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

Lei SU; Qianqian ZHANG; Jun GONG

doi:10.1007/s00343-018-6326-3

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

Updated：2023-05-19

- Development and evaluation of specific PCR primers targeting the ribosomal DNA-internal transcribed spacer (ITS) region of peritrich ciliates in environmental samples
- Chinese Journal of Oceanology and Limnology Vol. 36, Issue 3, Pages: 818-826(2018)
- Affiliations：
  
  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
- Author bio：
  
  GONG Jun, jgong@yic.ac.cn
- Funds：
- DOI：10.1007/s00343-018-6326-3
  CLC：
- Received：08 December 2016，
  
  Accepted：03 February 2017，
  
  Online First：28 March 2017，
  
  Published：2018-05
- Accepted：
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Lei SU, Qianqian ZHANG, Jun GONG. Development and evaluation of specific PCR primers targeting the ribosomal DNA-internal transcribed spacer (ITS) region of peritrich ciliates in environmental samples[J]. Chinese Journal of Oceanology and Limnology, 2018, 36(3): 818-826.
DOI：

Lei SU, Qianqian ZHANG, Jun GONG. Development and evaluation of specific PCR primers targeting the ribosomal DNA-internal transcribed spacer (ITS) region of peritrich ciliates in environmental samples[J]. Chinese Journal of Oceanology and Limnology, 2018, 36(3): 818-826. DOI： 10.1007/s00343-018-6326-3.

摘要

Abstract

Peritrich ciliates are highly diverse and can be important bacterial grazers in aquatic ecosystems. Morphological identifications of peritrich species and assemblages in the environment are time-consuming and expertise-demanding. In this study

two peritrich-specific 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 specificity 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 28S rDNA

and ITS+D1 region co-varied with

and generally more variable than

the V9 region of 18S rDNA in peritrichs. The newly designed specific primers thus provide additional tools to study the molecular diversity

community composition

and phylogeography of these ecologically important protists in different systems.

关键词

Keywords

references

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