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Identification of Halloween Genes and RNA Interference-Mediated Functional Characterization of a Halloween Gene shadow in Plutella xylostella

机译：小菜蛾万圣节基因阴影的万圣节基因鉴定和RNA干扰介导的功能表征

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Ecdysteroids play an essential role in controlling insect development and reproduction. Their pathway is regulated by a group of enzymes called Halloween gene proteins. The relationship between the Halloween genes and ecdysteroid synthesis has yet to be clearly understood in diamondback moth, Plutella xylostella (L.), a worldwide Lepidoptera pest attacking cruciferous crops and wild plants. In this study, complete sequences for six Halloween genes, neverland (nvd), shroud (sro), spook (spo), phantom (phm), disembodied (dib), shadow (sad), and shade (shd), were identified. Phylogenetic analysis revealed a strong conservation in insects, including Halloween genes of P. xylostella that was clustered with all other Lepidoptera species. Three Halloween genes, dib, sad, and shd were highly expressed in the adult stage, while nvd and spo were highly expressed in the egg and pupal stages, respectively. Five Halloween genes were highly expressed specifically in the prothorax, which is the major site of ecdysone production. However, shd was expressed predominantly in the fat body to convert ecdysone into 20-hydroxyecdysone. RNAi-based knockdown of sad, which is involved in the last step of ecdysone biosynthesis, significantly reduced the 20E titer and resulted in a longer developmental duration and lower pupation of fourth-instar larvae, as well as caused shorter ovarioles and fewer fully developed eggs of P. xylostella. Furthermore, after the knockdown of sad, the expression levels of Vg and VgR genes were significantly decreased by 77.1 and 53.0%. Meanwhile, the number of eggs laid after 3 days was significantly reduced in sad knockdown females. These results suggest that Halloween genes may play a critical role in the biosynthesis of ecdysteroids and be involved in the development and reproduction of P. xylostella. Our work provides a solid basis for understanding the functional importance of these genes, which will help to screening potential genes for pest management of P. xylostella.
机译：蜕皮类固醇在控制昆虫的发育和繁殖中起着至关重要的作用。它们的途径受一组称为万圣节基因蛋白的酶调节。万圣节基因与蜕皮甾类合成之间的关系尚未在小菜蛾小菜蛾（Plutella xylostella（L.））中清楚地理解，小菜蛾是一种全世界鳞翅目害虫，攻击十字花科作物和野生植物。在这项研究中，确定了六个万圣节基因的完整序列，其中包括梦幻岛（nvd），裹尸布（sro），幽灵（spo），幻影（phm），无肉（dib），阴影（sad）和阴影（shd）。系统发育分析表明，昆虫具有很强的保护力，其中包括与所有其他鳞翅目物种聚集在一起的小菜蛾万圣节基因。在成年期高表达三个万圣节基因dib，sad和shd，而在卵期和p期分别高表达nvd和spo。五个万圣节基因在胸廓中高度表达，而胸廓是蜕皮激素产生的主要部位。但是，shd主要在脂肪体内表达，可以将蜕皮激素转化为20-羟基蜕皮激素。蜕皮激素生物合成的最后一步涉及的基于RNAi的sad敲除可显着降低20E滴度，并导致更长的发育时间和更低的四龄幼虫化脓率，并导致更短的卵泡和更少的完全发育的卵的 P。小菜蛾。此外，敲除 sad 后， Vg 和 VgR 基因的表达水平显着降低了77.1％和53.0％。同时， sad 击倒雌性小鼠3天后产卵的数量显着减少。这些结果表明，万圣节基因可能在蜕皮类固醇的生物合成中起关键作用，并参与 P的发育和繁殖。小菜蛾。我们的工作为理解这些基因的功能重要性提供了坚实的基础，这将有助于筛选潜在的基因以用于 P的害虫管理。小菜蛾。

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