摘要
柑橘叶斑驳病毒(Citrus leaf blotch virus,CLBV)在陕西省栽培猕猴桃中发生普遍。为监测CLBV发生情况,本研究建立了CLBV的实时荧光定量PCR(Real-time fluorescent quantitative polymerase chain reaction,RT-qPCR)检测方法。该方法特异性强,可准确检测目的病毒,标准曲线斜率为-3.378,决定系数R~2=0.997 9,扩增效率为97.7%,比普通RT-PCR灵敏度高100倍,可用于猕猴桃植株CLBV的批量检测或低丰度病毒样本(如猕猴桃休眠枝条)的检测。为苗木携带CLBV病毒的早期诊断、果园病毒病预测预报和防控奠定了基础。
During the virus detection of kiwifruit cultivars in Shaanxi province,Citrus leaf blotch virus was found to occur at high frequencies. In order to detect CLBV accurately,a specific RT-qPCR assay based on SYBR Green I fluorescent dye was established. The results showed that the RT-q PCR assay had good specificity and sensitivity for CLBV detection. The slope of the standard curve and determination coefficient R~2 were-3.378 and 0.997 9,and the amplification efficiency reached 97.7%,which was 100 times more sensitive than conventional RT-PCR. This method was suited to batch inspection of field samples and low-abundance virus samples( such as dormant shoots of kiwifruit). The RT-qPCR assay laid the foundation for the early diagnosis of CLBV-infected seedlings,forecast and control of CLBV in orchard.
引文
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