The regulation of lipid metabolism by a hypothetical P-loop NTPase and its impact on fecundity of the brown planthopper.

BACKGROUND: Insect fecundity can be regulated by multiple genes in several important signaling pathways which form an extremely complicated regulatory network. However, there are still many genes that have significant impact on insect fecundity but their action mode are still unknown. METHODS: Quantitative real-time PCR (qRT-PCR), immunofluorescence and western blot were used to study the expression profile of Nl23867 in the brown planthopper, Nilaparvata lugens. RNA interference (RNAi), RNA-seq and isobaric tags for relative and absolute quantification (iTRAQ) were performed to investigate the action mode of Nl23867 in the regulation of fecundity. High performance liquid chromatography (HPLC) analysis was performed to detect the fatty acid contents. RESULTS: We show that knockdown of Nl23867, a gene encoding a hypothetical P-loop NTPase, significantly decreased fecundity of N. lugens. Underdeveloped ovaries, fewer eggs laid and reduction in vitellogenin (Vg) protein expression were observed after RNAi knockdown of Nl23867, and most of the affected genes and pathways are fatty acid metabolism-related. We further determined that Nl23867 directly impacts the palmitic acid biosynthesis by regulating the expression of palmitoyl-protein thioesterase (PPT), subsequently affecting the content of total lipids in N. lugens. CONCLUSIONS: Nl23867 regulates the fecundity of N. lugens by modulating the biosynthetic pathway of palmitic acid and affecting lipid metabolism during vitellogenesis and oocyte development. GENERAL SIGNIFICANCE: The presented study pioneers the exploration into how a function-unknown gene takes part in the regulation of fecundity in an insect, and will contribute to the construction of gene regulatory network for insect fecundity.

Functional screen for microRNAs of Nilaparvata lugens reveals that targeting of glutamine synthase by miR-4868b regulates fecundity.

Insect fecundity is regulated by the interaction of genotypes and the environment. MicroRNAs (miRNAs) also act in insect development and reproduction by regulating genes involved in these physiological processes. Although hundreds of insect miRNAs have been identified, the biological roles of most remain poorly understood. Here, we used a multi-algorithm approach for miRNA target prediction in 3'UTRs of fecundity-related genes in the brown planthopper (BPH) Nilaparvata lugens and identified 38 putative miRNAs targeting 9 fecundity-related genes. High-ranked miRNAs were selected for target validation. Using a dual luciferase reporter assay in S2 cells, we experimentally verified N. lugens glutamine synthetase (NlGS) as an authentic target of microRNA-4868b (miR-4868b). In the females, NlGS protein expression was down-regulated after injection of a miR-4868b mimic but up-regulated after injection of a miR-4868b inhibitor. In addition, overexpression of miR-4868b reduced fecundity, and disrupted ovary development and Vg expression in N. lugens. These findings showed that miR-4868b is involved in regulating N. lugens fecundity by targeting NlGS. Moreover, this study may lead to better understanding of the fecundity of this important agricultural insect pest.