RNA interference (RNAi) is a highly specific gene-silencing mechanism that can cause rapid insect mortality when essential genes are targeted. RNAi is being developed as a tool for integrated pest management of some crop pests. Here we focus on an aggressive forest pest that kills extensive tracts of pine forests, the southern pine beetle (SPB), Dendroctonus frontalis. We sought to identify reference genes for quantitative real-time PCR (qPCR) and validate RNAi responses in SPB by mortality and gene silencing analysis. Using an adult beetle feeding bioassay for oral ingestion of dsRNA, we measured the expression and demonstrated knockdown of target genes as well as insect mortality after ingestion of target genes. Our study validates reference genes for expression analyses and demonstrates highly effective RNAi responses in SPB, with RNAi response to some target dsRNAs causing 100% beetle mortality after ingestion.
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This work was supported by the University of Kentucky, the USDA Forest Service Forest Health Research and Education Center, and the Kentucky Agricultural Experiment Station under McIntire-Stennis 2351197000.
This is publication number 18-08-070 from the Kentucky Agricultural Experimental Station and is published with the approval of the director.
Supplementary information accompanies this paper at https://doi.org/10.1038/s41598-019-42072-6.
Kyre, Bethany R.; Rodrigues, Thais B.; and Rieske, Lynne K., "RNA Interference and Validation of Reference Genes for Gene Expression Analyses Using qPCR in Southern Pine Beetle, Dendroctonus frontalis" (2019). Entomology Faculty Publications. 186.