Peng Lu, Wang Qing, Zou Ming-Min, Qin Yu-Dong, Vasseur Liette, Chu Li-Na, Zhai Yi-Long, Dong Shi-Jie, Liu Li-Li, He Wei-Yi, Yang Guang, You Min-Sheng
State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fujian Agriculture and Forestry University, Fuzhou, China.
Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, China.
Front Physiol. 2020 Jan 23;10:1585. doi: 10.3389/fphys.2019.01585. eCollection 2019.
The vitellogenin receptor (VgR) belongs to the low-density lipoprotein receptor (LDLR) gene superfamily and plays an indispensable role in Vg transport, yolk deposition, and oocyte development. For this reason, it has become a promising target for pest control. The involvement of in Vg transport and reproductive functions remains unclear in diamondback moths, (L.), a destructive pest of cruciferous crops. Here, we cloned and identified the complete cDNA sequence of , which encoded 1805 amino acid residues and contained four conserved domains of LDLR superfamily. was mainly expressed in female adults, more specifically in the ovary. VgR protein also showed the similar expression profile with the transcript. CRISPR/Cas9-mediated knockout created a homozygous mutant of with 5-bp-nucleotide deletion in the . The expression deficiency of VgR protein was detected in the ovaries and eggs of mutant individuals. Vg protein was still detected in the eggs of the mutant individuals, but with a decreased expression level. However, transcripts were not significantly affected by the knockout. Knockout of resulted in shorter ovarioles of newly emerged females. No significant difference was detected between wild and mutant individuals in terms of the number of eggs laid in the first 3 days after mating. The loss of gene resulted in smaller and whiter eggs and lower egg hatching rate. This study represents the first report on the functions of in Vg transport, ovary development, oviposition, and embryonic development of using CRISPR/Cas9 technology. This study lays the foundation for understanding molecular mechanisms of reproduction, and for making use of as a potential genetic-based molecular target for better control of the
卵黄原蛋白受体（VgR）属于低密度脂蛋白受体（LDLR）基因超家族，在卵黄原蛋白运输、卵黄沉积和卵母细胞发育中发挥着不可或缺的作用。因此，它已成为害虫防治的一个有前景的靶点。在十字花科作物的毁灭性害虫小菜蛾（Plutella xylostella (L.)）中，VgR在卵黄原蛋白运输和生殖功能中的作用仍不清楚。在此，我们克隆并鉴定了小菜蛾VgR的完整cDNA序列，其编码1805个氨基酸残基，并包含LDLR超家族的四个保守结构域。PxVgR主要在雌成虫中表达，更具体地说是在卵巢中表达。VgR蛋白也显示出与PxVgR转录本相似的表达谱。CRISPR/Cas9介导的PxVgR敲除产生了PxVgR的纯合突变体，其在PxVgR中有5个碱基对的核苷酸缺失。在突变个体的卵巢和卵中检测到VgR蛋白的表达缺陷。在突变个体的卵中仍检测到Vg蛋白，但表达水平降低。然而，PxVgR转录本不受PxVgR敲除的显著影响。PxVgR的敲除导致新羽化雌虫的卵巢小管变短。在交配后的前3天，野生个体和突变个体之间产卵数量没有显著差异。PxVgR基因的缺失导致卵更小、更白，且孵化率更低。本研究首次报道了利用CRISPR/Cas9技术研究PxVgR在小菜蛾卵黄原蛋白运输、卵巢发育、产卵和胚胎发育中的功能。本研究为理解小菜蛾繁殖的分子机制以及将PxVgR作为潜在的基于基因的分子靶点以更好地控制小菜蛾奠定了基础。
