携带eGFP的ZYMV侵染性克隆的构建及其侵染性

Construction of ZYMV infectious clone carrying eGFP and its infectivity

小西葫芦黄花叶病毒(zucchini yellow mosaic virus,ZYMV)是瓜类作物主要病毒之一,严重威胁瓜类产业的可持续发展。本研究在ZYMV甜瓜分离物CH-87侵染性克隆基础上,利用同源重组策略在NIb和CP编码序列之间插入了eGFP,命名为pXT1-ZYMV-eGFP。将pXT1-ZYMV-eGFP转化农杆菌并接种西瓜和甜瓜,结果发现pXT1-ZYMV-eGFP可以系统侵染西瓜和甜瓜,引起典型的花叶症状,在紫外灯下发病植株呈现绿色荧光。经摩擦接种证实,该克隆接种后所产生的病毒子代可以稳定的传播繁殖。研究表明携带eGFP的ZYMV侵染性克隆成功构建,可用于该病毒致病性及病毒与寄主互作的相关研究。

The disease caused by zucchini yellow mosaic virus(ZYMV)infection poses a serious threat to the sustainable production of important cucurbits(watermelon and melon)in China.In order to better promote the research on ZYMV pathogenicity and resistance,we used homologous recombination strategy to construct the eGFP-carrying infectious clone of ZYMV melon isolate CH-87,named pXT1-ZYMV-eGFP.The infectivity was tested on watermelon and melon plants by using agrobacterium-mediated method.The results showed that the clone could systemically infect watermelon and melon by inducing typical mosaic symptoms.Also,the infected plants were able to successfully express eGFP which made them fluorescent under UV light.Virus progeny produced from the clone was confirmed to be biologically active and could be mechanically transmitted to watermelon and melon plants.It suggests that ZYMV infectious cDNA clone carrying eGFP was successfully constructed,and it could be used in the study of viral pathogenesis and virus-host interactions.