山东农业科学2012，Vol.44Issue(2)：9-14,6.
玉米叶面微生物DNA提取方法及PCR引物的筛选
Screening of DNA Extraction Methods and PCR Primers for Microorganisms from Maize Leaves
摘要
Abstract
Traditional isolation and culture method for microorganisms had been found to have limitations in reflecting the whole genome information of phyllospheric microorganisms and have been gradually replaced by molecular ecology methods. The high - quality, large - fragment and unbiased DNA is the important basis for studying phyllospheric microorganisms at molecular level. In this paper, four methods, improved Sambrook method, CTAB method, improved chemical method and Kit method, were adopted respectively to extract the total DNA of microorganisms from maize leaves in early, middle and late growth periods and then the size and output of DNA fragments were evaluated synthetically. The 16S rDNA were amplified by primers 357/518 and 984/1387, meanwhile, the primers NS1/fungi, ITS1 -F/ITS2, NS1/AM1, EF4/fung5 and SSU -0817/ SSU -1196 were used for the amplification of 18S rDNA. The results showed that all of the four methods could get appropriate fragments for PCR, but the output and purity of DNA were different between different methods and the best was Kit method. Except the improved chemical method, the other methods could get products of PCR. The primers 984/1387 and ITS1 - F/ITS2 were the best for amplifying 16S rDNA and 18S rDNA re-spectively.关键词
玉米/叶面微生物/DNA提取/PCR/16SrDNA/18SrDNAKey words
Maize/ Phyllospheric microorganisms/ DNA extraction/ PCR/ 16S rDNA/18S rDNA分类
生物科学引用本文复制引用
赵辉,李世国,孙红炜,李凡,田晓燕,颜世磊,路兴波..玉米叶面微生物DNA提取方法及PCR引物的筛选[J].山东农业科学,2012,44(2):9-14,6.基金项目
农业部转基因生物新品种培育重大专项(2009ZX08011-020B) （2009ZX08011-020B）
