Development and evaluation of specific PCR primers targeting the ribosomal DNA-internal transcribed spacer(ITS)region of peritrich ciliates in environmental samples  

作　　者：SU Lei ZHANG Qianqian GONG Jun 苏蕾;张倩倩;龚骏(Laboratory of Microbial Ecology and Matter Cycles,Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences Yantai 264003,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]Laboratory of Microbial Ecology and Matter Cycles,Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai 264003,China [2]University of Chinese Academy of Sciences,Beijing 100049,China

出　　处：《Journal of Oceanology and Limnology》2018年第3期818-826,共9页海洋湖沼学报（英文）

基　　金：Supported by the National Natural Science Foundation of China(Nos.31572255,41522604,31301867);the Strategic Priority Research Program of CAS(No.XDA11020702);the Science and Technology Development Program of Yantai(No.2014ZH073)

摘　　要：Peritrich ciliates are highly diverse and can be important bacterial grazers in aquatic ecosystems. Morphological identifi cations of peritrich species and assemblages in the environment are time-consuming and expertise-demanding. In this study, two peritrich-specifi c PCR primers were newly designed to amplify a fragment including the internal transcribed spacer(ITS) region of ribosomal rDNA from environmental samples. The primers showed high specifi city in silico, and in tests with peritrich isolates and environmental DNA. Application of these primers in clone library construction and sequencing yielded exclusively sequences of peritrichs for water and sediment samples. We also found the ITS1, ITS2, ITS, D1 region of 28 S rDNA, and ITS+D1 region co-varied with, and generally more variable than, the V9 region of 18 S rDNA in peritrichs. The newly designed specifi c primers thus provide additional tools to study the molecular diversity, community composition, and phylogeography of these ecologically important protists in dif ferent systems.Peritrich ciliates are highly diverse and can be important bacterial grazers in aquatic ecosystems. Morphological identifi cations of peritrich species and assemblages in the environment are time-consuming and expertise-demanding. In this study, two peritrich-specifi c PCR primers were newly designed to amplify a fragment including the internal transcribed spacer（ITS） region of ribosomal rDNA from environmental samples. The primers showed high specifi city in silico, and in tests with peritrich isolates and environmental DNA. Application of these primers in clone library construction and sequencing yielded exclusively sequences of peritrichs for water and sediment samples. We also found the ITS1, ITS2, ITS, D1 region of 28 S rDNA, and ITS＋D1 region co-varied with, and generally more variable than, the V9 region of 18 S rDNA in peritrichs. The newly designed specifi c primers thus provide additional tools to study the molecular diversity, community composition, and phylogeography of these ecologically important protists in dif ferent systems.

关 键 词：Ciliophora Peritrichia clone library internal transcribed spacer（ITS） rDNA specific PCR PRIMERS 

分 类 号：X524[环境科学与工程—环境工程]