Comparative morphological, ecological, and molecular studies of Aspergillus versicolor (Vuill.) tiraboschi strains isolated from different ecotopes

Cultural, morphological, ecological, and trophic properties (growth at different temperatures and on various organic substrates), as well as molecular and genetic peculiarities of Aspergillus versicolor (Vuill.) Tiraboschi strains of different origins, were determined. The strains were isolated from different ecotopes (upper horizons of modern soils of several geographic regions, ancient soils and peat, and permafrost). No essential distinctions in cultural and morphological properties were revealed between the strains. Strains obtained from peat of the Aleutian Islands were characterized by the highest radial rates of colony growth. Some variations in the ITS loci of rDNA were observed in strains isolated from different ecotopes; the distinctions were most pronounced (1.7%) in the strain isolated from 100 000-year-old permafrost.     

Exometabolites of the fungal isolates (Genus <Emphasis Type="Italic">Penicillium</Emphasis>, Section <Emphasis Type="Italic">Chrysogena</Emphasis>) from low-temperature ecotopes

Exometabolites of 22 strains of the genus Penicillium, section Chrysogena isolated from low-temperature ecotopes of various geographical regions were analyzed. The ecotopes included permafrost deposits, frozen volcanic ash, a fossil horse, cryopeg, and water from an Antarctic lake. The studied strains were found to contain exometabolites belonging to the groups of penicillins (penicillin G), chrysogines (chrysogine, 3-acetyl-quinazolin-4(3H)-one, 2-pyruvoylaminobenzamide, 2-(2-hydroxypropionylamino)-benzamide, and questiomycin A), roquefortines (3,12-dihydroroquefortine, roquefortine, glandicolines A and B, and meleagrine), xanthocillins (xanthocillin X), and simple tryptophan derivatives (N-acetyltriptamine and indoleacetic acid). In five P. chrysogenum strains and three P. nalgiovense strains, a correlation was found between exometabolite spectra and morphological characteristics of the cultures isolated from modern ecotopes. For other strains species, identification was based on morphological features due to the absence of biosynthesis of penicillin G on the major chemotaxonomic markers for these species.     

Comparison of the adaptive potential of the Arthrobacter oxydans and Acinetobacter lwoffii isolates from permafrost sedimentary rock and the analogous collection strains

A comparative study was conducted on the adaptive mechanisms of the strains Arthrobacter oxydans K14 and Acinetobacter lwoffii EK30A isolated from permafrost subsoil sediments and of those of the analogous collection strains (Ac-1114 Type and BSW-27, respectively). In each pair of the strains compared, the strains differed in terms of (i) growth-related, physiological, and biochemical properties; (ii) resistance to stress factors; (iii) capacity for generation of dormant forms (DFs) under growth arrest conditions, and (iv) intrapopulation production of phase variants. The strains isolated from permafrost displayed a lower growth rate but were more resistant to repeated freezing-thawing treatment than the collection strains. Under the same growth conditions, the permafrost strains formed larger numbers of cystlike anabiotic DFs, extraordinarily small cells, and forms that became nonculturable during long-term storage. Resuscitation of the nonculturable forms resulted in a 2- to-7-fold increase in the percentage of FISH-detectable metabolically active cells. The permafrost strains were also distinguished by increased genome lability. This facilitated their dissociation into intrapopulation variants with phenotypically distinct colonial and morphological properties and different antibiotic resistance. The phenotypic variability was more prominent in Arthrobacter (for which it was not reported previously) than in Acinetobacter. In the populations produced by plating the dormant bacterial forms, the qualitative and quantitative characteristics of the phase variant spectra varied depending on the formation conditions and the composition of the solid media used for the plating. Thus, the permafrost isolates of A. oxydans and Ac. lwoffii were distinguished from their collection analogs by a more manifest adaptive potential including stress resistance, the intensity of DF generation under growth arrest conditions, and increased intrapopulation variability.     

Isolation and characterization of nitrogen-fixing bacteria of the genus Azospirillum from the soil of a Sphagnum peat bog

The 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.     

Bacteria Antagonistic to Pseudomonas tolaasii and their Control of Brown Blotch of the Cultivated Mushroom Agaricus bisporus

S ummary : Pseudomonas tolaasii was isolated from casing peat of healthy and diseased mushroom beds, compost of diseased mushroom beds and from soils round a mushroom farm. It was not isolated from fresh peat or compost from healthy mushroom beds. Three bacteria antagonistic to Ps. tolaasii were isolated from soil and peat. These were a nonfluorescent Pseudomonas sp. (closest to Ps. multivorans ) from soil; and strains of Ps. fluorescens and Enterobacter aerogenes from peat. When the antagonists and the pathogen were added in the ratio of 8 × 10⁷: 10⁶ cells/ml to unsterilized peat and applied to mushroom trays, infection of mushroom sporophores by the pathogen was effectively controlled. In vitro studies failed to show lysis or growth inhibition of Ps. tolaasii by the antagonists.     

Soil organic matter molecular composition and state of decomposition in three locations of the European Arctic

Increased mineralization of the organic matter (OM) stored in permafrost is expected to constitute the largest additional global warming potential from terrestrial ecosystems exposed to a warmer climate. Chemical composition of permafrost OM is thought to be a key factor controlling the sensitivity of decomposition to warming. Our objective was to characterise OM from permafrost soils of the European Arctic: two mineral soils—Adventdalen, Svalbard, Norway and Vorkuta, northwest Russia—and a “palsa” (ice-cored peat mound patterning in heterogeneous permafrost landscapes) soil in Neiden, northern Norway, in terms of molecular composition and state of decomposition. At all sites, the OM stored in the permafrost was at an advanced stage of decomposition, although somewhat less so in the palsa peat. By comparing permafrost and active layers, we found no consistent effect of depth or permafrost on soil organic matter (SOM) chemistry across sites. The permafrost-affected palsa peat displayed better preservation of plant material in the deeper layer, as indicated by increasing contribution of lignin carbon to total carbon with depth, associated to decreasing acid (Ac) to aldehyde (Al) ratio of the syringyl (S) and vanillyl (V) units, and increasing S/V and contribution of plant-derived sugars. By contrast, in Adventdalen, the Ac/Al ratio of lignin and the Alkyl C to O-alkyl C ratio in the NMR spectra increased with depth, which suggests less oxidized SOM in the active layer compared to the permafrost layer. In Vorkuta, SOM characteristics in the permafrost profile did not change substantially with depth, probably due to mixing of soil layers by cryoturbation. The composition and state of decomposition of SOM appeared to be site-specific, in particular bound to the prevailing organic or mineral nature of soil when attempting to predict the SOM proneness to degradation. The occurrence of processes such as palsa formation in organic soils and cryoturbation should be considered when up-scaling and predicting the responses of OM to climate change in arctic soils.     

The disappearance of relict permafrost in boreal north America: Effects on peatland carbon storage and fluxes

Boreal peatlands in Canada have harbored relict permafrost since the Little Ice Age due to the strong insulating properties of peat. Ongoing climate change has triggered widespread degradation of localized permafrost in peatlands across continental Canada. Here, we explore the influence of differing permafrost regimes (bogs with no surface permafrost, localized permafrost features with surface permafrost, and internal lawns representing areas of permafrost degradation) on rates of peat accumulation at the southernmost limit of permafrost in continental Canada. Net organic matter accumulation generally was greater in unfrozen bogs and internal lawns than in the permafrost landforms, suggesting that surface permafrost inhibits peat accumulation and that degradation of surface permafrost stimulates net carbon storage in peatlands. To determine whether differences in substrate quality across permafrost regimes control trace gas emissions to the atmosphere, we used a reciprocal transplant study to experimentally evaluate environmental versus substrate controls on carbon emissions from bog, internal lawn, and permafrost peat. Emissions of CO₂ were highest from peat incubated in the localized permafrost feature, suggesting that slow organic matter accumulation rates are due, at least in part, to rapid decomposition in surface permafrost peat. Emissions of CH₄ were greatest from peat incubated in the internal lawn, regardless of peat type. Localized permafrost features in peatlands represent relict surface permafrost in disequilibrium with the current climate of boreal North America, and therefore are extremely sensitive to ongoing and future climate change. Our results suggest that the loss of surface permafrost in peatlands increases net carbon storage as peat, though in terms of radiative forcing, increased CH₄ emissions to the atmosphere will partially or even completely offset this enhanced peatland carbon sink for at least 70 years following permafrost degradation.     

Viability, diversity and composition of the bacterial community in a high Arctic permafrost soil from Spitsbergen, Northern Norway

The viable and non-viable fractions of the bacterial community in a 2347-year-old permafrost soil from Spitsbergen were subjected to a comprehensive investigation using culture-independent and culture-dependent methods. LIVE/DEAD BacLight staining revealed that 26% of the total number of bacterial cells were viable. Quantitatively, aerobic microcolonies, aerobic colony-forming units and culturable anaerobic bacteria comprised a minor fraction of the total number of viable bacteria, which underlines the necessity for alternative cultivation approaches in bacterial cryobiology. Sulfate reduction was detected at temperatures between -2 degrees C and 29 degrees C while methanogenesis was not detected. Bacterial diversity was high with 162 operational taxonomic units observed from 800 16S rDNA clone sequences. The 158 pure cultures isolated from the permafrost soil affiliated with 29 different bacterial genera, the majority of which have not previously been isolated from permafrost habitats. Most of the strains isolated were affiliated to the genera Cellulomonas and Arthrobacter and several of the pure cultures were closely related to bacteria reported from other cryohabitats. Characterization of viable bacterial communities in permafrost soils is important as it will enable identification of functionally important groups together with the as yet undescribed adaptations that bacteria have evolved for surviving subzero temperatures for millennia.     

Survival ofFrankia strains introduced into soil

Factors affecting the establishment of Alnus/Frankia symbioses were studied partly by following the survival ofFrankia strains exposed to different soil conditions, and partly by investigating the effect of pH on nodulation. TwoFrankia strains were used, both of the Sp⁻ type (sporangia not formed in nodules). One of the strains sporulated heavily, while the other formed mainly hyphae. The strains originated fromAlnus incana root nodules growing in soils of pH 3.5 and 5.0. The optimum pH for their growth in pure culture was found to be 6.7 and 6.2, respectively. The strains were introduced into twoFrankia-free soils, peat and fine sand. Their survival, measured as the persistance of nodulation capacity using the plant infection technique, was followed for 14 months. The survival curves of the strains were similar despite the morphological differences between the strains in pure culture. The nodulation capacities declined over time both at 14 and 22°C. Survival was better in soils limed to a pH above 6 than in soils at their original pH (peat 2.9, fine sand 4.2). The effect of pH on nodule formation in Alnus seedlings by theFrankia strains was studied in liquid culture. The number of nodules increased linearly within the pH range studied (3.5–5.8). No nodules were formed at pH 3.5.     

Hot spots for nitrous oxide emissions found in different types of permafrost peatlands

Recent findings on large nitrous oxide (N₂O) emissions from permafrost peatlands have shown that tundra soils can support high N₂O release, which is on the contrary to what was thought previously. However, field data on this topic have been very limited, and the spatial and temporal extent of the phenomenon has not been known. To address this question, we studied N₂O dynamics in two types of subarctic permafrost peatlands, a peat plateau in Russia and three palsa mires in Finland, including also adjacent upland soils. The peatlands studied have surfaces that are uplifted by frost (palsas and peat plateaus) and partly unvegetated as a result of wind erosion and frost action. Unvegetated peat surfaces with high N₂O emissions were found from all the studied peatlands. Very high N₂O emissions were measured from peat circles at the Russian site (1.40±0.15 g N₂O m^?2 yr^?1). Elevated, sparsely vegetated peat mounds at the same site had significantly lower N₂O release. The N₂O emissions from bare palsa surfaces in Northern Finland were highly variable but reached high rates, similar to those measured from the peat circles. All the vegetated soils studied had negligible N₂O release. At the bare peat surfaces, the large N₂O emissions were supported by the absence of plant N uptake, the low C : N ratio of the peat, the relatively high gross N mineralization rate and favourable moisture content, together increasing availability of mineral N for N₂O production. We hypothesize that frost heave is crucial for high N₂O emissions, since it lifts the peat above the water table, increasing oxygen availability and making it vulnerable to the the physical processes that may remove the vegetation cover. In the future, permafrost thawing may change the distribution of wet and dry surfaces in permafrost peatlands, which will affect N₂O emissions.     

Isolation and characterization of nitrogen-fixing bacteria of the genus <Emphasis Type="Italic">Azospirillum</Emphasis> from the soil of a <Emphasis Type="Italic">Sphagnum</Emphasis> peat bog

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.     

八株芽孢杆菌菌株的分类及固氮活性的测定

从8个省市的不同土壤分离了200个菌株,筛选出8株（JR₁,JR₂,JR₅,ZZ₃,ZZ₁₂,ZZ₁₆,M₁,K₄）能在无氮培养基上生长良好的典型的芽抱杆菌,其中6株（JR₁,JR₂,JR₄和ZZ₃,ZZ₁₂,ZZ     

A new yeast species,Candida aurita sp. nov., from oligotrophic bogs of Western Siberia

Five anamorphous yeast strains of ascomycetous affinity with a specific mode of budding were isolated from raised bog soils of Bakcharskoe Bog (Tomsk oblast). According to their morphological and physiological properties, these strains belong to the genus Candida but differ from all species described previously. The level of DNA-DNA homology with species similar in the assimilation spectrum was as low as 7%. Based on these data, the new species Candida aurita sp. nov. is described.     

Decade of experimental permafrost thaw reduces turnover of young carbon and increases losses of old carbon,without affecting the net carbon balance

Thicker snowpacks and their insulation effects cause winter‐warming and invoke thaw of permafrost ecosystems. Temperature‐dependent decomposition of previously frozen carbon (C) is currently considered one of the strongest feedbacks between the Arctic and the climate system, but the direction and magnitude of the net C balance remains uncertain. This is because winter effects are rarely integrated with C fluxes during the snow‐free season and because predicting the net C balance from both surface processes and thawing deep layers remains challenging. In this study, we quantified changes in the long‐term net C balance (net ecosystem production) in a subarctic peat plateau subjected to 10 years of experimental winter‐warming. By combining ²¹⁰Pb and ¹⁴Cdating of peat cores with peat growth models, we investigated thawing effects on year‐round primary production and C losses through respiration and leaching from both shallow and deep peat layers. Winter‐warming and permafrost thaw had no effect on the net C balance, but strongly affected gross C fluxes. Carbon losses through decomposition from the upper peat were reduced as thawing of permafrost induced surface subsidence and subsequent waterlogging. However, primary production was also reduced likely due to a strong decline in bryophytes cover while losses from the old C pool almost tripled, caused by the deepened active layer. Our findings highlight the need to estimate long‐term responses of whole‐year production and decomposition processes to thawing, both in shallow and deep soil layers, as they may contrast and lead to unexpected net effects on permafrost C storage.     

Temperature and peat type control CO2 and CH4 production in Alaskan permafrost peats

Controls on the fate of ~277 Pg of soil organic carbon (C) stored in permafrost peatland soils remain poorly understood despite the potential for a significant positive feedback to climate change. Our objective was to quantify the temperature, moisture, organic matter, and microbial controls on soil organic carbon (SOC) losses following permafrost thaw in peat soils across Alaska. We compared the carbon dioxide (CO₂) and methane (CH₄) emissions from peat samples collected at active layer and permafrost depths when incubated aerobically and anaerobically at ?5, ?0.5, +4, and +20 °C. Temperature had a strong, positive effect on C emissions; global warming potential (GWP) was >3× larger at 20 °C than at 4 °C. Anaerobic conditions significantly reduced CO₂ emissions and GWP by 47% at 20 °C but did not have a significant effect at ?0.5 °C. Net anaerobic CH₄ production over 30 days was 7.1 ± 2.8 μg CH₄‐C gC^?1 at 20 °C. Cumulative CO₂ emissions were related to organic matter chemistry and best predicted by the relative abundance of polysaccharides and proteins (R² = 0.81) in SOC. Carbon emissions (CO₂‐C + CH₄‐C) from the active layer depth peat ranged from 77% larger to not significantly different than permafrost depths and varied depending on the peat type and peat decomposition stage rather than thermal state. Potential SOC losses with warming depend not only on the magnitude of temperature increase and hydrology but also organic matter quality, permafrost history, and vegetation dynamics, which will ultimately determine net radiative forcing due to permafrost thaw.     

Microbial Respiration in Arctic Upland and Peat Soils as a Source of Atmospheric Carbon Dioxide

Knowledge on soil microbial respiration (SMR) rates and thus soil-related CO₂ losses from Arctic soils is vital because of the crucial importance of this ecosystem within the global carbon (C) cycle and climate system. Here, we measured SMR from various habitats during the growing season in Russian subarctic tundra by applying two different approaches: ¹⁴C partitioning approach and root trenching. The variable habitats encompassed peat and mineral soils, bare and vegetated surfaces and included both dry and moist ones. The field experiment was complemented by laboratory studies to measure bioavailability of soil carbon and identify sources of CO₂. Differences in bioavailability of soils, measured in the laboratory as basal soil respiration rates, were generally greater than inter-site differences in SMR rates measured in situ, suggesting secondary constraints at field conditions, such as soil C content. There was a tendency towards lower SMR in vegetated peat plateaus compared to upland mineral tundra (on average 137 vs. 185 g CO₂ m^?2 growing season^?1, respectively), but no significant differences were found. Surprisingly, the bare surfaces (peat circles) with 3500-year-old C at the surface exhibited about the largest SMR among all sites as shown by both methods. This was related to the general development of peat plateaus in the region, and uplifting of deeper peat with high C content to the surface during the genesis of peat circles. This observation is particularly relevant for decomposition of deeper peat in vegetated peat plateaus, where soil material similar to the bare surfaces can be found. The data indicate that the large stocks of C stored in permafrost peatlands are principally available for decomposition despite old age.     

A New Yeast Species,Candida aurita sp. nov., from Oligotrophic Bogs of Western Siberia

Five anamorphous yeast strains of ascomycetous affinity with a specific mode of budding were isolated from high bog soils of the Bakcharskoe Bog (Tomsk oblast). According to their morphological and physiological properties, these strains belong to the genus Candida but differ from all species described previously. The level of DNA–DNA homology with species similar in the assimilation spectrum was as low as 7%. Based on these data, the new species Candida aurita sp. nov. is described.     

Production of mycophenolic acid by fungi of the genus Penicillium link

Out of 36 strains of fungi of the genus Penicillium, some of which were isolated from ancient permafrost soils, 14 strains synthesized mycophenolic acid (MPA). Maximal (over 500 mg/l) accumulation of MPA in culture liquid was observed in P. brevicompactum strains (VKM F-457, VKM F-477, and VKM F-1150). This was the first study to detect MPA in representatives of the species P. rugulosum; in three strains of this species (VKM FW-665, VKM FW-717, and VKM FW-733), the level of MPA accumulation exceeded 300 mg/l. The time course of the synthesis of MPA by the P. rugulosum strain VKM FW-733 was studied. It was shown that the synthesis of this metabolite was dramatically intensified at the stationary growth phase (ten days).     

Characterization of new phototrophic heliobacteria and their habitats

Specific enrichment culture methods were used to isolate new phototrophic heliobacteria (anoxygenic phototrophic bacteria containing bacteriochlorophyll g) from various natural samples. A survey of terrestrial and aquatic habitats yielded heliobacteria only from soils, in particular rice soils, and from certain hot springs. Thirteen nonthermophilic and 7 thermophilic (capable of growth above 50) strains of heliobacteria were isolated from such habitats and characterized as to their basic cellular and nutritional properties. Phylogenetic studies of four strains showed them to be related to known species of heliobacteria. It is concluded that, unlike phototrophic purple and green bacteria, heliobacteria are primarily (if not exclusively) terrestrial, except for hot spring species. This suggests that the ecology of heliobacteria is fundamentally different from that of other anoxyphototrophs.     

Permafrost thaw causes large carbon loss in boreal peatlands while changes to peat quality are limited

Rapid, ongoing permafrost thaw of peatlands in the discontinuous permafrost zone is exposing a globally significant store of soil carbon (C) to microbial processes. Mineralization and release of this peat C to the atmosphere as greenhouse gases is a potentially important feedback to climate change. Here we investigated the effects of permafrost thaw on peat C at a peatland complex in western Canada. We collected 15 complete peat cores (between 2.7 and 4.5 m deep) along four chronosequences, from elevated permafrost peat plateaus to saturated thermokarst bogs that thawed up to 600 years ago. The peat cores were analysed for peat C storage and peat quality, as indicated by decomposition proxies (FTIR and C/N ratios) and potential decomposability using a 200-day aerobic laboratory incubation. Our results suggest net C loss following thaw, with average total peat C stocks decreasing by ~19.3 ± 7.2 kg C m⁻² over <600 years (~13% loss). Average post-thaw accumulation of new peat at the surface over the same period was ~13.1 ± 2.5 kg C m⁻². We estimate ~19% (±5.8%) of deep peat (>40 cm below surface) C is lost following thaw (average 26 ± 7.9 kg C m⁻² over <600 years). Our FTIR analysis shows peat below the thaw transition in thermokarst bogs is slightly more decomposed than peat of a similar type and age in permafrost plateaus, but we found no significant changes to the quality or lability of deeper peat across the chronosequences. Our incubation results also showed no increase in C mineralization of deep peat across the chronosequences. While these limited changes in peat quality in deeper peat following permafrost thaw highlight uncertainty in the exact mechanisms and processes for C loss, our analysis of peat C stocks shows large C losses following permafrost thaw in peatlands in western Canada.