Methane (CH 4 ) emission by carbon-rich cryosols at the high latitudes in Northern Hemisphere has been studied extensively. In contrast, data on the CH 4 emission potential of carbon-poor cryosols is limited, despite their spatial predominance. This work employs CH 4 flux measurements in the field and under laboratory conditions to show that the mineral cryosols at Axel Heiberg Island in the Canadian high Arctic consistently consume atmospheric CH 4 . Omics analyses present the first molecular evidence of active atmospheric CH 4 -oxidizing bacteria (atmMOB) in permafrost-affected cryosols, with the prevalent atmMOB genotype in our acidic mineral cryosols being closely related to Upland Soil Cluster α. The atmospheric (atm) CH 4 uptake at the study site increases with ground temperature between 0°C and 18°C. Consequently, the atm CH 4 sink strength is predicted to increase by a factor of 5-30 as the Arctic warms by 5-15°C over a century. We demonstrate that acidic mineral cryosols are a previously unrecognized potential of CH 4 sink that requires further investigation to determine its potential impact on larger scales. This study also calls attention to the poleward distribution of atmMOB, as well as to the potential influence of microbial atm CH 4 oxidation, in the context of regional CH 4 flux models and global warming.

A bacterial strain Rhodococcus imtechensis RKJ300 (= MTCC 7085(T) = JCM 13270(T)) was isolated from pesticide-contaminated soil of Punjab by the enrichment technique on minimal medium containing 4-nitrophenol. Strain RKJ300 is capable of utilizing 4-nitrophenol, 2-chloro-4-nitrophenol, and 2,4-dinitrophenol as sole sources of carbon and energy. The strain involved both oxidative and reductive catabolic mechanisms for initial transformation of these compounds. In the case of 2-chloro-4-nitrophenol, colorimetric analysis indicated that nitrite release was followed by stoichiometric elimination of chloride ions. Experiments using whole cells and cell-free extracts showed chlorohydroquinone and hydroquinone as the intermediates of 2-chloro-4-nitrophenol degradation. This is the first report of degradation on 2-chloro-4-nitrophenol by a bacterium under aerobic condition to the best of our knowledge. However, pathways for degradation of 4-nitrophenol and 2,4-dinitrophenol were similar to those reported in other strains of Rhodococcus. Laboratory-scale soil microcosm studies demonstrated that the organism was capable of degrading a mixture of nitrophenols simultaneously, indicating its applicability toward in situ bioremediation of contaminated sites. The fate of the augmented strain as monitored by the plate-counting method and hybridization technique was found to be fairly stable throughout the period of microcosm experiments.

The total community genomic DNA (gDNA) from permafrost was extracted using four commercial DNA extraction kits. The gDNAs were compared using quantitative real‐time PCR (qPCR) targeting 16S rRNA genes and bacterial diversity analyses obtained via 454 pyrosequencing of the 16S rRNA (V3 region) amplified in single or nested PCR. The FastDNA® SPIN (FDS) Kit provided the highest gDNA yields and 16S rRNA gene concentrations, followed by MoBio PowerSoil® (PS) and MoBio PowerLyzer™ (PL) kits. The lowest gDNA yields and 16S rRNA gene concentrations were from the Meta‐G‐Nome™ (MGN) DNA Isolation Kit. Bacterial phyla identified in all DNA extracts were similar to that found in other soils and were dominated by Actinobacteria, Firmicutes, Gemmatimonadetes, Proteobacteria, and Acidobacteria. Weighted UniFrac and statistical analyses indicated that bacterial community compositions derived from FDS, PS, and PL extracts were similar to each other. However, the bacterial community structure from the MGN extracts differed from other kits exhibiting higher proportions of easily lysed β‐ and γ‐Proteobacteria and lower proportions of Actinobacteria and Methylocystaceae important in carbon cycling. These results indicate that gDNA yields differ between the extraction kits, but reproducible bacterial community structure analysis may be accomplished using gDNAs from the three bead‐beating lysis extraction kits.

Application of microbial metabolic potential (bioremediation) is accepted as an environmentally benign and economical measure for decontamination of polluted environments. Bioremediation methods are generally categorized into ex situ and in situ bioremediation. Although in situ bioremediation methods have been in use for two to three decades, they have not yet yielded the expected results. Their limited success has been attributed to reduced ecological sustainability under environmental conditions. An important determinant of sustainability of in situ bioremediation is pollutant bioavailability. Microbial chemotaxis is postulated to improve pollutant bioavailability significantly; consequently, application of chemotactic microorganisms can considerably enhance the performance of in situ degradation. The environmental fate of degradative microorganisms and the ecological consequence of intervention constitute other important descriptors for the efficiency and sustainability of bioremediation processes. Integrative use of culture-dependent, culture-independent methods (e.g. amplified rDNA restriction analysis, terminal restriction fragment length polymorphism, denaturing/thermal gradient gel electrophoresis, phospholipid fatty acid, etc.), computational and statistical analyses has enabled successful monitoring of the above aspects. The present review provides a detailed insight into some of the key factors that affect the efficiency of in situ bioremediation along with a comprehensive account of the integrative approaches used for assessing the ecological sustainability of processes. The review also discusses the possibility of developing suicidal genetically engineered microorganisms for optimized and controlled in situ bioremediation.

Polycyclic aromatic hydrocarbons (PAHs) are compounds of intense public concern due to their persistence in the environment and potentially deleterious effects on human, environmental and ecological health. The clean up of such contaminants using invasive technologies has proven to be expensive and more importantly often damaging to the natural resource properties of the soil, sediment or aquifer. Bioremediation, which exploits the metabolic potential of microbes for the clean-up of recalcitrant xenobiotic compounds, has come up as a promising alternative. Several approaches such as improvement in PAH solubilization and entry into the cell, pathway and enzyme engineering and control of enzyme expression etc. are in development but far from complete. Successful application of the microorganisms for the bioremediation of PAH-contaminated sites therefore requires a deeper understanding of the physiology, biochemistry and molecular genetics of potential catabolic pathways. In this review, we briefl y summarize important strategies adopted for PAH bioremediation and discuss the potential for their improvement.

Previous studies investigating organic-rich tundra have reported that increasing biodegradation of Arctic tundra soil organic carbon (SOC) under warming climate regimes will cause increasing CO 2 and CH 4 emissions. Organic-poor, mineral cryosols, which comprise 87% of Arctic tundra, are not as well characterized. This study examined biogeochemical processes of 1 m long intact mineral cryosol cores (1-6% SOC) collected in the Canadian high Arctic. Vertical profiles of gaseous and aqueous chemistry and microbial composition were related to surface CO 2 and CH 4 fluxes during a simulated spring/summer thaw under light versus dark and in situ versus water saturated treatments. CO 2 fluxes attained 0.8 ± 0.4 mmol CO 2 m À2 h À1 for in situ treatments, of which 85 ± 11% was produced by aerobic SOC oxidation, consistent with field observations and metagenomic analyses indicating aerobic heterotrophs were the dominant phylotypes. The Q 10 values of CO 2 emissions ranged from 2 to 4 over the course of thawing. CH 4 degassing occurred during initial thaw; however, all cores were CH 4 sinks at atmospheric concentration CH 4 . Atmospheric CH 4 uptake rates ranged from À126 ± 77 to À207 ± 7 nmol CH 4 m À2 h À1 with CH 4 consumed between 0 and 35 cm depth.Metagenomic and gas chemistry analyses revealed that high-affinity Type II methanotrophic sequence abundance and activity were highest between 0 and 35 cm depth. Microbial sulfate reduction dominated the anaerobic processes, outcompeting methanogenesis for H 2 and acetate. Fluxes, microbial community composition, and biogeochemical rates indicate that mineral cryosols of Axel Heiberg Island act as net CO 2 sources and atmospheric CH 4 sinks during summertime thaw under both in situ and water saturated states.

Microbial degradation studies have pointed toward the occurrence of two distinct PNP catabolic pathways in Gram positive and Gram negative bacteria. The former involves 4-nitrocatechol (4-NC), 1,2,4-benzenetriol (BT), and maleylacetate (MA) as major degradation intermediates, whereas the later proceeds via formation of 1,4-benzoquinone (BQ) and hydroquinone (HQ). In the present study we identified a Gram negative organism viz. Burkholderia sp. strain SJ98 that degrades PNP via 4NC, BT, and MA. A 6.89 Kb genomic DNA fragment of strain SJ98 that encompasses seven putatively identified ORFs (orfA, pnpD, pnpC, orfB, orfC, orfD, and orfE) was cloned. PnpC is benzenetriol dioxygenase belonging to the intradiol dioxygenase superfamily, whereas PnpD is identified as maleylacetate reductase, a member of the Fe-ADH superfamily showing NADH dependent reductase activity. The in vitro activity assays carried out with purified pnpC and pnpD (btd and mar) gene products transformed BT to MA and MA to beta-ketoadipate, respectively. The cloning, sequencing, and characterization of these genes along with the functional PNP degradation studies ascertained the involvement of 4-NC, BT, and MA as degradation intermediates of PNP pathway in this strain. This is one of the first conclusive reports for 4-NC and BT mediated degradation of PNP in a Gram negative organism.

Burkholderia sp. strain SJ98 (DSM 23195) was previously isolated and characterized for degradation and co-metabolic transformation of a number nitroaromatic compounds. In the present study, we evaluated its metabolic activity on chlorinated nitroaromatic compounds (CNACs). Results obtained during this study revealed that strain SJ98 can degrade 2-chloro-4-nitrophenol (2C4NP) and utilize it as sole source of carbon, nitrogen, and energy under aerobic conditions. The cells of strain SJ98 removed 2C4NP from the growth medium with sequential release of nearly stoichiometric amounts of chloride and nitrite in culture supernatant. Under aerobic degradation conditions, 2C4NP was transformed into the first intermediate that was identified as p-nitrophenol by high-performance liquid chromatography, LCMS-TOF, and GC-MS analyses. This transformation clearly establishes that the degradation of 2C4NP by strain SJ98 is initiated by "reductive dehalogenation"; an initiation mechanism that has not been previously reported for microbial degradation of CNAC under aerobic conditions.