【摘要】 将本室合成、克隆的马铃薯卷叶病毒（ＰｏｔａｔｏＬｅａｆｒｏｌＶｉｒｕｓ，ＰＬＲＶ）中国分离株的基因间隔区（ｉｎｔｅｒｇｅｎｉｃｓｅｑｕｅｎｃｅ，ＩＳ）双链ｃＤＮＡ以正、反向两种方式分别构建于转化载体ｐＲＯＫ２中，通过致瘤农杆菌介导，以马铃薯叶圆片为转化材料，转化马铃薯栽培品种Ｄｅｓｉｒｅ，获得了转基因植株。卡那霉素抗性分析和ＰＣＲ检测目的基因，证明ＰＬＲＶＩＳ双链ｃＤＮＡ已经整合到转基因马铃薯的染色体基因组中。将转基因植株移栽网棚用蚜虫接种ＰＬＲＶ，观察症状并用酶联免疫吸附测定（ＥＬＩＳＡ）检测转基因植株中ＰＬＲＶ含量。结果表明，表达ＰＬＲＶＩＳ正意和反意ＲＮＡ的转基因植株，接种病毒后表现无症状或症状轻微，ＰＬＲＶ平均滴度均较未转基因对照植株低。表达正意ＲＮＡ的转基因植株ＰＬＲＶ滴度降低４３％～７２％，表达反意ＲＮＡ的转基因植株ＰＬＲＶ滴度降低７２％～８６％，由此可见，表达ＰＬＲＶＩＳ反意ＲＮＡ的转基因马铃薯对ＰＬＲＶ抗性较强。更多还原

【Abstract】 The intergenic sequence (IS) cDNA of Potato Leafroll Virus (PLRV) Chinese isolate was constructed into transformation binary vector pROK2 in positive or negative direction, then introduced into the commercial potato cultivar Desiree via Agrobacterium tumefaciens mediated gene transfer. Kanamycin resistance, PCR amplification of transgenic lines showed that intergenic sequence of PLRV Ch was integrated into the genome of potato plants. The transgenic lines were transplanted into the screen house, then inoculated with PLRV by viruliferous Myzus persicae Sulz. After inoculation, the symptom development was observed and the virus titer was detected by ELISA. Results showed that the transgenic plants displayed slight or no symptom. The average PLRV titer in the transgenic lines was lower than the untransformed plants. The transgenic lines expressing sense RNA reduced PLRV titers about 43%～72% in comparison with untransformed plants, while the transgenic lines expressing antisense RNA reduced about 72%～86%. It showed that the antisense RNA confered higher resistance to PLRV.更多还原