Esson Kaitlin C, Lin Xueju, Kumaresan Deepak, Chanton Jeffrey P, Murrell J Colin, Kostka Joel E
School of Biology, Georgia Institute of Technology, Atlanta, Georgia, USA.
School of Earth and Environment, University of Western Australia, Crawley, WA, Australia.
Appl Environ Microbiol. 2016 Apr 4;82(8):2363-2371. doi: 10.1128/AEM.03640-15. Print 2016 Apr.
The objective of this study was to characterize metabolically active, aerobic methanotrophs in an ombrotrophic peatland in the Marcell Experimental Forest, in Minnesota. Methanotrophs were investigated in the field and in laboratory incubations using DNA-stable isotope probing (SIP), expression studies on particulate methane monooxygenase (pmoA) genes, and amplicon sequencing of 16S rRNA genes. Potential rates of oxidation ranged from 14 to 17 μmol of CH4g dry weight soil(-1)day(-1) Within DNA-SIP incubations, the relative abundance of methanotrophs increased from 4% in situ to 25 to 36% after 8 to 14 days. Phylogenetic analysis of the(13)C-enriched DNA fractions revealed that the active methanotrophs were dominated by the genera Methylocystis(type II;Alphaproteobacteria),Methylomonas, and Methylovulum(both, type I;Gammaproteobacteria). In field samples, a transcript-to-gene ratio of 1 to 2 was observed for pmoA in surface peat layers, which attenuated rapidly with depth, indicating that the highest methane consumption was associated with a depth of 0 to 10 cm. Metagenomes and sequencing of cDNA pmoA amplicons from field samples confirmed that the dominant active methanotrophs were Methylocystis and Methylomonas Although type II methanotrophs have long been shown to mediate methane consumption in peatlands, our results indicate that members of the genera Methylomonas and Methylovulum(type I) can significantly contribute to aerobic methane oxidation in these ecosystems.
本研究的目的是对明尼苏达州马塞尔实验森林中一个雨养泥炭地的代谢活跃的好氧甲烷氧化菌进行特征描述。利用DNA稳定同位素探测(SIP)、颗粒甲烷单加氧酶(pmoA)基因的表达研究以及16S rRNA基因的扩增子测序,在野外和实验室培养中对甲烷氧化菌进行了研究。氧化的潜在速率范围为14至17 μmol CH4 g干重土壤(-1)天(-1)。在DNA-SIP培养中,甲烷氧化菌的相对丰度从原位的4%增加到8至14天后的25%至36%。对(13)C富集的DNA组分进行系统发育分析表明,活跃的甲烷氧化菌主要由甲基孢囊菌属(II型;α-变形菌纲)、甲基单胞菌属和甲基卵菌属(均为I型;γ-变形菌纲)主导。在野外样本中,表层泥炭层中pmoA的转录本与基因比率为1至2,随深度迅速衰减,表明最高的甲烷消耗与0至10厘米的深度相关。对野外样本的宏基因组和cDNA pmoA扩增子测序证实,占主导地位的活跃甲烷氧化菌是甲基孢囊菌属和甲基单胞菌属。虽然长期以来已证明II型甲烷氧化菌介导泥炭地中的甲烷消耗,但我们的结果表明,甲基单胞菌属和甲基卵菌属(I型)的成员可对这些生态系统中的好氧甲烷氧化做出显著贡献。