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为了探讨SYBR Green Ⅰ实时定量RT-PCR技术在甜樱桃病毒粒子定量分析中的应用前景,以复合感染李属坏死环斑病毒(Prunus necrotic ringspot virus,PNRSV)、李矮缩病毒(Prune dwarf vi-rus,PDV)、樱桃病毒A(Cherry virus A,CVA)、樱桃小果病毒-2(Little cherry virus-2,LChV-2)的甜樱桃“红灯”Prunus avium cv.Red Lamp植株为研究对象,采用相对定量方法,分析各病毒的外壳蛋白基因的表达,用以指示病毒的增殖量。在花、幼叶、功能叶、衰老叶中均能检测到4个基因,但各基因表达量在各器官中存在差异。PNRSV-CP与CVA-CP表达模式相似,功能叶中明显高于其它器官,衰老叶中急剧降低。PDV-CP与LChV2-CP表达模式类似,幼叶中的表达量较高,功能叶片中较低。PNRSV-CP在花、功能叶中的表达显著高于其它3个病毒基因。LChV2-CP在各器官中的表达量均低于其余3个基因。该方法适用于植物组织内多种甜樱桃病毒增殖量的分析。
In order to explore the application of SYBR Green Ⅰ real-time quantitative RT-PCR in the quantitative analysis of sweet cherry virions, we used Prunus necrotic ringspot virus (PNRSV), Prune dwarf vi- rus, PDV), Cherry virus A (CVA), Cherry blight-2 (LChV-2) The subjects, the relative quantitative method, the analysis of the coat protein gene expression of each virus to indicate the proliferation of the virus. Four genes were detected in flowers, young leaves, functional leaves and aged leaves. However, the expression of each gene differed among organs. PNRSV-CP and CVA-CP expression pattern is similar, functional leaves were significantly higher than other organs, the sharp decline in the aging leaves. The expression patterns of PDV-CP and LChV2-CP were similar, with higher expression in young leaves and lower in functional leaves. The expression of PNRSV-CP in flowers and functional leaves was significantly higher than the other three viral genes. The expression levels of LChV2-CP in various organs were lower than the other three genes. The method is suitable for the analysis of proliferation of sweet cherry viruses in plant tissues.