Methane oxidation by microorganisms inhabiting aerobic soils is a key process involved in the regulation of the concentration of this significant greenhouse gas in the atmosphere; however, the microorganisms responsible for this process remain unknown. Three stable methane-oxidizing cultures were isolated from samples of forest soils (FS) and agricultural soils (AS) of Moscow oblast, as well as from soil samples collected from a Belgian agrocenosis (BS). The obtained enrichment cultures exhibit a high affinity for methane; their k_m values range from 54.2 to 176.8 nM CH₄ and are comparable to those of aerobic soils. Analysis of the fragments of the ribosomal (16S rRNA) and functional (pmoA) genes of methanotrophs by PCR— DGGE and cloning demonstrated the presence of bacteria belonging to the genera Methylocystis in FS, Methylosinus in AS and BS, and Methylocella in BS. It was established that Methylocystis and Methylosinus detected in the enrichment cultures contain the genes encoding the synthesis of the active center of two membrane-bound particulate methane monooxygenases; it is likely that one of these genes (pmoA2) is responsible for the capacity of these microorganisms for oxidation of atmospheric methane.

A novel species is proposed for a high-affinity methanotrophic representative of the genus Methylocystis . Strain FS^T was isolated from a weakly acidic (pH 5.3) mixed forest soil of the southern Moscow area. Cells of FS^T are aerobic, Gram-negative, non-motile, curved coccoids or short rods that contain an intracytoplasmic membrane system typical of type-II methanotrophs. Only methane and methanol are used as carbon sources. FS^T grew at a temperature range of 4–37 °C (optimum 25–30 °C) and a pH range of 4.5 to 7.5 (optimum pH 6.0–6.5). The major fatty acids were C_{18  :  1}ω8c, C_{18  :  1}ω7c and C_{18  :  0}; the major quinone as Q-8. FS^T displays 16S rRNA gene sequences similarity to other taxonomically recognized members of the genus Methylocystis, with Methylocystis hirsuta CSC1^T (99.6 % similarity) and Methylocystis rosea SV97^T (99.3 % similarity) as its closest relatives. The genome comprises 3.85 Mbp and has a DNA G+C content of 62.6 mol%. Genomic analyses and DNA–DNA relatedness with genome-sequenced members of the genus Methylocystis demonstrated that FS^T could be separated from its closest relatives. FS^T possesses two particulate methane monooxygenases (pMMO): low-affinity pMMO1 and high-affinity pMMO2. In laboratory experiments, it was demonstrated that FS^T might oxidize methane at atmospheric concentration. The genome contained various genes for nitrogen fixation, polyhydroxybutyrate synthesis, antibiotic resistance and detoxification of arsenic, cyanide and mercury. On the basis of genotypic, phenotypic and chemotaxonomic characteristics, it is proposed that the isolate represents a novel species, Methylocystis silviterrae sp. nov. The type strain is FS^T (=KCTC 82935^T=VKM B-3535^T).

The flux rates of carbon dioxide, methane, and nitrous oxide in the soils on autonomous, transitional, transitional-accumulative, and accumulative positions of a catena on the Oka River’s right bank (Moscow oblast) were assessed using the chamber method. The lowest rate of C-CO₂ emission (18.8–29.8 mg/m² per hour) was found for the gray forest soil in the autonomous position, and the highest rate (52.4–66.1 mg/m² per hour) was found for the alluvial meadow soil of the accumulative landscape. In the summer, the uptake of methane from the atmosphere exceeded its release from the soil at all the points of the catena (9–38 μg/m² per hour). The highest rate of the C-CH₄ uptake was observed for the soil in the transitional position. In the fall, the soils in the autonomous, transitional, and transitional-accumulative positions served as a sink of C-CH₄, and the soil of the accumulative position was a source of methane emission. The rate of the N-N₂O emission from the catena soils increased when going from the autonomous position to the accumulative one (0.41–11.2 μg/m² per hour). The spatial variation of the C-CO₂, C-CH₄, and N-N₂O fluxes within the catena was 33, 172, and 138%, respectively. The upper (0- to 10-cm) soil layer made the major contribution to the emission of carbon dioxide. This soil layer was characterized by its C-CH₄ uptake, and the emission of methane was typical for the deeper (0- to 20-cm) layer. The layers deeper than 10 and 20 cm emitted more N-N₂O than the surface layer.

He presence of nitrogen-fixing bacteria of the genus Azospirillum in the soils of acidic raised Sphagnum bogs is revealed for the first time. Three Azospirillum strains, B2, B21, and B22, were isolated as a component of methane-oxidizing enrichment cultures, whereas attempts to isolate them directly from peat samples have failed. The results of comparative analysis of the nucleotide sequences of 16S rRNA genes, DNA-DNA hybridization, and the analysis of the sequences of the functional genes encoding nitrogenase and ribulose-1, 5-bisphosphate carboxylase reveal that all the newly obtained strains can be classified as Azospirillum lipoferum. Yet, unlike A. lipoferum, the isolates do not require biotin and utilize sucrose, inositol, and glycerol for growth. The cell morphology of strain B2 differs from that of the type strain and strains B21 and B22. The results obtained indicate the variability of morphological, physiological, and biochemical properties in closely related Azospirillum strains and suggest the existence of metabolic relationships between methanotrophic bacteria and the representatives of the genus Azospirillum under peat bog conditions.

Human activities such as land management and global warming have great impact on the environment. Among changes associated with the global warming, rising methane emission is a serious concern. Therefore, we assessed methane oxidation activity and diversity of aerobic methanotrophic bacteria in eight soil types (both unmanaged and agricultural) distributed across the European part of Russia. Using a culture-independent approach targeting pmoA gene, we provide the first baseline data on the diversity of methanotrophs inhabiting most typical soil types. The analysis of pmoA clone libraries showed that methanotrophic populations in unmanaged soils are less diverse than in agricultural areas. These clone sequences were placed in three groups of, so far, uncultured methanotrophs: USC-gamma, cluster I, and pmoA/amoA cluster, which are believed to be responsible for atmospheric methane oxidation in upland soils. Agricultural soils harbored methanotrophs related to genera Methylosinus, Methylocystis, Methylomicrobium, Methylobacter, and Methylocaldum. Despite higher numbers of detected molecular operational taxonomic units (MOTUs), managed soils showed decreased methane oxidation rates as observed in both in situ and laboratory experiments. Our results also suggest that soil restoration may have a positive effect on methane consumption by terrestrial ecosystems.