绿芫菁成虫触角中气味结合蛋白和化学感受蛋白基因的鉴定和表达分析

doi:10.16380/j.kcxb.2022.08.001

昆虫学报 ›› 2022, Vol. 65 ›› Issue (8): 927-936.doi: 10.16380/j.kcxb.2022.08.001

• 研究论文 • 下一篇

绿芫菁成虫触角中气味结合蛋白和化学感受蛋白基因的鉴定和表达分析

刘盼静¹, 魏洪义², 郭坤¹, 马广源¹, 张涛^1,*

(1. 河北省农林科学院植物保护研究所, 河北保定 071000; 2. 江西农业大学昆虫研究所, 南昌 330045)

出版日期:2022-08-20 发布日期:2022-09-16

Identification and expression profiling of genes of odorant-binding proteins and chemosensory proteins in adult antennae of Lytta caraganae (Coleoptera: Meloidae)

LIU Pan-Jing¹, WEI Hong-Yi², GUO Kun¹, MA Guang-Yuan¹, ZHANG Tao^1,*

(1. Plant Protection Institute, Hebei Academy of Agriculture and Forestry Sciences, Baoding, Hebei 071000, China; 2. Institute of Entomology, Jiangxi Agricultural University, Nanchang 330045, China)

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

摘要/Abstract

摘要： 【目的】建立绿芫菁Lytta caraganae成虫触角转录组数据库，并挖掘和鉴定其成虫触角中嗅觉识别相关的气味结合蛋白(odorantbinding proteins, OBPs)和化学感受蛋白(chemosensory proteins, CSPs)基因。【方法】采用Illumina HiSeq平台对绿芫菁成虫触角进行转录组分析，比对NR, NT, KO, Pfam, Swiss-Prot, GO和KOG数据库进行基因注释，根据注释信息筛选绿芫菁OBP和CSP基因；利用ClustalX 1.83和MEGA 7.0软件分别分析OBP和CSP基因结构特点及进化关系；通过qRT-PCR技术测定绿芫菁雌雄成虫触角中OBP和CSP基因的表达量。【结果】从绿芫菁成虫触角转录组中共获得51 028条transcripts和41 998条unigenes；基因注释结果表明，在NR数据库中匹配度最高(87.3%)的物种是赤拟谷盗Tribolium castaneum；筛选到22个OBP基因和7个CSP基因，序列比对结果显示，有13个LcarOBPs包含6个保守的半胱氨酸残基，属于classic OBPs；系统进化树表明，绿芫菁的OBPs和LcarCSPs分别与眼斑沟芫菁Hycleus cichorii和大斑沟芫菁H. phaleratus的OBPs和CSPs序列氨基酸一致性最高，表明进化关系最近；qRT-PCR结果显示，2个LcarOBPs基因和2个LcarCSP基因在绿芫菁雄成虫触角中高表达，10个LcarOBP基因和2个LcarCSP基因在雌成虫触角中高表达。【结论】本研究首次鉴定了绿芫菁成虫触角中的OBP和CSP基因，为进一步研究绿芫菁嗅觉识别机制提供了理论基础。

关键词: 绿芫菁, 触角转录组, 嗅觉识别, 气味结合蛋白, 化学感受蛋白

Abstract: 【Aim】 This study aims to establish the adult antennal transcriptome database of Lytta caraganae, and explore and identify genes of olfactory-related odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) in its adult antennae. 【Methods】 Transcriptome analysis of adult antennae of L. caraganae was performed on Illumina HiSeq platform. Assembled genes were annotated by alignment against public databases NR, NT, KO, Pfam, Swiss-Prot, GO and KOG. OBP and CSP genes of L. caraganae were screened according to the annotation results. Structural characteristics and evolutionary relationship of OBP and
CSP genes were analyzed by ClustalX 1.83 and MEGA 7.0 software, respectively. The expression levels of OBP and CSP genes in female and male adult antennae of of L. caraganae were determined by qRTPCR. 【Results】 A total of 51 028 transcripts and 41 998 unigenes were obtained from the adult antennal transcriptome of L. caraganae. Gene annotation results showed that L. caraganae genes have the highest match (87.3%) to those of Tribolium castaneum. Twenty-wo OBP genes and seven CSP genes were screened. Sequence alignment result showed that 13 LcarOBPs are classified into classic OBPs， which contain six conserved cysteine residues. Phylogenetic tree showed that LcarOBPs and LcarCSPs show the highest amino acid sequence identity with OBPs and CSPs of Hycleus cichorii and H. phaleratus, respectively, indicating the closest evolutionary relationship. qRT-PCR results showed that two LcarOBP genes and two LcarCSP genes were highly expressed in the male adult antennae of L. caraganae, while ten LcarOBP genes and two LcarCSP genes highly expressed in the female adult antennae. 【Conclusion】 The OBP and CSP genes in adult antennae of L. caraganae have been identified for the first time, providing a theoretical basis for further study on the mechanism of olfactory recognition in L. caraganae.

Key words: Lytta caraganae, antennal transcriptome, olfactory recognition, odorant-binding protein, chemosensory protein

刘盼静, 魏洪义, 郭坤, 马广源, 张涛. 绿芫菁成虫触角中气味结合蛋白和化学感受蛋白基因的鉴定和表达分析[J]. 昆虫学报, 2022, 65(8): 927-936.

LIU Pan-Jing, WEI Hong-Yi, GUO Kun, MA Guang-Yuan, ZHANG Tao. Identification and expression profiling of genes of odorant-binding proteins and chemosensory proteins in adult antennae of Lytta caraganae (Coleoptera: Meloidae)[J]. Acta Entomologica Sinica, 2022, 65(8): 927-936.

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绿芫菁成虫触角中气味结合蛋白和化学感受蛋白基因的鉴定和表达分析

Identification and expression profiling of genes of odorant-binding proteins and chemosensory proteins in adult antennae of Lytta caraganae (Coleoptera: Meloidae)

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