番木瓜eIF4E和eIFiso4E基因嵌合hpRNA载体一步快速构建及沉默效果的研究

One-step Rapid Construction and Gene Silencing Effect of Chimeric Hairpin RNA Vector of Papaya eIF4E and eIFiso4E

番木瓜环斑病毒(Papaya ringspot virus,PRSV)严重威胁番木瓜种植业的发展,且目前没有十分有效的防治办法。病毒侵染植物依赖寄主因子的协助,真核翻译起始因子4E(eukaryotic initiation factor 4E,eIF4E)是多种RNA病毒侵染植物的必需因子。以番木瓜eIF4E家族基因为研究对象,构建同时干扰其eIF4E和eIFiso4E基因的发卡RNA(hairpin RNA,hpRNA)载体,并将其导入到番木瓜叶肉原生质中。通过荧光实时定量检测发现,番木瓜中eIF4E和eIFiso4E基因的表达量分别下降了49.8%和67.6%,这为进一步研究番木瓜eIF4E家族基因对PRSV侵染的影响以及利用RNA干扰技术靶向植物基因的病毒防治新策略提供理论和实践依据。

The devastating disease caused by Papaya ringspot virus（PRSV） is a serious threat to the development of papaya industry. At present, no effective strategies can prevent PRSV. The infection of plant virus depends on the assistance of host factors. Eukaryotic initiation factor 4E（elF4E） has been found to involve in the interaction of plant and virus, and is a necessary factor for plants infecfed by a variety of RNA viruses. Based on it, a chimeric hairpin RNA （hpRNA） vector containing papaya elF4E and elFiso4E fragments was successfully constructed, and had been transferred into papaya mesophyll protoplast. After detected by realime RT-PCR, the elF4E and elFiso4E gene expression were decreased by 49.8% and 67.6%, respectively. The results will help to study the effect of elF4E family genes on the PRSV infection, and provide theoretical basis for the new strategy of anti-virus by targeting plant genes with RNAi technology.