrid derived from a Cry1Ac protein, or a VIP3Aa protein or a toxic fragment thereof. 
     
     
         65 . The method of  claim 62 , wherein said insecticidal protein has at least 98% sequence identity to the amino acid sequence of SEQ ID NO. 2. 
     
     
         66 . The method of  claim 62 , wherein said insecticidal protein is an insecticidal protein having 5-10 amino acids added, replaced or deleted in the sequence of SEQ ID No. 2 without changing the insecticidal activity of the protein.