| 土壤微生物多样性研究的新方法 |
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| 引用本文： | 马万里,Josquin Tibbits,Mark Adams. 土壤微生物多样性研究的新方法[J]. 土壤学报, 2004, 41(1): 103-107 |
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| 作者姓名： | 马万里 Josquin Tibbits Mark Adams |
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| 作者单位： | 1. 内蒙古师范大学生物系,生物多样性保护研究中心,呼和浩特,010022
2. Forest Science Center, University of Melboume,Creswick, VIC3363,Australia |
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| 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 RFLP已成为国外深入研究土壤微生物多样性的理想方法之一
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| 关 键 词： | 土壤微生物多样性 16S核糖体DNA 真菌核糖体DNA的ITS TRFLP分析 |
| 收稿时间： | 2002-10-21 |
| 修稿时间： | 2003-05-10 |
A NEW METHOD FOR RESEARCH ON SOIL MICROBIAL DIVERSITY |
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| Ma Wanli,Josquin Tibbits and Mark Adams. A NEW METHOD FOR RESEARCH ON SOIL MICROBIAL DIVERSITY[J]. Acta Pedologica Sinica, 2004, 41(1): 103-107 |
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| Authors: | Ma Wanli Josquin Tibbits Mark Adams |
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| Affiliation: | Department of Biology, Inner Mongolia Normal University, Hohhot 010022, China;Forest Science Center, University of Melbourne, Creswick, VIC 3363, Australia;Forest Science Center, University of Melbourne, Creswick, VIC 3363, Australia |
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| Abstract: | 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 |
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| Keywords: | Soil microbial diversity 16S ribosomal DNA ITS of r ibosomal DNA T RFLP |
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