烟粉虱MED隐种14-3-3基因克隆及时空表达谱分析

doi:10.16380/j.kcxb.2024.05.001

昆虫学报 ›› 2024, Vol. 67 ›› Issue (5): 595-602.doi: 10.16380/j.kcxb.2024.05.001

• 研究论文 • 下一篇

烟粉虱MED隐种14-3-3基因克隆及时空表达谱分析

韦焕文^1,2,3, 王培^2,3, 陈建斌^2,3, 杜娇^2,3, 张德咏^1,2,3, 刘勇^1,2,3,*, 史晓斌^1,2,3,*

(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)

出版日期:2024-05-20 发布日期:2024-06-20

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

WEI Huan-Wen^1,2,3, WANG Pei^2,3, CHEN Jian-Bin^2,3, DU Jiao^2,3, ZHANG De-Yong^1,2,3, LIU Yong^1,2,3,*, SHI Xiao-Bin^1,2,3,*

(1. Longping Branch, College of Biology, Hunan University, Changsha 410125, China; 2. The Yuelushan Laboratory, Changsha 410125, China; 3. Institute of Plant Protection, Hunan Academy of Agricultural Sciences, Changsha 410125, China)

Online:2024-05-20 Published:2024-06-20

摘要/Abstract

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。

关键词: 烟粉虱, 基因克隆, 14-3-3蛋白, 生物信息学, 时空表达

Abstract: 【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. tabaciMED 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 of α-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 for subsequent studies on the molecular function of 14-3-3 proteins.
Key words: Bemisia tabaci; gene cloning; 14-3-3 protein; bioinformatics; spatiotemporal expression

韦焕文, 王培, 陈建斌, 杜娇, 张德咏, 刘勇, 史晓斌. 烟粉虱MED隐种14-3-3基因克隆及时空表达谱分析[J]. 昆虫学报, 2024, 67(5): 595-602.

WEI Huan-Wen, WANG Pei, CHEN Jian-Bin, DU Jiao, ZHANG De-Yong, LIU Yong, SHI Xiao-Bin. Cloning and spatiotemporal expression profiling of the 14-3-3 genes in Bemisia tabaci MED (Hemiptera: Aleyrodidae)[J]. Acta Entomologica Sinica, 2024, 67(5): 595-602.

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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)

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