绿豆象气味结合蛋白基因的克隆及表达分析

doi:10.16380/j.kcxb.2021.08.004

摘要/Abstract

摘要：

【目的】对绿豆象Callosobruchus chinensis气味结合蛋白(odorant binding proteins, OBPs)基因进行克隆、鉴定和组织表达分析，为研究OBPs在绿豆象嗅觉感受过程中的功能奠定基础。【方法】基于绿豆象触角转录组数据，通过RT-PCR克隆绿豆象6个OBP基因并进行生物信息学分析；通过qRT-PCR分析OBP基因在绿豆象雌雄成虫头(不含触角)、触角、腹、足和翅各组织中的表达情况。【结果】获得了6个绿豆象OBP基因的开放阅读框，命名为CchiOBP1-CchiOBP6(GenBank登录号: MN832700-MN832703, MN901841-MN901842)；CchiOBP5为一段C端不完整的Minus-C OBP，其余均属于完整的Classical OBPs，预测6个CchiOBPs均含有信号肽。系统发育分析表明CchiOBP1, CchiOBP2和CchiOBP5与叶甲科(Chrysomelidae)昆虫OBPs的亲缘关系较近，CchiOBP3, CchiOBP4和CchiOBP6与天牛科(Cerambycidae)昆虫OBPs的亲缘关系较近。qRT-PCR结果表明，6个CchiOBPs基因在绿豆象成虫的触角、头(不含触角)、腹、翅和足部均有不同的表达量，CchiOBP1-4和CchiOBP6在雌雄成虫触角中均呈现高表达，且极显著高于在其他组织中的。CchiOBP5在雌成虫触角和头部(不含触角)中呈现高表达，但在雄成虫中表现为在足部的表达量显著高于在其他组织中的，在触角中的表达量最低。【结论】确定了绿豆象6个CchiOBPs基因的核苷酸和氨基酸序列组成，其中有5个CchiOBPs基因在绿豆象雌雄成虫触角中高表达，推测其在绿豆象嗅觉识别寄主植物过程中发挥重要作用。

关键词: 绿豆象, 嗅觉系统, 气味结合蛋白, 基因克隆, 组织表达

Abstract:

【Aim】This study aims to clone and identify odorant-binding protein (OBP) genes in the adzuki bean seed weevil, Callosobruchus chinensis and to assay their tissue-specific expression profiles in C. chinensis adults, so as to to lay a foundation for studying the function of OBPs in olfactory perception in C. chinensis. 【Methods】Based on the antenna transcriptome data of C. chinensis, six OBP genes in C. chinensis were cloned by RT-PCR and analyzed by bioinformatics. The expression levels of OBP genes in tissues including head (excluding antennae), antennae, abdomen, legs and wings of female and male adults of C. chinensis were analyzed by qRT-PCR. 【Results】 The open reading frames of six OBP genes from C. chinensis were obtained, and named as CchiOBP1-CchiOBP6 (GenBank accession number: MN832700-MN832703, MN901841-MN901842). CchiOBP5 is a segment of Minus-C OBP with incomplete C-terminus, and the others belong to complete classical OBPs. It was predicted that all the six CchiOBPs contain signal peptides. Phylogenetic analysis showed that CchiOBP1, CchiOBP2 and CchiOBP5 were closely related to OBPs of Chrysomelidae, and CchiOBP3, CchiOBP4 and CchiOBP6 were closely related to OBPs of Cerambycidae. The qRT-PCR results showed that six CchiOBP genes were differentially expressed in the antennae, head (excluding antennae), abdomen, wings and legs of C. chinensis adults. CchiOBP1-4 and CchiOBP6 were highly expressed in antennae of female and male adults of C. chinensis, and their expression levels in antennae were extremely significantly higher than those in the other adult tissues. CchiOBP5 was highly expressed in female antennae and head (excluding antennae), but in males it showed significantly higher expression level in legs than in other adult tissues, with the lowest expression level in antennae. 【Conclusion】 The nucleotide and amino acid sequences of six CchiOBP genes of C. chinensis have been determined, among which five CchiOBP genes are highly expressed in the antennae of female and male adults of C. chinensis, suggesting their important roles in olfactory recognition of host plants.

Key words: Callosobruchus chinensis, olfactory system, odorant-binding protein, gene cloning, tissue-specific expression

王宏民, 张静, 张冲, 庄国动, 郑海霞, 张仙红. 绿豆象气味结合蛋白基因的克隆及表达分析[J]. 昆虫学报, 2021, 64(8): 920-928.

WANG Hong-Min, ZHANG Jing, ZHANG Chong, ZHUANG Guo-Dong, ZHENG Hai-Xia, ZHANG Xian-Hong. Cloning and expression profiling of odorant binding protein genes in Callosobruchus chinensis (Coleoptera: Bruchidae)[J]. Acta Entomologica Sinica, 2021, 64(8): 920-928.

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绿豆象气味结合蛋白基因的克隆及表达分析

Cloning and expression profiling of odorant binding protein genes in Callosobruchus chinensis (Coleoptera: Bruchidae)

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