转病毒移动蛋白及复制酶基因烟草的协生和重组风险分析

 对转化番茄花叶病毒 (ToMV)、烟草花叶病毒 (TMV)移动蛋白 (MP)基因和黄瓜花叶病毒P1株系(CMV P1)部分复制酶基因的 3种转基因烟草 ,分别接种马铃薯X病毒 (PVX)、马铃薯Y病毒 (PVY)、CMVRB株系 (CMV RB)和TMV。症状观察和ELISA测定发现 ,各病毒在转基因烟草和非转基因烟草上的症状及病毒浓度没有显著差异 ,表明这些病毒在转基因烟草上没有协生作用发生。对转化ToMVMP基因和转化CMV P1部分复制酶基因的转基因烟草 ,分别接种TMV和CMV RB ,定期进行生物学、ELISA和免疫捕获反转录PCR(IC RT PCR)检测 ,在 16个月的连续测定中未发现病毒重组现象。

Risk Assessment of Synergism and Recombination on the Transgenic Tobacco Plants Containing Viral Movement Protein and Replicase Genes

The transgenic tobacco plants transformed with movement protein genes of tomato mosaic virus (ToMV) and tobacco mosaic virus (TMV) and partial replicase gene of cucumber mosaic virus P1 isolate (CMV P1), respectively, were inoculated with PVX, PVY, TMV and CMV isolate RB, separately. Symptom observation showed that there were no differences in symptom between transgenic and non transgenic tobaccos. ELISA also illustrated that the virus concentrations in transgenic plants were similar to those in non transgenic tobaccos, indicating that no synergism is found in these plants. The transgenic tobacco plants with movement protein gene of ToMV or partial replicase gene of CMV P1 were inoculated with TMV and CMV RB, respectively. The local or systemic infected leaves were then used for elucidation of the possible virus recombination in transgenic plants with biological infectivity test, ELISA and immuno capture RT PCR. Within 16 months, no recombination has been found between transformed genes and inoculated virus genomes.