Ammonia-oxidizing archaea (AOA) are among the most abundant and ubiquitous microorganisms in the ocean, exerting primary control on nitrification and nitrogen oxides emission. Although united by a common physiology of chemoautotrophic growth on ammonia, a corresponding high genomic and habitat variability suggests tremendous adaptive capacity. Here, we compared 44 diverse AOA genomes, 37 from species cultivated from samples collected across diverse geographic locations and seven assembled from metagenomic sequences from the mesopelagic to hadopelagic zones of the deep ocean. Comparative analysis identified seven major marine AOA genotypic groups having gene content correlated with their distinctive biogeographies. Phosphorus and ammonia availabilities as well as hydrostatic pressure were identified as selective forces driving marine AOA genotypic and gene content variability in different oceanic regions. Notably, AOA methylphosphonate biosynthetic genes span diverse oceanic provinces, reinforcing their importance for methane production in the ocean. Together, our combined comparative physiological, genomic, and metagenomic analyses provide a comprehensive view of the biogeography of globally abundant AOA and their adaptive radiation into a vast range of marine and terrestrial habitats.

We investigated the quantities and phylogenies of amoA genes of ammonia-oxidizing archaea (AOA) belonging to Crenarchaeota and ammonia-oxidizing bacteria (AOB) belonging to Betaproteobacteria in water columns and deep-ocean cold seep sediment in the northeastern Japan Sea with a competitive PCR (cPCR) assay. Water samples were collected at depths of 2000 m and 2956 m. Sediment samples were collected where white bacterial mats had developed. The cPCR analysis revealed five to ten times more AOA than betaproteobacterial AOB in both the water columns and sediment. The abundance of the crenarchaeotal amoA gene was estimated at 6×10 1 and 3×10 2 copies ml −1 in the water columns at depths of 2000 m and 2956 m, and 1×10 8 and 1×10 7 copies g −1 in pelagic brown sediment and black sediment, respectively. Most archaeal amoA clones from water column at 2000 m fell into the Deep Marine Group. Most archaeal amoA clones from pelagic brown sediment were less closely related to known environmental clones. Moreover, incubation experiments revealed nitrite production at 4°C and 10°C. The results indicate that psychrophilic AOA and AOB may be responsible for nitrification in the deep-ocean region of the northeastern Japan Sea.

Nitrite-oxidizing bacteria (NOB) are responsible for the second step of nitrification in natural and engineered ecosystems. The recently discovered genus Nitrotoga belongs to the Betaproteobacteria and potentially has high environmental importance. Although environmental clones affiliated with Nitrotoga are widely distributed, the limited number of cultivated Nitrotoga spp. results in a poor understanding of their ecophysiological features. In this study, we successfully enriched the nonmarine cold-adapted Nitrotoga sp. strain AM1 from coastal sand in an eelgrass zone and investigated its physiological characteristics. Multistep-enrichment approaches led to an increase in the abundance of AM1 to approximately 80% of the total bacterial population. AM1 was the only detectable NOB in the bacterial community. The 16S rRNA gene sequence of AM1 was 99.6% identical to that of "Candidatus Nitrotoga arctica," which was enriched from permafrost-affected soil. The highest nitrogen oxidation rate of AM1 was observed at 16°C. The half-saturation constant (K m ) and the generation time were determined to be 25 M NO 2 Ϫ and 54 h, respectively. The nitrite oxidation rate of AM1 was stimulated at concentrations of Ͻ30 mM NH 4 Cl but completely inhibited at 50 mM NH 4 Cl. AM1 can grow well under specific environmental conditions, such as low temperature and in the presence of a relatively high concentration of free ammonia. These results help improve our comprehension of the functional importance of Nitrotoga.IMPORTANCE Nitrite-oxidizing bacteria (NOB) are key players in the second step of nitrification, which is an important process of the nitrogen cycle. Recent studies have suggested that the organisms of the novel NOB genus Nitrotoga were widely distributed and played a functional role in natural and engineered ecosystems. However, only a few Nitrotoga enrichments have been obtained, and little is known about their ecology and physiology. In this study, we successfully enriched a Nitrotoga sp. from sand in a shallow coastal marine ecosystem and undertook a physiological characterization. The laboratory experiments showed that the Nitrotoga enrichment culture could adapt not only to low temperature but also to relatively high concentrations of free ammonia. The determination of as-yet-unknown unique characteristics of Nitrotoga contributes to the improvement of our insights into the microbiology of nitrification.KEYWORDS Nitrospira, Nitrotoga, ammonia, coastal sand, cultivation, enrichment, microbial communities, nitrification, nitrite-oxidizing bacteria, physiology I n nitrification, ammonia is oxidized into nitrate via nitrite by phylogenetically different chemolithoautotrophic microorganisms, and this is an integral part of the global nitrogen cycle. The reactions are catalyzed by ammonia-oxidizing bacteria (AOB),

The complete nucleotide sequence of the RNA coliphage GA, a group II phage, is presented. The entire genome comprises 3466 bases. Three large open reading frames were identified, which correspond to the maturation protein gene (390 amino acids), the coat protein gene (129 amino acids) and the replicase beta-subunit protein gene (531 amino acids). In addition, untranslated regions occur at the 5' (135 bases) and 3' (122 bases) ends of the molecule. Two intercistronic untranslated regions occur between the cistrons for the maturation and coat proteins, and between the coat and beta-subunit proteins. We have compared the nucleotide sequence of GA RNA with the published sequence of MS2 RNA, and show that they are related. The comparative structures of two important regulatory regions are presented; the coat protein binding site which is involved in translational repression of the replicase beta-subunit protein gene, and a hairpin in a region proximal to the lysis protein gene.

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Ammonia-oxidizing archaea (AOA) are generally cultivated at ammonium concentrations of less than 2 mM. The physiology and abundance in the environment of AOA suggest an important role in the nitrogen cycle. We report here a novel marine ammonia-oxidizing crenarchaeote, strain NM25 belonged to 'Candidatus Nitrosopumilus', that was enriched from coastal sand of an eelgrass zone and grew in a medium containing 15 mM ammonium at 30°C. A phylogenetic analysis based on the 16S rRNA gene revealed this crenarchaeote was related to the ammonia-oxidizing archaeon 'Candidatus Nitrosopumilus maritimus' strain SCM1, with 98.5% identity. The ammonia monooxygenase subunit A (amoA) gene of strain NM25 was less closely related to that of known cultivable AOA (>95%) and environmental clones (>97%). This finding suggests the existence of AOA adapted to high ammonium-containing environments.

Seasonal changes in the abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) within the sand of an eelgrass (Zostera marina) zone were examined by a quantitative PCR of both crenarchaeotal and betaproteobacterial ammonia monooxygenase alpha subunit (amoA) genes together with temperature and concentrations of ammonium, nitrite, and nitrate from May 2007 to June 2008 at Tanoura Bay, Shizuoka, Japan. The abundance of both amoAs in the sand between May and June 2007 and between January and March 2008 was 1.5 to 2 orders of magnitude higher than the 10 4 copies g −1 of estimated amoA between September and December. Archaeal amoA was more diverse than betaproteobacterial amoA. Betaproteobacterial amoA clone libraries were dominated by Nitrosospiralike sequence types. An incubation experiment was conducted with sands collected in February 2008 and community structure was analyzed based on reverse-transcribed amoAs. RNA was extracted from sand incubated for 12 days at 30°C, 17 days at 20°C, and 80 days at 10°C. Different amoA clones were detected from in situ sand and incubated sand. This study reveals clear evidence of seasonal change in the abundance of AOA and AOB within the sand of an eelgrass zone.