分子技术在湿地微生物群落解析中的应用

The application of molecular techniques to characterize the microbial communities in constructed wetland

人工湿地的研究和应用近年来受到了广泛重视。微生物是人工湿地系统的重要组成部分,其群落结构对于湿地的净化功能的发挥具有重要影响。与传统的微生物分析技术相比,分子生物学技术在解析人工湿地微生物群落结构时无需纯培养,具有高效、快速、简便的特点,使其广泛应用于环境微生物的研究。文章综述了近年来在聚合酶链反应技术(Polymerase Chain Reaction,PCR)基础上发展起来的几种新的分子生物技术,包括PCR-DGGE、LH-PCR、T-RFLP、PCR-SSCP和ARDRA,以及其在人工湿地微生物研究中的应用现状。通过这些分子技术,可以分析湿地处理特定废水过程中微生物的数量、丰度、多样性及优势种;鉴定湿地中特定功能菌群(如氨氧化细菌、反硝化细菌、除硫菌等)的数量、活动分布、空间变更及与污染物去除的关系;判断各种系统条件(如不同基质、植物、水力负荷等)的设置对微生物多样性和稳定性的影响。最后,对分子技术在湿地领域的未来发展进行了展望。

The studies and applications of constructed wetland have become a hot topic in recent years.Microorganism is one of the most important factors in constructed wetland system.The structure of microbial communities have great influence on the purifying capacity of constructed wetland.Compared with traditional microbial analysis technologies,molecular biological techniques have many advantages,such as cultivation-independent,efficient,fast and simple,which have been widely used to identify environmental microorganisms.In the paper,the molecular biological techniques based on polymerase chain reaction（PCR）,such as PCR-DGGE,LH-PCR,T-RFLP,PCR-SSCP and ARDRA,were introduced.The mechanisms and research advances of the molecular techniques were also summarized.By using these techniques,the microbial number,density,diversity and dominant species in the constructed wetland systems treating with specific wastewater（such as swine wastewater,landfill leachates,etc.） can be analyzed;the relationship between the number,activity distribution,space shifting of specific microbial functional group（such as ammonia-oxidizing bacteria,denitrifiers,sulfate reducers,etc.） and the removal rates of main pollutants can be identified;the influence on microbial diversity and stability under different conditions（such as different substrate,wetland plants,hydraulic loading,etc.） can be estimated.Finally,the research prospects for the molecular techniques in the constructed wetland were also proposed.