论文部分内容阅读
用RT-PCR扩增黄瓜花叶病毒M株系(CMV-M)全长基因组cDNA,成功构建CMV-M RNA2和RNA3侵染性克隆后,与CMV-Fny基因组RNA交换得到3个假重组型病毒(F1M2F3、F1F2M3、F1M2M3)。用F1M2F3、F1F2M3、F1M2M3分别侵染白肋烟,产生坏死环斑、轻微绿斑驳、明脉、黄白化和叶尖线性化等症状。根据假重组型病毒和野生型病毒的表观症状,分析引起各种症状的关键因子,初步判定:CP基因是诱导花叶症状的关键因子,CMV-Fny RNA2是诱导叶尖线性化的关键因子,CMV-M RNA2是诱导叶尖坏死斑关键因子。实时荧光定量PCR结果显示:野生株CMV-M、CMV-Fny和假重组体F1M2F3、F1F2M3、F1M2M3侵染烟草后引起的症状差异与病毒基因组RNA累积没有直接关系。
The CMV-M full-length genomic cDNA was amplified by RT-PCR. CMV-M RNA2 and RNA3 infectious clones were successfully constructed and then exchanged with CMV-Fny genomic RNA to obtain three pseudotypes Virus (F1M2F3, F1F2M3, F1M2M3). Infection of burley tobacco with F1M2F3, F1F2M3 and F1M2M3, respectively, resulted in necrotic ring spot, mild green mottle, clear veins, yellowish whitening and tip linearization. Based on the apparent symptoms of pseudotype and wild-type virus, the key factors causing various symptoms were analyzed. It was initially determined that CP gene was the key factor inducing mosaic symptoms and CMV-Fny RNA2 was the key factor inducing tip linearization , CMV-M RNA2 is the key factor inducing necrosis of the tip. Real-time PCR results showed that there was no direct relationship between the symptoms of CMV-M, CMV-Fny wild-type and fake recombinant F1M2F3, F1F2M3 and F1M2M3 infected tobacco and the accumulation of viral genomic RNA.