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Chinese Journal of Applied Ecology ›› 2021, Vol. 32 ›› Issue (7): 2615-2622.doi: 10.13287/j.1001-9332.202107.032

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Quantitative stable isotope probing technique and its applications in microbial ecology

ZOU Wen-xuan1,2, SHEN Ju-pei1,2*, ZHANG Li-mei1,2, HU Ang3, WANG Jian-jun4, HE Ji-zheng5   

  1. 1Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China;
    2University of Chinese Academy of Sciences, Beijing 100049, China;
    3Hunan Agricultural University, Changsha 410128, China;
    4Nanjing Institute of Geography and Limnology, Chinese Academy of Sciences, Nanjing 210008, China;
    5Fujian Normal University, Fuzhou 350007, China
  • Received:2020-09-25 Revised:2021-03-16 Online:2021-07-15 Published:2022-01-15
  • Contact: *jpshen@rcees.ac.cn
  • Supported by:
    Program of Intergovernmental Cooperation in Science and Technology (2017YFE-0109800)and the National Natural Science Foundation of China(41930756).

Abstract: Quantitative stable isotope probing (qSIP) is a powerful tool, which links microbial taxon with functional metabolism in ecosystems and quantitatively determines the metabolic activity or growth rate of individual microbial taxa exposed to isotope tracers in the environment. qSIP technique employs quantitative PCR, high-throughput sequencing and stable isotope probing (SIP) techniques. The procedure involves adding labeled substrates to environmental samples for cultivation, separating labeled heavy fraction from unlabeled light fraction via isopycnic ultracentrifugation, making absolute quantification and sequencing analysis for microbial populations in all fractions, and then quantifying the isotope abundance of DNA involved in uptake and transformation based on the DNA density curve of unlabeled treatment and GC content. Here, we reviewed the rationale, data analysis and application of qSIP in microbial ecology, and discussed the existing problems and prospects of qSIP.

Key words: isotope labelling, microbial growth rate, high-throughput sequencing, microbial function, microbial taxon