CARD-FISH技术及其在微生物生态学研究中的应用

Catalyzed Reporter Deposition-Fluorescence in situ Hybridization(CARD-FISH) and Its Application in Microbial Ecology Study

催化报告沉积荧光原位杂交技术(Catalyzed reporter deposition-fluorescence in situ hybridization,CARDFISH)是基于传统的FISH技术发展而来,由于其较高的灵敏度及稳定性,可以检测微生物的rRNA、mRNA和DNA上的目标基因等,获得环境微生物的群落及功能信息,现已成为微生物生态学研究领域中的重要技术手段。近些年,CARD-FISH与同位素示踪技术、纳米二次离子质谱技术(Nano SIMS)、扫描电子显微镜(SEM)、流式细胞仪等技术方法的联合使用,不仅可以研究复杂环境中微生物的物种组成、数量及其高分辨形态学信息,而且可以获得微生物在单细胞水平的生理代谢信息及其活性,对在单细胞水平认识原位环境微生物的生理生态功能具有重要意义。本文重点介绍了CARD-FISH的技术路线和要点,并探讨CARD-FISH与相关技术联用在环境微生物生态学研究中的应用及进展。

Catalyzed reporter deposition-fluorescence in situ hybridization（CARD-FISH） is developed based on traditional FISH. With its high sensitivity and stability,that could determine rRNA,mRNA,DNA,other target genes etc. of microorganisms to obtain the community structure and function information of microorganism in the environment,now it has become an important technical means in environmental microbial ecology research sphere. Recently,the combination uses of CARD-FISH,isotopic tracer technique,scanning electron microscope/energy dispersive X-ray spectroscopy（SEM/EDX）,nano-meter secondary ion mass spectrometry（nano SIMS）,and flow cytometer etc. not only could study the component,quantity,and high resolution morphology information of microorganisms in complicated conditions,but also could obtain the physiological and metabolic information and activity of microorganisms in single cells level,these have important significance to understand the physiological and ecological functions of environmental microorganisms in situ. In this paper the priority is given to the introduction of technical route and the essentials of CARD-FISH,and the investigation of the application and advances in combination uses of CARD-FISH and related technologies on the studies of the environmental microorganism ecology.