T_RFLP技术及其在硝化细菌群落分析中的应用

T_RFLP是建立在PCR基础上的,一种不依赖于传统培养方法的微生物生态学的研究方法.具有快速、灵敏的特点.自1997年首次被报道以来,T_RFLP技术已广泛应用于菌种鉴定、群落对比分析、群落中系统发育种群多样性的评估等领域,并成为环境微生物群落结构分析的强有力工具之一.目前T_RFLP在国内的应用较少,硝化细菌的群落分析上还未见报道.但作为一种研究微生物群落结构特征的理想方法,将会得到广泛地应用.本文主要介绍了T_RFLP的基本原理,概括了在微生物群落分析上的应用,阐述了硝化细菌传统研究的局限性及T_RFLP在硝化细菌群落结构分析上的应用前景.

T_RFLP Technique and Its Application on Community Analysis of Nitrifying Bacteria

Terminal restriction fragment length polymorphism (T_RFLP) analysis is a culture- independent approach for analyzing microbial community in environment. It bases on PCR technology, and its process includes DNA extraction of environmental samples, amplification of genes encoding the 16S rRNA, 18S rRNA or enzymes with fluorescently labeled primers, the restriction enzyme digestion of PCR products, capillary electrophoresis and the analysis of T_RFLP profile. It has been proved to be powerful applied on microbial community in environment since developed in 1997. Currently, T_RFLP rarely applied in China, and it has no applications on microbial community analysis of nitrifying bacteria. In this article, the fundamental principle of this technique and the recent applications of T_RFLP on microbial community are summarized; in addition, it illustrates the confinements of conventional culture-dependent of nitrifying bacteria and the foreground of T_RFLP applying on microbial community structure analysis of nitrifying bacteria.