库尔勒香梨上ACLSV的RT—PCR检测

RT-PCR Detection of ACLSV in Korla's Xiangli Pear Variety

以库尔勒香梨叶片和皮层为材料通过改进提取方法提取总RNA,获得了较完整的RNA,在此基础上进行了反转录和PCR扩增。并进行了库尔勒香梨上ACLSV(Apple chlorotic leaf sopt virus)的RT-PCR检测,建立了该病毒的RT-PCR检测体系。用此体系扩增得到了ACLSV一个长约为358bp的片段。

In order to obtain high purity and integrality of RNA from the fresh leaf and bark of Korla's Xiangli pear variety infected with ACLSV(Apple chlorotic leqf sopt virus) , the extraction was improved by added 1% CTAH, 2% PVP, 2% 2-mercaptoethanol (added just before use) to the extraction buffer and lowed the concentration of ethanol to deposit amylose. Using total RNA as template, reverse transcriptase-polymerase reaction (RT-PCR) was performed, and RT-PCR detection technology of ACLSV in Korla's Xiangli pear variety was reported in this paper. PCR amplification conditions were optimized , and the optimum RT-PCR system of ACLSV was developed. A 358 bp nucleotide fragment of ACLSV was obtained by using the optimum system.