酵母pac1基因介导对葡萄B病毒的抗性

为明确广谱性抗病毒基因—酵母pac1基因对葡萄B病毒(Grapevine virus B,GVB)的抗性效果,通过农杆菌介导的遗传转化,将pac1基因导入西方烟37B,对转基因植株进行PCR鉴定及Southern blot分析,通过病毒摩擦接种观察症状以及实时荧光定量RT-PCR检测植株体内病毒含量,并对转基因植株抗病性进行初步鉴定。结果表明,目的基因pac1成功导入并整合至西方烟37B基因组,共获得10个转基因株系。不同株系的T1代烟草中阳性植株比例为16.7%~72.7%,表明目的基因可成功遗传到子代。接种病毒后转基因植株普遍延迟发病,但后期症状与非转基因对照相似,其中仅1个转基因株系B6具有不表现典型症状等抗性反应。接种植株病毒含量检测中,所有转基因植株均检测到病毒存在,但表现为抗病的B6株系中病毒含量显著低于非转基因对照,表明该转基因植株虽不能完全抵抗GVB侵染,但对GVB具有耐病性。

Resistance to Grapevine virus B mediated by Schizosaccharmyces pombe pac1 gene

To determine the resistance effect of a broad-spectrum antiviral gene (Schizosaccharmyces pombe pac1) to Grapevine virus B (GVB), the pac1 gene was introduced into Nicotiana occidentalis 37B via Agrobacterium tumefaciens-mediated transformation. The transgenic plants were analyzed by PCR and Southern blot. The resistance of transgenic plants was preliminarily evaluated by symptom observation and virus concentration analysis in inoculated transgenic plants by real-time quantitative RT-PCR. The results showed that the target gene was successfully introduced and integrated into the tobacco genome, and ten transgenic lines were obtained. 16.7%-72.7% T₁ plants in different transgenic lines were positive by PCR detection, indicating that the target gene was successfully inherited by the progeny. After inoculation, the viral symptom development was delayed in most transgenic plants, but the symptom was similar with that of the non-transgenic control in the later stage. Only one transgenic line (B6) showed resistance without typical symptoms. GVB could be detected in all inoculated transgenic plants, but the virus concentration in transgenic line B6 was significantly lower than that of non-transgenic controls. The results indicated that the transgenic tobacco B6 showed a tolerant resistance to GVB although it could not entirely resist GVB infection.