番茄抗黄叶卷曲病相关基因Ty-6的克隆、序列分析及表达特性

doi:10.13560/j.cnki.biotech.bull.1985.2019-1042

摘要/Abstract

摘要： 进一步从分子水平探究番茄黄叶卷曲病,扩展其抗病基因库。通过克隆番茄黄叶卷曲病相关基因Ty-6,利用生物信息学方法对其编码蛋白一级结构、二级结构及三级结构进行序列分析,构建进化树,最后通过感染黄叶卷曲病毒,检测Ty-6基因在番茄不同时期不同组织中的表达特性。Ty-6基因共编码305个氨基酸,其编码产物属于碱性蛋白,二级结构以无规则卷曲为主,预测蛋白定位于细胞质膜上;且含有51个磷酸化位点与39个糖基化位点;启动子区主要以TATA-box为主,且含多个光响应、茉莉酸、水杨酸以及脱落酸等诱导基因表达的元件;系统发育分析结果表明其与马铃薯的亲缘关系最近;q-PCR显示在感染病毒后叶中该基因的表达量明显高于对照,且具组织表达特异性。Ty-6基因的研究不仅扩展了育种者可用的Ty基因工具包,也提供了一种抗病的可行思路。

关键词: 番茄, 黄叶卷曲病, Ty-6, 生物信息学, 表达特性

Abstract: This work aims to further explore the yellow leaf curl disease of tomato at the molecular level and expand its disease resistance gene pool. The experiment was carried out by cloning the gene Ty-6 related to tomato yellow leaf curl disease,then using bioinformatics to conduct sequence analysis of its primary,secondary and tertiary structures,further constructing a phylogenetic tree,and finally detecting the expression characteristics of the gene Ty-6 in different tissues at different stages after infecting yellow leaf curl virus. The results showed that the gene Ty-6 encoded 305 amino acids,the encoded product belonged to the basic protein,and the secondary structure was dominated by random coils. The protein was predicted to be localized on the plasma membrane;and it contained 51 phosphorylation sites and 39 glycosylation sites. The promoter region was mainly TATA-box,and contained multiple elements of light response,jasmonic acid,salicylic acid and abscisic acid to induce gene expression. Additionally,phylogenetic tree results indicated that its affinity relation with potato was the closest. q-PCR results demonstrated that the expression level of this gene in the leaves after infection was significantly higher than that of the control group,and it expressed tissue specially. The results of studying gene Ty-6 not only expand Ty gene toolkit availability to breeders,and also provide a feasible approach of resisting disease.

Key words: tomato, yellow leaf curl disease, Ty-6, bioinformatics, expression characteristics

朱晓林, 魏小红, 王宝强, 王贤, 张朝阳. 番茄抗黄叶卷曲病相关基因Ty-6的克隆、序列分析及表达特性[J]. 生物技术通报, 2020, 36(7): 40-47.

ZHU Xiao-lin, WEI Xiao-hong, WANG Bao-qiang, WANG Xian, ZHANG Chao-yang. Cloning,Sequence Analysis and Expression Characteristics of Gene Ty-6 Related to Tomato Resistance to Yellow Leaf Curl Disease[J]. Biotechnology Bulletin, 2020, 36(7): 40-47.

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