Department of Microbial Ecology, Netherlands Institute of Ecology (NIOO-KNAW), P.O. Box 40, 6666 ZG Heteren, Netherlands.
Appl Environ Microbiol. 2010 Oct;76(20):6769-77. doi: 10.1128/AEM.00343-10. Epub 2010 Aug 20.
The bacterial phylum Acidobacteria has a widespread distribution and is one of the most common and diverse phyla in soil habitats. However, members of this phylum have often been recalcitrant to cultivation methods, hampering the study of this presumably important bacterial group. In this study, we used a cultivation-independent metagenomic approach to recover genomic information from soilborne members of this phylum. A soil metagenomic fosmid library was screened by PCR targeting acidobacterial 16S rRNA genes, facilitating the recovery of 17 positive clones. Recovered inserts appeared to originate from a range of Acidobacteria subdivisions, with dominance of subdivision 6 (10 clones). Upon full-length insert sequencing, gene annotation identified a total of 350 open reading frames (ORFs), representing a broad range of functions. Remarkably, six inserts from subdivision 6 contained a region of gene synteny, containing genes involved in purine de novo biosynthesis and encoding tRNA synthetase and conserved hypothetical proteins. Similar genomic regions had previously been observed in several environmental clones recovered from soil and marine sediments, facilitating comparisons with respect to gene organization and evolution. Comparative analyses revealed a general dichotomy between marine and terrestrial genes in both phylogeny and G+C content. Although the significance of this homologous gene cluster across subdivision 6 members is not known, it appears to be a common feature within a large percentage of all acidobacterial genomic fragments recovered from both of these environments.
酸杆菌门是一种广泛分布的细菌门,是土壤生境中最常见和最多样化的门之一。然而,该门的成员往往难以通过培养方法进行培养,这阻碍了对这个推测上很重要的细菌群的研究。在这项研究中,我们使用了一种非培养的宏基因组方法,从土壤来源的该门成员中回收基因组信息。通过针对酸杆菌 16S rRNA 基因的 PCR 筛选土壤宏基因组 fosmid 文库,促进了 17 个阳性克隆的回收。回收的插入片段似乎来源于酸杆菌门的多个细分群,其中以第 6 细分群(10 个克隆)为主导。经过全长插入片段测序,基因注释共鉴定出 350 个开放阅读框(ORFs),代表了广泛的功能。值得注意的是,第 6 细分群的 6 个插入片段包含一个基因同线性区域,其中包含参与嘌呤从头合成的基因,并编码 tRNA 合成酶和保守的假设蛋白。以前在从土壤和海洋沉积物中回收的几个环境克隆中已经观察到类似的基因组区域,便于在基因组织和进化方面进行比较。比较分析显示,海洋和陆地基因在系统发育和 G+C 含量上存在普遍的二分法。虽然这个同源基因簇在第 6 细分群成员中的意义尚不清楚,但它似乎是从这两个环境中回收的大量酸杆菌基因组片段的一个共同特征。