生物技术通报 ›› 2017, Vol. 33 ›› Issue (10): 26-32.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0550

• 特约综述 • 上一篇    下一篇

环境中微生物原位检测方法研究进展

宋伟凤, 李明聪, 高峥   

  1. 山东农业大学生命科学学院 作物生物学国家重点实验室,泰安271000
  • 收稿日期:2017-07-02 出版日期:2017-10-29 发布日期:2017-10-29
  • 作者简介:宋伟凤,女,研究方向:微生物生态;E-mail:songweifeng9719@qq.com,李明聪为共同第一作者
  • 基金资助:
    国家自然科学基金项目(41306150),山东省优秀中青年科学家科研奖励基金(BS2012HZ011),河口海岸学国家重点实验室开放课题(SKLEC-KF201603)

Research Progress on in situ Detection Methods of Microorganisms

SONG Wei-feng, LI Ming-cong, GAO Zheng   

  1. School of Life Science,Shandong Agricultural University,State Key Laboratory of Crop Biology,Tai’an 271000
  • Received:2017-07-02 Online:2017-10-29 Published:2017-10-29
  • Contact: 高峥,男,博士,副教授,研究方向:微生物生态与环境微生物学;E-mail:gaozheng@sdau.edu.cn

摘要: 微生物是生态系统物质循环和能量流动的主要参与者,在生态系统中起着重要的作用。但在现有技术条件下可培养微生物所占比例极小,限制了微生物资源的开发利用。目前有许多方法,可以避开微生物不可培养的问题,直接对微生物进行原位检测。对此,将前人关于微生物生态的原位检测研究方法进行了综述,方便以后对这些方法的合理应用。分别从DNA水平、RNA水平和蛋白质水平,介绍了对应的原位微生物检测方法(如BrdU标记、DNA-SIP、FISH和环境转录物组等),比较了它们的优缺点,并介绍了如何将这些方法与目前流行的高通量测序、单细胞测序等技术相结合来捕获原位活性微生物组等。同时,还对这些方法的特点进行了比较,使得人们可以更清楚地了解在不同场景下对不同方法的选择。这些经过改进的新兴方法及其与其它方法的结合使用将有助于解决微生物生态学研究中出现过或即将出现的很多问题。地球上的各种生态系统复杂而庞大,包含的微生物种群也各有差异。各种原位检测方法对微生物生理生态做出了更加真实有效的描述,必将成为研究微生物生态的有力手段。

关键词: 微生物原位检测, BrdU标记, DNA-SIP, 荧光原位杂交, 宏基因组, 转录物组, 生物正交反应

Abstract: As major participants in ecosystem material cycle and energy flow,microorganisms play an important role in the ecosystem. However,the proportion of the cultivable microorganism under the existing technology is very small,which limits the exploit of microbial resources. At present,there are a number of approaches that can avoid the problem of uncultured microorganisms,which are designed to study in situ microbial activity. Regarding this,we summarized some research methods of studying in situ microbial ecology,allowing it convenient to reasonably use these methods in the future. This article introduces the corresponding microbial detection methods of BrdU-labeling,DNA-SIP,fluorescence in situ hybridization(FISH),and environmental transcriptome from DNA level,RNA level and protein level,respectively,and compares their advantages and disadvantages. It also introduces how to apply these methods combined with popular high-throughput sequencing and single cell sequencing technology to capture the in situ activity of microbial groups. At the same time,comparing the characteristics of these methods,so that we can more clearly understand the choice of different methods under different scenarios. These modified methods combined with other methods will be conducive to solve many have-been or will-happen problems in the study of microbial ecology. The ecosystems on the earth are complex and huge,in which the microbial populations vary. A variety of in situ detection methods have made a more realistic and effective description for the physiology and ecology of microorganisms,which will become a powerful tool for the study of microorganisms.

Key words: microbial detection in sit, BrdU-labeling, DNA-SIP, fluorescence in situ hybridization, metagenome, transcriptome, BONCAT