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以茄子花叶病病叶为试材,经差速离心结合聚乙二醇(PEG—MW6000)沉淀法抽提病毒粒子,SDS-酚法提取病毒RNA,通过逆转录酶合成互补DNA(cDNA)。以此cDNA-RNA 杂交体为模板,用一对烟草花叶病毒特异性引物进行多聚酶链反应(Polymerase Chain Reaction;PCR),扩增产物经1%agarose-EB 电泳检测,产生了±200bp 的特异性核酸片断。从而表明沈阳地区茄子花叶病系由烟草花叶病毒引起,是我国茄子病毒病新的毒源。同时,通过实验证实,PCR 是一种敏感、快速、特异性强的植物病毒检测手段。
Virus particles were extracted by differential centrifugation and polyethylene glycol (PEG-MW6000) precipitation method. Virus RNA was extracted by SDS-phenol method and complementary DNA (cDNA) was synthesized by reverse transcriptase. . Using this cDNA-RNA hybrid as a template, Polymerase Chain Reaction (PCR) was performed with a pair of tobacco mosaic virus-specific primers. The amplified product was detected by 1% agarose-EB electrophoresis to produce a ± 200 bp specific Nucleic acid fragments. This indicated that the eggplant mosaic disease in Shenyang is caused by tobacco mosaic virus, which is a new source of eggplant virus disease in our country. At the same time, experiments confirmed that PCR is a sensitive, rapid and specific plant virus detection method.