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RNA interference (RNAi) based approaches can potentially be used to control insect pests. These approaches may depend on the usage of microRNA (miRNA) or double stranded RNA (dsRNA) mediated gene knockdown, which likely involves proteins that regulate these pathways, such as Argonaute 1 (Ago1), Argonaute 2 (Ago2), Dicer 1 (Dcr1), Dicer 2 (Dcr2), and Drosha in insects. We previously performed functional characterization of  Ago2  and  Dcr2  of western corn rootworm (WCR),  Diabrotica virgifera virgifera  (Coleoptera: Chrysomelidae) and observed that knockdown of  Ago2  and  Dcr2  ameliorated the lethal effect induced by the dsRNA-mediated knockdown of an essential gene in WCR, thereby confirming the involvement of Ago2 and Dcr2 in the dsRNA pathway. In the current study, we identified and characterized additional members of the  Argonaute  and  Dicer  gene families, namely  Ago1 ,  Ago3 ,  Aubergine , and  Dcr1 , in a previously developed WCR transcriptome. We also identified a  Drosha  homolog in the same transcriptome. We evaluated the impacts on WCR adult fitness associated with the dsRNA-mediated knockdown of  Ago1 ,  Ago2 ,  Dcr1 ,  Dcr2 , and  Drosha  genes. Among these putative RNAi pathway genes, only the knockdown of  Ago1  incurred significant fitness costs such as reduced survival and oviposition rate, as well as decreased egg viability. The present study, to our knowledge, represents the first report showing that  Ago1  is critical to the survival of insect adults. Our findings suggest that  Ago1  plays an essential role in broader life stages of an insect than previously thought. Importantly, since fitness costs were not observed, downregulation or loss of function of RNAi pathway genes such as  Ago2  or  Dcr2  may confer resistance to pest control measures that rely on the normal functions of these genes. However, the precise roles of these genes under field conditions (i.e., in the presence of possible viral pathogens) requires further investigation.

Distinct fitness costs associated with the knockdown of RNAi pathway genes in western corn rootworm adults