Vacuolar ATPase Subunit H Is Essential For The Survival And Molting Of Locusta Migratoria Manilensis

Vacuolar (H+)-ATPase is a multisubunit proton pump and widely distribute in theplasma membrane and the endomembrane system. Vacuolar (H+)-ATPase functions asan electrogenic pump, transporting protons from the cytoplasm to the extracellular fluidto generate cell-negative membrane voltage. The V-ATPase subunit H, encoded byVhasfd, is required for V-ATPase activity. It has been reported that silent the subunit ofA or E of V-ATPase will impact the target organisms. But little is known about thefunction of subunit H on individual insects.The oriental migratory locust (L. migratoria manilensis), a member of theorthopterans and a representative member of hemimetabolous insects, is one of the mostimportant agricultural pests of prairies and croplands. In China, infestations of L.migratoria manilensis are extremely destructive and they are mainly controlled byorganophosphate (OP) insecticides. However, the insects have developed resistance toOP, and subsequently, its efficiency has decreased. For this reason, new controlstrategies of locust need to be developed, such as pesticides aimed at new targets, or thecreation of transgenic plants that express dsRNA against specific insect genes. Thus, theidentification of genes that are essential for locust growth and development couldprovide potential novel targets for insect control.In this study,①The gene encoding V-ATPase subunit H from Locusta migratoria manilensiswas cloned, and designated as Lm-Vhasfd. The complete cDNA sequence is2,018bp, with an open reading frame encoding515amino acid residues.②Semi-quantitative PCR showed that Lm-Vhasfd transcription is high in thehemolymph, midgut, trunk and leg, but relatively low in the fat body and headtissues.③Injection with a specific double-strand RNA (dsRNA) led to a significantdecrease in Lm-Vhasfd mRNA, V-ATPase enzyme activity and ATPconcentration.④Bioassays showed that silencing Lm-Vhasfd led to the death of individuals andvarious molting defects. The accumulative mortality of the RNAi mutant11days post-injection was96.7%, which was conspicuously higher than that seenin wild type locusts. These RNAi phenotypes demonstrate that Lm-Vhasfd is essential for the growth and molting of L. migratoria manilensis.