Laodephax striatellus could not only damage rice directly,but also works as media to spread Rice Stripe Disease and Rice Black Streaked Dwarf Disease.To explore techniques for controlling Laodephax striatellus and rice virus diseases caused by it,RNAi vector was efficiently and rapidly constructed by a two-step method.One pair specific primer was de-signed for RACK gene from Laodephax striatellus to amplify 200 bp interference fragment,and then cloned it into entry vec-tor pENTR.Then the RNAi fragment was recombined into destination vector pANDA through LR reaction to form RNAi vector pANDA-RACK,which was transformed into rice cultivar Wulingjing No.1 mediated by Agrobacterium tumefaciens EHA105.Twenty transgenic plants were generated and identified by PCR,which plays a foundation against Laodephax stria-tellus and its transmitted virus diseases in rice.%灰飞虱不仅直接危害水稻,还是水稻条纹叶枯病和黑条矮缩病的传播媒介.为了探索防治灰飞虱及由灰飞虱引起的水稻病毒病的方法,本研究利用两步法快速高效构建灰飞虱RNAi载体,并应用于水稻遗传转化.针对灰飞虱的激活性蛋白激酶C受体( RACK)基因设计了一对特异性引物,通过PCR扩增一段200 bp的干扰片段,将干扰片段克隆到入门载体pENTR,再经过LR反应使入门载体上的干扰片段整合入目的载体pANDA,形成RNAi载体pANDA-RACK,通过农杆菌EHA105介导的水稻转化法转入武陵粳1号,获得20株经鉴定为阳性的转基因植株,为防控灰飞虱及其传播的水稻病毒病提供了珍贵材料.
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