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  4. 海岸带生物学与生物资源利用重点实验室
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Revealing the Diversity and Quantity of Peritrich Ciliates in Environmental Samples Using Specific Primer-based PCR and Quantitative PCR

文献类型：期刊论文

作者Liu, Xihan1,3; Gong, Jun1,2
刊名MICROBES AND ENVIRONMENTS
出版日期2012-12-07
卷号27期号:4页码:497-503
关键词Ciliophora Peritricha qPCR specific primers SSU rRNA gene copy number
ISSN号1342-6311
产权排序[Liu, Xihan; Gong, Jun] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Lab Environm Microbiol, Yantai 264003, Peoples R China; [Gong, Jun] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Key Lab Coastal Zone Environm Proc, Yantai 264003, Peoples R China; [Liu, Xihan] Ocean Univ China, Inst Evolut & Marine Divers, Qingdao 266003, Peoples R China
通讯作者Gong, J (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, Lab Environm Microbiol, Yantai 264003, Peoples R China.
中文摘要Peritrichs are a diverse, ecologically important ciliate group usually with a complex life cycle. To date, the community of the peritrichs has been investigated by using morphology-based methods such as living observation and silver staining. Here we show a molecular approach for characterizing the diversity and quantity of free-living peritrichs in environmental samples. We newly designed four peritrich-specific primers targeting 18S rRNA genes that allow clone library construction, screening and analysis. A quantitative real-time PCR (qPCR) assay was developed to quantify peritrichs in environmental samples by using rDNA copy number as an indicator. DNA extracted from four water samples of contrasting environmental gradients was analysed. The results showed that the peritrich community was differentiated among these samples, and that the diversity decreased with the increase of water salinity. The qPCR results are consistent with the library sequence analysis in terms of quantity variations from sample to sample. The development of peritrich-specific primers, for the first time, for conventional PCR and qPCR assays, provides useful molecular tools for revealing the diversity and quantity of peritrich ciliates in environmental samples. Also, our study illustrates the potential of these molecular tools to ecological studies of other ciliate groups in diverse environments.
英文摘要Peritrichs are a diverse, ecologically important ciliate group usually with a complex life cycle. To date, the community of the peritrichs has been investigated by using morphology-based methods such as living observation and silver staining. Here we show a molecular approach for characterizing the diversity and quantity of free-living peritrichs in environmental samples. We newly designed four peritrich-specific primers targeting 18S rRNA genes that allow clone library construction, screening and analysis. A quantitative real-time PCR (qPCR) assay was developed to quantify peritrichs in environmental samples by using rDNA copy number as an indicator. DNA extracted from four water samples of contrasting environmental gradients was analysed. The results showed that the peritrich community was differentiated among these samples, and that the diversity decreased with the increase of water salinity. The qPCR results are consistent with the library sequence analysis in terms of quantity variations from sample to sample. The development of peritrich-specific primers, for the first time, for conventional PCR and qPCR assays, provides useful molecular tools for revealing the diversity and quantity of peritrich ciliates in environmental samples. Also, our study illustrates the potential of these molecular tools to ecological studies of other ciliate groups in diverse environments.
学科主题Biotechnology & Applied Microbiology; Microbiology
研究领域[WOS]Biotechnology & Applied Microbiology ; Microbiology
关键词[WOS]RIBOSOMAL-RNA GENE ; REAL-TIME PCR ; MOLECULAR CHARACTERIZATION ; CILIOPHORA PERITRICHIA ; BIOFILM COMMUNITIES ; PLANKTONIC CILIATE ; NORTH CHINA ; OLIGOHYMENOPHOREA ; PROTOZOA ; QUANTIFICATION
收录类别SCI
资助信息One Hundred Talent Program of CAS; Natural Science Foundation of China [40976099, 41176143]; FANEDD [2007B27]
原文出处http://dx.doi.org/10.1264/jsme2.ME12056
语种英语
WOS记录号WOS:000312162800021
公开日期2013-03-08
源URL[http://ir.yic.ac.cn/handle/133337/6137]  
专题烟台海岸带研究所_海岸带生物学与生物资源利用所重点实验室
作者单位1.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Lab Environm Microbiol, Yantai 264003, Peoples R China
2.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Key Lab Coastal Zone Environm Proc, Yantai 264003, Peoples R China
3.Ocean Univ China, Inst Evolut & Marine Divers, Qingdao 266003, Peoples R China
推荐引用方式
GB/T 7714		 Liu, Xihan,Gong, Jun. Revealing the Diversity and Quantity of Peritrich Ciliates in Environmental Samples Using Specific Primer-based PCR and Quantitative PCR[J]. MICROBES AND ENVIRONMENTS,2012,27(4):497-503.
APA Liu, Xihan,&Gong, Jun.(2012).Revealing the Diversity and Quantity of Peritrich Ciliates in Environmental Samples Using Specific Primer-based PCR and Quantitative PCR.MICROBES AND ENVIRONMENTS,27(4),497-503.
MLA Liu, Xihan,et al."Revealing the Diversity and Quantity of Peritrich Ciliates in Environmental Samples Using Specific Primer-based PCR and Quantitative PCR".MICROBES AND ENVIRONMENTS 27.4(2012):497-503.

入库方式： OAI收割

来源：烟台海岸带研究所

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