Henosepilachna vigintioctopunctata is one of the most serious insect pests to a large number of nightshades and cucurbits. RNA interference (RNAi) triggered by double-stranded RNA (dsRNA) offers a reduced risk approach to control the beetle. Identification of amenable target genes and determination of appropriate life stage for dsRNA treatment are two critical steps in order to improve RNAi efficiency. In the present paper, we identified three vATPase genes, namely HvvATPaseC, HvvATPaseE and HvvATPaseH. We found that the three transcripts were widely expressed in the eggs, first- to fourth-instar larvae, prepupae, pupae and adults. They were abundantly transcribed in the hindgut and Malpighian tubules, in contrast to the epidermis and fat body. Three days' ingestion of dsvATPaseC, dsvATPaseE and dsvATPaseH by the fourth-instar larvae significantly decreased corresponding transcript level by 90.1, 88.9 and 97.2%, greatly reduced larval fresh weight by 28.0, 29.9 and 28.0%, and caused 66.7, 100 and 78.7% larval lethality respectively. Comparably, 3 days' exposure of the third-instar larvae to dsvATPaseC significantly reduced HvvATPaseC mRNA level by 89.5%, decreased approximately 80% of the larval fresh weight, and killed 100% of the treated larvae. Therefore, the three vATPase genes, especially HvvATPaseE, are potential amenable target genes and young larvae are more susceptible to dsRNA. Our findings will enable the development of the dsRNA-based pesticide to control H. vigintioctopunctata.

中文翻译：

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液泡型H+-ATP酶基因的RNAi导致Henosepilachna vigintioctopunctata生长延迟和蜕皮缺陷


八点金龟子是对大量茄属植物和葫芦科植物最严重的害虫之一。由双链 RNA (dsRNA) 触发的 RNA 干扰 (RNAi) 提供了一种降低风险的方法来控制甲虫。鉴定合适的靶基因和确定 dsRNA 治疗的适当生命阶段是提高 RNAi 效率的两个关键步骤。在本文中，我们鉴定了三个 vATPase 基因，即HvvATP酶C , HvvATP酶E和HvvATP酶H 。我们发现这三个转录本在卵、一至四龄幼虫、预蛹、蛹和成虫中广泛表达。与表皮和脂肪体相比，它们在后肠和马尔皮基小管中大量转录。三天摄入ds vATP酶C , ds vATP酶E和DS vATP酶H四龄幼虫的相应转录水平显着降低90.1%、88.9%和97.2%，幼虫鲜重大幅降低28.0%、29.9%和28.0%，幼虫致死率分别为66.7%、100%和78.7%。相比之下，三龄幼虫暴露于 ds 3 天vATP酶C显着减少HvvATP酶C mRNA 水平降低了 89.5%，幼虫鲜重减少了约 80%，并杀死了 100% 的处理幼虫。因此，这三个 vATPase 基因，特别是HvvATP酶E ，是潜在的合适靶基因，幼虫更容易受到 dsRNA 的影响。我们的研究结果将使基于 dsRNA 的农药的开发能够控制H. vigintioctopunctata 。