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基于培养法和分子指纹技术的土壤细菌群落多样性调查

Diversity surveys of soil bacterial community by cultivation--based methods and molecular fingerprinting techniques.

作者信息

Luo Hai-feng, Qi Hong-yan, Zhang Hong-xun

机构信息

Department of Environ-Biotechnology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.

出版信息

J Environ Sci (China). 2004;16(4):581-4.

Abstract

By combining the cultivation methods with molecular fingerprinting techniques, the diversity surveys of soil bacterial community in 13 areas of China were carried out. The cultivable heterotrophic diversity was investigated by colony morphology on solid LB medium. Genetic diversity was measured as bands on denaturing gradient gel electrophoresis(DGGE) by the extraction and purification of the total soil DNA, and amplification of bacterial 16S rDNA fragments by polymerase chain reaction (PCR). The Shannon-Wiener indices of diversity (H), richness (S) and evenness (E(H)) were employed to estimate the diversity of soil bacterial community. The results showed that there was an obvious diversification existed in soil from the different areas. However, the genetic diversity estimated by PCR-DGGE can provide more comprehensive information on bacterial community than the cultivation-based methods. Therefore, it is suggested to combine the traditional methods with genetic fingerprinting techniques to survey and estimate soil bacterial diversity.

摘要

通过将培养方法与分子指纹技术相结合,对中国13个地区的土壤细菌群落进行了多样性调查。通过在固体LB培养基上的菌落形态研究可培养异养生物的多样性。通过提取和纯化土壤总DNA,并通过聚合酶链反应(PCR)扩增细菌16S rDNA片段,以变性梯度凝胶电泳(DGGE)上的条带为指标测量遗传多样性。利用香农-维纳多样性指数(H)、丰富度(S)和均匀度(E(H))来估计土壤细菌群落的多样性。结果表明,不同地区的土壤存在明显的多样性。然而,与基于培养的方法相比,通过PCR-DGGE估计的遗传多样性能够提供关于细菌群落更全面的信息。因此,建议将传统方法与遗传指纹技术相结合来调查和估计土壤细菌多样性。

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