青海湖河源湿地甲烷氧化菌群落对温度升高的响应

甲烷氧化菌对甲烷的氧化能力显著影响全球甲烷排放总量。研究河源湿地甲烷氧化菌群落对温度升高的响应有助于深入探究河源湿地生态系统的甲烷循环机制。采用高通量测序方法研究了温度升高后青海湖河源湿地甲烷氧化菌群落特征的变化。青海湖河源湿地甲烷氧化菌的优势菌门为变形菌门, 增温处理没有显著影响河源湿地甲烷氧化菌的群落多样性。河源湿地甲烷氧化菌的群落结构对温度升高响应明显, 增温显著增加了属水平优势菌群(相对丰度 > 0.1%)的相对丰度。LEfSe分析表明, 增温处理组与自然对照组共存在23个差异菌群, 增温显著增加了甲基球菌属的相对丰度, 显著降低了甲基杆菌属的相对丰度。河源湿地甲烷氧化菌群落功能与碳、氮代谢过程密切相关。整体而言, 河源湿地甲烷氧化菌的群落结构对温度升高较为敏感, 部分菌群的相对丰度变化显著。     

青海湖3种类型高寒湿地甲烷氧化菌群落特征

【背景】甲烷氧化菌在维持湿地生态系统碳平衡方面发挥着重要作用,青海湖高寒湿地具有十分重要的生态地位,但目前有关该地区甲烷氧化菌的研究相对较少。【目的】探究不同类型高寒湿地土壤甲烷氧化菌的群落特征与驱动因素。【方法】以青海湖流域内的小泊湖沼泽湿地、鸟岛湖滨湿地、瓦颜山河源湿地为研究对象,通过高通量测序技术对土壤甲烷氧化菌进行检测。【结果】3种不同类型高寒湿地土壤甲烷氧化菌的优势菌门均为变形菌门(Proteobacteria)。鸟岛湖滨湿地与瓦颜山河源湿地的甲烷氧化菌α多样性存在显著差异(P<0.05),而小泊湖沼泽湿地与二者的甲烷氧化菌α多样性的差异不显著(P>0.05)。LEfSe分析表明,不同类型高寒湿地共存在40个差异菌群,尤以瓦颜山河源湿地差异菌群数量最多,从门到属水平均存在显著差异。冗余分析(redundancy analysis,RDA)表明,甲烷氧化菌菌群变化的主要驱动因子为土壤温度、土壤水分、电导率。【结论】整体而言,青海湖3种类型高寒湿地土壤理化性质及甲烷氧化菌群落多样性均存在差异,且部分菌群的相对丰度具有显著性差异(P<0.05)。     

土壤中甲烷厌氧氧化菌多样性的分子检测

甲烷厌氧氧化作用是减少海洋底泥甲烷释放的重要生物地球化学过程,然而在陆地生态系统中甲烷厌氧氧化作用及其功能菌群的生态功能仍然不确定。对甲烷厌氧氧化菌多样性的研究可为减少甲烷排放提供重要科学依据。与传统的分离培养方法比较,分子检测方法是一种更为快速和高效的研究手段,可直接和全面的反映参与甲烷厌氧氧化作用的功能微生物。以DNA分子标记物为研究对象,重点探讨三类主要的分子标记基因,即16S rRNA,mcr A和pmo A,所采用的相关探针和引物信息,同时从定性和定量两个角度综述土壤甲烷厌氧氧化菌的多样性研究的主要进展,最后提出厌氧甲烷氧化菌多样性研究中存在的一些问题和相应的解决思路。     

稻鱼共生系统的土壤产甲烷和甲烷氧化微生物群落

稻鱼共生对稻田甲烷(CH₄)排放产生明显影响，但稻鱼共生是否影响与CH₄排放相关的产甲烷菌和甲烷氧化菌仍有待阐明。本研究以全球重要农业文化遗产——青田稻鱼系统为例，通过田间试验，比较研究了水稻单作系统(RM)、无饲料投放的稻鱼共生系统(RFN)和有饲料投放的稻鱼共生系统(RFF)水稻和田鱼的产量、土壤碳氮磷含量以及产甲烷和甲烷氧化微生物的特征。结果表明，RFF的水稻产量和土壤碳氮增量均显著高于RM。荧光定量PCR分析表明，稻鱼共生(RFN和RFF)的产甲烷菌和甲烷氧化菌丰度显著高于RM,且RFF的产甲烷菌丰度显著高于RFN。Illumina Miseq测序分析表明，稻鱼共生显著影响产甲烷菌群落结构，但对甲烷氧化菌群落结构的影响不显著；对于不同代谢类型的产甲烷菌，稻鱼共生(RFN和RFF)氢营养型产甲烷菌的丰度显著高于RM,且RFF的乙酸营养型产甲烷菌丰度显著高于RFN;对于不同代谢类型的甲烷氧化菌，RFN和RFF对类型Ⅰ的甲烷氧化菌丰度影响均不显著；RFF中类型Ⅱ的甲烷氧化菌丰度显著高于RM和RFN。可见，稻鱼共生可明显影响产甲烷菌和甲烷氧化...     

长期不同施肥下水稻土甲烷氧化能力及甲烷氧化菌多样性的变化

稻田内源甲烷的氧化是稻田甲烷减排的重要途径。而甲烷氧化菌是土壤中甲烷氧化的主要施动者,在长期不同施肥条件下,土壤微生物群落的演变是否影响到土壤甲烷氧化菌群落结构及其活性,进而影响到田土壤CH4向大气的实际排放强度还不清楚。为此,选择太湖地区一个长期肥料试验的稻田土壤为研究对象,分析长期不同肥料施用对土壤甲烷氧化能力的影响及其与土壤中甲烷氧化菌群落结构变化的可能关系。结果表明,长期不同的施肥措施下稻田土壤对甲烷的氧化能力产生了明显差异,伴随着土壤中甲烷氧化菌（MOBI和MOBII）的基因群落多样性的显著变化。长期单一施用氮肥为主的化肥显著降低了土壤对甲烷的氧化能力,同时显著降低了稻田土壤甲烷氧化菌的多样性和丰富度;不同施肥下甲烷氧化菌多样性的变化与土壤的甲烷氧化能力的变化趋势相一致。因此,研究显示长期不同施肥处理下甲烷氧化菌群落结构的改变可能是引起水稻土甲烷氧化能力变化的一个主要因素,有机无机配合施用可以明显降低稻田土壤甲烷的大气释放潜能。但长期不同施肥处理下甲烷氧化菌活性的变化还有待于进一步研究。     

闽江河口区淡水和半咸水潮汐沼泽湿地土壤产甲烷菌多样性

为认识盐度对河口潮汐沼泽湿地土壤产甲烷菌的影响,应用PCR-RFLP技术及测序分析对闽江河口区淡水-半咸水盐度梯度上分布的4个短叶茳芏潮汐沼泽湿地土壤产甲烷菌群落结构进行研究。闽江河口区短叶茳芏潮汐沼泽湿地土壤产甲烷菌群落结构受盐度影响明显,位于下洋洲和塔礁洲的短叶茳芏潮汐淡水沼泽湿地土壤产甲烷菌的香农-威纳多样性指数值分别为2.81和2.65,位于蝙蝠洲和鳝鱼滩的短叶茳芏潮汐半咸水沼泽湿地土壤产甲烷菌香农-威纳多样性指数值分别仅为2.33和2.27。系统发育分析表明:短叶茳芏沼泽湿地土壤产甲烷菌类群主要有甲烷杆菌目(Methanobacteriales),包括Methanobacterium、Methanobrevibacter和Methanobacteriaceae;甲烷微菌目(Methanomicrobiales),主要有Methanoregula,以及甲烷八叠球菌目(Methanosarcinales),主要有Methanosarcina和Methanococcoides。闽江河口区短叶茳芏潮汐淡水沼泽湿地土壤主要的优势产甲烷菌有Methanoregula、Methanosarcina和Methanobacterium,而短叶茳芏潮汐半咸水沼泽湿地土壤主要的优势产甲烷菌则转化为仅以Methanoregula为主。     

互花米草入侵对滩涂湿地甲烷排放的影响

在浙江省台州市附近滩涂湿地设置3个不同互花米草入侵密度梯度,即仅有本土植物样地、互花米草与本土植物混生样地和互花米草单优群落样地,研究互花米草入侵对滩涂湿地CH₄排放的影响.结果表明: 3个样地CH₄排放通量为0.68～5.88 mg·m^-2·h^-1,CH₄排放通量随着互花米草入侵梯度的增加而显著升高,互花米草单优群落样地CH₄排放通量分别为本土植物样地和混生样地的8.7和2.3倍.互花米草入侵显著提高了产甲烷菌数量、产甲烷潜力、甲烷氧化菌数量、甲烷氧化潜力、植物生物量、土壤有机碳含量和土壤pH,降低了土壤全氮含量.CH₄排放通量与土壤全氮呈显著负相关,与产甲烷菌数量、产甲烷潜力、甲烷氧化菌数量、甲烷氧化潜力、植物生物量和土壤pH呈显著正相关.互花米草的入侵提高了滩涂湿地植物群落生物量和土壤pH,促进了产甲烷菌数量和产甲烷潜力,从而提高了滩涂湿地的CH₄排放.     

湿地反硝化型甲烷厌氧氧化研究进展

甲烷是最重要的温室气体之一,其单分子温室效应是CO2的298倍。湿地是甲烷重要的排放源,也是氮素的源和汇。微生物参与湿地碳、氮转化的生物地球化学循环过程,湿地CH4是土壤淹水条件下微生物厌氧降解有机质而产生,微生物又可以通过反硝化型甲烷厌氧氧化过程(DAMO)降低湿地甲烷的排放,对缓解全球温室效应具有重要作用。本文系统介绍了DAMO过程机理、功能微生物Methylomirabilis oxyfera菌群特性、分布以及土壤DAMO过程的检测方法和DAMO过程的影响因素,并对未来更多的湿地DAMO微生物的发现,尤其是对稻田湿地DAMO过程的相关研究提出展望,以期推动该领域更深入的研究,为稻田湿地甲烷排放量的估算及制定合理的减排措施提供科学依据。     

冻土甲烷循环微生物群落及其对全球变化的响应

冻土是陆地生态系统中最容易受到全球气候变化影响的碳库,既发挥着碳源又起着碳汇的作用。人们非常关注贮存于冻土中有机碳的最终归宿,是因为全球气候变暖会加快冻土的解冻,释放更多的温室气体（二氧化碳和甲烷）到大气中,从而进一步加剧温室效应。据估计每年从北半球冻原陆地生态系统释放进入大气的甲烷约占全球自然界释放甲烷总量的25%。研究证实冻土生物源甲烷的产生和消耗分别由耐(嗜)低温的产甲烷菌(methanogens)和甲烷氧化菌(methanotrophs)介导。鉴于冻土甲烷循环对全球甲烷平衡的显著作用以及在冻土生物地球化学循环中的重要功能,对介导冻土甲烷循环的产甲烷菌和甲烷氧化菌的研究将有助于更好地评估冻土生态系统对全球气候变化的响应和影响,本文就冻土甲烷循环过程、产甲烷菌、甲烷氧化菌的群落结构、活动、生态功能及其对气候和环境变化的响应机制的最新研究进行综述,以期为我国开展冻土甲烷循环机理研究提供支持。     

DGGE揭示红壤稻田以产甲烷菌为主的稳定古菌群落结构特征

尽管古菌在农田土壤生态系统物质、能量循环中的作用尚未系统揭示,但已有研究表明产甲烷古菌和氨氧化古菌等在甲烷排放、固氮等方面具有重要生态功能。本研究采用优化的变性梯度凝胶电泳(DGGE)与克隆、序列测定相结合的方法,基于古菌16S V4~V5区分析我国水稻种植最常见的红壤地区稻田古菌在水稻生长不同时期的群落结构及多样性。在采集自江西省南昌地区的稻田土壤中,共检测到2个门、4个纲及5个目的古菌。作为主要优势类群,产甲烷菌纲(Methanomicrobia)丰度达49.15%;甲烷八迭球菌目(Methanosarcinales)丰度达39.45%。而氨氧化古菌属(Nitrosotalea)丰度也达6.07%。水稻分蘖期、灌浆期及成熟期稻田土壤古菌DGGE电泳条带数量及序列高度相似,分蘖期、灌浆期香农多样性指数未检测到具有统计学意义的差异(p=0.534)。成熟期香农多样性指数略高于分蘖期(p=0.017)和灌浆期(p=0.009),分析可能主要由土壤含水显著下降导致。红壤稻田的古菌群落结构在水稻生长期内能够基本保持稳定,是稻田古菌与人类耕作活动的长期共进化造成的。产甲烷菌中的优势类群——甲烷八迭球菌在水稻土壤中究竟是以消耗乙酸还是利用氢气/二氧化碳合成甲烷或两种代谢方式转换条件等方面研究,将有助于建立红壤水稻可持续耕作及红壤稻田酸化修复等的新方法。总之,稻田等农田生态系统中古菌的生态功能研究具有重要的理论意义和潜在应用价值,应予以高度重视。     

The impact of aridification and vegetation type on changes in the community structure of methane-cycling microorganisms in Japanese wetland soils

Over the years, the wetlands covered by Sphagnum in Bibai, Japan have been turning into areas of aridity, resulting in an invasion of Sasa into the bogs. Yet little is known about the methane-cycling microorganisms in such environments. In this study, the methanotrophic, methanogenic, and archaeal community structures within these two types of wetland vegetation were studied by phylogenetic analysis targeting particulate methane monooxygenase (pmoA), methyl coenzyme M reductase (mcrA), and the archaeal 16S rRNA gene. The pmoA library indicated that Methylomonas and Methylocystis predominated in the Sphagnum-covered and Sasa-invaded areas, respectively. The mcrA and 16S rRNA libraries indicated that Methanoregula were abundant methanogens in the Sphagnum-covered area. In the Sasa-invaded area, by contrast, mcrA genes were not detected, and no 16S rRNA clones were affiliated with previously known methanogens. Because the Sasa-invaded area still produced methane, of the various uncultured populations detected, novel euryarchaeotal lineages are candidate methane producers.     

Diversity of functional genes of methanogens, methanotrophs and sulfate reducers in deep-sea hydrothermal environments

To contribute to the identification of methanogens, methanotrophs and sulfate-reducing bacteria (SRB) in microbial communities from the 13 degrees N (East Pacific Rise) and Rainbow (Mid-Atlantic Ridge) hydrothermal vent fields, we investigated the diversity of mcrA, pmoA and dsrAB genes sequences. Clone libraries were obtained using DNA isolated from fragments of diffuse vents, sediment and in situ samplers. The clones were categorized by restriction fragment length polymorphism, and representatives of each group were sequenced. Sequences were related to that of hyperthermophilic (order Methanopyrales and family Methanocaldococcaceae), thermophilic and mesophilic (family Methanococcaceae) methanogens, thermophilic (proposed genus 'Methylothermus') and mesophilic type I methanotrophs, and hyperthermophilic (order Archaeoglobales), thermophilic (order Thermodesulfobacteriales) and mesophilic (family Desulfobulbaceae) SRB. Several of the obtained sequences were distantly related to the genes of cultivated organisms, providing evidence of the existence of novel lineages in the three functional groups. This study provides for the first time an insight into the diversity of several functional genes of deep-sea hydrothermal system microorganisms.     

Diversity of the particulate methane monooxygenase gene in methanotrophic samples from different rice field soils in China and the Philippines

Methanotrophic bacteria play a crucial role in regulating the emission of CH4 from rice fields into the atmosphere. We investigated the CH4 oxidation activity together with the diversity of methanotrophic bacteria in ten rice field soils from different geographic locations. Upon incubation of aerated soil slurries under 7% CH4, rates of CH4 oxidation increased after a lag phase of 1-4 days and reached values of 3-10 micromol d(-1) g-dw(-1) soil. The methanotrophic community was assayed by retrieval of the pmoA gene which encodes the a subunit of the particulate methane monooxygenase. After extraction of DNA from actively CH4-oxidizing soil samples and PCR-amplification of the pmoA, the community was analyzed by Denaturant Gradient Gel Electrophoresis (DGGE) and Terminal Restriction Fragment Length Polymorphism (T-RFLP). DGGE bands were excised, the pmoA re-amplified, sequenced and the encoded amino acid sequence comparatively analyzed by phylogenetic treeing. The analyses allowed the detection of pmoA sequences related to the following methanotrophic genera: the type-I methanotrophs Methylobacter, Methylomicrobium, Methylococcus and Methylocaldum, and the type-II methanotrophs Methylocystis and Methylosinus. T-RFLP analysis detected a similar diversity, but type-II pmoA more frequently than DGGE. All soils but one contained type-II in addition to type-I methanotrophs. Type-I Methylomonas was not detected at all. Different combinations of methanotrophic genera were detected in the different soils. However, there was no obvious geographic pattern of the distribution of methanotrophs.     

Heavy-machinery traffic impacts methane emissions as well as methanogen abundance and community structure in oxic forest soils

Temperate forest soils are usually efficient sinks for the greenhouse gas methane, at least in the absence of significant amounts of methanogens. We demonstrate here that trafficking with heavy harvesting machines caused a large reduction in CH(4) consumption and even turned well-aerated forest soils into net methane sources. In addition to studying methane fluxes, we investigated the responses of methanogens after trafficking in two different forest sites. Trafficking generated wheel tracks with different impact (low, moderate, severe, and unaffected). We found that machine passes decreased the soils' macropore space and lowered hydraulic conductivities in wheel tracks. Severely compacted soils yielded high methanogenic abundance, as demonstrated by quantitative PCR analyses of methyl coenzyme M reductase (mcrA) genes, whereas these sequences were undetectable in unaffected soils. Even after a year after traffic compression, methanogen abundance in compacted soils did not decline, indicating a stability of methanogens here over time. Compacted wheel tracks exhibited a relatively constant community structure, since we found several persisting mcrA sequence types continuously present at all sampling times. Phylogenetic analysis revealed a rather large methanogen diversity in the compacted soil, and most mcrA gene sequences were mostly similar to known sequences from wetlands. The majority of mcrA gene sequences belonged either to the order Methanosarcinales or Methanomicrobiales, whereas both sites were dominated by members of the families Methanomicrobiaceae Fencluster, with similar sequences obtained from peatland environments. The results show that compacting wet forest soils by heavy machinery causes increases in methane production and release.     

闽江口芦苇沼泽湿地土壤产甲烷菌群落结构的垂直分布

应用PCR-RFLP技术及测序分析对闽江口芦苇湿地土壤产甲烷菌群落结构的垂直分布特征进行了研究。在构建的6个克隆文库中,每个克隆文库随机挑选100个克隆进行菌落PCR验证,共得到591个阳性克隆。PCR产物经限制性内切酶MspⅠ进行RFLP分析后得到37个不同的分类操作单元(OTUs)。对37个克隆子进行了序列测定,与GenBank数据库中的序列进行比对,最近相似性在91%—99%之间。RFLP分析和系统发育分析表明,闽江口芦苇湿地土壤中产甲烷菌群落包括3大类群:甲烷杆菌目(Methanobacteriales)、甲烷微菌目(Methanomirobiales)和甲烷八叠球菌目(Methanosarcinales)。不同土壤深度中产甲烷菌群落的分布呈现出不同的特征。土壤表层(0—10 cm)优势产甲烷菌类群为Methanoregula,约占76%;10—20 cm土层主要的产甲烷菌类群为Methanolinea和Methanoregula,分别约占23%和29%;20—30cm土层优势的产甲烷菌类群为Methanolinea,约占66%。Shannon指数(H’)和Simpson多样性指教(D)表明,10—20cm土层产甲烷菌多样性高于土壤表层(0—10 cm)和20—30 cm土层。37个测序OTUs中有26个OTUs属于不可培养的产甲烷菌序列,表明闽江口芦苇湿地土壤中存在大量不可培养的产甲烷菌。     

Quantification of mcrA by fluorescent PCR in methanogenic and methanotrophic microbial communities

A quantitative fluorogenic PCR method for detecting methanogenic and methanotrophic orders was established using a refined primer set for the methyl coenzyme M reductase subunit A gene (mcrA). The method developed was applied to several microbial communities in which diversity and abundance of methanogens or anaerobic methanotrophs (ANMEs) was identified by 16S rRNA gene clone analysis, and strong correlations between the copy numbers of mcrA with those of archaeal 16S rRNA genes in the communities were observed. The assay can be applied to detecting and assessing the abundance of methanogens and/or ANMEs in anoxic environments that could not be detected by 16S rRNA gene sequence analyses.     

湿地松和马尾松人工林土壤甲烷代谢微生物群落的结构特征

土壤甲烷代谢微生物影响甲烷的产生和氧化,然而关于林型对土壤中甲烷代谢微生物群落结构影响的研究较少。采用基因芯片GeoChip 3.0研究了湿地松人工林和马尾松人工林土壤甲烷代谢微生物群落结构特征。结果如下,(1)两种林型的甲烷代谢微生物群落结构存在极显著差异(P=0.008),林型能解释其34.9%的变异;(2)产甲烷菌(包含甲基辅酶M还原α亚基基因mcrA的微生物)的优势菌群发生了变化,湿地松人工林的的优势菌为Methanocorpusculum labreanum Z,马尾松人工林的优势菌群除Methanocorpusculum labreanum Z外,还包括产甲烷古菌和Methanosarcina mazei Gol;(3)甲烷营养菌(包含甲烷单加氧酶基因pmoA基因的微生物)的优势菌为Ⅱ型,有3种不可培养细菌只在湿地松人工林检测到,在马尾松人工林中未检测到;(4)mcrA基因丰度或同源基因数量与土壤容重正相关,与土壤粘粒含量呈显著负相关;pmoA基因信号强度或多样性指数与土壤全碳含量、全磷含量和速效氮含量显著正相关。总之,相比本地种马尾松人工林,引进种湿地松人工林的土壤甲烷代谢微生物群落结构发生了显著变化。     

Detection of methanotroph diversity on roots of submerged rice plants by molecular retrieval of pmoA, mmoX, mxaF, and 16S rRNA and ribosomal DNA, including pmoA-based terminal restriction fragment length polymorphism profiling

The diversity of methanotrophic bacteria associated with roots of submerged rice plants was assessed using cultivation-independent techniques. The research focused mainly on the retrieval of pmoA, which encodes the alpha subunit of the particulate methane monooxygenase. A novel methanotroph-specific community-profiling method was established using the terminal restriction fragment length polymorphism (T-RFLP) technique. The T-RFLP profiles clearly revealed a more complex root-associated methanotrophic community than did banding patterns obtained by pmoA-based denaturing gradient gel electrophoresis. The comparison of pmoA-based T-RFLP profiles obtained from rice roots and bulk soil of flooded rice microcosms suggested that there was a substantially higher abundance of type I methanotrophs on rice roots than in the bulk soil. These were affiliated to the genera Methylomonas, Methylobacter, Methylococcus, and to a novel type I methanotroph sublineage. By contrast, type II methanotrophs of the Methylocystis-Methylosinus group could be detected with high relative signal intensity in both soil and root compartments. Phylogenetic treeing analyses and a set of substrate-diagnostic amino acid residues provided evidence that a novel pmoA lineage was detected. This branched distinctly from all currently known methanotrophs. To examine whether the retrieval of pmoA provided a complete view of root-associated methanotroph diversity, we also assessed the diversity detectable by recovery of genes coding for subunits of soluble methane monooxygenase (mmoX) and methanol dehydrogenase (mxaF). In addition, both 16S rRNA and 16S ribosomal DNA (rDNA) were retrieved using a PCR primer set specific to type I methanotrophs. The overall methanotroph diversity detected by recovery of mmoX, mxaF, and 16S rRNA and 16S rDNA corresponded well to the diversity detectable by retrieval of pmoA.     

Microbial diversity in sediments associated with a shallow methane seep in the tropical Timor Sea of Australia reveals a novel aerobic methanotroph diversity

This study examined the diversity of Bacteria, Archaea and in particular aerobic methanotrophs associated with a shallow (84 m) methane seep in the tropical Timor Sea, Australia. Seepage of thermogenic methane was associated with a large carbonate hardground covered in coarse carbonate-rich sediments and various benthic organisms such as solitary corals. The diversity of Bacteria and Archaea was studied by analysis of cloned 16S rRNA genes, while aerobic methanotrophic bacteria were quantified using real-time PCR targeting the α-subunit of particulate methane monooxygenase ( pmoA ) genes and diversity was studied by analysis of cloned pmoA genes. Phylogenetic analysis of bacterial and archaeal 16S rRNA genes revealed diverse and mostly novel phylotypes related to sequences previously recovered from marine sediments. A small number of bacterial 16S rRNA gene sequences were related to aerobic methanotrophs distantly related to the genera Methylococcus and Methylocaldum . Real-time PCR targeting pmoA genes showed that the highest numbers of methanotrophs were present in surface sediments associated with the seep area. Phylogenetic analysis of pmoA sequences revealed that all phylotypes were novel and fell into two large clusters comprised of only marine sequences distantly related to the genera Methylococcus and Methylocaldum that were clearly divergent from terrestrial phylotypes. This study provides evidence for the existence of a novel microbial diversity and diverse aerobic methanotrophs that appear to constitute marine specialized lineages.