番茄中受TYLCV诱导上调表达基因tetraspanin3的功能分析

[目的]本文旨在研究tetraspanin3基因在抗番茄黄化曲叶病毒(TYLCV)病中的作用及对番茄生长发育的影响.[方法]通过MEG 5.0软件构建进化树,分析tetraspanin3基因的进化;采用实时荧光定量PCR(RT-qPCR)检测tetraspanin3在番茄各个器官中的表达量及对TYLCV诱导的响应;通过病毒诱导基因沉默技术(VIGS)将番茄中的tetraspanin3沉默,测定基因沉默后番茄的抗病性并观测沉默植株表型.[结果]构建的进化树显示:番茄的tetraspanin3基因与葡萄同源基因在一个独立的进化分支上.tetraspanin3基因在各器官中均有表达,在茎、老叶、花中表达量较高,在根、果实中表达量较低,在新叶中的表达量处于中间水平.tetraspanin3受TYLCV诱导后上调表达.沉默tetraspanin3后的植株出现了叶片扭曲,并且在接种TYLCV后,TYLCV病毒量降低50%以上.[结论]tetraspanin3与番茄叶片的生长发育有关,并且对TYLCV侵染番茄植株起正调控作用.
[Objectives]The research was conducted to study the effect of tetraspanin3 on the resisitance to Tomato yellow leaf curl virus(TYLCV)and development of tomato. [Methods]The phylogenetic tree of tetranspnin3 was constructed by the MEG 5.0 software to analyse the evolution of tetraspanin3;the expression pattern of tetraspanin3 in different organs and the response of tetraspanin3 during TYLCV infection were determined by RT-qPCR;the virus induced gene silencing(VIGS)technology was used to silence the tetraspanin3 gene and the resistance to TYLCV was tested in silenced plant and the phenotypes of silenced plants was watched. [Results]tetraspanin3 and homologous gene in grape were in a distinct branch;tetraspanin3 gene was expressed in all organs,the higher expression in stem,the old leaf,flower and the lower expression in root,fruit,and the expression in the new leaf was in medium level,tetraspanin3 gene was up-regulated during TYLCV infection. The tetraspanin3-silenced plant had distorted leaves,and had an over 50% decline in the quantity of TYLCV after vaccinating the TYLCV. [Conclusions]tetraspanin3 gene has an effect on the development of tomato leaves and acts as a positive regulator of the infection of TYLCV to tomato plants