沃尔巴克氏菌调控小茶尺蠖和灰茶尺蠖杂交卵孵化

doi:10.16380/j.kcxb.2022.08.008

昆虫学报 ›› 2022, Vol. 65 ›› Issue (8): 999-1009.doi: 10.16380/j.kcxb.2022.08.008

沃尔巴克氏菌调控小茶尺蠖和灰茶尺蠖杂交卵孵化

王志博¹, 刘永健², 白家赫¹, 张欣欣¹, 肖强^1,*

(1. 中国农业科学院茶叶研究所, 农业农村部茶叶质量安全控制重点实验室, 杭州310008; 2. 中国科学院分子植物卓越创新中心, 上海 200031)

出版日期:2022-08-20 发布日期:2022-09-16

Wolbachia regulates the egg hatching of hybrids of Ectropis obliqua and E. grisescens (Lepidoptera: Geometridae)

WANG Zhi-Bo¹, LIU Yong-Jian², BAI Jia-He¹, ZHANG Xin-Xin¹, XIAO Qiang^1,*

(1. Key Laboratory of Tea Quality and Safety Control, Ministry of Agriculture and Rural Affairs, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310008, China; 2. Center for Excellence in Molecular Plant Science, Chinese Academy of Sciences, Shanghai 200031, China)

Online:2022-08-20 Published:2022-09-16

摘要/Abstract

摘要： 【目的】明确沃尔巴克氏菌Wolbachia对茶尺蠖两近缘种杂交卵孵化的影响及相关机制。【方法】使用2.5 mg/mL四环素浸液处理1 min后的新鲜茶树叶片连续饲喂灰茶尺蠖Ectropis grisescens幼虫3代，建立不携带Wolbachia的灰茶尺蠖种群。通过携带和不携带Wolbachia的灰茶尺蠖成虫分别与小茶尺蠖E. obliqua成虫进行杂交，探究Wolbachia对二者杂交卵孵化的影响。同时利用iTRAQ技术检测并分析携带和不携带Wolbachia的灰茶尺蠖精子蛋白的差异。【结果】与自然携带Wolbachia的灰茶尺蠖相比，去除Wolbachia后的灰茶尺蠖与小茶尺蠖杂交卵孵化率从3.92%显著提高到56.20%，表明Wolbachia诱导胞
质不亲合。精子蛋白差异分析显示，携带和不携带Wolbachia的灰茶尺蠖的精子蛋白中，共有128个蛋白显著差异表达，其中KEGG数据库注释到45个差异蛋白富集到鞘脂类代谢、唾液分泌、溶酶体、鞘脂信号通路和鞘糖脂生物合成等106个通路中，其中鞘脂类代谢通路中与神经酰胺合成相关的神经酰胺酶和磷酸酯酶显著上调表达。【结论】共生菌Wolbachia通过胞质不亲和作用介导了茶尺蠖两近缘种间的合子前生殖隔离，导致杂交卵的孵化率显著下降；Wolbachia可能通过鞘脂代谢通路实现了对雄性精子蛋白的修饰。

关键词: 茶尺蠖, 近缘种, 沃尔巴克氏菌, 杂交卵, 胞质不亲和, 精子蛋白

Abstract: 【Aim】 The objective of this study is to reveal the impacts of Wolbachia on the egg hatching of hybrids of two sibling tea geometrids and the underlying mechanism. 【Methods】 A subset of Ectropis grisescens larvae were fed with fresh tea leaves soaked in 2.5 mg/mL tetracycline solution for 1 min to remove Wolbachia. E. grisescens populations without Wolbachia were established after being fed with tetracycline-treated tea leaves for three generations. E. obliqua adults were hybridized with E. grisescens adults with and without Wolbachia to produce hybrids for the investigation of the impacts of Wolbachia on the egg hatching. The iTRAQ technique was used to detect and analyze the differences in sperm proteins between E. grisescens with and without Wolbachia. 【Results】 When Wolbachia was removed from E. grisescens, the egg hatching rates of hybrids between E. grisescens and E. obliqua were significantly increased from 3.92% to 56.20%, indicating that Wolbachia induced cytoplasmic incompatibility. Results of differential sperm proteins analysis showed that a total of 128 proteins were confirmed to have significantly different expression levels in sperms of E. grisescens with and without Wolbachia. According to the KEGG database, 45 differential proteins were enriched in 106 pathways, including sphingolipid metabolism, salivary secretion, lysosome, sphingolipid signaling, sphingolipid biosynthesis, etc. Among them the expression levels of ceramidases and phosphatases related to ceramide synthesis in the sphingolipid metabolic pathway were significantly up-regulated. 【Conclusion】 The symbiotic bacterium, Wolbachia, mediates the prezygotic reproductive isolation between the two sibling tea geometrids through cytoplasmic incompatibility, leading to a significant decrease in the egg hatching rate of hybrids. The observed cytoplasmic incompatibility may be attributable to the modification of the male sperm proteins caused by the impacts of Wolbachia on the sphingolipid metabolism pathway.

Key words: Tea geometrids, sibling species; Wolbachia, hybrid eggs, cytoplasmic incompatibility, sperm protein

王志博, 刘永健, 白家赫, 张欣欣, 肖强. 沃尔巴克氏菌调控小茶尺蠖和灰茶尺蠖杂交卵孵化[J]. 昆虫学报, 2022, 65(8): 999-1009.

WANG Zhi-Bo, LIU Yong-Jian, BAI Jia-He, ZHANG Xin-Xin, XIAO Qiang. Wolbachia regulates the egg hatching of hybrids of Ectropis obliqua and E. grisescens (Lepidoptera: Geometridae)[J]. Acta Entomologica Sinica, 2022, 65(8): 999-1009.

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沃尔巴克氏菌调控小茶尺蠖和灰茶尺蠖杂交卵孵化

Wolbachia regulates the egg hatching of hybrids of Ectropis obliqua and E. grisescens (Lepidoptera: Geometridae)

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