Revealing the Diversity and Quantity of Peritrich Ciliates in Environmental Samples Using Specific Primer-based PCR and Quantitative PCR
Liu, Xihan1,3; Gong, Jun1,2
2012-12-07
发表期刊MICROBES AND ENVIRONMENTS
ISSN1342-6311
卷号27期号:4页码:497-503
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
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.; 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.
通讯作者Gong, J (reprint author), Chinese Acad Sci, Yantai Inst Coastal Zone Res, Lab Environm Microbiol, Yantai 264003, Peoples R China.
关键词Ciliophora Peritricha Qpcr Specific Primers Ssu Rrna Gene Copy Number
作者部门海岸带生物学与生物资源利用所重点实验室
学科领域Biotechnology & Applied Microbiology ; Microbiology
项目资助者One Hundred Talent Program of CAS; Natural Science Foundation of China [40976099, 41176143]; FANEDD [2007B27]
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收录类别SCI
关键词[WOS]RIBOSOMAL-RNA GENE ; REAL-TIME PCR ; MOLECULAR CHARACTERIZATION ; CILIOPHORA PERITRICHIA ; BIOFILM COMMUNITIES ; PLANKTONIC CILIATE ; NORTH CHINA ; OLIGOHYMENOPHOREA ; PROTOZOA ; QUANTIFICATION
文章类型Article
语种英语
资助项目Liu, Xihan; Gong, Jun
WOS研究方向Biotechnology & Applied Microbiology ; Microbiology
WOS记录号WOS:000312162800021
引用统计
被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.yic.ac.cn/handle/133337/6137
专题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
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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.
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