Psychrophilic pucciniomycotinous yeasts inhabit snowfields and glacial ecosystems worldwide, yet their ecological role remains unclear. We isolated a clonal strain of Phenoliferia psychrophenolica (LCC-F-001-001) from an alpine red snowfield. Its 42-Mbp genome contains 11,523 genes, including 37 ice-binding protein genes, the highest number recorded in fungi, mainly acquired through horizontal transfers. This yeast tolerates freezing, grows optimally at 10 °C and forms pseudohyphae above 15 °C. Known to assimilate phenol and small metabolites, we found LCC-F-001-001 also hydrolyzes carotenoid and phenolic pigments from snow and glacier ice algae. LCC-F-001-001 genome encodes ~ 500 carbohydrate-active enzymes, ranking among the most catalytically versatile sequenced Microbotryomycetes, alongside a related permafrost species. P. psychrophenolica and related fungi are therefore key players in snow and ice ecosystems, capable of degrading organic molecules in snowfields and glaciers, likely during algal bloom declines, and connecting the carbon cycle between cryospheric environments and soils through their exceptional metabolic adaptability.

• MeSH
• Basidiomycota * genetika   metabolismus   fyziologie   MeSH
• ekosystém * MeSH
• fylogeneze MeSH
• genom fungální MeSH
• led MeSH
• ledový příkrov * mikrobiologie   MeSH
• sníh * mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• led MeSH

Melting snow and glacier surfaces host microalgal blooms in polar and mountainous regions. The aim of this study was to determine the dominant taxa at the species level in the European Arctic and the Alps. A standardized protocol for amplicon metabarcoding using the 18S rRNA gene and ITS2 markers was developed. This is important because previous biodiversity studies have been hampered by the dominance of closely related algal taxa in snow and ice. Due to the limited resolution of partial 18S rRNA Illumina sequences, the hypervariable ITS2 region was used to further discriminate between the genotypes. Our results show that red snow was caused by the cosmopolitan Sanguina nivaloides (Chlamydomonadales, Chlorophyta) and two as of yet undescribed Sanguina species. Arctic orange snow was dominated by S. aurantia, which was not found in the Alps. On glaciers, at least three Ancylonema species (Zygnematales, Streptophyta) dominated. Golden-brown blooms consisted of Hydrurus spp. (Hydrurales, Stramenophiles) and these were mainly an Arctic phenomenon. For chrysophytes, only the 18S rRNA gene but not ITS2 sequences were amplified, showcasing how delicate the selection of eukaryotic 'universal' primers for community studies is and that primer specificity will affect diversity results dramatically. We propose our approach as a 'best practice'.

• Klíčová slova
• ITS2 secondary structure, cryosphere, eDNA, glacier ice algae, next generation sequencing, snow algae, species delimitation,
• MeSH
• Chlorophyceae * genetika   MeSH
• Chlorophyta * genetika   MeSH
• geny rRNA MeSH
• ledový příkrov MeSH
• RNA ribozomální 18S genetika   MeSH
• sníh MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• RNA ribozomální 18S MeSH

In this study, we used microscopic, spectroscopic, and molecular analysis to characterize endolithic colonization in gypsum (selenites and white crystalline gypsum) from several sites in Sicily. Our results showed that the dominant microorganisms in these environments are cyanobacteria, including: Chroococcidiopsis sp., Gloeocapsopsis pleurocapsoides, Gloeocapsa compacta, and Nostoc sp., as well as orange pigmented green microalgae from the Stephanospherinia clade. Single cell and filament sequencing coupled with 16S rRNA amplicon metagenomic profiling provided new insights into the phylogenetic and taxonomic diversity of the endolithic cyanobacteria. These organisms form differently pigmented zones within the gypsum. Our metagenomic profiling also showed differences in the taxonomic composition of endoliths in different gypsum varieties. Raman spectroscopy revealed that carotenoids were the most common pigments present in the samples. Other pigments such as gloeocapsin and scytonemin were also detected in the near-surface areas, suggesting that they play a significant role in the biology of endoliths in this environment. These pigments can be used as biomarkers for basic taxonomic identification, especially in case of cyanobacteria. The findings of this study provide new insights into the diversity and distribution of phototrophic microorganisms and their pigments in gypsum in Southern Sicily. Furthemore, this study highlights the complex nature of endolithic ecosystems and the effects of gypsum varieties on these communities, providing additional information on the general bioreceptivity of these environments.

• Klíčová slova
• cyanobacteria, endoliths, green algae, gypsum, metagenomics, phototrophs,
• Publikační typ
• časopisecké články MeSH

Snow algae blooms often form green or red coloured patches in melting alpine and polar snowfields worldwide, yet little is known about their biology, biogeography, and species diversity. We investigated eight isolates collected from red snow in northern Norway, using a combination of morphology, 18S rRNA gene and internal transcribed spacer 2 (ITS2) genetic markers. Phylogenetic and ITS2 rRNA secondary structure analyses assigned six isolates to the species Raphidonema nivale, Deuterostichococcus epilithicus, Chloromonas reticulata, and Xanthonema bristolianum. Two novel isolates belonging to the family Stichococcaceae (ARK-S05-19) and the genus Chloromonas (ARK-S08-19) were identified as potentially new species. In laboratory cultivation, differences in the growth rate and fatty acid profiles were observed between the strains. Chlorophyta were characterized by abundant C18:3n-3 fatty-acids with increases in C18:1n-9 in the stationary phase, whilst Xanthonema (Ochrophyta) was characterized by a large proportion of C20:5n-3, with increases in C16:1n-7 in the stationary phase. In a further experiment, lipid droplet formation was studied in C. reticulata at the single-cell level using imaging flow cytometry. Our study establishes new cultures of snow algae, reveals novel data on their biodiversity and biogeography, and provides an initial characterization of physiological traits that shape natural communities and their ecophysiological properties.

• Klíčová slova
• 18S rRNA, ITS2 rRNA, fatty acids, imaging flow cytometry, microalgae, phylogeny,
• MeSH
• Chlorophyceae * MeSH
• Chlorophyta * genetika   MeSH
• fylogeneze MeSH
• lipidy MeSH
• mikrobiota * genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Norsko MeSH
• Názvy látek
• lipidy MeSH

Seasonally slowly melting mountain snowfields are populated by extremophilic microalgae. In alpine habitats, high-light sensitive, green phytoflagellates are usually observed in subsurface layers deeper in the snowpack under dim conditions, while robust orange to reddish cyst stages can be seen exposed on the surface. In this study, uncommon surface green snow was investigated in the High Tatra Mountains (Slovakia). The monospecific community found in the green surface bloom consisted of vegetative Chloromonas cells (Volvocales, Chlorophyta). Molecular data demonstrated that the field sample and the strain isolated and established from the bloom were conspecific, and they represent a new species, Chloromonas kaweckae sp. nov., which is described based on the morphology of the vegetative cells and asexual reproduction and on molecular analyses of the strain. Cells of C. kaweckae accumulated approximately 50% polyunsaturated fatty acids, which is advantageous at low temperatures. In addition, this new species performed active photosynthesis at temperatures close to the freezing point showed a light compensation point of 126 ± 22 μmol photons · m-2 · s-1 and some signs of photoinhibition at irradiances greater than 600 μmol photons · m-2 · s-1 . These data indicate that the photosynthetic apparatus of C. kaweckae could be regarded as adapted to relatively high light intensities, otherwise unusual for most flagellate stages of snow algae.

• Klíčová slova
• biodiversity, cryoflora, environmental sample, fatty acids, fluorometry, vegetative stages,
• MeSH
• Chlorophyceae * MeSH
• Chlorophyta * fyziologie   MeSH
• fotosyntéza fyziologie   MeSH
• nízká teplota MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Slovenská republika MeSH

UNLABELLED: Red snow caused by spherical cysts can be found worldwide, while an orange snow phenomenon caused by spherical cells is restricted to (Sub-)Arctic climates. Both bloom types, occurring in the same localities at Svalbard, were compared ecophysiologically. Using a combination of molecular markers and light- and transmission electron microscopy, cells were identified as Sanguina nivaloides and Sanguina aurantia (Chlorophyceae). In search for reasons for a cosmopolitan vs. a more restricted distribution of these microbes, significant differences in fatty acid and pigment profiles of field samples were found. S. aurantia accumulated much lower levels of polyunsaturated fatty acids (21% vs. 48% of total fatty acids) and exhibited lower astaxanthin-to-chlorophyll-a ratio (2-8 vs. 12-18). These compounds play an important role in adaptation to extreme conditions at the snow surface and within snow drifts. Accordingly, the performance of photosystem II showed that one third to nearly half of the photosynthetic active irradiation was sufficient in S. aurantia, compared to S. nivaloides, to become light saturated. Furthermore, formation of plastoglobules observed in S. nivaloides but missing in S. aurantia may contribute to photoprotection. The rapid light curves of the two species show to a certain extent the shade-adapted photosynthesis under the light conditions at Svalbard (high α-value 0.16 vs. 0.11, low saturation point I k 59 vs. 86). These results indicate significant physiological and ultrastructural differences of the two genetically closely related cryoflora species, but the reasons why S. aurantia has not been found at conditions outside (Sub-)Arctic climate types remain unknown. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s00300-020-02778-0) contains supplementary material, which is available to authorised users.

• Klíčová slova
• Arctic, Astaxanthin, Chlamydomonas nivalis, Cryoflora, Green algae, Polyunsaturated fatty acid,
• Publikační typ
• časopisecké články MeSH

Snow algae are photosynthetic microbes that inhabit the melting snow surface in alpine and polar regions. We analyzed the pigment and species composition of colored snow collected on Mt. Tateyama in Japan during the melting seasons of 2015 and 2016. High-performance liquid chromatographic analyses of the pigments extracted from the colored snow showed that their composition varied within the study area and were classified into four types: Type A (astaxanthin-monoester dominant), Type B (medium astaxanthin-monoester content), Type C (abundant primary carotenoids and free-astaxanthin), and Type D (abundant primary carotenoids and astaxanthin diesters). Types A and B were most commonly observed in the study area, whereas Types C and D appeared only at specific sites. Analysis of the 18S ribosomal RNA (18S rRNA) gene revealed six major amplicon sequence variants (ASVs) of snow algae, belonging to the Sanguina, Chloromonas, and Chlainomonas groups. The relative abundance of the algal ASVs showed that Sanguina was dominant (>48%) in both Types A and B, suggesting that the difference in astaxanthin abundance between the two types was caused by the production of pigments in the algal cells. The algal community structures of Types C and D differed from those of Types A and B, indicating that the primary carotenoids and astaxanthin diesters were derived from certain algal species in these types. Therefore, astaxanthin-rich Sanguina algae mostly induced the red snow that appeared widely in this alpine area; however, they were partially dominated by Chloromonas or Chlainomonas algae, causing different pigment compositions.

• Klíčová slova
• 18S rRNA, Chloromonas, Sanguina, astaxanthin, phylogenetic analysis, pigment composition, red snow, snow algae,
• Publikační typ
• časopisecké články MeSH

Melting polar and alpine ice surfaces frequently exhibit blooms of dark pigmented algae. These microbial extremophiles significantly reduce the surface albedo of glaciers, thus accelerating melt rates. However, the ecology, physiology and taxonomy of cryoflora are not yet fully understood. Here, a Swiss and an Austrian glacier dominated either by filamentous Ancylonema nordenskioeldii or unicellular Mesotaenium berggrenii var. alaskanum, were sampled. Molecular analysis showed that both species are closely related, sharing identical chloroplast morphologies (parietal-lobed for Ancylonema vs. axial plate-like for Mesotaenium sensu stricto), thus the unicellular species was renamed Ancylonema alaskana. Moreover, an ecophysiological comparison of the two species was performed: pulse-amplitude modulated (PAM) fluorometry confirmed that they have a high tolerance to elevated solar irradiation, the physiological light preferences reflected the conditions in the original habitat; nonetheless, A. nordenskioeldii was adapted to higher irradiances while the photosystems of A. alaskana were able to use efficiently low irradiances. Additionally, the main vacuolar polyphenol, which effectively shields the photosystems, was identical in both species. Also, about half of the cellular fatty acids were polyunsaturated, and the lipidome profiles dominated by triacylglycerols were very similar. The results indicate that A. alaskana is physiologically very similar and closely related but genetically distinct to A. nordenskioeldii.

• Klíčová slova
• Mesotaeniaceae, cryoflora, fatty acids, lipidome, photosynthesis, phylogeny, polyphenols, supraglacial communities,
• Publikační typ
• časopisecké články MeSH

Melting mountainous snowfields are populated by extremophilic microorganisms. An alga causing orange snow above timberline in the High Tatra Mountains (Poland) was characterised using multiple methods examining its ultrastructure, genetics, life cycle, photosynthesis and ecophysiology. Based on light and electron microscopy and ITS2 rDNA, the species was identified as Chloromonas krienitzii (Chlorophyceae). Recently, the taxon was described from Japan. However, cellular adaptations to its harsh environment and details about the life cycle were so far unknown. In this study, the snow surface population consisted of egg-shaped cysts containing large numbers of lipid bodies filled presumably with the secondary carotenoid astaxanthin. The outer, spiked cell wall was shed during cell maturation. Before this developmental step, the cysts resembled a different snow alga, Chloromonas brevispina. The remaining, long-lasting smooth cell wall showed a striking UV-induced blue autofluorescence, indicating the presence of short wavelengths absorbing, protective compounds, potentially sporopollenin containing polyphenolic components. Applying a chlorophyll fluorescence assay on intact cells, a significant UV-A and UV-B screening capability of about 30 and 50%, respectively, was measured. Moreover, intracellular secondary carotenoids were responsible for a reduction of blue-green light absorbed by chloroplasts by about 50%. These results revealed the high capacity of cysts to reduce the impact of harmful UV and high visible irradiation to the chloroplast and nucleus when exposed at alpine snow surfaces during melting. Consistently, the observed photosynthetic performance of photosystem II (evaluated by fluorometry) showed no decline up to 2100 μmol photons m-2 s-1. Cysts accumulated high contents of polyunsaturated fatty acids (about 60% of fatty acids), which are advantageous at low temperatures. In the course of this study, C. krienitzii was found also in Slovakia, Italy, Greece and the United States, indicating a widespread distribution in the Northern Hemisphere.

• Klíčová slova
• UV-A radiation, UV-B radiation, astaxanthin, chlorophyll fluorescence, cysts, photosynthesis, polyunsaturated fatty acids, snow algae,
• Publikační typ
• časopisecké články MeSH

Melting summer snow in the Austrian Alps exhibited a yellowish bloom that was mainly comprised of an unidentified unicellular chrysophyte. Molecular data (18S rRNA and rbcL genes) showed a close relationship to published sequences from an American pond alga formerly identified as Kremastochrysis sp. The genera Kremastochrysis and Kremastochrysopsis are morphologically distinguished by the number of flagella observed with the light microscope, and therefore we assigned the Austrian snow alga and an American pond alga to the genus Kremastochrysopsis. Transmission and scanning electron microscopy revealed that swimming cells had two flagella oriented in opposite directions, typical for the Hibberdiales. Molecular phylogenetic analyses showed that both new species were closely related to Hibberdia. Kremastochrysopsis ocellata, the type species and only known species, has two chloroplasts per cell and the zoospores have red eyespots. Our two organisms had only a single chloroplast and no zoospore eyespot, but their gene sequences differed substantially. Therefore, we described two new species, Kremastochrysopsis austriaca sp. nov and Kremstochrysopsis americana sp. nov. When grown in culture, both taxa showed a characteristic hyponeustonic growth (hanging below the water surface), whereas older immotile cells grew at the bottom of the culture vessel. Ecologically, Kremastochrysopsis austriaca sp. nov., which caused snow discolorations, had no close phylogenetic relationships to other psychrophilic chrysophytes, for example, Chromulina chionophilia, Hydrurus sp., and Ochromonas-like flagellates.

• Klíčová slova
• Chromophyton, Kremastochrysis, psychrophilic, snow algae, substitutional saturation,
• MeSH
• chloroplasty * MeSH
• Chrysophyceae * MeSH
• fylogeneze MeSH
• RNA ribozomální 16S MeSH
• RNA ribozomální 18S MeSH
• sekvenční analýza DNA MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Rakousko MeSH
• Názvy látek
• RNA ribozomální 16S MeSH
• RNA ribozomální 18S MeSH