Methods for genetic control of insect infestations in plants and compositions thereof

Abstract

The present invention relates to control of pest infestation by inhibiting one or more biological functions. The invention provides methods and compositions for such control, By feeding one or more recombinant double stranded RNA molecules provided by the invention to the pest, a reduction in pest infestation is obtained through suppression of gene expression. The invention is also directed to methods for making transgenic plants that express the double stranded RNA molecules, and to particular combinations of transgenic pesticidal agents for use in protecting plants from pest infestation.

1 . A recombinant polynucleotide comprising a nucleic acid sequence selected from the group consisting of:
 (a) a polynucleotide comprising a nucleic acid sequence of SEQ ID NO: 819;   (b) a polynucleotide that hybridizes to a nucleic acid sequence of SEQ ID NO: 819 under wash conditions of 5×SSC, 50% formamide and 42° C. for 10 minutes;   (c) a polynucleotide comprising at least 70% sequence identity to a nucleic acid sequence of SEQ ID NO: 819;   (d) a fragment of at least 21 contiguous nucleotides of a nucleic acid sequence of SEQ ID NO: 819, wherein ingestion by a coleopteran plant pest of a double stranded ribonucleotide sequence comprising at least one strand that is complementary to said fragment inhibits the growth of said pest; and   (e) a complement of the sequence of (a), (b), (c) or (d);   
       wherein the nucleic acid sequence is operably linked to a heterologous promoter. 
     
     
         2 - 3 . (canceled) 
     
     
         4 . The recombinant polynucleotide of  claim 1 , defined as comprised on a plant transformation vector. 
     
     
         5 . A double stranded ribonucleotide sequence produced from the expression of a polynucleotide according to  claim 1 , wherein ingestion of said ribonucleotide sequence by a coleopteran plant pest inhibits the growth of said pest. 
     
     
         6 . The double stranded ribonucleotide sequence of  claim 5 , defined as produced by preparing a recombinant polynucleotide sequence comprising a first, a second and a third polynucleotide sequence, wherein the first polynucleotide sequence comprises the recombinant polynucleotide of  claim 1 , wherein the third polynucleotide sequence is linked to the first polynucleotide sequence by the second polynucleotide sequence, and wherein the third polynucleotide sequence is substantially the reverse complement of the first polynucleotide sequence such that the first and the third polynucleotide sequences hybridize when transcribed into a ribonucleic acid to form the double stranded ribonucleotide molecule stabilized by the linked second ribonucleotide sequence. 
     
     
         7 . The double stranded ribonucleotide sequence of  claim 5 , wherein ingestion of the polynucleotide sequence by the pest inhibits the expression of a nucleotide sequence substantially complementary to said polynucleotide sequence. 
     
     
         8 . A cell transformed with the polynucleotide of  claim 1 . 
     
     
         9 . The cell of  claim 8 , defined as a prokaryotic cell. 
     
     
         10 . The cell of  claim 8 , defined as a eukaryotic cell. 
     
     
         11 . The cell of  claim 8 , defined as a plant or bacterial cell. 
     
     
         12 . A plant transformed with the polynucleotide of  claim 1 . 
     
     
         13 . A seed of the plant of  claim 12 , wherein the seed comprises the polynucleotide. 
     
     
         14 . (canceled) 
     
     
         15 . The plant of  claim 12 , wherein said polynucleotide is expressed in a cell of the plant as a double stranded ribonucleotide sequence and ingestion of an insect pest inhibitory amount of said double stranded ribonucleotide sequence in a diet inhibits the pest from further feeding on said diet. 
     
     
         16 . The plant of  claim 15 , wherein the insect pest is selected from the group consisting of  Diabrotica virgifera, Diabrotica virgifera virgifera, Diabrotica virgifera zea, Diabrotica balteata, Diabrotica barberi, Diabrotica viridula, Diabrotica speciosa , and  Diabrotica undecimpunctata.    
     
     
         17 . The plant of  claim 15 , wherein ingestion of the insect pest inhibitory amount of the double stranded ribonucleotide sequence stunts the growth of the pest. 
     
     
         18 . A commodity product produced from a plant according to  claim 12 , wherein said commodity product comprises a detectable amount of the polynucleotide of  claim 1  or a ribonucleotide expressed therefrom. 
     
     
         19 - 34 . (canceled) 
     
     
         35 . The plant of  claim 12 , wherein said plant is a corn plant. 
     
     
         36 . The seed of  claim 13 , wherein said seed is corn seed. 
     
     
         37 . The cell according to  claim 11 , wherein said plant cell is a corn plant cell. 
     
     
         38 . A cell of the plant of  claim 35 , wherein said cell comprises the polynucleotide.