Henosepilachna vigintioctopunctata, a major threat to commercial potato production, is sensitive to RNA interference (RNAi), a gene-silencing mechanism triggered by double stranded RNA (dsRNA) matching a specific target gene sequence. Vacuolar ATPases (vATPases) are ATP-driven proton pumps involved in numerous biological processes. A holoenzyme vATPase consists of at least 14 different subunits and accessory proteins. Previous results reveal that silencing vATPaseB decreases survival rate, reduces food consumption and inhibits development in H. vigintioctopunctata. In this report, we identified two other subunit genes (vATPasea and vATPased) and compared the RNAi efficacy of vATPasea and vATPased with that of vATPaseB. We found the highest expression level of HvvATPasea, HvvATPaseB and HvvATPased occurred in the hindgut, followed by those in the foregut, Malpighian tubules and midgut, and lowest in the epidermis and fat body. When the fourth-instar larvae were allowed to ingest corresponding dsRNA for three days, the relative transcript levels of HvvATPasea, HvvATPaseB and HvvATPased were significantly decreased by 79%, 83% and 73%, respectively. Knockdown of HvvATPasea, HvvATPaseB and HvvATPased caused 50%, 85% and 100% larval lethality. After the third-instar larvae had exposed to corresponding dsRNA for three days, the levels of HvvATPasea and HvvATPased significantly reduced by 91% and 96% respectively. All the treated larvae were dead. Our results demonstrate that RNAi efficiencies varied among different vATPase subunit genes and various development stages. Moreover, our data imply the feasibility of RNAi as an alternative method for controlling this critical potato pest.

Henosepilachna vigintioctopunctata是商业马铃薯生产的主要威胁，它对RNA干扰（RNAi）敏感，RNAi是由与特定目标基因序列匹配的双链RNA（dsRNA）触发的基因沉默机制。液泡ATPase（vATPase）是ATP驱动的质子泵，涉及许多生物过程。全酶vATPase由至少14个不同的亚基和辅助蛋白组成。以前的研究结果表明，沉默vATPaseB降低存活率，降低了食品消费和抑制发展H.瓢虫。在本报告中，我们鉴定了其他两个亚基基因（vATPasea和vATPased），并比较了vATPasea和vATP与vATPaseB相同。我们发现HvvATPasea，HvvATPaseB和HvvATPased的表达水平最高的发生在后肠，其次是前肠，马尔皮根小管和中肠，而最低的发生在表皮和脂肪体。当四龄幼虫摄入相应的dsRNA 3天后，HvvATPasea，HvvATPaseB和HvvATPased的相对转录水平分别显着降低了79％，83％和73％。击倒HvvATPasea，HvvATPaseB和HvvATP造成50％，85％和100％的幼虫致死率。三龄幼虫暴露于相应的dsRNA三天后，HvvATPasea和HvvATPas的水平分别显着降低了91％和96％。所有处理过的幼虫都死了。我们的结果表明，RNAi效率在不同的vATPase亚基基因和不同的发育阶段之间有所不同。此外，我们的数据暗示了RNAi作为控制这种重要马铃薯害虫的替代方法的可行性。