稻田土壤DNA的提取及nifA基因的PCR扩增  

Extraction of DNA from Soils of Paddy Fields and PCR Amplification of nifA Gene

作　　者：魏志琴[1] 宋培勇[1] 杨秀荣[1] 马莉莉 

机构地区：[1]遵义师范学院生物系,贵州遵义563002 [2]湄潭求是高级中学,贵州湄潭564100

出　　处：《贵州农业科学》2011年第11期126-128,132,共4页Guizhou Agricultural Sciences

基　　金：贵州省科学技术基金项目"贵州石漠化环境中宏基因组文库的构建"(20082103)

摘　　要：为弄清茅贡米土壤微生物群落结构和生物固氮等代谢活动机制,采用试剂盒、SDS高盐和土壤预处理+SDS高盐3种土壤DNA提取方法,以从贵州茅贡米基地采集的土样提取的宏基因组DNA为模板,进行nifA基因的PCR扩增,比较了3种土壤DNA提取方法的效果。结果表明,试剂盒提取的DNA扩增到约200bp和900bp的nifA基因片段,SDS高盐法、预处理+SDS高盐法提取的DNA仅扩增到约200bp的nifA基因片段,土壤预处理对提高DNA纯度有一定的效果;3种方法都可以从土壤中提取到约23kb大小的DNA片段,但DNA得率和纯度存在一定的差异。Three methods of DNA extraction from soils of paddy fields for production of Maogong rice variety were compared by PCR amplification of nifA gene taking DNA of metagenome as the template to study the community structure of soil microorganism and metabolic mechanism of biological nitrogen fixation.The results showed that the nifA gene segments with 200 bp and 900 bp could be amplified from the extracted DNA by a kit,but the nifA gene segments with 200 bp were amplified from the extracted DNA by SDS and pre-treatment ＋ SDS methods only.The soil pretreatment had the certain effect on improvement of DNA purity.The DNA segments with about 23 kb were extracted from soils of paddy fields for production of Maogong rice variety by three tested extraction methods,but there was difference in DNA acquirement rate and purity between three extraction methods.

关 键 词：茅贡米 稻田土壤 DNA提取方法 宏基因组 NIFA基因 聚合酶链式反应 

分 类 号：S151.93[农业科学—土壤学]