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      Isolation of ‘ Candidatus Nitrosocosmicus franklandus’, a novel ureolytic soil archaeal ammonia oxidiser with tolerance to high ammonia concentration

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          Abstract

          Studies of the distribution of ammonia oxidising archaea (AOA) and bacteria (AOB) suggest distinct ecological niches characterised by ammonia concentration and pH, arising through differences in substrate affinity and ammonia tolerance. AOA form five distinct phylogenetic clades, one of which, the ‘ Nitrososphaera sister cluster’, has no cultivated isolate. A representative of this cluster, named ‘ Candidatus Nitrosocosmicus franklandus’, was isolated from a pH 7.5 arable soil and we propose a new cluster name: ‘Nitrosocosmicus’. While phylogenetic analysis of amoA genes indicates its association with the Nitrososphaera sister cluster, analysis of 16S rRNA genes provided no support for a relative branching that is consistent with a ‘sister cluster’, indicating placement within a lineage of the order Nitrososphaerales. ‘ Ca. N. franklandus’ is capable of ureolytic growth and its tolerances to nitrite and ammonia are higher than in other AOA and similar to those of typical soil AOB. Similarity of other growth characteristics of ‘ Ca. N. franklandus’ with those of typical soil AOB isolates reduces support for niche differentiation between soil AOA and AOB and suggests that AOA have a wider physiological diversity than previously suspected. In particular, the high ammonia tolerance of ‘ Ca. N. franklandus’ suggests potential contributions to nitrification in fertilised soils.

          Abstract

          This study describes ecophysiological characteristics of the first cultivated representative of a soil archaeal group performing an important biogeochemical process, ammonia oxidation, with significance for niche specialisation and archaeal phylogeny.

          Abstract

          Graphical Abstract Figure.

          This study describes ecophysiological characteristics of the first cultivated representative of a soil archaeal group performing an important biogeochemical process, ammonia oxidation, with significance for niche specialisation and archaeal phylogeny.

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          The influence of soil pH on the diversity, abundance and transcriptional activity of ammonia oxidizing archaea and bacteria.

          Autotrophic ammonia oxidation occurs in acid soils, even though laboratory cultures of isolated ammonia oxidizing bacteria fail to grow below neutral pH. To investigate whether archaea possessing ammonia monooxygenase genes were responsible for autotrophic nitrification in acid soils, the community structure and phylogeny of ammonia oxidizing bacteria and archaea were determined across a soil pH gradient (4.9-7.5) by amplifying 16S rRNA and amoA genes followed by denaturing gradient gel electrophoresis (DGGE) and sequence analysis. The structure of both communities changed with soil pH, with distinct populations in acid and neutral soils. Phylogenetic reconstructions of crenarchaeal 16S rRNA and amoA genes confirmed selection of distinct lineages within the pH gradient and high similarity in phylogenies indicated a high level of congruence between 16S rRNA and amoA genes. The abundance of archaeal and bacterial amoA gene copies and mRNA transcripts contrasted across the pH gradient. Archaeal amoA gene and transcript abundance decreased with increasing soil pH, while bacterial amoA gene abundance was generally lower and transcripts increased with increasing pH. Short-term activity was investigated by DGGE analysis of gene transcripts in microcosms containing acidic or neutral soil or mixed soil with pH readjusted to that of native soils. Although mixed soil microcosms contained identical archaeal ammonia oxidizer communities, those adapted to acidic or neutral pH ranges showed greater relative activity at their native soil pH. Findings indicate that different bacterial and archaeal ammonia oxidizer phylotypes are selected in soils of different pH and that these differences in community structure and abundances are reflected in different contributions to ammonia oxidizer activity. They also suggest that both groups of ammonia oxidizers have distinct physiological characteristics and ecological niches, with consequences for nitrification in acid soils.
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            amoA-based consensus phylogeny of ammonia-oxidizing archaea and deep sequencing of amoA genes from soils of four different geographic regions

            Ammonia-oxidizing archaea (AOA) play an important role in nitrification and many studies exploit their amoA genes as marker for their diversity and abundance. We present an archaeal amoA consensus phylogeny based on all publicly available sequences (status June 2010) and provide evidence for the diversification of AOA into four previously recognized clusters and one newly identified major cluster. These clusters, for which we suggest a new nomenclature, harboured 83 AOA species-level OTU (using an inferred species threshold of 85% amoA identity). 454 pyrosequencing of amoA amplicons from 16 soils sampled in Austria, Costa Rica, Greenland and Namibia revealed that only 2% of retrieved sequences had no database representative on the species-level and represented 30–37 additional species-level OTUs. With the exception of an acidic soil from which mostly amoA amplicons of the Nitrosotalea cluster were retrieved, all soils were dominated by amoA amplicons from the Nitrososphaera cluster (also called group I.1b), indicating that the previously reported AOA from the Nitrosopumilus cluster (also called group I.1a) are absent or represent minor populations in soils. AOA richness estimates on the species level ranged from 8–83 co-existing AOAs per soil. Presence/absence of amoA OTUs (97% identity level) correlated with geographic location, indicating that besides contemporary environmental conditions also dispersal limitation across different continents and/or historical environmental conditions might influence AOA biogeography in soils.
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              Cleavage of parallel-stranded DNA duplex by peplomycin metal complexes.

              Peplomycin-mediated degradation of parallel-stranded (ps) duplex was investigated. It was found that Co- and Fe-peplomycins degraded ps DNA duplex by 4'-hydrogen abstraction at 5'-GPy (pyrimidine) site in a similar manner to that of antiparallel B-DNA. While the orientation of two strands of ps and B-form DNA duplexes are reversed, peplomycin metal complex can bind to ps DNA duplex to cause oxidative DNA damage. These results indicate that peplomycin metal complex mainly interacts with one strand which is damaged.
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                Author and article information

                Journal
                FEMS Microbiol Ecol
                FEMS Microbiol. Ecol
                femsec
                femsec
                FEMS Microbiology Ecology
                Oxford University Press
                0168-6496
                1574-6941
                13 March 2016
                May 2016
                13 March 2016
                : 92
                : 5
                : fiw057
                Affiliations
                Institute of Biological and Environmental Sciences, Cruickshank Building, St Machar Drive, University of Aberdeen, Aberdeen AB24 3UU, UK
                Author notes
                [* ] Corresponding author: Institute of Biological and Environmental Sciences, University of Aberdeen, Cruickshank Building, St Machar Drive, Aberdeen AB24 3UU, UK. Tel: +0044-1224-273254; E-mail: j.prosser@ 123456abdn.ac.uk
                []Joint first authors.
                [] Present address: Environmental Microbial Genomics Group, Laboratoire Ampere, École Centrale de Lyon, Université de Lyon, 69134 Ecully cedex, France.
                Article
                10.1093/femsec/fiw057
                4830249
                26976843
                e1026a1d-0df7-4c93-bc85-268a2feb4b77
                © FEMS 2016.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 09 March 2016
                : 23 December 2015
                Page count
                Pages: 10
                Categories
                Research Article
                Custom metadata
                May 2016

                Microbiology & Virology
                thaumarchaeota,nitrososphaera sister cluster,soil,ammonia inhibition,nitrosocosmicus,nitrification

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