表达病毒CP基因的转基因南方烟草的攻毒试验

Challenge Inoculation Tests with Viruses on the Southern Transgenic Tobacco Expressed CMV,TMV CP Genes

利用抗 Ｃ Ｍ Ｖ、 Ｔ Ｍ Ｖ 的抗血清对转 Ｃ Ｍ Ｖ、 Ｔ Ｍ Ｖ Ｃ Ｐ 基因的南方烟草的６ 个株系即：２ Ｔ６ ，２ Ｔ２０ ，２ Ｔ２５ ，２ Ｃ３１ ， ２ Ｃ３２ 和２ Ｔ Ｃ１００ 进行了 Ｅ Ｌ Ｉ Ｓ Ａ 和 Ｗｅｓｔｅｒｎ ｂｌｏｔ 分析， 证明了外源基因已在转基因烟草中得到表达． 经组织培养技术， 将表达外源基因的转基因烟草进行快繁， 然后进行温室的攻毒试验， 收集各抗病株系转基因烟草的套袋自交种子， 进行 Ｆ１ 代植株的大田攻毒试验． 温室和大田攻毒试验都表明， 转 Ｃ Ｍ Ｖ、 Ｔ Ｍ Ｖ Ｃ Ｐ 基因的烟草能够抵抗病毒的侵袭， 且转 Ｔ Ｍ Ｖ Ｃ Ｐ 基因烟草的抗病能力较好， 可能是植物体内 Ｔ Ｍ Ｖ Ｃ Ｐ

ELISA and Western Blot analyses showed that foreign CMV, TMV CP genes expressed in six lines of southern transgenic tobacco KY 14, i e ,2T6, 2T20, 2T25, 2C31, 2C32 and 2TC100, where T means the transformation with TMV CP gene and C with CMV CP gene by the use of antiserum against CMV,TMV CP The R 0 transgenic tobacco expressed CP genes were propagated with plant tissue culture and then challenge inoculated with CMV or/and TMV in greenhouse, according the transformation with CMV or/and TMV CP genes Self crossed seeds were collected from transgenic tobacco that were resistant against viruses and their F 1 progeny plants were challenge inoculated with viruses in field in the same way as in greenhouse These challenge inoculation tests demonstrated that transgenic plants with CMV, TMV CP genes were resistant to infection by viruses and the ability of resistance against virus infection in TMV transgenic tobacco was better than that in CMV transgenic tobacco, which may be caused by the larger synthesis of TMV CP in TMV transgenic plants