Stable-isotope probing as a tool in microbial ecology
Stable-isotope probing as a tool in microbial ecology
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DOI：
10.1038/35001054
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发表时间：
2000-02-10
期刊：
影响因子：
64.8
通讯作者：
Murrell, JC
中科院分区：
文献类型：
--
作者：
Radajewski, S;Ineson, P;Murrell, JC
微生物负责驱动地球上元素的地球化学循环。尽管它们的重要性和巨大的多样性（1），参与任何特定过程的微生物的分类身份通常仅限于已分离和培养的小部分微生物群。最近，分子生物学方法与生物标志物中稳定同位素丰度的结合提供了一种独立于培养的方法，将细菌的身份与其在环境中的功能联系起来（2，3）。在这里，我们表明，C-13-DNA，在生长过程中产生的代谢不同的微生物群体的C-13-富集的碳源，可以解决从C-12- DNA的密度梯度离心。从目标微生物组分离的DNA可以通过基因探测和序列分析在分类学和功能上表征。这种技术的应用，以调查土壤中的甲醇利用微生物表现出参与的成员的两个不同的真细菌群的α-变形菌和Acidobacterium谱系。因此，稳定同位素探测提供了一种强有力的新技术，用于识别在接近原位发生的条件下积极参与特定代谢过程的微生物。
Microorganisms are responsible for driving the biogeochemical cycling of elements on Earth. Despite their importance and vast diversity(1), the taxonomic identity of the microorganisms involved in any specific process has usually been confined to that small fraction of the microbiota that has been isolated and cultivated. The recent coupling of molecular biological methods with stable-isotope abundance in biomarkers has provided a cultivation-independent means of linking the identity of bacteria with their function in the environment(2,3). Here we show that C-13-DNA, produced during the growth of metabolically distinct microbial groups on a C-13-enriched carbon source, can be resolved from C-12- DNA by density-gradient centrifugation. DNA isolated from the target group of microorganisms can be characterized taxonomically and functionally by gene probing and sequence analysis. Application of this technique to investigate methanol-utilizing microorganisms in soil demonstrated the involvement of members of two phylogenetically distinct groups of eubacteria; the alpha-proteobacterial and Acidobacterium lineages. Stable-isotope probing thus offers a powerful new technique for identifying microorganisms that are actively involved in specific metabolic processes under conditions which approach those occurring in situ.