改良RNA提取法及樱桃PDV和PNRSV的RT-PCR检测(英文)

Modified RNA extraction from field woody plants for the routine detection of PDV and PNRSV in cherry by RT-PCR

介绍了一种从木本植物组织中获得高质量RNA的快速、简单和高效的核酸提取方法。该方法是基于核酸的二氧化硅捕获,避免了使用苯酚、氯仿等有机溶剂。利用该方法从樱桃组织中提取的总RNA用RT-PCR技术检测PDV,PNRSV均获得成功。从感病植株的一年生枝的叶片、韧皮部及芽组织中扩增出了预期的目的片段,即172和449bp,而健康组织中无此扩增带。该法提取的总RNA用于RT-PCR技术检测,其敏感性至少与商业出售的QiagenRNeasy提取试剂盒相当,但简单经济。

An efficient and effective procedure for the extraction of high-quality RNA from woody plants without the use of phenol,organic solvents,or alcohol precipitation is described,which is based on silica capture.The method described has been successfully used for the detection of PDV and PNRSV in cherry by RT-PCR assay using DNA primers for the viral coat protein region.The expected sizes of the amplified products were 172 and 449 bp.Samples from bark,leaves and buds were used.Detection of viral RNA in samples of total plant RNA prepared using this method was found to be as sensitive as the methods previously described using the commercially available Qiagen's RNeasy extraction kit.