Binga Erik K, Lasken Roger S, Neufeld Josh D
Department of Biology, University of Waterloo, Waterloo, Ontario, Canada.
ISME J. 2008 Mar;2(3):233-41. doi: 10.1038/ismej.2008.10. Epub 2008 Feb 7.
Microbial ecology is a field that applies molecular techniques to analyze genes and communities associated with a plethora of unique environments on this planet. In the past, low biomass and the predominance of a few abundant community members have impeded the application of techniques such as PCR, microarray analysis and metagenomics to complex microbial populations. In the absence of suitable cultivation methods, it was not possible to obtain DNA samples from individual microorganisms. Recently, a method called multiple displacement amplification (MDA) has been used to circumvent these limitations by amplifying DNA from microbial communities in low-biomass environments, individual cells from uncultivated microbial species and active organisms obtained through stable isotope probing incubations. This review describes the development and applications of MDA, discusses its strengths and limitations and highlights the impact of MDA on the field of microbial ecology. Whole genome amplification via MDA has increased access to the genomic DNA of uncultivated microorganisms and low-biomass environments and represents a 'power tool' in the molecular toolbox of microbial ecologists.
微生物生态学是一个应用分子技术来分析与地球上众多独特环境相关的基因和群落的领域。过去，低生物量以及少数丰富群落成员的主导地位阻碍了诸如聚合酶链式反应（PCR）、微阵列分析和宏基因组学等技术在复杂微生物群体中的应用。在缺乏合适培养方法的情况下，无法从单个微生物中获取DNA样本。最近，一种称为多重置换扩增（MDA）的方法已被用于克服这些限制，该方法通过扩增低生物量环境中微生物群落的DNA、未培养微生物物种的单个细胞以及通过稳定同位素探测培养获得的活性生物体的DNA来实现。本综述描述了MDA的发展和应用，讨论了其优点和局限性，并强调了MDA对微生物生态学领域的影响。通过MDA进行的全基因组扩增增加了对未培养微生物和低生物量环境基因组DNA的获取，并且是微生物生态学家分子工具箱中的一种“有力工具”。
