Geology defines microbiome structure and composition in nunataks and valleys of the Sør Rondane Mountains, East Antarctica.

Understanding the relation between terrestrial microorganisms and edaphic factors in the Antarctic can provide insights into their potential response to environmental changes. Here we examined the composition of bacterial and micro-eukaryotic communities using amplicon sequencing of rRNA genes in 105 soil samples from the Sør Rondane Mountains (East Antarctica), differing in bedrock or substrate type and associated physicochemical conditions. Although the two most widespread taxa (Acidobacteriota and Chlorophyta) were relatively abundant in each sample, multivariate analysis and co-occurrence networks revealed pronounced differences in community structure depending on substrate type. In moraine substrates, Actinomycetota and Cercozoa were the most abundant bacterial and eukaryotic phyla, whereas on gneiss, granite and marble substrates, Cyanobacteriota and Metazoa were the dominant bacterial and eukaryotic taxa. However, at lower taxonomic level, a distinct differentiation was observed within the Cyanobacteriota phylum depending on substrate type, with granite being dominated by the Nostocaceae family and marble by the Chroococcidiopsaceae family. Surprisingly, metazoans were relatively abundant according to the 18S rRNA dataset, even in samples from the most arid sites, such as moraines in Austkampane and Widerøefjellet ("Dry Valley"). Overall, our study shows that different substrate types support distinct microbial communities, and that mineral soil diversity is a major determinant of terrestrial microbial diversity in inland Antarctic nunataks and valleys.

Mineral substrate quality determines the initial soil microbial development in front of the Nordenskiöldbreen, Svalbard.

Substrate geochemistry is an important factor influencing early microbial development after glacial retreat on nutrient-poor geological substrates in the High Arctic. It is often difficult to separate substrate influence from climate because study locations are distant. Our study in the retreating Nordenskiöldbreen (Svalbard) is one of the few to investigate biogeochemical and microbial succession in two adjacent forefields, which share the same climatic conditions but differ in their underlying geology. The northern silicate forefield evolved in a classical chronosequence, where most geochemical and microbial parameters increased gradually with time. In contrast, the southern carbonate forefield exhibited high levels of nutrients and microbial biomass at the youngest sites, followed by a significant decline and then a gradual increase, which caused a rearrangement in the species and functional composition of the bacterial and fungal communities. This shuffling in the early stages of succession suggests that high nutrient availability in the bedrock could have accelerated early soil succession after deglaciation and thereby promoted more rapid stabilization of the soil and production of higher quality organic matter. Most chemical parameters and bacterial taxa converged with time, while fungi showed no clear pattern.

Metagenomic Analysis of Antarctic Biocrusts Unveils a Rich Range of Cold-Shock Proteins.

Microorganisms inhabiting Antarctic biocrusts develop several strategies to survive extreme environmental conditions such as severe cold and drought. However, the knowledge about adaptations of biocrusts microorganisms are limited. Here, we applied metagenomic sequencing to study biocrusts from east Antarctica. Biocrusts were dominated by cyanobacteria, actinobacteria and proteobacteria. Furthermore, the results provided insights into the presence and abundance of cold shock proteins (Csp), cold shock domain A proteins (CsdA), and antifreeze proteins (AFP) in these extreme environments. The metagenomic analysis revealed a high number of CsdA across the samples. The majority of the Csp recorded in the studied biocrusts were Csp A, C, and E. In addition, CsdA, Csp, and AFP primarily originated from proteobacteria and actinobacteria.

Microbial community composition of terrestrial habitats in East Antarctica with a focus on microphototrophs.

The Antarctic terrestrial environment harbors a diverse community of microorganisms, which have adapted to the extreme conditions. The aim of this study was to describe the composition of microbial communities in a diverse range of terrestrial environments (various biocrusts and soils, sands from ephemeral wetlands, biofilms, endolithic and hypolithic communities) in East Antarctica using both molecular and morphological approaches. Amplicon sequencing of the 16S rRNA gene revealed the dominance of Chloroflexi, Cyanobacteria and Firmicutes, while sequencing of the 18S rRNA gene showed the prevalence of Alveolata, Chloroplastida, Metazoa, and Rhizaria. This study also provided a comprehensive assessment of the microphototrophic community revealing a diversity of cyanobacteria and eukaryotic microalgae in various Antarctic terrestrial samples. Filamentous cyanobacteria belonging to the orders Oscillatoriales and Pseudanabaenales dominated prokaryotic community, while members of Trebouxiophyceae were the most abundant representatives of eukaryotes. In addition, the co-occurrence analysis showed a prevalence of positive correlations with bacterial taxa frequently co-occurring together.

Biocrusts from Iceland and Svalbard: Does microbial community composition differ substantially?

A wide range of microorganisms inhabit biocrusts of arctic and sub-arctic regions. These taxa live and thrive under extreme conditions and, moreover, play important roles in biogeochemical cycling. Nevertheless, their diversity and abundance remain ambiguous. Here, we studied microbial community composition in biocrusts from Svalbard and Iceland using amplicon sequencing and epifluorescence microscopy. Sequencing of 16S rRNA gene revealed the dominance of Chloroflexi in the biocrusts from Iceland and Longyearbyen, and Acidobacteria in the biocrusts from Ny-Ålesund and South Svalbard. Within the 18S rRNA gene sequencing dataset, Chloroplastida prevailed in all the samples with dominance of Trebouxiophyceae in the biocrusts from Ny-Ålesund and Embryophyta in the biocrusts from the other localities. Furthermore, cyanobacterial number of cells and biovolume exceeded the microalgal in the biocrusts. Community compositions in the studied sites were correlated to the measured chemical parameters such as conductivity, pH, soil organic matter and mineral nitrogen contents. In addition, co-occurrence analysis showed the dominance of positive potential interactions and, bacterial and eukaryotic taxa co-occurred more frequently together.

Annual Cycle of Mat-Forming Filamentous Alga Tribonema cf. minus (Stramenopiles, Xanthophyceae) in Hydro-Terrestrial Habitats in the High Arctic Revealed By Multiparameter Fluorescent Staining.

The filamentous microalga Tribonema sp. (Stramenopiles, Xanthophyceae) plays an important role in shallow water polar (streams and seepages) and seasonally cold habitats in temperate regions (ponds). In these habitats, freezing and desiccation, and thus freeze-thawing and drying-rewetting cycles, are frequent. These regions produce visible biomass and are important components of low temperature-adapted communities. We characterized the annual cycles of a Tribonema cf. minus population in two habitats (seepage and stream) in the High Arctic, Svalbard. Seasonality, locality, and their combination (particularly changing environmental conditions) together with cultivation conditions of strains significantly affected their morphological characteristics. Morphological changes following hardening processes related to preparation for the winter period (transition from vegetative cells to akinete and/or pre-akinete) were recorded. Over the year, positive water temperatures (warmest 13.3°C) occurred for 5 months while negative (lowest temperature was -17.4°C) lasted for 7 months. In winter, there were two melt periods. Vitality staining protocol showed a high number of viable (77.4% and 53.8%) and dormant cells (1.7% and 4.1%; capable of growth and reproduction once suitable conditions return) in the winter seepage and stream, respectively. NPQ and OJIP chlorophyll fluorescence parameters revealed several hours recovery of photosynthesis (both field and control samples). During recovery, only minor or mild stress on photosynthesis was detected. FV /FM values (the photosynthetic efficiency of photosystem II in a dark-adapted state) in all field and control samples varied around 0.4. Tribonema cf. minus is capable of surviving winter Arctic conditions (perennial strategy).

Annual Cycle of Freshwater Diatoms in the High Arctic Revealed by Multiparameter Fluorescent Staining.

Diatoms (Bacillariophyceae) are important primary producers in a wide range of hydro-terrestrial habitats in polar regions that are characterized by many extreme environmental conditions. Nevertheless, how they survive periods of drought and/or freeze remains unknown. A general strategy of microorganisms to overcome adverse conditions is dormancy, but morphologically distinct diatom resting stages are rare. This study aimed to evaluate the annual cycle of freshwater diatoms in the High Arctic (Central Spitsbergen) and provide an insight into their physiological cell status variability. The diversity and viability of diatom cells were studied in samples collected five times at four study sites, tracing the key events for survival (summer vegetative season, autumn dry-freezing, winter freezing, spring melting, summer vegetative season [again]). For viability evaluation, a multiparameter fluorescent staining was used in combination with light microscopy and allowed to reveal the physiological status at a single-cell level. The proportions of the cell categories were seasonally and locality dependent. The results suggested that a significant portion of vegetative cells survive winter and provide an inoculum for the following vegetative season. The ice thickness significantly influenced spring survival. The thicker the ice layer was, the more dead cells and fewer other stages were observed. The influence of the average week max-min temperature differences in autumn and winter was not proven.

Phylogenetic evaluation of the genus Nostoc and description of Nostoc neudorfense sp. nov., from the Czech Republic.

Cyanobacterial strain ARC8 was isolated from seepage coming into the river Dracice, Frantiskov, Czech Republic, and was characterized using a polyphasic approach. Strain ARC8 showed a typical Nostoc-like morphology and in-depth morphological characterization indicated that it is a member of the genus Nostoc. Furthermore, in the 16S rRNA gene phylogeny inferred using Bayesian inference, maximum likelihood and neighbour joining methods, strain ARC8 clustered within the Nostoc sensu stricto clade. The phylogenetic distance and the positioning of strain ARC8 also indicated that it is a member of the genus Nostoc. Furthermore, the rbcL gene phylogeny along with the 16S-23S ITS secondary structure analysis also supported the findings from the 16S rRNA gene tree. In accordance with the International Code of Nomenclature for Algae, Fungi and Plants we describe a novel species of Nostoc with the name Nostoc neudorfense sp. nov.

Ecophysiological Features of Polar Soil Unicellular Microalgae1.

Due to their ecological, physiological, and molecular adaptations to low and varying temperatures, as well as varying seasonal irradiances, polar non-marine eukaryotic microalgae could be suitable for low-temperature biotechnology. Adaptations include the synthesis of compounds from different metabolic pathways that protect them against stress. Production of biological compounds and various biotechnological applications, for instance, water treatment technology, are of interest to humans. To select prospective strains for future low-temperature biotechnology in polar regions, temperature and irradiance of growth requirements (Q10 and Ea of 10 polar soil unicellular strains) were evaluated. In terms of temperature, three groups of strains were recognized: (i) cold-preferring where temperature optima ranged between 10.1 and 18.4°C, growth rate 0.252 and 0.344 · d-1 , (ii) cold- and warm-tolerating with optima above 10°C and growth rate 0.162-0.341 · d-1 , and (iii) warm-preferring temperatures above 20°C and growth rate 0.249-0.357 · d-1 . Their light requirements were low. Mean values Q10 for specific growth rate ranged from 0.7 to 3.1. The lowest Ea values were observed on cold-preferring and the highest in the warm-preferring strains. One strain from each temperature group was selected for PN and RD measurements. The PN :RD ratio of the warm-preferring strains was less affected by temperature similarly as Q10 and Ea. For future biotechnological applications, the strains with broad temperature tolerance (i.e., the group of cold- and warm-tolerating and warm-preferring strains) will be most useful.

Marked Succession of Cyanobacterial Communities Following Glacier Retreat in the High Arctic.

Cyanobacteria are important colonizers of recently deglaciated proglacial soil but an in-depth investigation of cyanobacterial succession following glacier retreat has not yet been carried out. Here, we report on the successional trajectories of cyanobacterial communities in biological soil crusts (BSCs) along a 100-year deglaciation gradient in three glacier forefields in central Svalbard, High Arctic. Distance from the glacier terminus was used as a proxy for soil age (years since deglaciation), and cyanobacterial abundance and community composition were evaluated by epifluorescence microscopy and pyrosequencing of partial 16S rRNA gene sequences, respectively. Succession was characterized by a decrease in phylotype richness and a marked shift in community structure, resulting in a clear separation between early (10-20 years since deglaciation), mid (30-50 years), and late (80-100 years) communities. Changes in cyanobacterial community structure were mainly connected with soil age and associated shifts in soil chemical composition (mainly moisture, SOC, SMN, K, and Na concentrations). Phylotypes associated with early communities were related either to potentially novel lineages (< 97.5% similar to sequences currently available in GenBank) or lineages predominantly restricted to polar and alpine biotopes, suggesting that the initial colonization of proglacial soil is accomplished by cyanobacteria transported from nearby glacial environments. Late communities, on the other hand, included more widely distributed genotypes, which appear to establish only after the microenvironment has been modified by the pioneering taxa.

Chloromonas svalbardensis n. sp. with Insights into the Phylogroup Chloromonadinia (Chlorophyceae).

The traditional green algal genus Chloromonas accommodates mesophilic, cold-tolerant and cold-adapted microorganisms. In this paper, we studied a new strain isolated from a wet hummock meadow in the High Arctic. We used morphological, ultrastructural and molecular data to assess the taxonomic position and phylogenetic relationships of the new isolate. The observed morphological features generally corresponded to the cold-tolerant Chloromonas characteristics. However, ellipsoidal or wide ellipsoidal vegetative cells, a massive parietal cup-shaped chloroplast with a number of continuously connected lobes, a thick cell wall, a prominent hemispherical papilla and the anterior position of an oblong or round eyespot distinguished the alga from all previously described Chloromonas species. Analyses of rbcL and 18S rRNA genes showed that the new strain formed an independent lineage within a clade containing mesophilic and psychrotolerant Chloromonas species. Comparisons of secondary structure models of a highly variable ITS2 rDNA marker supported a separate species identity of the new isolate. Considering the morphological and molecular differences from its relatives, a new psychrotolerant species, Chloromonas svalbardensis, is proposed. Further, our results demonstrated the paraphyletic origin of Chloromonas within Chloromonadinia with genetically, morphologically and ecologically well-defined clades. We discuss a scenario of a possible Chloromonas split and revision.

Cyanobacteria inhabiting biological soil crusts of a polar desert: Sør Rondane Mountains, Antarctica.

Molecular and morphological methods were applied to study cyanobacterial community composition in biological soil crusts (BSCs) from four areas (two nunataks and two ridges) in the Sør Rondane Mountains, Antarctica. The sampling sites serve as control areas for open top chambers (OTCs) that were put in place in 2010 at the time of sample collection and will be compared with BSC samples taken from the OTCs in the future. Cyanobacterial cell biovolume was estimated using epifluorescence microscopy, which revealed the dominance of filamentous cyanobacteria in all studied sites except the Utsteinen ridge, where unicellular cyanobacteria were the most abundant. Cyanobacterial diversity was studied by a combination of molecular fingerprinting methods based on the 16S rRNA gene (denaturing gradient gel electrophoresis (DGGE) and 454 pyrosequencing) using cyanobacteria-specific primers. The number of DGGE sequences obtained per site was variable and, therefore, a high-throughput method was subsequently employed to improve the diversity coverage. Consistent with previous surveys in Antarctica, both methods showed that filamentous cyanobacteria, such as Leptolyngbya sp., Phormidium sp. and Microcoleus sp., were dominant in the studied sites. In addition, the studied localities differed in substrate type, climatic conditions and soil parameters, which probably resulted in differences in cyanobacterial community composition. Furthermore, the BSC growing on gneiss pebbles had lower cyanobacterial abundances than BSCs associated with granitic substrates.

Chloromonas arctica sp. nov., a psychrotolerant alga from snow in the High Arctic (Chlamydomonadales, Chlorophyta).

With the advent of molecular phylogenetic methods, it has become possible to assess the bioversity of snow algae more accurately. In this study, we focused on a morphological, ultrastructural and taxonomic description of a new Chloromonas-like alga isolated from snow in the High Arctic (Svalbard). Light and transmission electron microscopy revealed broad ellipsoidal or ellipsoidal-cylindrical, occasionally spherical cells with a chloroplast without a pyrenoid, an inconspicuous eyespot and a papilla. The size difference and the aforementioned morphological traits clearly distinguished the alga from its closest counterparts within the genus Chloromonas. Moreover, we were able to cultivate the alga at both 5 and 20 °C, revealing the psychrotolerant nature of the strain. Phylogenetic analyses of the plastid rbcL and nuclear 18S rRNA gene showed that the alga is nested within a clade containing a number of psychrotolerant strains within the Chloromonadinia phylogroup (Chlorophyceae). In the rbcL phylogeny, the alga formed an independent lineage, sister to the freshwater species Chloromonas paraserbinowii. Comparisons of secondary structure models of a highly variable ITS2 rDNA marker showed support for a distinct species identity for the new strain. The ITS2 secondary structure of the new isolate differed from the closest matches 'Chlamydomonas' gerloffii and Choloromonas reticulata by three and five compensatory base changes, respectively. Considering the morphological and molecular differences from its closest relatives, a new psychrotolerant species from the Arctic, Choromonas arctica sp. nov., is proposed.

Identity, ecology and ecophysiology of planktic green algae dominating in ice-covered lakes on James Ross Island (northeastern Antarctic Peninsula).

The aim of this study was to assess the phylogenetic relationships, ecology and ecophysiological characteristics of the dominant planktic algae in ice-covered lakes on James Ross Island (northeastern Antarctic Peninsula). Phylogenetic analyses of 18S rDNA together with analysis of ITS2 rDNA secondary structure and cell morphology revealed that the two strains belong to one species of the genus Monoraphidium (Chlorophyta, Sphaeropleales, Selenastraceae) that should be described as new in future. Immotile green algae are thus apparently capable to become the dominant primary producer in the extreme environment of Antarctic lakes with extensive ice-cover. The strains grew in a wide temperature range, but the growth was inhibited at temperatures above 20 °C, indicating their adaptation to low temperature. Preferences for low irradiances reflected the light conditions in their original habitat. Together with relatively high growth rates (0.4-0.5 day-1) and unprecedently high content of polyunsaturated fatty acids (PUFA, more than 70% of total fatty acids), it makes these isolates interesting candidates for biotechnological applications.

Widespread green algae Chlorella and Stichococcus exhibit polar-temperate and tropical-temperate biogeography.

Chlorella and Stichococcus are morphologically simple airborne microalgae, omnipresent in terrestrial and aquatic habitats. The minute cell size and resistance against environmental stress facilitate their long-distance dispersal. However, the actual distribution of Chlorella- and Stichococcus-like species has so far been inferred only from ambiguous morphology-based evidence. Here we contribute a phylogenetic analysis of an expanded SSU and ITS2 rDNA sequence dataset representing Chlorella- and Stichococcus-like species from terrestrial habitats of polar, temperate and tropical regions. We aim to uncover biogeographical patterns at low taxonomic levels. We found that psychrotolerant strains of Chlorella and Stichococcus are closely related with strains originating from the temperate zone. Species closely related to Chlorella vulgaris and Muriella terrestris, and recovered from extreme terrestrial environments of polar regions and hot deserts, are particularly widespread. Stichococcus strains from the temperate zone, with their closest relatives in the tropics, differ from strains with the closest relatives being from the polar regions. Our data suggest that terrestrial Chlorella and Stichococcus might be capable of intercontinental dispersal; however, their actual distributions exhibit biogeographical patterns.

Bacterial and eukaryotic biodiversity patterns in terrestrial and aquatic habitats in the Sør Rondane Mountains, Dronning Maud Land, East Antarctica.

The bacterial and microeukaryotic biodiversity were studied using pyrosequencing analysis on a 454 GS FLX+ platform of partial SSU rRNA genes in terrestrial and aquatic habitats of the Sør Rondane Mountains, including soils, on mosses, endolithic communities, cryoconite holes and supraglacial and subglacial meltwater lenses. This inventory was complemented with Denaturing Gradient Gel Electrophoresis targeting Chlorophyta and Cyanobacteria. OTUs belonging to the Rotifera, Chlorophyta, Tardigrada, Ciliophora, Cercozoa, Fungi, Bryophyta, Bacillariophyta, Collembola and Nematoda were present with a relative abundance of at least 0.1% in the eukaryotic communities. Cyanobacteria, Proteobacteria, Bacteroidetes, Acidobacteria, FBP and Actinobacteria were the most abundant bacterial phyla. Multivariate analyses of the pyrosequencing data revealed a general lack of differentiation of both eukaryotes and prokaryotes according to habitat type. However, the bacterial community structure in the aquatic habitats was dominated by the filamentous cyanobacteria Leptolyngbya and appeared to be significantly different compared with those in dry soils, on mosses, and in endolithic habitats. A striking feature in all datasets was the detection of a relatively large amount of sequences new to science, which underscores the need for additional biodiversity assessments in Antarctic inland locations.

Annual Cycles of Two Cyanobacterial Mat Communities in Hydro-Terrestrial Habitats of the High Arctic.

Cyanobacteria form extensive macroscopic mats in shallow freshwater environments in the High Arctic and Antarctic. In these habitats, the communities are exposed to seasonal freezing and desiccation as well as to freeze-thawing and drying-rewetting cycles. Here, we characterized the annual cycles of two Phormidium communities in very shallow seepages located in central Svalbard. We observed the structure of the communities and the morphology, ultrastructure, metabolic activity, and viability of filaments and single cells. The communities overwintered as frozen mats, which were formed by long filaments enclosed in thick multilayered polysaccharide sheaths. No morphologically and/or ultrastructurally distinct spore-like cells were produced for surviving the winter, and the winter survival of the communities was not provided by a few resistant cells, which did not undergo visible morphological and ultrastructural transformations. Instead, a high proportion of cells in samples (85%) remained viable after prolonged freezing. The sheaths were the only morphological adaption, which seemed to protect the trichomes from damage due to freezing and freeze-associated dehydration. The cells in the overwintering communities were not dormant, as all viable cells rapidly resumed respiration after thawing, and their nucleoids were not condensed. During the whole vegetative season, defined by the presence of water in a liquid state, the communities were constantly metabolically active and contained <1% of dead and injured cells. The morphology and ultrastructure of the cells remained unaltered during observations throughout the year, except for light-induced changes in thylakoids. The dissemination events are likely to occur in spring as most of the trichomes were split into short fragments (hormogonia), a substantial proportion of which were released into the environment by gliding out of their sheaths, as well as by cracking and dissolving their sheaths. The short fragments subsequently grew longer and gradually produced new polysaccharide sheaths.

Diversity and dispersal capacities of a terrestrial algal genus Klebsormidium (Streptophyta) in polar regions.

The distribution of microbial eukaryotes (protists) has been frequently discussed during the last two decades. The ubiquity hypothesis assumes the lack of latitudinal gradients in protist diversity due to their unlimited global dispersal. In this study, we examined the diversity and distribution of the very common, globally distributed green algal genus Klebsormidium across climatic zones, focusing on the polar regions. We tested whether (i) there is comparable diversity among the polar and temperate regions, and (ii) whether a spatial genetic differentiation occurs at the global scale. We collected a total of 58 Arctic, Antarctic and temperate strains, and genetically characterized them by sequencing the rbcL gene and two highly variable chloroplast markers. Our analyses revealed the presence of two different distribution patterns which are supposed to characterize both macroorganisms and protists. On the one hand, we demonstrated unlimited dispersal and intensive gene flow within one of the inferred lineages (superclade B). On the other hand, the majority of Klebsormidium clades showed rather a limited distribution. In addition, we detected a significant decrease of species richness towards the poles i.e. the macroecological pattern typical for macroorganisms. Species within a single protist genus may thus exhibit highly contrasting distribution patterns, based on their dispersal capabilities, which are usually shaped by both intrinsic and extrinsic factors.