桃树李矮缩病的RT-PCR检测

比较了4种桃叶片RNA提取方法,建立了RT-PCR桃李矮缩病检测体系。结果表明凯基TRIzol试剂法简便快捷,提取的RNA质量较好,可用于RT-PCR。RT-PCR最适反应体系为MgCl21.5 mmol/L,引物0.5μmol/L,模板1.0μg/50μL,退火温度为43℃。采集桃主产区北京、陕西、山东、河北、河南、新疆以及江苏的徐州、无锡、南京等地带叶芽的桃枝样品作为试材,利用RT-CR进行李矮缩病检测。结果显示,在北京大久保、山东春雪、陕西美引15#和江苏南京霞晖7号上发现PDV。

Detection of PDV in Peach by RT-PCR

The RNAs of peach leaves were extracted by 4 kinds of methods.RT-PCR detection system was established.As a result,a rapid and simple method-Kaiji TRizol kit was selected for the extraction of total RNA from peach.The RNA can be used for RT-PCR.The best RT-PCR system is MgCl2 1.5 mmol/L,primer 0.5 μmol/L,cDNA 1.0 μg/50μL,anneal temperature 43 ℃.The peaches come from Beijing,Shaanxi,Shandong,Hebei,Henan,Xinjiang and Jiangsu province were detected by RT-PCR.PDV was found in Beijing Okubo,Shandong Chunxue,Shaanxi Meiyin 15# and Nanjing Xiahui 7#.