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	Real-time PCR for quantification of aerobic anoxygenic phototrophic bacteria based on pufM gene in marine environment
	Du HL ; Jiao NZ ; Hu YH ; Zeng YH
	2006-02
关键词	aerobic anoxygenic phototrophic bacteria genetic diversity pufM real-time quantitative PCR
英文摘要	The abundance of aerobic anoxygenic phototrophic bacteria (AAPB), a new functional group that plays important roles in marine carbon cycling, is determined frequently by infrared epifluorescence microscopic analysis (IREM) or high-performance liquid chromatography (HPLC) based on detecting BChl a (bacteriochlorophyll a) fluorescence signal at 880 run. Unfortunately, the fluorescence signal is often influenced by environmental variables and physiological state of cell. Here we developed a real-time quantitative PCR (qPCR) assay based on pufM gene to specifically quantify AAPB in marine environments. High specificity and sensitivity for estimation of AAPB abundance were revealed by analysis of amplification products, melting curves and target sequences. The phylogenetic tree indicated that this primer set is suitable for a wide genetic diversity of AAPB, including alpha-3, alpha-4 Proteobacteria and clones of unclear taxonomic position. In contrast, no amplicon was obtained from green non-sulphur bacteria and oxygenic phototrophic bacteria such as Cyanobacterial genomic DNA. The melting behavior could indicate predominant phenotypes in AAPB community in addition to validating the products of qPCR. The AAPB was estimated to range from 1.3 X 10(4) cell/ml to 3.4 X 10(5) cell/ml in our 10 tested water samples by this qPCR assay. Further investigations on the abundance distribution of AAPB in marine environments using the qPCR assay may provide new insight into their ecological functions. (c) 2005 Elsevier B.V. All rights reserved.
语种	英语
出版者	ELSEVIER SCIENCE BV, PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/1307]
专题	海洋环境－已发表论文
推荐引用方式 GB/T 7714	Du HL,Jiao NZ,Hu YH,et al. Real-time PCR for quantification of aerobic anoxygenic phototrophic bacteria based on pufM gene in marine environment[J],2006.
APA	Du HL,Jiao NZ,Hu YH,&Zeng YH.(2006).Real-time PCR for quantification of aerobic anoxygenic phototrophic bacteria based on pufM gene in marine environment..
MLA	Du HL,et al."Real-time PCR for quantification of aerobic anoxygenic phototrophic bacteria based on pufM gene in marine environment".(2006).

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