Control of coleopteran pests using RNA molecules
A molecular and nucleic acid molecule technology, applied in the field of pest control, can solve the problem of inability to provide coleopteran pest mortality and other problems
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[0190] The present invention may be further described by reference to the following specific examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise indicated. Example 1. Identification of RNAi gene targets in western corn root firefly beetle
example 1
[0190] The present invention may be further described by reference to the following specific examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise indicated. Example 1. Identification of RNAi gene targets in western corn root firefly beetle
[0191] This example describes the cloning and sequencing of RNAi target genes and coding sequences from Rhizoctonia insects.
[0192] Preparation and Sequencing of Pyrosequencing Library of Western Maize Rhizome
[0193] Whole body neonatal Western corn rootworm (WCR )) Transcriptome sequencing. The resulting determined reads (i.e., short fragments of nucleic acid sequence) were trimmed and assembled into contigs using the MIRA assembler (see, e.g., Chevreux et al. 2004 Genome Res. 14:1147-1159, incorporated by reference here).
[0194] Identification of the most important target genes from Rhizoctonia genus
[0195] The assembled contigs were compared by BLAST to k...
example 2
[0204] Example 2.dsRNA (4 kinds of concentrations) against the activity of western corn root firefly beetle-DRC
[0205] This example describes the testing of the biological activity of dsRNAs of the invention against western corn root beetle (WCR).
[0206] Starting from 1 μg dsRNA / well, the above 30 dsRNA molecules were tested for toxicity against WCR in a 10-fold dilution series in laboratory bioassays. Bioassays were performed using the RNA-treated artificial diet method. Briefly, molten artificial feed modified from the feed of Marrone et al. 1985 (J. Econ. Entomol. [Journal of Economic Entomol] 78:290-293) was poured into each well of a 48-well plate and allowed to solidify . Synthetic dsRNA molecules were diluted to appropriate concentrations such that 20 μl of the solution was added to the surface of the feed in half of the wells of the 48-well plate, with final coverage concentrations of 1 μg, 0.1 μg, 0.01 μg and 0.001 μg per well. Add one or two WCR larvae to each...
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