番茄双链RNA绑定蛋白(SlDRB)基因家族鉴定及抗TYLCV防御反应分析  

Identification of Double RNA Binding Protein(DRB)Gene Family and Analysis of Defense Response to TYLCV in Tomato

作　　者：黄鑫 方远鹏 岳宁波 张龙 吴丹 李云洲 HUANG Xin;FANG Yuanpeng;YUE Ningbo;ZHANG Long;WU Dan;LI Yunzhou(College of Agriculture,Guizhou University,Guiyang,Guizhou 550025)

机构地区：[1]贵州大学农学院,贵州贵阳550025

出　　处：《核农学报》2022年第2期291-301,共11页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金项目(32060679);贵州大学培育项目(贵大培育[2019]52号、黔科合平台人才[2017]5788-28);贵州大学人才引进科研项目(贵大人基合字[2017]50号)。

摘　　要：番茄生产是现代蔬菜生产的主导产业,因经济价值高,成为乡村振兴的优势产业。然而番茄生产大多面临番茄黄化曲叶病毒(TYLCV)和其他病毒的严重威胁。最新研究发现双链RNA结合蛋白(DRB)在植物RNA干扰(RNAi)抗病毒通路中发挥重要作用。为探究番茄DRB(SlDRB)基因抗TYLCV防御反应,本研究通过生物信息学方法鉴定并分析番茄DRB基因的基础性质,包括DRB基因家族进化树、基因结构和基因染色体上的位置、保守基序、组织特异性表达等,同时利用实时荧光定量(qRT-PCR)技术检测SlDRB基因家族对TYLCV的防御反应。结果表明,番茄含有8个SlDRB基因家族成员,分布于6条染色体上,其中3个SlDRB基因位于1号染色体。番茄DRB家族可分出3个亚族,即DRB2/3、DRB6/7和DRB1/4/5/8,不同的亚族内含子数量和蛋白结构分布有所差异。亚细胞定位结果表明SlDRB1、SlDRB5、SlDRB6、SlDRB7和SlDRB8定位于胞外,SlDRB2和SlDRB3定位于细胞质,SlDRB4定位于细胞核。不同SlDRB基因的组织特异性表达差异明显,但是在叶片组织中大多数SlDRB基因表达相对适中,在根、花、果组织中多数基因表达水平相对较高。所有SlDRB基因在番茄受TYLCV侵染后表达水平均有升高,根据表达趋势分为四类:第一类基因SlDRB7、SlDRB8,表达水平持续上调;第二类基因SlDRB2、SlDRB3,在病毒接种第7天表达上调,在第14天下调;第三类基因SlDRB1、SlDRB4,仅在病毒侵染7 d表达上调;第四类基因SlDRB5、SlDRB6,仅在病毒侵染14 d表达上调。本研究在番茄上鉴定了8个SlDRB基因,并明确了该基因参与植株抗病毒的防御反应,为研究SlDRB在番茄RNAi抗病毒中的功能与作用奠定了基础。Tomato is the leading industry of modern vegetables,due to its high economic value,it is also an advantageous industry for rural revitalization.However,tomato production has faced a serious threat from Tomato yellow leaf curl virus(TYLCV).RNA interference(RNAi)is an important antiviral mechanism in plants.Recent studies indicated that double-stranded RNA-binding proteins(DRB)played an important role in the plant RNAi pathway.In order to the identificate of tomato DRB genes defense responses against TYLCV.In this study,bioinformatics methods were used to identify and analyze the basic properties of tomato DRB genes,including the evolutionary tree of DRB gene family,gene structure and gene location,conservative motifs,tissue-specific expression,etc.In addition,real-time fluorescent quantitative PCR(Quantitative Real-time PCR,qRT-PCR)technology was used to detect the expression of tomato DRB(SlDRB)gene family after infected with TYLCV.The results showed that tomato contains 8 SlDRB genes,and those 8 members are distributed on six chromosomes,of which 3 SlDRB genes are on chromosome 1.Tomato DRB family can be divided into 3 subfamilies,namely DRB2/3,DRB6/7,DRB1/4/5/8,and the number of introns and protein structure distribution of different subfamilies are different.The subcellular localization results showed that SlDRB1,SlDRB5,SlDRB6,SlDRB7 and SlDRB8 were localized in the extracellular,SlDRB2 and SlDRB3 were localized in the cytoplasm and SlDRB4 in nucleus.More importantly,these SlDRB genes were tissue-specific expressed obviously.The expression level in leaf tissues was relatively moderate,and most genes have the highest expression levels in root,flower,and fruit tissues.The expression level of all SlDRB genes were increased after plants infected with TYLCV.According to the expression trend,these genes can be divided into four categories.The first type of genes,SlDRB7 and SlDRB8,had continually increased expression.The second type of genes SlDRB2 and SlDRB3,the expression of which were up-regulated at 7 days after

关 键 词：番茄 抗病毒 双链RNA结合蛋白(DRB) 生物信息学 表达分析 

分 类 号：S436.412.1[农业科学—农业昆虫与害虫防治]