烟粉虱MED隐种14-3-3基因克隆及时空表达谱分析  

Cloning and spatiotemporal expression profiling of the 14-3-3 genes in Bemisia tabaci MED(Hemiptera:Aleyrodidae)

作　　者：韦焕文 王培 陈建斌[2,3] 杜娇 张德咏[1,2,3] 刘勇 史晓斌 WEI Huan-Wen;WANG Pei;CHEN Jian-Bin;DU Jiao;ZHANG De-Yong;LIU Yong;SHI Xiao-Bin(Longping Branch,College of Biology,Hunan University,Changsha 410125,China;The Yuelushan Laboratory,Changsha 410125,China;Institute of Plant Protection,Hunan Academy of Agricultural Sciences,Changsha 410125,China)

机构地区：[1]湖南大学生物学院隆平分院,长沙410125 [2]岳麓山实验室,长沙410125 [3]湖南省农业科学院植物保护研究所,长沙410125

出　　处：《昆虫学报》2024年第5期595-602,共8页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(32272535,32030088);湖南省青年科技人才项目(2022RC1149);大宗蔬菜产业体系项目(CARS-23-D-02)。

摘　　要：【目的】真核生物中,14-3-3蛋白是一类可与多种蛋白互作的调节蛋白,能够参与信号转导、免疫反应、生长发育和胁迫响应等。本研究旨在克隆烟粉虱Bemisia tabaci MED隐种的14-3-3基因的全长cDNA序列,了解该基因编码的蛋白特征和该基因的时空表达模式。【方法】使用RT-PCR技术克隆烟粉虱MED隐种的14-3-3基因全长cDNA序列,通过生物信息学软件和在线网站分析14-3-3基因的生物学特性;使用RT-qPCR测定14-3-3基因在烟粉虱MED隐种不同发育阶段(卵、1-4龄若虫和成虫)、雌雄成虫以及雌成虫头、胸和腹中的表达量。【结果】克隆并鉴定了烟粉虱MED隐种14-3-3基因的两个亚型:Bt14-3-3 epsilon(GenBank登录号:XM_019046102.1)和Bt14-3-3 zeta(GenBank登录号:XM_019057395.1),开放阅读框(ORFs)分别长771和744 bp,分别编码256和247个氨基酸,编码的蛋白是无跨膜螺旋区和信号肽的亲水性蛋白,其二级结构主要由α旋螺旋组成。系统发育树分析表明,Bt14-3-3 epsilon与褐飞虱Nilaparvata lugens、温带臭虫Cimex lectularius和茶翅蝽Halyomorpha halys的14-3-3 epsilon聚为一支,同源性较高;Bt14-3-3 zeta与褐飞虱的14-3-3 zeta的亲缘关系更近。RT-qPCR结果表明,Bt14-3-3 epsilon和Bt14-3-3 zeta在烟粉虱MED隐种的卵、雌成虫和雌成虫腹部中的表达量较高。【结论】明确了烟粉虱MED隐种14-3-3基因的两个亚型的全长序列、编码蛋白特征及时空表达特点,为后续研究14-3-3蛋白的分子功能奠定了基础。【Aim】14-3-3 proteins are a class of regulatory proteins found in eukaryotic organisms,which can be involved in signal transduction,immune response,growth and development,and stress response.The aim of this study is to clone the full-length cDNA sequences of the 14-3-3 genes in Bemisia tabaci MED,and understand the characteristics of the proteins encoded by 14-3-3 genes and the spatiotemporal expression patterns of 14-3-3 genes.【Methods】The full-length cDNA sequences of 14-3-3 genes of B.tabaci MED were cloned by RT-PCR,and their biological properties were analyzed by bioinformatics software and online website.RT-qPCR was used to determine the expression levels of 14-3-3 genes in different developmental stages(egg,1st-4th instar nymphs and adult),in adult male and female,and in the head,thorax and abdomen of female adult of B.tabaci MED.【Results】Two subtypes of the 14-3-3 gene of B.tabaci MED were cloned and characterized:Bt14-3-3 epsilon(GenBank accession no.:XM_019046102.1)and Bt14-3-3 zeta(GenBank accession no.:XM_019057395.1).The open reading frames(ORFs)of Bt14-3-3 epsilon and Bt14-3-3 zeta were 771 and 744 bp,encoding 256 and 247 amino acids,respectively.The proteins encoded by Bt14-3-3 epsilon and Bt14-3-3 zeta were hydrophilic proteins without transmembrane helical region and signal peptide,and their secondary structure mainly consisted ofα-helices.Phylogenetic tree analysis showed that Bt14-3-3 epsilon was clustered into one cluster with 14-3-3 epsilon proteins of Nilaparvata lugens,Cimex lectularius and Halyomorpha halys,sharing higher homology,while Bt14-3-3 zeta was more closely related to 14-3-3 zeta of N.lugens.RT-qPCR results showed that Bt14-3-3 epsilon and Bt14-3-3 zeta had higher expression levels in the egg,female adults and abdomen of female adults of B.tabaci MED.【Conclusion】The full-length sequence,characteristics of the coded proteins and spatiotemporal expression of two subtypes of the 14-3-3 gene of B.tabaci MED have been clarified.The results of this study provide a basis

关 键 词：烟粉虱 基因克隆 14-3-3蛋白 生物信息学 时空表达 

分 类 号：Q966[生物学—昆虫学]