【作者】 牛建新； 刘连科；

【Author】 Niu Jinxin and Liu Lianke ( Department of Horticulture & Forestry Engineering, Agricultural college of Xinjiang Shihezi University, Xinjiang Shihezi 832003)

【机构】 石河子大学农学院园艺园林工程系；

【摘要】 以苹果新鲜叶片和皮层为材料,改进总RNA的提取方法,获得纯度高、完整性好的总RNA,在此基础上进行了反转录和PCR扩增。检测到了苹果褪绿叶斑病毒(ACLSV)和苹果茎痘病毒(ASPV),建立了ACLSV、ASPV的RT-PCR检测程序,并且得到应用。更多还原

【Abstract】 In order to obtain high purity and integrality total RNA from fresh leaf and bark of apple infected with apple chlorotic leaf spot vims (ACLSV) and apple stem pitting virus (ASPV) , the extraction was improved. PVP powder was added to when leaf or bark was powered in order to eliminate hydroxyben-zene, and CTAB/NaCl solution was used to deposit amylose. Using total RNA as template, reverse tran-scriptase - polymerase reaction (RT - PCR) was performed. PCR amplification conditions were optimized, and the optimum RT - PCR systems of two viruses were developed. A 358bp fragment of ACLSV was obtained based on the optimum system and a 316bp fragment of ASPV.更多还原