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

A terrestrial green alga was isolated at Iceland, and the strain (SAG 2627) was described for its morphology and phylogenetic position and tested for biotechnological capabilities. Cells had a distinctly curved, crescent shape with conical poles and a single parietal chloroplast. Phylogenetic analyses of 18S rDNA and rbcL markers placed the strain into the Trebouxiophyceae (Chlorophyta). The alga turned out to belong to an independent lineage without an obvious sister group within the Trebouxiophyceae. Based on morphological and phylogenetic data, the strain was described as a new genus and species, Thorsmoerkia curvula gen. et sp. nov. Biomass was generated in column reactors and subsequently screened for promising metabolites. Growth was optimized by pH-regulated, episodic CO2 supplement during the logarithmic growth-phase, and half of the biomass was thereafter exposed to nitrogen and phosphate depletion. The biomass yield reached up to 53.5 mg L-1 day-1. Fatty acid (FA) production peaked at 24 mg L-1 day-1 and up to 83% of all FAs were unsaturated. At the end of the log phase, approximately 45% of dry mass were lipids, including eicosapentaenoic acid. Carotenoid production reached up to 2.94 mg L-1 day-1 but it was halted during the stress phase. The N-linked glycans of glycoproteins were assessed to reveal chemotaxonomic patterns. The study demonstrated that new microalgae can be found at Iceland, potentially suitable for applied purposes. The advantage of T. curvula is its robustness and that significant amounts of lipids are already accumulated during log phase, making a subsequent stress exposure dispensable.

• Klíčová slova
• Biodiversity, Iceland, Polyunsaturated fatty acids, Semi-terrestrial algae, Taxonomy, Trebouxiophyceae,
• 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 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

Slowly melting snowfields in mountain and polar regions are habitats of snow algae. Orange blooms were sampled in three European mountain ranges. The cysts within the blooms morphologically resembled those of Chloromonas nivalis (Chlorophyceae). Molecular and morphological traits of field and cultured material showed that they represent a new species, Chloromonas hindakii sp. nov. The performance of photosystem II was evaluated by fluorometry. For the first time for a snow alga, cyst stages collected in a wide altitudinal gradient and the laboratory strain were compared. The results showed that cysts were well adapted to medium and high irradiance. Cysts from high light conditions became photoinhibited at three times higher irradiances (600 µmol photons m-2 s-1) than those from low light conditions, or likewise compared to cultured flagellates. Therefore, the physiologic light preferences reflected the conditions in the original habitat. A high content of polyunsaturated fatty acids (about 60% of total lipids) and the accumulation of the carotenoid astaxanthin was observed. They are regarded as adaptations to cope with extreme environmental conditions of snow that include low temperatures, freeze-thaw cycles, and variable light intensity. The intraspecific ability of adaptation of the photosynthetic apparatus to different irradiance regimes seems to be advantageous for thriving in different snow habitats.

• Klíčová slova
• astaxanthin, cryoflora, cysts, environmental sample, fatty acids, photosynthesis,
• Publikační typ
• časopisecké články MeSH

Melting snowfields in polar and alpine regions often exhibit a red and orange colouration caused by microalgae. The diversity of these organisms is still poorly understood. We applied a polyphasic approach using three molecular markers and light and electron microscopy to investigate spherical cysts sampled from alpine mountains in Europe, North America and South America as well as from both polar regions. Molecular analyses revealed the presence of a single independent lineage within the Chlamydomonadales. The genus Sanguina is described, with Sanguina nivaloides as its type. It is distinguishable from other red cysts forming alga by the number of cell wall layers, cell size, cell surface morphology and habitat preference. Sanguina nivaloides is a diverse species containing a total of 18 haplotypes according to nuclear ribosomal DNA internal transcribed spacer 2, with low nucleotide divergence (≤3.5%). Based on molecular data we demonstrate that it has a cosmopolitan distribution with an absence of geographical structuring, indicating an effective dispersal strategy with the cysts being transported all around the globe, including trans-equatorially. Additionally, Sanguina aurantia is described, with small spherical orange cysts often clustered by means of mucilaginous sheaths, and causing orange blooms in snow in subarctic and Arctic regions.

• Klíčová slova
• Chlamydomonas nivalis, haplotype network, red snow, snow algae,
• MeSH
• Chlorophyta klasifikace   genetika   fyziologie   MeSH
• ekosystém MeSH
• fylogeneze MeSH
• fylogeografie MeSH
• mezerníky ribozomální DNA MeSH
• Rhodophyta MeSH
• sníh mikrobiologie   MeSH
• zmrazování MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH
• Jižní Amerika MeSH
• Severní Amerika MeSH
• Názvy látek
• mezerníky ribozomální DNA MeSH

Long-lasting, slowly melting snowfields in mountainous regions are frequently populated by specialised microalgae whose diversity is still vastly underestimated. Cysts causing sub-surficial green snow were collected in the Austrian Alps, Tyrol, and morphologically accorded to the snow alga Scotiella cryophila sensu Chodat, initially described from Switzerland. The cytology and photobiology of this population were investigated to understand mechanisms of adaptation to the harsh habitat. Cysts of S. cryophila K-1 had secondary cell walls with pronounced rib-like surface structures and contained several small spherical plastids. The cytoplasm was dominated by lipid bodies, which developed reddish secondary pigmentation. Partial life cycle observations showed that daughter cells lacked structured cell walls. Cysts performed active photosynthesis at temperature conditions close to the freezing point and were photoinhibited at irradiances greater than 70 μmol m-2 s-1. This corresponded exactly to habitat conditions 20 to 40 cm below the snow surface. Phylogenetic analyses using 18S rDNA, rbcL and ITS2 rDNA sequences indicated that S. cryophila K-1 is related to Chloromonas, known to contain several snow algae. The taxon forms an independent lineage and is clearly genetically distinct from the type strain of Chloromonas rosae var. psychrophila from North America that is supposed to have morphologically identical cysts. For a taxonomic treatment including a species assignment of S. cryophila K-1 from Europe within Chloromonas, flagellates will have to be cultivated from cysts or from acquired field material for a detailed morphological description. Acquisition and genetic analysis of cysts that resemble S. cryophila from America could elucidate their relationship to European samples.

• Klíčová slova
• Cryoflora, Cryospheric algae, Cysts, Extremophiles, Fluorometry, Ultrastructure,
• Publikační typ
• časopisecké články MeSH