Abstract
The brown marmorated stink bug (BMSB) is native to Asia and recently invaded the USA. RNA interference (RNAi) is a gene silencing mechanism in which the introduction of double-stranded RNA (dsRNA) inhibits gene function by degrading target mRNA. In dsRNA stability assays, the dsRNases present in the hemolymph and salivary gland secretions of BMSB showed lower activity than those in the hemolymph of Heliothis virescens. We evaluated six housekeeping genes (18S rRNA, EF1-α, Actin, Ubiquitin, 60S RP and β-Tubulin) across dsRNA treatments (injection and feeding) in nymphs and adults of BMSB and identified 18S rRNA and 60S RP as the best genes to use as a reference in reverse-transcriptase quantitative PCR (RT-qPCR). Homologs of 13 genes that were shown to function as effective RNAi targets in other insects were identified and evaluated by injecting dsRNA targeting these homologs into BMSB adults. Five out of 13 dsRNAs tested caused more than 70% mortality by seven days after injection of dsRNA. Feeding dsRNA targeting five of these genes (IAP, ATPase, SNF7, GPCR, and PPI) to nymphs caused more than 70% mortality by three of the five dsRNAs tested. These data suggest that feeding dsRNA causes target gene knockdown and mortality in BMSB.
Highlights
The brown marmorated stink bug (BMSB) is native to Asia and recently invaded the USA
To identify target genes that could be used in the development of RNA interference (RNAi)-based control methods for BMSB, we identified homologs of 13 genes that worked well as RNAi targets in other insects and screened them by injecting double-stranded RNA (dsRNA) targeting these genes in adult BMSB
To determine if the dsRNase activity in the hemolymph of BMSB could be detrimental to RNAi in this insect, we compared dsRNase activity in the hemolymph of BMSB and tobacco budworm (TBW) (Lepidoptera)
Summary
Marmorated Stink Bug: Identification of target genes and Received: 7 November 2017 Accepted: 15 February 2018 Published: xx xx xxxx reference genes for RT-qPCR. Feeding dsRNA targeting five of these genes (IAP, ATPase, SNF7, GPCR, and PPI) to nymphs caused more than 70% mortality by three of the five dsRNAs tested. Some published reports showed successful silencing of target genes by delivering dsRNA to homopteran insects by injection, feeding or through transgenic plants[18,19,20]. Knockdown of genes coding for Juvenile hormone acid O-methyltransferase (JHAMT) and vitellogenin (Vg) in BMSB nymphs after orally delivering dsRNA targeting these genes through green beans was reported recently[22]. In these studies, a significant mortality of BMSB after knockdown of these genes was not observed.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
