Methods for genetic control of plant pest infestation and compositions thereof

Abstract

The present invention is directed to controlling plant pest infestation, and particularly plant nematode infestation, by inhibiting one or more biological functions in the plant pest. The invention discloses methods and compositions for use in controlling plant pest infestation by providing one or more different recombinant double stranded RNA molecules in the diet of the pest in order to achieve a reduction in pest infestation through suppression of pest gene expression. The invention is also directed to methods for making transgenic plants that express the double stranded RNA molecules, to methods for detecting cells comprising the disclosed sequences, and to methods for detecting the disclosed sequences in biological samples.

What is claimed is: 
     
         1 . An isolated  Heterodera glycines  nucleotide sequence selected from the group consisting of SEQ ID NO:22219, SEQ ID NO:45701, and SEQ ID NO:52890. 
     
     
         2 . An isolated nucleotide sequence selected from the group consisting of SEQ ID NO:45701 and SEQ ID NO:52890 encoding a polypeptide, wherein said polypeptide is isolated from  Heterodera  species. 
     
     
         3 . A method for suppressing a peptide coding sequence in a  Heterodera glycines  pest, comprising selecting a target polynucleotide sequence of a  Heterodera glycines  pest genome comprising at least from about 24 contiguous nucleotides, expressing the polynucleotide sequence as an RNA sequence that forms a double stranded RNA structure in a plant cell, and providing said plant cell in the diet of said  Heterodera glycines  pest, wherein uptake of the contents of said cell by said pest results in the morbidity and/or mortality of said pest, and wherein said target polynucleotide sequence is selected from the group consisting of SEQ ID NO:22219, SEQ ID NO:45701, and SEQ ID NO:52890. 
     
     
         4 . The method of  claim 3 , wherein said RNA sequence comprises contiguous nucleotides from at least two or more target polynucleotide sequences from two or more different coding sequences of a  Heterodera glycines  pest. 
     
     
         5 . The method of  claim 4 , wherein each of said two or more different coding sequences are isolated from different  Heterodera glycines  cDNA sequences. 
     
     
         6 . A method for controlling  Heterodera glycines  infection in a soybean plant comprising
 a) transforming a soybean plant cell with a DNA construct that expresses a dsRNA molecule in said soybean cell; and   b) regenerating said transformed soybean plant cell into a fertile transgenic soybean plant that exhibits resistance to infection by  Heterodera glycines;      wherein said DNA construct comprises a polynucleotide sequence comprising at least about 24 contiguous nucleotides selected from the group consisting of SEQ ID NO:22219, SEQ ID NO:45701, and SEQ ID NO:52890.   
     
     
         7 . The method of  claim 6 , wherein said DNA construct comprises at least one selectable marker providing herbicide tolerance to the transgenic soybean plant. 
     
     
         8 . The method of  claim 7 , wherein said transgenic soybean plant exhibits tolerance to the herbicide glyphosate. 
     
     
         9 . A plasmid library comprising SCN genome sequences wherein said library is ATCC Patent Deposit No. PTA-6583 deposited on Feb. 15, 2005. 
     
     
         10 . An isolated and purified plasmid selected from the library of  claim 9 . 
     
     
         11 . A nucleotide sequence selected from the group consisting of SEQ ID NO:1-SEQ ID NO:45568, and the complement thereof, isolated from a plasmid in the library of  claim 9 . 
     
     
         12 . A vcDNA sequence selected from the group consisting of SEQ ID NO:45569-SEQ ID NO:97729, wherein said vcDNA sequence is isolated from a plasmid in the library of  claim 9 . 
     
     
         13 . An amino acid sequence selected from the group consisting of SEQ ID NO:119146-SEQ ID NO:121220 encoded by the vcDNA of  claim 12 . 
     
     
         14 . A transgenic soybean plant comprising the double stranded structure of  claim 3 . 
     
     
         15 . A seed produced from the plant of  claim 14 , wherein said seed comprises said structure. 
     
     
         16 . A plant grown from the seed of  claim 15 .