Taubert Martin, Grob Carolina, Howat Alexandra M, Burns Oliver J, Dixon Joanna L, Chen Yin, Murrell J Colin
School of Environmental Sciences, University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ, UK.
Department of Aquatic Geomicrobiology, Friedrich Schiller University Jena, Dornburger Str. 159, Jena, 07743, Germany.
Environ Microbiol. 2015 Oct;17(10):3937-48. doi: 10.1111/1462-2920.12896. Epub 2015 Jun 25.
The xoxF gene, encoding a pyrroloquinoline quinone-dependent methanol dehydrogenase, is found in all known proteobacterial methylotrophs. In several newly discovered methylotrophs, XoxF is the active methanol dehydrogenase, catalysing the oxidation of methanol to formaldehyde. Apart from that, its potential role in methylotrophy and carbon cycling is unknown. So far, the diversity of xoxF in the environment has received little attention. We designed PCR primer sets targeting clades of the xoxF gene, and used 454 pyrosequencing of PCR amplicons obtained from the DNA of four coastal marine environments for a unique assessment of the diversity of xoxF in these habitats. Phylogenetic analysis of the data obtained revealed a high diversity of xoxF genes from two of the investigated clades, and substantial differences in sequence composition between environments. Sequences were classified as being related to a wide range of both methylotrophs and non-methylotrophs from Alphaproteobacteria, Betaproteobacteria and Gammaproteobacteria. The most prominent sequences detected were related to the family Rhodobacteraceae, the genus Methylotenera and the OM43 clade of Methylophilales, and are thus related to organisms that employ XoxF for methanol oxidation. Furthermore, our analyses revealed a high degree of so far undescribed sequences, suggesting a high number of unknown bacterial species in these habitats.
编码吡咯喹啉醌依赖性甲醇脱氢酶的xoxF基因存在于所有已知的变形菌甲基营养菌中。在几种新发现的甲基营养菌中,XoxF是活性甲醇脱氢酶,催化甲醇氧化为甲醛。除此之外,其在甲基营养和碳循环中的潜在作用尚不清楚。到目前为止,环境中xoxF的多样性很少受到关注。我们设计了针对xoxF基因分支的PCR引物组,并对从四个沿海海洋环境的DNA中获得的PCR扩增子进行454焦磷酸测序,以独特地评估这些栖息地中xoxF的多样性。对所得数据的系统发育分析表明,在两个被研究的分支中xoxF基因具有高度多样性,并且不同环境之间的序列组成存在实质性差异。序列被分类为与来自α-变形菌纲、β-变形菌纲和γ-变形菌纲的广泛的甲基营养菌和非甲基营养菌相关。检测到的最突出的序列与红杆菌科、嗜甲基菌属和嗜甲基单胞菌目的OM43分支相关,因此与利用XoxF进行甲醇氧化的生物体相关。此外,我们的分析揭示了大量迄今未描述的序列,表明这些栖息地中存在大量未知细菌物种。