Characterization of C1-metabolizing prokaryotic communities in methane seep habitats at the Kuroshima Knoll, southern Ryukyu Arc, by analyzing pmoA, mmoX, mxaF, mcrA, and 16S rRNA genes

Samples from three submerged sites (MC, a core obtained in the methane seep area; MR, a reference core obtained at a distance from the methane seep; and HC, a gas-bubbling carbonate sample) at the Kuroshima Knoll in the southern Ryuku arc were analyzed to gain insight into the organisms present and the processes involved in this oxic-anoxic methane seep environment. 16S rRNA gene analyses by quantitative real-time PCR and clone library sequencing revealed that the MC core sediments contained abundant archaea (approximately 34% of the total prokaryotes), including both mesophilic methanogens related to the genus Methanolobus and ANME-2 members of the Methanosarcinales, as well as members of the delta-Proteobacteria, suggesting that both anaerobic methane oxidation and methanogenesis occurred at this site. In addition, several functional genes connected with methane metabolism were analyzed by quantitative competitive-PCR, including the genes encoding particulate methane monooxygenase (pmoA), soluble methane monooxygenase (mmoX), methanol dehydrogenese (mxaF), and methyl coenzyme M reductase (mcrA). In the MC core sediments, the most abundant gene was mcrA (2.5 x 10(6) copies/g [wet weight]), while the pmoA gene of the type I methanotrophs (5.9 x 10(6) copies/g [wet weight]) was most abundant at the surface of the MC core. These results indicate that there is a very complex environment in which methane production, anaerobic methane oxidation, and aerobic methane oxidation all occur in close proximity. The HC carbonate site was rich in gamma-Proteobacteria and had a high copy number of mxaF (7.1 x 10(6) copies/g [wet weight]) and a much lower copy number of the pmoA gene (3.2 x 10(2) copies/g [wet weight]). The mmoX gene was never detected. In contrast, the reference core contained familiar sequences of marine sedimentary archaeal and bacterial groups but not groups specific to C1 metabolism. Geochemical characterization of the amounts and isotopic composition of pore water methane and sulfate strongly supported the notion that in this zone both aerobic methane oxidation and anaerobic methane oxidation, as well as methanogenesis, occur.     

Microbial diversity of active layer and permafrost in an acidic wetland from the Canadian High Arctic

The abundance and structure of archaeal and bacterial communities from the active layer and the associated permafrost of a moderately acidic (pH < 5.0) High Arctic wetland (Axel Heiberg Island, Nunavut, Canada) were investigated using culture- and molecular-based methods. Aerobic viable cell counts from the active layer were ～100-fold greater than those from the permafrost (2.5 × 10(5) CFU·(g soil dry mass)(-1)); however, a greater diversity of isolates were cultured from permafrost, as determined by 16S rRNA gene sequencing. Isolates from both layers demonstrated growth characteristics of a psychrotolerant, halotolerant, and acidotolerant community. Archaea constituted 0.1% of the total 16S rRNA gene copy number and, in the 16S rRNA gene clone library, predominantly (71% and 95%) consisted of Crenarchaeota related to Group I. 1b. In contrast, bacterial communities were diverse (Shannon's diversity index, H = ～4), with Acidobacteria constituting the largest division of active layer clones (30%) and Actinobacteria most abundant in permafrost (28%). Direct comparisons of 16S rRNA gene sequence data highlighted significant differences between the bacterial communities of each layer, with the greatest differences occurring within Actinobacteria. Comparisons of 16S rRNA gene sequences with those from other Arctic permafrost and cold-temperature wetlands revealed commonly occurring taxa within the phyla Chloroflexi, Acidobacteria, and Actinobacteria (families Intrasporangiaceae and Rubrobacteraceae).     

Methanogenic communities in permafrost-affected soils of the Laptev Sea coast, Siberian Arctic, characterized by 16S rRNA gene fingerprints

Permafrost environments in the Arctic are characterized by extreme environmental conditions that demand a specific resistance from microorganisms to enable them to survive. In order to understand the carbon dynamics in the climate-sensitive Arctic permafrost environments, the activity and diversity of methanogenic communities were studied in three different permafrost soils of the Siberian Laptev Sea coast. The effect of temperature and the availability of methanogenic substrates on CH4 production was analysed. In addition, the diversity of methanogens was analysed by PCR with specific methanogenic primers and by denaturing gradient gel electrophoresis (DGGE) followed by sequencing of DGGE bands reamplified from the gel. Our results demonstrated methanogenesis with a distinct vertical profile in each investigated permafrost soil. The soils on Samoylov Island showed at least two optima of CH4 production activity, which indicated a shift in the methanogenic community from mesophilic to psychrotolerant methanogens with increasing soil depth. Furthermore, it was shown that CH4 production in permafrost soils is substrate-limited, although these soils are characterized by the accumulation of organic matter. Sequence analyses revealed a distinct diversity of methanogenic archaea affiliated to Methanomicrobiaceae, Methanosarcinaceae and Methanosaetaceae. However, a relationship between the activity and diversity of methanogens in permafrost soils could not be shown.     

Impacts of warming and fertilization on nitrogen-fixing microbial communities in the Canadian High Arctic

The impacts of simulated climate change (warming and fertilization treatments) on diazotroph community structure and activity were investigated at Alexandra Fiord, Ellesmere Island, Canada. Open Top Chambers, which increased growing season temperatures by 1-3 degrees C, were randomly placed in a dwarf-shrub and cushion-plant dominated mesic tundra site in 1995. In 2000 and 2001 20N:20P2O5:20K2O fertilizer was applied at a rate of 5 gm(-2) year(-1). Estimates of nitrogen fixation rates were made in the field by acetylene reduction assays (ARA). Higher rates of N fixation were observed 19-35 days post-fertilization but were otherwise unaffected by treatments. However, moss cover was significantly positively associated with ARA rate. NifH gene variants were amplified from bulk soil DNA and analyzed by terminal restriction fragment length polymorphism analysis. Non-metric multidimensional scaling was used to ordinate treatment plots in nifH genotype space. NifH gene communities were more strongly structured by the warming treatment late in the growing season, suggesting that an annual succession in diazotroph community composition occurs.     

Seasonal and annual variability in the quality of important forage plants on Banks Island,Canadian High Arctic

Abstract. In vitro acid‐pepsin digestibility (IVDMD), crude protein (CP), fibre, lignin, and energy content were measured for a variety of forage plants collected annually from Banks Island over five summers and three winters from 1993–1998. Summer samples were collected during mid‐June (start of growing season), mid‐July (peak of growing season), and mid‐late August (senescence). Winter samples were collected in early (November), mid‐ (February), and late‐ (April/May) winter. Samples, collected in areas of both high and low muskox density, included Carex aquatilis, unidentified Carex, Salixarctica, Dryasintegrifolia, Cassiope tetragona, Saxifraga spp., Astragalus spp., Oxytropis spp., lichen, and grass. Seasonal dynamics in forage quality during the growing season were similar to those reported elsewhere in the arctic and high arctic and were consistent across years. However, there were significant year effects in lignin, fibre, and energy content of forages and the crude protein (CP) content of C. aquatilis in winter, indicating annual differences in the quality of forage available to herbivores. The quality of forages on Banks Island was similar between areas subjected to different densities of muskox (ca. 1.6–1.9 versus 0.3 ‐ 0.4/km²) implying that quality was not affected by these grazing intensities. The Banks Island high arctic ecosystem supports an abundance of herbivores. It has been hypothesized that this is because forage quality and/or quantity are superior on Banks Island than elsewhere in the high arctic. Our results regarding forage quality are equivocal. Although the maximum CP content of forages from Banks Island was generally higher than reported elsewhere in the arctic and high arctic, CP content reported elsewhere fell within the interannual range reported from Banks Island. Fibre and energy content of forages from Banks Island were similar to slightly lower than elsewhere in the arctic and high arctic. Such comparisons must be considered in light of the interannual variability in quality we report.     

Modeling density dependence and climatic disturbances in caribou: a case study from the Bathurst Island complex, Canadian High Arctic

Peary caribou Rangifer tarandus pearyi is the northernmost subspecies of Rangifer in North America and endemic to the Canadian High Arctic. Because of severe population declines following years of unfavorable winter weather with ice coating on the ground or thicker snow cover, it is believed that density-independent disturbance events are the primary driver for Peary caribou population dynamics. However, it is unclear to what extent density dependence may affect population dynamics of this species. Here, we test for different levels of density dependence in a stochastic, single-stage population model, based on available empirical information for the Bathurst Island complex (BIC) population in the Canadian High Arctic. We compare predicted densities with observed densities during 1961–2001 under various assumptions of the strength of density dependence. On the basis of our model, we found that scenarios with no or very low density dependence led to population densities far above observed densities. For average observed disturbance regimes, a carrying capacity of 0.1 caribou km⁻² generated an average caribou density similar to that estimated for the BIC population over the past four decades. With our model we also tested the potential effects of climate change-related increases in the probability and severity of disturbance years, that is unusually poor winter conditions. On the basis of our simulation results, we found that, in particular, potential increases in disturbance severity (as opposed to disturbance frequency) may pose a considerable threat to the persistence of this species.     

Physical and Chemical Limnological Characteristics of 38 Lakes and Ponds on Bathurst Island,Nunavut, Canadian High Arctic

The limnological features that characterize the shallow ponds (<2 m deep) and lakes (>2 m deep) on Bathurst Island, Nunavut, Canada were examined through chemical analyses and multivariate statistical methods as part of a larger on‐going survey to document and monitor environmental changes in these remote and sensitive areas. All sites were relatively oligotrophic and alkaline. Nutrient ratios indicated that nitrogen could be limiting algal growth to a greater degree than phosphorus in over 63% of the sites. Principal components analysis (PCA) was used to explore the patterns of variation in the limnological dataset. The three dominant limnological gradients were: major ion content and dissolved organic carbon (DOC) levels along Axis 1; and pH along Axis 2.     

The effect of nutrient deposition on bacterial communities in Arctic tundra soil

The microbial communities of high‐latitude ecosystems are expected to experience rapid changes over the next century due to climate warming and increased deposition of reactive nitrogen, changes that will likely affect microbial community structure and function. In moist acidic tundra (MAT) soils on the North Slope of the Brooks Range, Alaska, substantial losses of C and N were previously observed after long‐term nutrient additions. To analyse the role of microbial communities in these losses, we utilized 16S rRNA gene tag pyrosequencing coupled with community‐level physiological profiling to describe changes in MAT bacterial communities after short‐ and long‐term nutrient fertilization in four sets of paired control and fertilized MAT soil samples. Bacterial diversity was lower in long‐term fertilized plots. The Acidobacteria were one of the most abundant phyla in all soils and distinct differences were noted in the distributions of Acidobacteria subgroups between mineral and organic soil layers that were also affected by fertilization. In addition, Alpha‐ and Gammaproteobacteria were more abundant in long‐term fertilized samples compared with control soils. The dramatic increase in sequences within the Gammaproteobacteria identified as Dyella spp. (order Xanthomonadales) in the long‐term fertilized samples was confirmed by quantitative PCR (qPCR) in several samples. Long‐term fertilization was also correlated with shifts in the utilization of specific substrates by microbes present in the soils. The combined data indicate that long‐term fertilization resulted in a significant change in microbial community structure and function linked to changes in carbon and nitrogen availability and shifts in above‐ground plant communities.     

Short-term impacts of active layer detachments on carbon exchange in a High Arctic ecosystem,Cape Bounty,Nunavut, Canada

Localized permafrost disturbances such as active layer detachments (ALDs) are increasing in frequency and severity across the Canadian Arctic impacting terrestrial ecosystem functioning. However, the contribution of permafrost disturbance-carbon feedbacks to the carbon (C) balance of Arctic ecosystems is poorly understood. Here, we explore the short-term impact of active layer detachments (ALDs) on carbon dioxide (CO₂) exchange in a High Arctic semi-desert ecosystem by comparing midday C exchange between undisturbed areas, moderately disturbed areas (intact islands of vegetation within an ALD), and highly disturbed areas (non-vegetated areas due to ALD). Midday C exchange was measured using a static chamber method between June 23 and August 8 during the 2009 and 2010 growing seasons. Results show that areas of high disturbance had significantly reduced gross ecosystem exchange and ecosystem respiration (R _E) compared to control and moderately disturbed areas. Moderately disturbed areas showed significantly enhanced net ecosystem exchange compared to areas of high disturbance, but were not significantly different from control areas. Disturbance did not significantly impact soil thermal, physical or chemical properties. According to average midday fluxes, ALDs as a whole (moderately disturbed areas: ?1.942 μmol m^?2 s^?1+ highly disturbed areas: 2.969 μmol m^?2 s^?1) were a small CO₂ source of 1.027 μmol m^?2 s^?1 which did not differ significantly from average midday fluxes in control areas 1.219 μmol m^?2 s^?1. The findings of this study provide evidence that the short-term impacts of ALDs on midday, net C exchange and soil properties in a High Arctic semi-desert are minimal.     

Diatoms and their relationship to environmental variables from lakes and ponds on Bathurst Island, Nunavut, Canadian High Arctic

Baseline and historical environmental data are sparse in the High Arctic, however diatom assemblages preserved in high arctic lake and pond sediment profiles can provide proxy data of past environmental changes. Ecological data, however, are still sparse. Diatom taxa preserved in the surficial sediment of lakes and ponds on Bathurst Island (75° 42 N, 97° 21 W), Nunavut, Canadian High Arctic, and their relationship to 34 environmental variables were examined using multivariate statistics. A total of 148 diatom taxa were identified from the surface sediments of 29 study sites. Five environmental variables, Fe³⁺, Total Phosphorus (Unfiltered) (TPU), Total Nitrogen (TN), Temperature (TEMP) and pH significantly (P0.05) accounted for most of the variation in the diatom assemblages. TN was also significantly correlated to other variables (e.g. TPF, DOC, POC). A CCA constrained to TN indicated that this variable best explained the species distributions, and a weighted-averaging (WA) model was developed to infer nutrient levels from the relative abundances of the 58 dominant taxa. Interestingly, previous limnological work indicated that nitrogen most likely limited algal production in some of these high arctic sites. This model may be used to quantitatively estimate past TN levels from diatom assemblages preserved in sediment cores from Bathurst Island, and may provide a means to track past environmental changes in the High Arctic.     

Impacts of seabird-derived nutrients on water quality and diatom assemblages from Cape Vera,Devon Island,Canadian High Arctic

Relationships between key phytoplankton attributes including Chl a-specific light absorption, pigment composition and concentration, photosynthesis, primary production and community structure were studied in two open shallow nutrient-poor coastal systems receiving similar amounts of sewage water. Both systems were significantly nitrogen limited. However, differences in wastewater treatment (primary vs secondary) and sewage dilution (50%) between the two systems caused a greater difference between systems than locally around the outflows. For both systems, water at the outlet had significantly lower water transparency caused by a 20% higher absorption by coloured dissolved organic matter. Nutrient concentrations were also elevated, gradually decreasing with distance north (governing current) of the outflows, causing higher abundance of nano-sized phytoplankton, higher content of carotenoid pigments, 20–50% higher Chl a-specific absorption coefficients and higher photosynthetic capacity. Although maximum rates of Chl a-normalised photosynthesis were strongly related to nitrate availability, no effects were found on the derived areal primary production or algal biomass suggesting that photosynthetic and optical parameters are more sensitive indicators of nutrient enrichment than biomass or productivity. Handling editor: Tasman Peter Crowe     

Limnology of 46 lakes and ponds on Banks Island, N.W.T., Canadian Arctic Archipelago

The goal of this study was to describe and quantify the physical and chemical limnological properties of 46 lakes and ponds on Banks Island, to explore the effects of ecoclimatic differences on the water chemistry of these study sites, and to establish baseline conditions for this previously unexplored limnological region, which could then be used in subsequent long-term environmental monitoring programs. A key finding was that the study sites on Banks Island represented a large nutrient concentration gradient from ultra-oligotrophic to hypereutrophic waters. In general, the study sites were relatively nutrient rich by Arctic standards (i.e. mean total nitrogen (TN_mean)=504.2μg/l and mean total phosphorus (TPU_mean)=18.0μg/l); concentrations that are amongst the highest of any previous limnological survey from similar latitudes. Dissolved organic carbon (DOC) concentrations were also some of the highest reported to date amongst all other Canadian Arctic island limnological surveys. These values reflect the milder climate, concentrated animal life and lushness of Banks Island, as compared to other Canadian Arctic Archipelago islands. Principal components analysis (PCA) separated sites along a conductivity/ionic and elevation gradient on Axis 1 (λ₁=0.343), and a metal (Fe, Zn, Al) and alkalinity-related (DIC, pH) gradient on the second axis (λ₂=0.187). Canonical variate analysis (CVA) was used to explore the classification of the study sites into Low Arctic, Mid Arctic or High Arctic designations based on their limnological characteristics.     

Ecology of terrestrial testate amoebae assemblages from coastal Lowlands on Devon Island (NWT,Canadian Arctic)

Summary Fifty-seven taxa belonging to 17 genera of testate rhizopods were recorded in samples of mosses, soils and lichens, collected from the Truelove and Sparbo Hardy Lowlands on the high arctic Devon Island (NWT, Canada). Different groups of habitats are characterized by particular assemblages of Testacea species, with the quantity and quality of the available water as important ecological parameters. The Assulina muscorum-Corythion dubium assemblage characterizes the most dry and acidic moss habitats, while the Paraquadrula irregularis assemblage is to be found in wet and rather neutral habitats. The sedge moss meadows with their more moderately coarse textured and poorly drained soils, bear a Plagiopyxis callida-Plagiopyxis declivis soil-dwelling testate amoebae assemblage. The Centropyxis minuta assemblage lives in the most coarsely textured soils. pH seems to play a subordinate role, not only in the mosses but also in the soils. Multivariate methods (cluster analysis and CANOCO) have been used to separate these different assemblages.     

Experimental manipulation of the winter surface ice layer: the effects on a High Arctic soil microarthropod community

The effect of patch size on the tolerance of the soil microarthropod population to an experimentally induced environmental catastrophe, a thick surface ice layer, was studied at a High Arctic site (78°55 'N, 11°53' E). Such an ice layer currently occurs infrequently; however, climate change models suggest that the occurrence of such an ice layer is likely to increase in frequency. The experimental approach was a factorial design with two patch sizes, an icing treatment and controls. A thin layer of natural ice was present even in the controls and this was treated as a covariate in the analysis. The soil microarthropod fauna at the experimental site consisted of five species of Collembola and seven species of oribatid mite. The experimental surface ice layer reduced the total number of the soil microarthropods studied by 50%; however, mortality differed between mites and Collembola and species within the two taxo-nomic groups. Mites were very resistant and showed no significant change, Collembola more sensitive (the populations of Hypogastrura tullbergi declined by 56% and Folsomia quadrioculata by 54'% in the iced treatment plots). The thin annual surface ice layer seemed to have an additional effect on H. tullbergi and the mite Lauroppia translamcllata. That such a thin ice layer could reduce survival was unexpected and could play an important role in determining the extreme patchy distribution of arctic soil animals.     

Development of a swine-specific fecal pollution marker based on host differences in methanogen mcrA genes

The goal of this study was to evaluate methanogen diversity in animal hosts to develop a swine-specific archaeal molecular marker for fecal source tracking in surface waters. Phylogenetic analysis of swine mcrA sequences compared to mcrA sequences from the feces of five animals (cow, deer, sheep, horse, and chicken) and sewage showed four distinct swine clusters, with three swine-specific clades. From this analysis, six sequences were chosen for molecular marker development and initial testing. Only one mcrA sequence (P23-2) showed specificity for swine and therefore was used for environmental testing. PCR primers for the P23-2 clone mcrA sequence were developed and evaluated for swine specificity. The P23-2 primers amplified products in P23-2 plasmid DNA (100%), pig feces (84%), and swine waste lagoon surface water samples (100%) but did not amplify a product in 47 bacterial and archaeal stock cultures and 477 environmental bacterial isolates and sewage and water samples from a bovine waste lagoon and a polluted creek. Amplification was observed in only one sheep sample out of 260 human and nonswine animal fecal samples. Sequencing of PCR products from pig feces demonstrated 100% similarity to pig mcrA sequence from clone P23-2. The minimal amount of DNA required for the detection was 1 pg for P23-2 plasmid, 1 ng for pig feces, 50 ng for swine waste lagoon surface water, 1 ng for sow waste influent, and 10 ng for lagoon sludge samples. Lower detection limits of 10(-6) g of wet pig feces in 500 ml of phosphate-buffered saline and 10(-4) g of lagoon waste in estuarine water were established for the P23-2 marker. This study was the first to utilize methanogens for the development of a swine-specific fecal contamination marker.     

The Physical and Chemical Limnology of 24 Ponds and One Lake from Isachsen,Ellef Ringnes Island,Canadian High Arctic

The limnology of freshwaters surrounding Isachsen, Ellef Ringnes Island, Nunavut was examined to determine the baseline physical and chemical limnological conditions present in the region. Sites were found to be circumneutral to slightly acidic, and were oligotrophic. Concentrations of most measured chemical variables were highly variable, with broad ranges that greatly exceeded those found in previous surveys conducted in the High Arctic. Ratios of nitrogen to phosphorus suggest that nitrogen may be the limiting nutrient for algal growth at the majority of sites. Principal Components Analysis (PCA) indicated that the major controls on water chemistry variability between sites were conductivity and related variables, and nutrients, explaining 36.5% and 26.5% of the variation in the dataset, respec‐tively.     

Metagenomic analysis of stress genes in microbial mat communities from Antarctica and the High Arctic

Polar and alpine microbial communities experience a variety of environmental stresses, including perennial cold and freezing; however, knowledge of genomic responses to such conditions is still rudimentary. We analyzed the metagenomes of cyanobacterial mats from Arctic and Antarctic ice shelves, using high-throughput pyrosequencing to test the hypotheses that consortia from these extreme polar habitats were similar in terms of major phyla and subphyla and consequently in their potential responses to environmental stresses. Statistical comparisons of the protein-coding genes showed similarities between the mats from the two poles, with the majority of genes derived from Proteobacteria and Cyanobacteria; however, the relative proportions differed, with cyanobacterial genes more prevalent in the Antarctic mat metagenome. Other differences included a higher representation of Actinobacteria and Alphaproteobacteria in the Arctic metagenomes, which may reflect the greater access to diasporas from both adjacent ice-free lands and the open ocean. Genes coding for functional responses to environmental stress (exopolysaccharides, cold shock proteins, and membrane modifications) were found in all of the metagenomes. However, in keeping with the greater exposure of the Arctic to long-range pollutants, sequences assigned to copper homeostasis genes were statistically (30%) more abundant in the Arctic samples. In contrast, more reads matching the sigma B genes were identified in the Antarctic mat, likely reflecting the more severe osmotic stress during freeze-up of the Antarctic ponds. This study underscores the presence of diverse mechanisms of adaptation to cold and other stresses in polar mats, consistent with the proportional representation of major bacterial groups.     

Physiological diversity of bacterial communities from different soil locations on Livingston Island, South Shetland archipelago, Antarctica

Terrestrial food webs of Antarctica are simple and dominated by microorganisms. Soil bacteria play an important role in nutrient cycling, yet little is known about their capacity to utilize different carbon sources and to participate in site nutrient turnover. Biolog EcoPlate? was applied to study the catabolic activity and physiological diversity of bacteria inhabiting the soil of moss, vascular plants, and fell field habitats from Livingston Island, Antarctica. Additionally, the number of oligotrophic and copiotrophic bacteria was counted by the agar plate method. Results indicated a lack of site-specific distribution of bacterial abundance, in contrast to bacterial catabolic activity and community level physiological profiles. Community level physiological profiles revealed a common capacity of soil bacteria to intensively utilize polyols, which are cryoprotectants widely produced by Antarctic organisms, as well as site-specific phenolic compounds (vegetated habitats), amino acids/amines (moss habitats), carbohydrates and carboxylic acids (fell field habitat). It was concluded that the physiology of soil bacteria is habitat specific concerning both the rate of catabolic activity and pattern of carbon source utilization.     

Hidden biodiversity in the Arctic – a study of soil seed banks at Disko Island,Qeqertarsuaq, West Greenland

Seed dispersal is a key process in plant community dynamics, and soil seed banks represent seed dispersal in time rather than in space. Despite their potential importance, seed bank dynamics in the Arctic are poorly understood. We investigated soil seed banks and corresponding plant community composition in three contrasting vegetation types in West Greenland, viz. dwarf shrub heaths, herb slopes and fell‐fields. Through germination testing, 31 species were documented in soil seed banks. All of these were herbaceous, while no dwarf‐shrub species, which represents the dominating growth form in the above‐ground vegetation, were emerging from the seed bank. Consequently, across vegetation types, the lowest similarity between seed bank and above‐ground vegetation was found in dwarf shrub heath. Nine plant species were exclusively found in seed bank, all of which were non‐clonal forbs. Seed bank size (total number of seeds) and species richness seemed to increase with the level of natural disturbance. Additionally, we examined the effect of different experimental light and temperature conditions on the quantity and diversity of germinating seeds. The difference in diversity in vegetation and seed bank at the species level will impact population dynamics, regeneration of vegetation after disturbances and its potential to respond to climate change.