摘要
传统的分离培养和鉴定土壤微生物方法所具有的困难性和局限性 ,是造成难以深入了解土壤微生物生态学特性和多样性组成方面的主要障碍。本文运用分子生物学技术 ,以澳大利亚两种主要森林类型的土壤微生物多样性研究为实例 ,介绍了从土壤中直接提取土壤微生物DNA的方法以及末端限制性酶切片段长度多态性 (T RFLP)分析的基本原理和方法。作者认为 ,用该方法提取的土壤真菌DNA的纯度高 ,完全适合PCR扩增和T RFLP分析的要求。T
The inability to culture most microbe from soil samples is a fundam ental obstacle to understanding soil microbial ecology and diversity Therefore , amplification of DNA from soil and a rapid,preferable DNA extraction method in v olving necessary purification is needed for rapid and thorough analysis of micro bial diversity in environmental samples The method extracting fungi DNA that i s applicable to two kinds of forest soil types in Australia is developed in this research,and result showed that extracted DNA purity and size is reasonable an d suitable for PCR amplification At the same time,a framework for soil microb ia l diversity analysis with T RFLP (Terminal restriction fragment length polymorp h ism),a recent molecular approach that can be used to assess subtle genetic dif f erences between DNA samples as well as provide insight into the structure and fu nction of microbial communities in soils,is presented here Generally,PCR a mpl ification of extracted DNA sample from soil is conducted by using a set of fluor escent tagged specific primers of the bacterial 16S rDNA or ITS of fungal rDNA, and then products are digested with a four base restriction endonuclease,as ma x imizing the resulting number of terminal fragments Finally the entire terminal fragments are detected and sized by the ABI automated sequencer The technique h as both high sensitivity and throughput making it an ideal choice for comparativ e community analyses
出处
《土壤学报》
CAS
CSCD
北大核心
2004年第1期103-107,共5页
Acta Pedologica Sinica
