定量稳定性同位素探针技术及其在微生物生态学研究中的应用

doi:10.13287/j.1001-9332.202107.032

应用生态学报 ›› 2021, Vol. 32 ›› Issue (7): 2615-2622.doi: 10.13287/j.1001-9332.202107.032

定量稳定性同位素探针技术及其在微生物生态学研究中的应用

邹文萱^1,2, 沈菊培^1,2*, 张丽梅^1,2, 胡盎³, 王建军⁴, 贺纪正⁵

¹中国科学院生态环境研究中心, 北京 100085;
²中国科学院大学, 北京 100049;
³湖南农业大学, 长沙 410128;
⁴中国科学院南京地理与湖泊研究所, 南京 210008;
⁵福建师范大学, 福州 350007

收稿日期:2020-09-25 修回日期:2021-03-16 出版日期:2021-07-15 发布日期:2022-01-15
通讯作者: *jpshen@rcees.ac.cn
作者简介:邹文萱,女,1997年生,硕士研究生.主要从事微生物生态与氮循环研究.E-mail:zouwenxuan19@mails.ucas.ac.cn
基金资助:
科技部政府间合作项目(2017YFE0109800)和国家自然科学基金项目(41930756)

Quantitative stable isotope probing technique and its applications in microbial ecology

ZOU Wen-xuan^1,2, SHEN Ju-pei^1,2*, ZHANG Li-mei^1,2, HU Ang³, WANG Jian-jun⁴, HE Ji-zheng⁵

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).

摘要/Abstract

摘要： 定量稳定性同位素探针技术(qSIP)是将生态系统中微生物分类性状与代谢功能联系起来的有效工具,能够定量测定特定环境中单个微生物类群暴露于同位素示踪剂后微生物代谢活动或生长速率。qSIP技术采用定量PCR与高通量测序技术并结合稳定同位素探针技术(SIP),通过向环境样品添加标记底物进行培养,提取微生物生物标记物,利用超高速等密度梯度离心将被同位素标记的重链核酸与未被标记的轻链核酸进行分离,并对所有组分微生物类群进行绝对定量和测序分析,基于GC含量和未标记处理DNA密度曲线量化参与吸收转化的DNA同位素丰度。本文重点阐述qSIP的技术原理、数据分析流程及其在微生物生态学研究中的应用进展,并对该技术存在的问题进行了分析和展望。

关键词: 同位素标记, 微生物生长速率, 高通量测序, 微生物功能, 微生物分类单元

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

邹文萱, 沈菊培, 张丽梅, 胡盎, 王建军, 贺纪正. 定量稳定性同位素探针技术及其在微生物生态学研究中的应用[J]. 应用生态学报, 2021, 32(7): 2615-2622.

ZOU Wen-xuan, SHEN Ju-pei, ZHANG Li-mei, HU Ang, WANG Jian-jun, HE Ji-zheng. Quantitative stable isotope probing technique and its applications in microbial ecology[J]. Chinese Journal of Applied Ecology, 2021, 32(7): 2615-2622.

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定量稳定性同位素探针技术及其在微生物生态学研究中的应用

Quantitative stable isotope probing technique and its applications in microbial ecology

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