title: Diversity and activity of methanotrophs in alkaline soil from a Chinese coal mine.
abstract: Culture-independent molecular biological techniques, including 16S rRNA gene and functional gene clone libraries and microarray analyses using pmoA (encoding a key subunit of particulate methane monooxygenase), were applied to investigate the methanotroph community structure in alkaline soil from a Chinese coal mine. This environment contained a high diversity of methanotrophs, including the type II methanotrophs Methylosinus/Methylocystis, type I methanotrophs related to Methylobacter/Methylosoma and Methylococcus, and a number of as yet uncultivated methanotrophs. In order to identify the metabolically active methane-oxidizing bacteria from this alkaline environment, DNA stable isotope probing (DNA-SIP) experiments using (13)CH(4) were carried out. This showed that both type I and type II methanotrophs were active, together with methanotrophs related to Methylocella, which had previously been found only in acidic environments. Methylotrophs, including Methylopila and Hyphomicrobium, were also detected in soil DNA and after DNA-SIP experiments. DNA sequence information on the most abundant, active methanotrophs in this alkaline soil will facilitate the design of oligonucleotide probes to monitor enrichment cultures when isolating key alkaliphilic methanotrophs from such environments.
authors.affiliation: Department of Chemical Engineering, Tsinghua University, Beijing, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Han
authors.forename: Yin
authors.initials: Y
authors.lastname: Chen
authors.forename: Guy
authors.initials: G
authors.lastname: Abell
authors.forename: Hao
authors.initials: H
authors.lastname: Jiang
authors.forename: Levente
authors.initials: L
authors.lastname: Bodrossy
authors.forename: Jiangang
authors.initials: J
authors.lastname: Zhao
authors.forename: J Colin
authors.initials: JC
authors.lastname: Murrell
authors.forename: Xin-Hui
authors.initials: XH
authors.lastname: Xing
mesh_terms: D044822:Biodiversity; D002681:China; D003031:Coal / Q000382:microbiology; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D015723:Gene Library; D007553:Isotope Labeling; D008697:Methane / Q000378:metabolism; D008772:Methylococcaceae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008906:Mining; D020411:Oligonucleotide Array Sequence Analysis; D010084:Oxidation-Reduction; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012987:Soil / Q000032:analysis*; D012988:Soil Microbiology
chemical_list: D003031:Coal; D015342:DNA Probes; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012987:Soil; D008697:Methane