土壤微生物分离新技术的研究进展
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湖南省研究生科研创新项目(CX2014B298)资助


Progress in cultivation research on soil microbes
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    摘要:

    土壤是环境微生物学研究中最具挑战性的环境,也是生物分子、遗传资源开发的热点区域。在“宏-组学”方法快速发展的今天,土壤微生物分离培养技术在微生物环境功能研究、代谢途径的阐明、特定功能的验证及基础实验和生产实践的应用等方面仍然发挥着重要作用。本文首先分析了大多数环境微生物不能被分离培养的几个原因。然后重点综述了近年来在土壤微生物分离培养技术方面的探索,主要包括生长培养基的优化和重新设计如培养基稀释分离、添加生长限制因子、更换凝固剂等,在延长培养时间、改善含氧量、降低培养温度等方面对生长条件进行改进,采用“扩散盒”、“原位培养陷阱”等技术对微生物进行天然环境原位培养,对环境微生物群体培养和共培养,利用“宏基因组学”“宏蛋白质组学”等非培养技术辅助分离等。最后,作者对未来土壤微生物分离研究的方向提出了自己的建议。

    Abstract:

    Soil is probably the most challenging of all natural environments for microbiologists, and the hotspot for discovering of novel biomolecules and genetic resources. Cultivation of environmental microorganisms was ignored and marginalized in the tremendous progress of “meta-omics” technologies. Nevertheless, by cultivation-based approaches, researchers can acquire novel microorganisms that are undetected by molecular methods, be allowed for the verification and testing of hypotheses of metabolic potential and function of novel genes determined by the meta-omic data and also do the microbial foundational research or industrial production. In the era of meta-omics, cultivation method is still useful in understanding the detailed metabolism and functions of those environmental organisms, and this makes the genetic manipulation, confirming certain functions, and the research of environmental function of microorganisms possible. The challenge is to bring these recalcitrant microorganisms into the laboratory for future exploration. This review summarizes the reasons why microbes are not growing in the lab such as. Then we focus on the advanced cultivation techniques. The methods of modifications to growth media include using low substrate concentrations, adding growth-limiting factors into media and changing gelling reagents. Modifications to growth conditions include extending incubation time, incubating in a optimum oxygen level, and using low-temperature strategies. The strategies of cultures in situ or cultures in simulated natural conditions include “growth chamber” and “trap for in situ cultivation”. The methods of community culture and co-culture also elevated the possibility of novel bacteria from environmental samples. Cultivation of soil microorganisms by assistance of culture-independent methods such as metagenomics and metaproteomics have been developed to improve the cultivating process. Finally, we propose the directions of isolation of soil microbes in the future.

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袁志辉,王 健,杨文蛟,吴永尧.土壤微生物分离新技术的研究进展[J].土壤学报,2014,51(6):1183-1191. DOI:10.11766/trxb201311270569 Yuan Zhihui, Wang Jian, Yang Wenjiao, Wu Yongyao. Progress in cultivation research on soil microbes[J]. Acta Pedologica Sinica,2014,51(6):1183-1191.

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  • 收稿日期:2013-11-27
  • 最后修改日期:2014-04-24
  • 录用日期:2014-06-26
  • 在线发布日期: 2014-08-26
  • 出版日期: