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| Vacuolar (H+)-ATPase (V-ATPase) functions as an electrogenic pump, transporting protons from the cytoplasm to the extracellular fluid to generate cell-negative membrane voltage. The V-ATPase subunit H, encoded by Vhasfd, is required for the V-ATPase activity. In this study, the gene encoding V-ATPase subunit H from Locusta migratoria manilensis was cloned, and designated as Lm-Vhasfd. The complete cDNA sequence is 2018 bp, with an open reading frame encoding 515 amino acid residues. Semi-quantitative PCR showed that Lm-Vhasfd transcription is high in hemolymph, midgut, trunk and leg, but relatively low in fat body and head tissues. Injection with a specific double strand RNA (dsRNA) led to a significant decrease in Lm-Vhasfd mRNA, V-ATPase enzyme activity and ATP concentration. Bioassays showed that silencing Lm-Vhasfd led to the death of individuals and various molting defects. The accumulative mortality of the RNAi mutant 11 days post-injection was 96.7%, conspicuously higher than that seen in wild type. These RNAi phenotypes demonstrate that Lm-Vhasfd is essential for the growth and molting of L. migratoria manilensis. |
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| Keywords:vacuolar-ATPase; Vhasfd; Locusta migratoria manilensis; RNA interference; double-stranded RNA. |
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