Cloning and RNAi-mediated three lethal genes that can be potentially used for Chilo suppressalis (Lepidoptera: Crambidae) management.

RNA interference (RNAi) has gained attention in recent years as a viable pest control strategy. Here, RNAi assays were performed to screen the potential functionality of genes in Chilo suppressalis, a serious pest of rice, and to determine their potential for developing a highly targeted molecular control approach. Potential homologs of NADH dehydrogenase (ND), glycerol 3-phosphate dehydrogenase (GPDH) and male specific lethal 3 (MSL3) were cloned from C. suppressalis, and their spatiotemporal gene expression evaluated. The expression of all three genes was higher in the pupal and adult stages than the larval stages and largely higher in the larval head compared to other tissues. Newly hatched larvae exhibited high mortalities and suppressed growth when fed bacteria producing double-stranded RNAs (dsRNAs) corresponding to the three target genes. This study provides insights into the function of ND, GPDH and MSL3 during C. suppressalis larval development and suggests that all may be candidate gene targets for C. suppressalis pest management.

Co-Expression Network Analysis: A Future Approach for Pest Control Target Discovery.

The striped stem borer (SSB, Chilo suppressalis Walker) is a major pest of rice worldwide. Double-stranded RNAs (dsRNAs) targeting essential genes can trigger a lethal RNA interference (RNAi) response in insect pests. In this study, we applied a Weighted Gene Co-expression Network Analysis (WGCNA) to diet-based RNA-Seq data as a method to facilitate the discovery of novel target genes for pest control. Nieman-Pick type c 1 homolog b (NPC1b) was identified as the gene with the highest correlation values to hemolymph cholesterol levels and larval size. Functional characterization of the gene supported CsNPC1b expression with dietary cholesterol uptake and insect growth. This study revealed the critical role of NPC1b for intestinal cholesterol absorption in lepidopteran insects and highlights the utility of the WGCNA approach for identifying new pest management targets.

The HSP/co-chaperone network in environmental cold adaptation of Chilo suppressalis.

Winter cold is one of the major environmental stresses for ectotherm species. Chilo suppressalis, a notorious lepidopteran pest of rice, has a wide geographic region that includes temperate zones with severe environmental conditions. Although C. suppressalis exhibits remarkable cold tolerance, its cold-adaptation mechanisms remain unclear. Here, we used bioinformatics approaches to evaluate transcript levels of genes comprising the C. suppressalis heat shock protein (HSP)/co-chaperone network in response to cold-induced stress. Using all such genes identified in the C. suppressalis genome, we experimentally examined the corresponding transcript levels under cold-acclimation or intermittent cold-shock stresses in diapause and non-diapausing larvae. In total, we identified 19 HSPs and 8 HSP co-chaperones in the C. suppressalis genome. Nine (hsp90, hsp75, hsp70, hsp40, small hsp, activator of 90 kDa heat shock protein ATPase-like, heat shock factor, heat shock factor binding protein 1-like and HSPB1-associated protein 1) were highly cold-inducible and likely comprise the principal cold-response HSP/co-chaperone network in C. suppressalis. We also found that transcriptional regulation of the HSP/co-chaperone networks response differs between cold-acclimation and short-term cold-shock. Moreover, activation of the HSP/co-chaperone network depends on the diapause state of overwintering larvae and cold acclimation may further increase larval cold tolerance. These results provide key new insights in the cold-adaptation mechanisms in C. suppressalis.

Nanomaterial-wrapped dsCYP15C1, a potential RNAi-based strategy for pest control against Chilo suppressalis.

BACKGROUND: Although the utility of double-stranded RNA (dsRNA)-mediated knockdown as an environmentally friendly pest management strategy has gained traction in recent years, its overall efficacy has been limited by poor stability and limited cellular uptake. Encapsulation of dsRNAs with various nanomaterials, however, has shown promise in overcoming these limitations. This study sought to investigate the biological efficacy of an oral dsRNA nanomaterial mixture targeting the CYP15C1 gene product in the economically important rice pest, Chilo suppressalis. RESULTS: A putative CYP15C1 ortholog was cloned from C. suppressalis midguts. The transcript is downregulated in fifth-instar larvae and is most highly expressed in heads. RNA interference (RNAi)-mediated knockdown of CsCYP15C1 was associated with significantly increased mortality. More importantly, feeding a dsRNA-nanomaterial mixture significantly increased larval mortality compared with feeding dsRNA alone. CONCLUSION: A critical role for CsCYP15C1 function in molting is supported by sequence similarity with known juvenile hormone epoxidases, its expression profile, and abnormal molting phenotypes associated with RNA-mediated knockdown. CsCYP15C1 is thus a prime target for controlling C. suppressalis. Furthermore, RNAi-mediated characterization of candidate gene function can be enhanced by incorporating an enveloping nanomaterial. © 2020 Society of Chemical Industry.