Compositions and methods for controlling diabrotica

Abstract

The present invention provides methods for controlling invertebrate pest infestations, for instance in plants, and related compositions and polynucleotides useful in such methods. More specifically, the present invention provides polynucleotides and methods of use thereof for modifying the expression of genes in an invertebrate pest, for instance through RNA interference.

1 .- 46 . (canceled) 
     
     
         47 . A method of causing mortality or stunting in  Diabrotica  species larvae comprising providing in the diet of  Diabrotica  species larvae at least one recombinant RNA comprising at least one silencing element essentially identical or essentially complementary to a target gene of said  Diabrotica  species larvae, wherein said target gene sequence is selected from the group consisting of SEQ ID NOs:1-450 or the complement thereof, and wherein ingestion of said recombinant RNA by said  Diabrotica  species larvae results in mortality or stunting in said  Diabrotica  species larvae. 
     
     
         48 . The method of  claim 47 , wherein said  Diabrotica  species is at least one selected from the group consisting of  Diabrotica balteata, Diabrotica barberi, Diabrotica beniensis, Diabrotica cristata, Diabrotica curvipustulata, Diabrotica dissimilis, Diabrotica elegantula, Diabrotica emorsitans, Diabrotica graminea, Diabrotica hispanolae, Diabrotica lemniscata, Diabrotica linsleyi, Diabrotica longicornis, Diabrotica milleri, Diabrotica nummularis, Diabrotica occlusa, Diabrotica porracea, Diabrotica scutellata, Diabrotica speciosa, Diabrotica tibialis, Diabrotica trifasciata, Diabrotica undecimpunctata, Diabrotica virgifera , and  Diabrotica viridula.    
     
     
         49 . The method of  claim 47 , wherein said  Diabrotica  species is at least one selected from the group consisting of  Diabrotica virgifera virgifera  (Western Corn Rootworm, WCR),  Diabrotica undecimpunctata howardii  (Southern Corn Rootworm, SCR),  Diabrotica barberi  (Northern Corn Rootworm, NCR),  Diabrotica virgifera zeae  (Mexican Corn Rootworm, MCR),  Diabrotica balteata  (Brazilian Corn Rootworm, BZR), or Brazilian Corn Rootworm complex (BCR) consisting of  Diabrotica viridula  and  Diabrotica speciosa.    
     
     
         50 . The method of  claim 47 , wherein said silencing element comprises at least 18 contiguous nucleotides with a sequence of about 95% to about 100% identity with a segment of equivalent length of said target gene. 
     
     
         51 .- 97 . (canceled) 
     
     
         98 . A method of providing a plant having improved resistance to a  Diabrotica  species infestation comprising expressing in said plant at least one polynucleotide comprising at least 18 contiguous nucleotides that are essentially identical or complementary to a segment of equivalent length of a DNA having a sequence selected from the group consisting of SEQ ID NOs:1-450. 
     
     
         99 . (canceled) 
     
     
         100 . The method of  claim 98 , wherein said plant is selected from the group consisting of maize, cucumber, squash, soybeans, and dry beans. 
     
     
         101 .- 102 . (canceled) 
     
     
         103 . The method of  claim 98 , wherein said  Diabrotica  species is at least one selected from the group consisting of  Diabrotica balteata, Diabrotica barberi, Diabrotica beniensis, Diabrotica cristata, Diabrotica curvipustulata, Diabrotica dissimilis, Diabrotica elegantula, Diabrotica emorsitans, Diabrotica graminea, Diabrotica hispanolae, Diabrotica lemniscata, Diabrotica linsleyi, Diabrotica longicornis, Diabrotica milleri, Diabrotica nummularis, Diabrotica occlusa, Diabrotica porracea, Diabrotica scutellata, Diabrotica speciosa, Diabrotica tibialis, Diabrotica trifasciata, Diabrotica undecimpunctata, Diabrotica virgifera , and  Diabrotica viridula.    
     
     
         104 . The method of  claim 98 , wherein said  Diabrotica  species is at least one selected from the group consisting of  Diabrotica virgifera virgifera  (Western Corn Rootworm, WCR),  Diabrotica undecimpunctata howardii  (Southern Corn Rootworm, SCR),  Diabrotica barberi  (Northern Corn Rootworm, NCR),  Diabrotica virgifera zeae  (Mexican Corn Rootworm, MCR),  Diabrotica balteata  (Brazilian Corn Rootworm, BZR), or Brazilian Corn Rootworm complex (BCR) consisting of  Diabrotica viridula  and  Diabrotica speciosa.    
     
     
         105 . The method of  claim 98 , wherein said expressing is by means of transgenic expression or transient expression. 
     
     
         106 . The method of  claim 98 , further comprising expression in said plant of at least one pesticidal agent selected from the group consisting of a patatin, a plant lectin, a phytoecdysteroid, a  Bacillus thuringiensis  insecticidal protein, a  Xenorhabdus  insecticidal protein, a  Photorhabdus  insecticidal protein, a  Bacillus laterosporous  insecticidal protein, and a  Bacillus sphearicus  insecticidal protein. 
     
     
         107 . The method of  claim 98 , wherein said identity is 100% identity. 
     
     
         108 . The plant having improved resistance to a  Diabrotica  species infestation, provided by the method of  claim 98 . 
     
     
         109 . Seed of the plant of  claim 98 . 
     
     
         110 . A recombinant DNA construct comprising a heterologous promoter operably linked to DNA comprising at least one segment of 18 or more contiguous nucleotides with a sequence of about 95% to about 100% identity with a segment of equivalent length of a DNA having a sequence selected from the group consisting of SEQ ID NOs:1-450 or the DNA complement thereof. 
     
     
         111 . The recombinant DNA construct of  claim 110 , wherein said heterologous promoter is selected from the group consisting of a promoter functional in a plant, a promoter functional in a prokaryote, a promoter functional in a fungal cell, and a baculovirus promoter. 
     
     
         112 . A recombinant vector comprising the recombinant DNA construct of  claim 110 . 
     
     
         113 . The recombinant vector of  claim 112 , wherein said recombinant vector is a recombinant plant virus vector or a recombinant baculovirus vector. 
     
     
         114 . (canceled) 
     
     
         115 . A plant chromosome or plastid comprising the recombinant DNA construct of  claim 110 . 
     
     
         116 . A transgenic plant cell having in its genome the recombinant DNA construct of  claim 110 . 
     
     
         117 . A transgenic plant comprising the transgenic plant cell of  claim 116 . 
     
     
         118 .- 195 . (canceled)