苹果轮纹病菌DNA提取方法的比较  

Comparison of three DNA extraction methods from Botryosphaeria berengriana f. sp. piricola

作　　者：苏平[1,2] 周增强[2] 朱建兰[1] 侯珲[2] 王丽[2] 

机构地区：[1]甘肃农业大学草业学院,甘肃兰州730070 [2]中国农业科学院郑州果树研究所,河南郑州450009

出　　处：《甘肃农业大学学报》2010年第6期99-104,共6页Journal of Gansu Agricultural University

基　　金：公益性行业(农业)科研专项(nyhyzx07-055)

摘　　要：采用尿素提取法、氯化苄提取法以及SDS-CTAB提取法对苹果轮纹病菌的基因组DNA进行了提取和比较.结果表明,3种方法均能提取到苹果轮纹病菌的基因组DNA.其中,SDS-CTAB法效率最高,提取的DNA质量最好,但是耗时较长;尿素提取法操作简单快速,但是提取的DNA质量较低;氯化苄提取法提取的DNA纯度最低,蛋白质污染严重.将所提取的DNA经ITS试验检测与电泳检测,证明3种方法提取的DNA扩增效果良好,均可满足该真菌rDNAITS分析的要求.Three extraction methods,urea,benzyl chloride and SDS-CTAB,were applied to extract the genomic DNA from Botryosphaeria berengriana f.sp.piricola.The results showed that all the three methods were capable of extracting the genomic DNA of the pathogen.The SDS-CTAB method had the highest quality DNA extraction and showed the most efficient; however,consumed longest extraction time.The urea method was the simplest and most convenient procedure,but had a lower quality extraction.The benzyl chloride method had a lowest quality and the extracted DNA contained a lot of protein impurities.The rDNA ITS analysis and electrophoresis analysis showed that the DNA extracted by all the three methods has fairly good quality for DNA amplification and satisfied the requirements for rDNA ITS analysis of the Botryosphaeria berengriana f.sp.piricola.

关 键 词：苹果轮纹病菌 基因组DNA DNA提取 rDNAITS 

分 类 号：S436.611[农业科学—农业昆虫与害虫防治]