不同利用方式对红壤坡地微生物多样性和硝化势的影响

采集了中国科学院桃源农业生态试验站红壤坡地农田、自然恢复林和茶园土壤样品,采用末端限制性酶切片段长度多态性分析(T.RFLP)技术分析土壤细菌、古菌、氨氧化细菌(AOB)和氨氧化古菌(AOA)的多样性,采用好气培养法测定不同土壤的硝化势,研究不同土地利用方式对微生物多样性和硝化势的影响.结果表明:土壤AOB和AOA多样性指数差异不显著,且在3种不同土地利用方式中呈现相同的趋势,均为农田=茶园>自然恢复林;通过RDA分析发现,不同利用方式造成土壤理化性状的改变是影响土壤AOA和AOB群落结构的主要原因;好气培养法测得不同土壤硝化势农田最高,茶园次之而自然恢复林最低;相关性分析显示,硝化势与细菌16S rRNA、AOA和AOB amoA基因多样性指数呈显著正相关,其中与AOA amoA基因关系最为密切;总体来说,红壤坡地不同利用方式改变了土壤细菌、古菌、AOA和AOB的多样性,土壤AOB和AOA积极参与了土壤的硝化过程,且AOA在氨氧化微生物群落生态功能中占有重要地位,AOA比AOB与硝化势的关系更为密切.     

南美白对虾养殖底泥中氨氧化细菌与氨氧化古菌多态性分析

【目的】本研究皆在了解虾养殖底泥中氨氧化细菌与氨氧化古菌群落多态性。【方法】以功能基因为基础,构建氨氧化细菌(AOB)与氨氧化古菌(AOA)的氨单加氧酶α亚基基因(amoA)克隆文库。利用限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)技术将克隆文库阳性克隆子进行归类分析分成若干个可操作分类单元(Operational Taxa Units,OTUs)。【结果】通过序列多态性分析,表明AOB amoA基因克隆文库中所有序列都属于变形杆菌门β亚纲(β-Proteobacteria)中的亚硝化单细胞菌属(Nitrosomonas)及Nitrosomonas-like,未发现亚硝化螺旋菌属(Nitrosospira)。AOA amoA基因克隆文库中只有一个OTU序列属于未分类的古菌(Unclassified-Archaea),其余序列都属于泉古菌门(Crenarchaeote)。AOA群落结构单一且存在一个绝对优势类群OTU3,其克隆子数目占克隆文库的57.45%。AOB和AOA amoA基因克隆文库分别包括13个OTUs和9个OTUs,其文库覆盖率分别为73.47%和90.43%。AOB amoA基因克隆文库的Shannon-Wiener指数、Evenness指数、Simpson指数、Richness指数均高于AOA。【结论】虾养殖塘底泥中存在氨氧化古菌的amoA基因,且多态性低于氨氧化细菌,表明氨氧化古菌在虾养殖塘底泥的氮循环中可能具有重要的作用。     

河口生态系统氨氧化菌生态学研究进展

由amoA基因编码的氨单加氧酶(AMO)所调控的氨氧化作用,是硝化作用的限速步骤和中心环节,而含有amoA基因的氨氧化细菌(AOB)和氨氧化古菌(AOA)多样性与环境因子关系密切,对缓解河口生态系统因人类活动造成的富营养化等环境问题具有特别重要的意义。水、陆和海交汇形成高度变异的具环境因子梯度的河口生态系统,是研究AOA和AOB生态学的天然实验室。河口AOA与AOB的群落组成、丰富度特征和生物有效性,与河口主要环境因子盐度、富营养化程度、植被、温度、碳、氮、硫、铁等,尤其是对盐度和富营养化有着较为强烈的响应。AOA和AOB多样性变化规律及其与河口特有的环境因子之间的相关性,应当是今后我国河口氨氧化菌研究的方向和重点。包括:(1)建立有效的氨氧化菌活性评价方法;(2)研究AOA的同化作用方式;(3)依据氨氧化菌分类和组成对河口环境变化的适应进化机制,建议可作为指示河口环境质量变化的生物标记;(4)将传统的分离培养方法与现代分子生物学研究方法相结合,筛选我国河口高效的氨氧化菌,并将其应用于生产。     

辉腾锡勒草原干涸湖泊中氨氧化微生物群落结构分析

【目的】以内蒙古辉腾锡勒草原九十九泉湿地为对象,研究湖泊干涸过程中氨氧化微生物的群落结构及其变化。【方法】通过MPN-PCR定量测定氨氧化古菌(AOA)和氨氧化细菌(AOB)的数量;构建amoA基因克隆文库,进行系统发育分析;结合土壤环境因子,探讨湿地退化过程中影响氨氧化微生物的潜在因素。【结果】依湖泊湿地退水梯度的不同样点中,有75%的样点AOB的数量高于AOA,AOB与AOA的数量比率为0.3-18.1。从湖心到湖岸草原带,AOA和AOB的数量有明显增加,但生物多样性呈降低趋势,二者没有呈现正相关。研究发现,AOB的数量与土壤中NH 4+-N的变化存在良好响应。系统发育分析显示,退化湖泊湿地AOA克隆序列均来自于泉古菌门(Crenarchaeota);AOB的amoA基因的克隆序列大部分与亚硝化单胞菌属(Nitrosomonas)有一定同源性,较少部分与亚硝化螺菌属(Nitrosospira)有一定同源性。【结论】湖泊退水过程增加了湿地土壤氨氧化微生物的数量,而氨氧化微生物的种群丰度有所降低。AOA和AOB群落对湖泊湿地的退化过程做出了响应,其中AOB的响应较为明显,氧化条件和土壤铵浓度的改变可能是促成这种响应的重要原因。     

华北典型旱地小麦土壤amoA基因的PCR-RFLP分析

通过构建氨氧化细菌(AOB )和氨氧化古菌(AOA)的氨氧化酶基因亚基A (amoA )克隆文库,并采用限制性片段长度多态性(Restriction Fragment Length Polymorphism, RFLP )技术分析了华北地区典型旱地冬小麦土壤中amoA 基因的多样性.采用MspI 和AfaI 两种限制性内切酶对amoA 基因克隆文库中阳性克隆子进行双酶切后,共得到了18 个氨氧化细菌的可操作分类单元(Operational Taxa Units, OTUs )和10 个氨氧化古菌可操作分类单元(Operational Taxa Units, OTUs ),其文库覆盖率分别达到92.9%和88.3%.氨氧化细菌的Shannon-Wiener 指数、丰富度指数、均匀度指数均高于氨氧化古菌.通过对文库中amoA细菌测序分析,所有的序列都属于Nitrosospira cluster 3 .而在氨氧化古菌中存在着一个绝对优势种群,它占到克隆文库的80 %,测序分析的结果表明,氨氧化古菌属于不可培养的泉古菌门.     

白洋淀湖滨湿地岸边带氨氧化古菌与氨氧化细菌的分布特性

摘要：本研究通过分子生物学分析方法,以amoA基因为标记,考察了氨氧化古菌（Ammonia-Oxidizing Archaea, AOA）和氨氧化细菌（Ammonia-Oxidizing Bacteria,AOB）在华北平原的白洋淀这一典型湖泊的湖滨湿地岸边带系统中的生物多样性和丰度分布。在前人的研究中,氨氧化古菌在海洋、原生态土壤和人为干扰土壤等环境中主导氨氧化过程的完成。但本研究发现,在湿地岸边带系统中氨氧化过程并不是完全由氨氧化古菌主导完成,即氨氧化古菌和氨氧化细菌在不同区域分别占据主导地位。根据主导微生物的不同,可以将湿地岸边带区域划分为陆相区、中间区和湖相区。在湿地岸边带陆相区,氨氧化古菌主导氨氧化过程,氨氧化古菌的amoA基因丰度是氨氧化细菌的526倍（AOA：1.23?108每克干土;AOB：2.34?105每克干土）;在岸边带湖相区,氨氧化细菌主导氨氧化过程,氨氧化古菌的amoA基因丰度只有氨氧化细菌的1/50倍（AOA：3.17?106每克干土;AOB：1.39?108每克干土）;在岸边带中间区,两种微生物对氨氧化过程的贡献相当,二者的amoA基因丰度也相当 (AOA：9.83?106, AOB：4.08?106)。研究还发现,湿地中间区的微生物生物多样性高于陆相区和湖相区。在湿地中间区,氨氧化古菌和氨氧化细菌的生物多样性都最高,分别有5和7个操作分类单元（OTUs）;相比之下,岸边带陆相区和湖相区的多样性依次降低,陆相区的氨氧化古菌和氨氧化细菌分别有3和6个操作分类单元,湖相区的氨氧化古菌和氨氧化细菌分别有2和6个分类单元。本研究的两个结论进一步反映了湿地岸边带极强的空间异质性。     

长期不同施肥制度对水稻土氨氧化微生物数量和群落结构的影响

由氨氧化微生物驱动的氨氧化过程是硝化作用的限速步骤,在土壤氮素循环过程中扮演着重要角色.以湖南省宁乡县长达30 a定位试验水稻土壤为研究对象,采用荧光定量PCR和Illumina MiSeq高通量测序分析方法,以amoA基因为靶标,研究了4种施肥制度[不施肥(CK)、化肥(CF)、70%化肥+30%有机肥(CFM₁)和40%化肥+60%有机肥(CFM₂)]水稻土壤氨氧化微生物的数量和群落结构变化.结果表明: 不同施肥处理氨氧化古菌(AOA)和氨氧化细菌(AOB) amoA基因拷贝数分别为3.09×10⁷～8.37×10⁷和1.04×10⁷～7.03×10⁷ copies·g^-1干土.施肥显著提高了AOA和AOB数量,但处理CFM₂中AOB数量与CK差异不显著.有机肥配施比例对AOB群落α多样性指数的影响强于AOA,处理CFM₁中AOA群落的多样性指数(Shannon)和AOB群落的丰富度指数(ACE和Chao1)均显著高于CK.奇古菌门和泉古菌门是AOA群落的优势门类群,占AOA amoA基因总序列的83.4%;亚硝化螺菌属、environmental_samples_norank、Bacteria_unclassified和Nitrosomonadales_unclassified是AOB群落的优势属类群,占AOB amoA基因总序列的97.8%.维恩分析结果显示,有机肥配施比例对AOB群落操作分类单元(OTU)数量的影响强于AOA,但对各处理共有AOA和AOB amoA基因序列条数的影响均较小.冗余分析结果显示,不同施肥处理AOB群落结构差异强于AOA,且所有土壤理化性质均与AOA和AOB群落结构存在显著相关关系.综上可知:有机肥配施比例显著改变了AOA和AOB数量、多样性和群落结构,配施30%有机肥时,AOA群落的Shannon指数最高,AOB群落数量、ACE和Chao1指数均最高.研究结果可为进一步探讨农业系统中氨氧化微生物对不同施肥制度的响应机制及其在氮素转化中的作用提供科学依据.     

亚高山/高山森林土壤有机层氨氧化细菌和氨氧化古菌丰度特征

为了解季节性冻融作用对川西亚高山/高山地区土壤氨氧化微生物群落的影响,采用qPCR技术,以氨单加氧酶基因的α亚基(amoA)为标记,在生长阶段、冻结阶段、融化阶段中的9个关键时期调查了该地区不同森林群落：岷江冷杉(Abies faxoniana)原始林(PF)、岷江冷杉(A. faxoniana)和红桦(Betula albosinensis)混交林(MF)、岷江冷杉次生林(SF)土壤有机层的氨氧化细菌(ammonia-oxidizing bacteria, AOB)和氨氧化古菌(ammonia-oxidizing archaea, AOA)丰度的特征。结果表明,三个森林群落土壤有机层中都具有相当数量的氨氧化细菌和古菌,均表现出从生长阶段至冻结阶段显著降低,在冻结阶段最低,但冻结阶段后显著增加,在融化阶段为全年最高的趋势。土壤氨氧化微生物类群结构(AOA/AOB)受负积温影响明显。冻结后期三个森林群落土壤负积温最大时,AOA数量明显高于AOB,但其他关键时期土壤氨氧化微生物类群结构与群落类型密切相关。高海拔的PF群落土壤有机层表现为AOA>AOB(冻结初期除外),低海拔的SF群落中表现为AOB>AOA(冻结后期除外),而MF群落则仅在融冻期和生长季节末期表现为AOB>AOA。这些结果为认识亚高山/高山森林及其相似区域的生态过程提供了一定的科学依据。     

缢蛏养殖池塘氨氧化微生物的季节变化特征

为探究缢蛏(Sinonovacula constricta)养殖池塘中氨氧化细菌(AOB)和氨氧化古菌(AOA)的季节变化及其环境因子相关性, 试验以amoA基因为分子标记, 利用荧光定量PCR技术对养殖池塘水体及沉积物中的AOA和AOB进行了研究。结果显示, 季节变化对于水体及沉积物中的AOA和AOB的amoA基因丰度有一定的影响, 不同季节的水体及沉积物中AOA含量均比AOB含量高一个数量级。其中水体中AOB在秋季显著高于其余三季(P<0.05); AOA在冬季最低, 且显著低于春秋两季(P<0.05)。沉积物中AOB和AOA均在冬季显著高于其余三季(P<0.05)。环境因子相关性分析结果表明: 水体中AOB丰度与氨氮成显著正相关(P<0.05), 与亚硝氮成显著负相关(P<0.05); AOA丰度与水温成正显著相关(P<0.05), 与硝氮成负显著相关(P<0.05)。沉积物中AOB丰度与溶解氧、硝氮、总氮及亚硝氮成显著正相关(P<0.05), 与水温成显著负相关(P<0.05); AOA丰度与溶解氧、总氮、硝氮成显著性正相关(P<0.05), 与水温成负显著相关(P<0.05)。多元线性回归方程结果显示, 缢蛏养殖池塘的AOA及AOB丰度与硝氮密切相关。研究表明, AOA在缢蛏养殖池塘氨氧化进程中占据主导地位, AOA和AOB丰度受盐度、温度、氮素浓度及溶解氧等环境因子的共同影响。     

东湖表层沉积物中氨氧化古菌和氨氧化细菌丰度及多样性研究

研究通过高通量测序和荧光定量PCR等分子生物学分析方法, 以氨单加氧酶基因(amoA)为分子标记, 研究了东湖表层沉积物中AOA和AOB的群落多样性、丰度及其与环境因子的关系。结果表明, 东湖沉积物AOA主要为Nitrosopumilus, 其群落结构与沉积物中总氮含量显著相关, 而AOB主要为Nitrosomonas, 群落结构与沉积物中总有机碳和总磷显著相关。此外, 不同季节AOA丰度均高于AOB, 且沉积物AOA数量与温度呈显著负相关, 但AOB丰度变化不明显。东湖沉积物中AOA可能主导了氨氧化过程。     

Comparative analysis of archaeal 16S rRNA and <Emphasis Type="Italic">amoA</Emphasis> genes to estimate the abundance and diversity of ammonia-oxidizing archaea in marine sediments

Considering their abundance and broad distribution, non-extremophilic Crenarchaeota are likely to play important roles in global organic and inorganic matter cycles. The diversity and abundance of archaeal 16S rRNA and putative ammonia monooxygenase alpha-subunit (amoA) genes were comparatively analyzed to study genetic potential for nitrification of ammonia-oxidizing archaea (AOA) in the surface layers (0-1 cm) of four marine sediments of the East Sea, Korea. After analysis of a 16S rRNA gene clone library, we found various archaeal groups that include the crenarchaeotal group (CG) I.1a (54.8%) and CG I.1b (5.8%), both of which are known to harbor ammonia oxidizers. Notably, the 16S rRNA gene of CG I.1b has only previously been observed in terrestrial environments. The 16S rRNA gene sequence data revealed a distinct difference in archaeal community among sites of marine sediments. Most of the obtained amoA sequences were not closely related to those of the clones retrieved from estuarine sediments and marine water columns. Furthermore, clades of unique amoA sequences were likely to cluster according to sampling sites. Using real-time PCR, quantitative analysis of amoA copy numbers showed that the copy numbers of archaeal amoA ranged from 1.1 x 10(7) to 4.9 x 10(7) per gram of sediment and were more numerous than those of bacterial amoA, with ratios ranging from 11 to 28. In conclusion, diverse CG I.1a and CG I.1b AOA inhabit surface layers of marine sediments and AOA, and especially, CG I.1a are more numerous than other ammonia-oxidizing bacteria.     

Contrasting spatiotemporal patterns and environmental drivers of diversity and community structure of ammonia oxidizers,denitrifiers, and anammox bacteria in sediments of estuarine tidal flats

The spatial and temporal patterns of diversity, community structure, and their drivers are fundamental issues in microbial ecology. This study aimed to investigate the relative importance of spatial and seasonal controls on the distribution of nitrogen cycling microbes in sediments of estuarine tidal flats, and to test the hypothesis that metals impact the distribution of nitrogen-cycling microbes in the coastal system. Two layers of sediment samples were collected from three estuarine tidal flats of Laizhou Bay in 2010 winter and 2011 summer. The alpha diversities (Shannon and Simpson indices) and community structure of ammonia oxidizing bacteria (AOB) and archaea (AOA), denitrifier and anammox bacteria (AMB) were revealed using denaturing gradient gel electrophoresis and clone library analysis of amoA, nosZ and 16S rRNA gene markers. We found that both AOB and AMB exhibited distinct seasonal patterns in either alpha diversity or community turnover; AOA had different alpha diversities in two layers, but neither spatial nor seasonal patterns were found for their community turnover. However, no distinct spatiotemporal pattern was observed for either diversity or community composition of nosZ-type denitrifiers. For correlations between alpha diversities and environmental factors, significant correlations were found between AOB and ammonium, temperature and As, between denitrifiers and nitrite, salinity and Pb, and between AMB and Pb, ratio of organic carbon to nitrogen, ammonium, pH and dissolved oxygen. Salinity and sediment grain size were the most important factors shaping AOB and AOA communities, respectively; whereas AMB community structure was mostly determined by temperature, dissolved oxygen, pH and heavy metals As and Cd. These results stress that ammonia oxidizers, denitrifiers and anammox bacteria have generally different distributional patterns across time and space, and heavy metals might have contributed to their differentiated distributions in coastal sediments.     

Abundance and composition of epiphytic bacterial and archaeal ammonia oxidizers of marine red and brown macroalgae

Ammonia-oxidizing bacteria (AOB) and archaea (AOA) are important for nitrogen cycling in marine ecosystems. Little is known about the diversity and abundance of these organisms on the surface of marine macroalgae, despite the algae's potential importance to create surfaces and local oxygen-rich environments supporting ammonia oxidation at depths with low dissolved oxygen levels. We determined the abundance and composition of the epiphytic bacterial and archaeal ammonia-oxidizing communities on three species of macroalgae, Osmundaria volubilis, Phyllophora crispa, and Laminaria rodriguezii, from the Balearic Islands (western Mediterranean Sea). Quantitative PCR of bacterial and archaeal 16S rRNA and amoA genes was performed. In contrast to what has been shown for most other marine environments, the macroalgae's surfaces were dominated by bacterial amoA genes rather than those from the archaeal counterpart. On the basis of the sequences retrieved from AOB and AOA amoA gene clone libraries from each algal species, the bacterial ammonia-oxidizing communities were related to Nitrosospira spp. and to Nitrosomonas europaea and only 6 out of 15 operational taxonomic units (OTUs) were specific for the host species. Conversely, the AOA diversity was higher (43 OTUs) and algal species specific, with 17 OTUs specific for L. rodriguezii, 3 for O. volubilis, and 9 for P. crispa. Altogether, the results suggest that marine macroalgae may exert an ecological niche for AOB in marine environments, potentially through specific microbe-host interactions.     

Effects of allylthiourea,salinity, and pH on ammonia/ammonium-oxidizing prokaryotes in mangrove sediment incubated in laboratory microcosms

Anaerobic ammonium-oxidizing (anammox) bacteria, aerobic ammonia-oxidizing archaea (AOA) and bacteria (AOB) are three groups of ammonia/ammonium-oxidizing prokaryotes (AOPs) involved in the biochemical nitrogen cycling. In this study, the effects of allylthiourea (ATU), pH, and salinity on these three groups from mangrove sediment were investigated through microcosm incubation in laboratory. ATU treatments (50, 100, and 500 mg L^?1) obviously affected the community structure of anammox bacteria and AOB, but only slightly for AOA. ATU began to inhibit anammox bacteria growth slightly from day 10, but had an obvious inhibition on AOA growth from the starting of the study. At 100 mg L^?1 of ATU or higher, AOB growth was inhibited, but only lasted for 5 days. The pH treatments showed that acidic condition (pH 5) had a slight effect on the community structure of anammox bacteria and AOA, but an obvious effect on AOB. Acidic condition promoted the growth of all groups of AOPs in different extent, but alkaline condition (pH 9) had a weak effect on AOB community structure and a strong effect on both anammox bacteria and AOA. Alkaline condition obviously inhibited anammox bacteria growth, slightly promoted AOA, and slightly promoted AOB in the first 20 days, but inhibited afterward. Salinity treatment showed that higher salinity (20 and 40?‰) resulted in higher anammox bacteria diversity, and both AOA and AOB might have species specificity to salinity. High salinity promoted the growth of both anammox bacteria and AOB, inhibited AOA between 5 and 10 days, but promoted afterward. The results help to understand the role of these microbial groups in biogeochemical nitrogen cycling and their responses to the changing environments.     

古尔班通古特沙漠生物土壤结皮对氨氧化微生物生态位的影响

生物结皮作为荒漠地表的重要覆被类型, 在荒漠生态系统的氮素循环中扮演重要角色。融雪期为古尔班通古特沙漠生物结皮的复苏和生长提供了充足的水分, 也成为该沙漠氮素固定和转化的重要时期, 但该时期生物结皮如何影响驱动氨氧化转化的微生物群落动态尚未明确。因此, 我们利用荧光定量PCR (fluorescent quantitative PCR, qPCR)方法分析融雪期生物结皮与去除结皮不同土层(0-2, 2-5, 5-10和10-20 cm)氨氧化菌群丰度特征, 结合潜在硝化速率和土壤理化参数, 探究融雪期生物结皮对荒漠土壤氮素转化作用。结果表明: 氨氧化古菌(ammonia-oxidizing archaea, AOA)是古尔班通古特沙漠土壤优势氨氧化菌, 生物结皮对0-2 cm层土壤中AOA、氨氧化细菌(ammonia-oxidizing bacteria, AOB) amoA基因丰度具有显著抑制作用(P < 0.01), 对10-20 cm层土壤中AOA amoA基因丰度具有显著促进作用(P < 0.01)。冗余分析(redundancy analysis, RDA)表明, AOA、AOB amoA基因丰度主要受土壤含水量和铵态氮含量的影响, 占总条件效应的54.90%。氨氧化速率分析发现, 去除生物结皮显著降低古尔班通古特沙漠土壤硝化作用潜力(P < 0.001), 证实生物结皮对荒漠土壤氮素转化具有重要的调控作用。综上所述, 古尔班通古特沙漠氨氧化微生物的分布规律受环境因子调控, 特别是生物结皮可以通过调节土壤含水量和铵态氮含量影响AOA和AOB的空间生态位分化, 促进沙漠土壤的硝化作用。     

Aquarium nitrification revisited: Thaumarchaeota are the dominant ammonia oxidizers in freshwater aquarium biofilters

Ammonia-oxidizing archaea (AOA) outnumber ammonia-oxidizing bacteria (AOB) in many terrestrial and aquatic environments. Although nitrification is the primary function of aquarium biofilters, very few studies have investigated the microorganisms responsible for this process in aquaria. This study used quantitative real-time PCR (qPCR) to quantify the ammonia monooxygenase (amoA) and 16S rRNA genes of Bacteria and Thaumarchaeota in freshwater aquarium biofilters, in addition to assessing the diversity of AOA amoA genes by denaturing gradient gel electrophoresis (DGGE) and clone libraries. AOA were numerically dominant in 23 of 27 freshwater biofilters, and in 12 of these biofilters AOA contributed all detectable amoA genes. Eight saltwater aquaria and two commercial aquarium nitrifier supplements were included for comparison. Both thaumarchaeal and bacterial amoA genes were detected in all saltwater samples, with AOA genes outnumbering AOB genes in five of eight biofilters. Bacterial amoA genes were abundant in both supplements, but thaumarchaeal amoA and 16S rRNA genes could not be detected. For freshwater aquaria, the proportion of amoA genes from AOA relative to AOB was inversely correlated with ammonium concentration. DGGE of AOA amoA genes revealed variable diversity across samples, with nonmetric multidimensional scaling (NMDS) indicating separation of freshwater and saltwater fingerprints. Composite clone libraries of AOA amoA genes revealed distinct freshwater and saltwater clusters, as well as mixed clusters containing both freshwater and saltwater amoA gene sequences. These results reveal insight into commonplace residential biofilters and suggest that aquarium biofilters may represent valuable biofilm microcosms for future studies of AOA ecology.     

PCR profiling of ammonia-oxidizer communities in acidic soils subjected to nitrogen and sulphur deposition

Communities of ammonia-oxidizing bacteria (AOB) were characterized in two acidic soil sites experimentally subjected to varying levels of nitrogen and sulphur deposition. The sites were an acidic spruce forest soil in Deepsyke, Southern Scotland, with low background deposition, and a nitrogen-saturated upland grass heath in Pwllpeiran, North Wales. Betaproteobacterial ammonia-oxidizer 16S rRNA and ammonia monooxygenase (amoA) genes were analysed by cloning, sequencing and denaturing gradient gel electrophoresis (DGGE). DGGE profiles of amoA and 16S rRNA gene fragments from Deepsyke soil in 2002 indicated no effect of nitrogen deposition on AOB communities, which contained both Nitrosomonas europaea and Nitrosospira. In 2003, only Nitrosospira could be detected, and no amoA sequences could be retrieved. These results indicate a decrease in the relative abundance of AOB from the year 2002 to 2003 in Deepsyke soil, which may be the result of the exceptionally low rainfall in spring 2003. Nitrosospira-related sequences from Deepsyke soil grouped in all clusters, including cluster 1, which typically contains only sequences from marine environments. In Pwllpeiran soil, 16S rRNA gene libraries were dominated by nonammonia oxidizers and no amoA sequences were detectable. This indicates that autotrophic AOB play only a minor role in these soils even at high nitrogen deposition.     

Mimicking the oxygen minimum zones: stimulating interaction of aerobic archaeal and anaerobic bacterial ammonia oxidizers in a laboratory‐scale model system

In marine oxygen minimum zones (OMZs), ammonia‐oxidizing archaea (AOA) rather than marine ammonia‐oxidizing bacteria (AOB) may provide nitrite to anaerobic ammonium‐oxidizing (anammox) bacteria. Here we demonstrate the cooperation between marine anammox bacteria and nitrifiers in a laboratory‐scale model system under oxygen limitation. A bioreactor containing ‘Candidatus Scalindua profunda’ marine anammox bacteria was supplemented with AOA (Nitrosopumilus maritimus strain SCM1) cells and limited amounts of oxygen. In this way a stable mixed culture of AOA, and anammox bacteria was established within 200 days while also a substantial amount of endogenous AOB were enriched. ‘Ca. Scalindua profunda’ and putative AOB and AOA morphologies were visualized by transmission electron microscopy and a C₁₈ anammox [3]‐ladderane fatty acid was highly abundant in the oxygen‐limited culture. The rapid oxygen consumption by AOA and AOB ensured that anammox activity was not affected. High expression of AOA, AOB and anammox genes encoding for ammonium transport proteins was observed, likely caused by the increased competition for ammonium. The competition between AOA and AOB was found to be strongly related to the residual ammonium concentration based on amoA gene copy numbers. The abundance of archaeal amoA copy numbers increased markedly when the ammonium concentration was below 30 μM finally resulting in almost equal abundance of AOA and AOB amoA copy numbers. Massive parallel sequencing of mRNA and activity analyses further corroborated equal abundance of AOA and AOB. PTIO addition, inhibiting AOA activity, was employed to determine the relative contribution of AOB versus AOA to ammonium oxidation. The present study provides the first direct evidence for cooperation of archaeal ammonia oxidation with anammox bacteria by provision of nitrite and consumption of oxygen.