JIA-2021-1973  小菜蛾绒毛膜基因chorions鉴定及RNA干扰介导的chorion-1功能分析

doi:10.1016/j.jia.2022.08.079

Journal of Integrative Agriculture ›› 2022, Vol. 21 ›› Issue (11): 3278-3292.DOI: 10.1016/j.jia.2022.08.079

JIA-2021-1973 小菜蛾绒毛膜基因chorions鉴定及RNA干扰介导的chorion-1功能分析

收稿日期:2021-11-17 接受日期:2022-04-18 出版日期:2022-11-01 发布日期:2022-04-18

Identification of chorion genes and RNA interference-mediated functional characterization of chorion-1 in Plutella xylostella

DONG Shi-jie^{1, 2, 3, 4, 5}^*, LIU Bo⁶^*, ZOU Ming-min^{1, 2, 3, 4, 5}, LIU Li-li^{1, 2, 3, 4, 5}, CAO Min-hui^{1, 2, 3, 4, 5}, HUANG Meng-qi^{1, 2, 3, 4, 5}, LIU Yan^{1, 2, 3, 4, 5}, Liette VASSEUR^{1, 2, 3, 4, 7}, YOU Min-sheng^{1, 2, 3, 4, 5}, PENG Lu^{1, 2, 3, 4, 5}

¹State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
²International Joint Research Laboratory of Ecological Pest Control, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
³Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
⁴Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops, Ministry of Agriculture, Fuzhou 350002, P.R.China
⁵Fujian Provincial Key Laboratory of Insect Ecology, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
⁶Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, P.R.China
⁷Department of Biological Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada

Received:2021-11-17 Accepted:2022-04-18 Online:2022-11-01 Published:2022-04-18
About author:DONG Shi-jie, E-mail: dongsj01@163.com; LIU Bo, E-mail: liubo03@caas.cn; Correspondence YOU Min-sheng, E-mail: msyou@fafu.edu.cn; PENG Lu, E-mail: pl526520@163.com * These authors contributed equally to this study.
Supported by:
This work was funded by the National Natural Science Foundation of China (32172404), the Natural Science Foundation of Fujian Province, China (2019J01666), the Fujian Agriculture and Forestry University Fund for Distinguished Young Scholars, China (xjq201903), and the “111” Program - Innovation Center for Ecologically Based Pest Management of Subtropical Crops, Fujian Agriculture and Forestry University, China.

摘要/Abstract

摘要：

绒毛膜形成期是昆虫卵子发生的最后阶段，在这一阶段，滤泡细胞通过合成分泌绒毛膜蛋白，转移至发育的卵母细胞表面沉积形成卵壳，为胚胎发育提供保护屏障。目前对绒毛膜基因家族的研究大多集中在模式昆虫，如家蚕和果蝇，在世界性害虫小菜蛾中仍缺少对绒毛膜基因家族的系统鉴定以及其功能分析。因此明确绒毛膜基因家族在小菜蛾基因组上的分布情况及其转录特征，解析绒毛膜基因在小菜蛾卵子发生过程，以及胚胎发育过程中的重要作用，可为小菜蛾的遗传调控提供潜在分子靶标。本研究分析鉴定了小菜蛾绒毛膜基因的数量及染色体定位、分子特征、进化关系及其启动子区的序列特性，基于转录组数据以及qPCR实验，分析了绒毛膜基因在不同龄期和不同组织的表达模式，并基于RNAi揭示了PxCho-1的生殖功能。在小菜蛾中一共鉴定得到15个绒毛膜基因，分为A、B两大类。不同类型绒毛膜基因以成对的方式分布在染色体上。部分绒毛膜基因对，共享一个双向启动子调控区。系统发育分析表明，A、B两类绒毛膜基因具有高度的保守性，并且在对应类别中，小菜蛾绒毛膜基因均具有物种特异性。不同龄期与组织的表达谱与qPCR分析均显示，绒毛膜基因主要在小菜蛾雌成虫中显著高表达，并且在卵黄完全沉积的卵巢内显著高表达，说明绒毛膜基因在小菜蛾雌成虫的生殖发育中具有重要作用，且主要作用于卵子发生后期。抑制PxCho-1基因转录虽然对卵黄沉积没有影响，但会导致卵子变小，孵化率急剧下降，同时导致卵子内卵壳柱状层的排列松弛，以及外卵壳绒毛变短。本研究为探索小菜蛾雌性生殖调控机制奠定了理论基础，有利于筛选潜在的小菜蛾遗传防控分子靶标。

Abstract:

Choriogenesis is the last step of insect oogenesis, a process by which the chorion polypeptides are produced by the follicular cells and deposited on the surface of oocytes in order to provide a highly specialized protective barrier to the embryo. The essential features of chorion genes have yet to be clearly understood in the diamondback moth, Plutella xylostella, a worldwide Lepidoptera pest attacking cruciferous crops and wild plants. In this study, complete sequences for 15 putative chorion genes were identified, and grouped into A and B classes. Phylogenetic analysis revealed that both classes were highly conserved and within each, branches are also species-specific. Chorion genes from each class were located in pairs on scaffolds of the P. xylostella genome, some of which shared the common promoter regulatory region. All chorion genes were highly specifically expressed in the P. xylostella adult females, mostly in the ovary with full yolk, which is a crucial period to build the shells of the eggs. RNAi-based knockdown of chorion-1, which is located on the Px_scaffold 6 alone, although had no effect on yolk deposition, resulted in smaller eggs and sharply reduced hatchability. Additionally, inhibition of PxCho-1 expression caused a less dense arrangement of the columnar layers, reduced exochorion roughness and shorter microvilli. Our study provides the foundation for exploring molecular mechanisms of female reproduction in P. xylostella, and for making use of chorion genes as the potential genetic-based molecular target to better control this economically important pest.

Key words: Plutella xylostella , chorion genes , RNAi , oogenesis , pest control

. JIA-2021-1973 小菜蛾绒毛膜基因chorions鉴定及RNA干扰介导的chorion-1功能分析[J]. Journal of Integrative Agriculture, 2022, 21(11): 3278-3292.

DONG Shi-jie, LIU Bo, ZOU Ming-min, LIU Li-li, CAO Min-hui, HUANG Meng-qi, LIU Yan, Liette VASSEUR, YOU Min-sheng, PENG Lu. Identification of chorion genes and RNA interference-mediated functional characterization of chorion-1 in Plutella xylostella[J]. Journal of Integrative Agriculture, 2022, 21(11): 3278-3292.

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JIA-2021-1973 小菜蛾绒毛膜基因chorions鉴定及RNA干扰介导的chorion-1功能分析

Identification of chorion genes and RNA interference-mediated functional characterization of chorion-1 in Plutella xylostella

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