Institute of Applied Biology, Shanxi University, Taiyuan, China.
College of Life Sciences, Shanxi University, Taiyuan, China.
Pest Manag Sci. 2022 Apr;78(4):1555-1566. doi: 10.1002/ps.6774. Epub 2022 Jan 20.
Vacuolar (H )-ATPase (V-ATPase) is a multi-subunit enzyme that hydrolyzes adenosine triphosphate (ATP) to transport protons across a cellular membrane, and it plays an important role in numerous biological processes, including in growth, development and immune responses. The c subunit of V-ATPase is a highly conserved subunit of the rotatory proteolipid ring that is required for binding and transporting protons. To date, there are only a few published reports on V-ATPase-c functions in insects.
We identified and characterized the V-ATPase-c gene in Locusta migratoria, one of the most destructive agricultural insect pests in the world. LmV-ATPase-c was predominately expressed in Malpighian tubules of nymphs, followed by the hindgut and ovary, while the other tissues showed relatively low expression levels. Silencing of LmV-ATPase-c caused severe molting defects in nymphs and a high mortality rate of > 90%. Histological staining and microscopic examination of sections from the abdominal cuticle revealed the absence of newly formed cuticle in nymphs that were injected with dsLmV-ATPase-c. In addition, silencing of LmV-ATPase-c transcript levels significantly impaired RNA interference (RNAi) efficiency of a reporter gene. By quantifying double-stranded RNA (dsRNA) amounts by quantitative polymerase chain reaction (PCR), we found that RNAi against LmV-ATPase-c provoked a dramatic accumulation of dsRNA in the endosomes of epidermal and midgut cells of Locusta migratoria.
Our results indicate that LmV-ATPase-c is indispensable for the formation of new cuticle during the molting process and has pivotal functions in dsRNA escape from endosomes. LmV-ATPase-c might be a valuable target for developing new strategies for insect pest management. © 2022 Society of Chemical Industry.
液泡（H + ）-ATP 酶（V-ATPase）是一种多亚基酶，可将三磷酸腺苷（ATP）水解为跨细胞膜运输质子，并在许多生物过程中发挥重要作用，包括生长、发育和免疫反应。V-ATPase 的 c 亚基是旋转蛋白脂质环的高度保守亚基，对于结合和运输质子是必需的。迄今为止，关于昆虫中 V-ATPase-c 功能的报道仅有少数。
我们鉴定并表征了世界上最具破坏性的农业害虫之一——飞蝗中的 V-ATPase-c 基因。LmV-ATPase-c 在若虫的马氏管中表达最为丰富，其次是后肠和卵巢，而其他组织的表达水平相对较低。LmV-ATPase-c 的沉默导致若虫蜕皮严重缺陷，死亡率超过 90%。对腹部表皮切片进行组织学染色和显微镜检查显示，注射 dsLmV-ATPase-c 的若虫缺乏新形成的表皮。此外，LmV-ATPase-c 转录本水平的沉默显著损害了报告基因的 RNA 干扰（RNAi）效率。通过定量聚合酶链反应（PCR）定量双链 RNA（dsRNA）的量，我们发现针对 LmV-ATPase-c 的 RNAi 会引起飞蝗表皮和中肠细胞的内体中 dsRNA 的大量积累。
我们的结果表明，LmV-ATPase-c 在蜕皮过程中形成新表皮是必不可少的，并且在 dsRNA 从内体逃逸中具有关键作用。LmV-ATPase-c 可能是开发新的害虫管理策略的有价值的靶标。© 2022 化学工业协会。
