Lepidopteran-Active Bacillus Thuringiensis Delta-Endotoxin Compositions and Methods of Use

Abstract

Disclosed are Bacillus thuringiensis strains comprising novel crystal proteins which exhibit insecticidal activity against lepidopteran insects. Also disclosed are novel B. thuringiensis genes and their encoded crystal proteins, as well as methods of making and using transgenic cells comprising the novel nucleic acid sequences of the invention.

1 - 52 . (canceled) 
     
     
         53 . An isolated nucleic acid segment having utility as a probe or primer, characterized as:
 (a) a nucleic acid segment comprises at least a 14 nucleotide long contiguous sequence that has the same sequence as, or is complementary to, a 14 nucleotide long contiguous DNA segment of SEQ ID NO: 1, SEQ ID NO:45 or SEQ ID NO:47; or   (b) a nucleic acid segment comprising at least a 14 nucleotide long contiguous sequence that specifically hybridizes to SEQ ID NO: 1, SEQ ID NO:45 or SEQ ID NO:47, or the complement thereof under stringent conditions.   
     
     
         54 . The isolated nucleic acid segment of  claim 53  consisting of the sequence as set forth in SEQ ID NO:60 or SEQ ID NO:61. 
     
     
         55 . A method for detecting a nucleic acid sequence encoding a Lepidopteran toxic crystal protein, comprising the steps of:
 (a) obtaining sample nucleic acids suspected of encoding a Lepidopteran toxic crystal protein;   (b) contacting said sample nucleic acids with the isolated nucleic acid segment of  claim 53  or  54  under conditions effective to allow hybridization of complementary nucleic acids; and   (c) detecting the hybridized complementary nucleic acids thus formed.   
     
     
         56 . The method of  claim 55 , wherein said nucleic acid segment comprises a detectable label and said hybridized complementary nucleic acids are detected by detecting said label. 
     
     
         57 . A nucleic acid detection kit comprising, in suitable container means, the nucleic acid segment of  claim 53  or  54  and a detection reagent. 
     
     
         58 . A purified antibody that specifically binds to the polypeptide of SEQ ID NO:2 or an insecticidal fragment thereof. 
     
     
         59 . The purified antibody of  claim 58  generated by using a polypeptide according to SEQ ID NO:2 as an immunogen. 
     
     
         60 . The purified antibody of  claim 59  produced by a hybridoma, wherein a polypeptide according to SEQ ID NO:2 is used to generate the hybridoma producing the antibody. 
     
     
         61 . A method for detecting a Lepidopteran toxic crystal protein in a biological sample, comprising the steps of:
 (a) obtaining a biological sample suspected of containing the protein;   (b) contacting the sample with an antibody in accordance with any one of  claims 58  to  60  under conditions effective to allow formation of complexes; and   (c) detecting the complexes so formed.   
     
     
         62 . An immunodetection kit comprising, in suitable container means, an antibody in accordance with any one of  claims 58  to  60 , and an immunodetection reagent. 
     
     
         63 . A transgenic plant event expressing the polypeptide of SEQ ID NO:2 or an insecticidal fragment thereof. 
     
     
         64 . The transgenic plant event of  claim 63 , further defined as a monocotyledonous plant, progeny or seed thereof. 
     
     
         65 . The transgenic plant event of  claim 64 , further defined as a corn, wheat, oat, rice, barley, turf grass, or pasture grass plant, progeny or seed thereof. 
     
     
         66 . The transgenic plant event of  claim 63 , further defined as a dicotyledonous plant, progeny or seed thereof. 
     
     
         67 . The transgenic plant event of  claim 66 , further defined as a legume, soybean, tobacco, tomato, potato, cotton, fruit, berry, vegetable or tree plant, progeny or seed thereof. 
     
     
         68 . An artificial diet for Lepidopteran insects, comprising an insecticidal protein as set forth in SEQ ID NO:2 or an insecticidal fragment thereof. 
     
     
         69 . The artificial diet of  claim 68 , wherein said insecticidal protein is prepared by a process comprising the steps of:
 a) culturing a  Bacillus thuringiensis  cell having the accession number NRRL B-21921 under conditions effective to produce an insecticidal protein; and   b) obtaining from said cell the insecticidal protein so produced.   
     
     
         70 . A method for killing Lepidopteran insects, comprising providing the artificial diet of  claim 68  or  69  to Lepidopteran insect larvae.