Silencing gut genes associated with the peritrophic matrix of Reticulitermes flavipes (Blattodea: Rhinotermitidae)
A F Sandoval-Mojica1, M E Scharf1
1Department of Entomology, Purdue University, West Lafayette, IN, USA.
Abstract:
The peritrophic matrix (PM) is a noncellular structure that lines the gut of most insects. Because of its close involvement in digestive processes and its role as a barrier against pathogens and toxins, the PM is an attractive target for pest management strategies. The objectives of this study were to (1) reduce the expression of a chitin synthase gene (Reticulitermes flavipes chitin synthase B, RfCHSB), a putative peritrophin [R. flavipes Protein with Peritrophin-A domain 1, (RfPPAD1)] and a confirmed peritrophin [R. flavipes Protein with Peritrophin-A domain 2 (RfPPAD2)] in R. flavipes by means of RNA interference, and (2) to evaluate the susceptibility of R. flavipes to termiticides and a bacterial pathogen, after silencing the target genes. Force feeding termites with 55 and 100 ng of long double-stranded RNAs (dsRNAs), targeting RfCHSB and RfPPAD2, respectively, resulted in the highest levels of transcript suppression. RfCHSB expression was reduced by 70%, whereas the transcript level of RfPPAD2 was decreased by 90%. Force feeding 100 ng/termite of a long RfPPAD1 dsRNA reduced the expression of the transcript by 30%. Challenging termites with imidacloprid, chlorantraniliprole and noviflumuron, after silencing RfCHSB, significantly increased termite mortality. Force feeding termites a dsRNA cocktail, targeting RfCHSB, RfPPAD1 and RfPPAD2, caused the highest significant increase in termite mortality after challenging the insects with imidacloprid. These results demonstrate the viability of the R. flavipes PM as a target in termite pest management.
Insights
Targeting the insect peritrophic matrix (PM) with RNA interference can enhance termiticide efficacy. Silencing key genes in the PM increased termite mortality when exposed to common termiticides, demonstrating PM as a viable pest management target.
Area of Science:
- Entomology
- Molecular Biology
- Pest Management
Background:
- The insect peritrophic matrix (PM) is crucial for digestion and acts as a barrier against toxins and pathogens.
- Its role in insect physiology makes the PM a promising target for developing novel pest control strategies.
Purpose of the Study:
- To investigate the feasibility of using RNA interference (RNAi) to reduce the expression of specific genes within the Reticulitermes flavipes peritrophic matrix.
- To assess the impact of silencing these genes on the susceptibility of R. flavipes to termiticides and a bacterial pathogen.
Main Methods:
- RNA interference (RNAi) was employed using double-stranded RNAs (dsRNAs) to silence chitin synthase (RfCHSB), RfPPAD1, and RfPPAD2 genes in R. flavipes.
- Gene expression levels were quantified post-dsRNA feeding.
- Termites with silenced genes were subsequently challenged with imidacloprid, chlorantraniliprole, and noviflumuron to evaluate mortality rates.
Main Results:
- Significant transcript suppression was achieved: 70% for RfCHSB, 90% for RfPPAD2, and 30% for RfPPAD1.
- Silencing RfCHSB notably increased termite mortality when exposed to imidacloprid, chlorantraniliprole, and noviflumuron.
- A dsRNA cocktail targeting all three genes resulted in the highest increase in mortality upon imidacloprid challenge.
Conclusions:
- The peritrophic matrix of R. flavipes is a viable and effective target for termite pest management strategies.
- RNAi-mediated silencing of PM genes can enhance the efficacy of existing termiticides, offering a new approach to termite control.
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