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

A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.

• Klíčová slova
• 18S rDNA, Algae phylogeny, Electron microscopy, Fatty acids, ITS, Strain FGS-001,
• MeSH
• biologické pigmenty analýza   MeSH
• biotechnologie MeSH
• Chlorophyta klasifikace   cytologie   genetika   MeSH
• druhová specificita MeSH
• feofytiny analýza   MeSH
• fylogeneze * MeSH
• karotenoidy analýza   MeSH
• kyselina alfa-linolenová analýza   MeSH
• mastné kyseliny analýza   MeSH
• mikrořasy klasifikace   cytologie   genetika   izolace a purifikace   MeSH
• ribozomální DNA MeSH
• RNA ribozomální 18S genetika   MeSH
• xanthofyly MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Norsko MeSH
• Názvy látek
• astaxanthine MeSH Prohlížeč
• biologické pigmenty MeSH
• feofytiny MeSH
• karotenoidy MeSH
• kyselina alfa-linolenová MeSH
• mastné kyseliny MeSH
• neoxanthin MeSH Prohlížeč
• ribozomální DNA MeSH
• RNA ribozomální 18S MeSH
• violaxanthin MeSH Prohlížeč
• xanthofyly 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

Melting snow fields are an extremophilic habitat dominated by closely related Chlamydomonadaceae (Chlorophyta). Microscopy-based classification of these cryophilic microalgae is challenging and may not reveal the true diversity. High-throughput sequencing (HTS) allows for a more comprehensive evaluation of the community. However, HTS approaches have been rarely used in such ecosystems and the output of their application has not been evaluated. Furthermore, there is no consensus on the choice for a suitable DNA marker or data processing workflow. We found that the correct placement of taxonomic strings onto OTUs strongly depends on the quality of the reference databases. We improved the assignments of the HST data by generating additional reference sequences of the locally abundant taxa, guided by light microscopy. Furthermore, a manual inspection of all automated OTU assignments, oligotyping of the most abundant 18S OTUs, as well as ITS2 secondary structure analyses were necessary for accurate species assignments. Moreover, the sole use of one marker can cause misleading results, either because of insufficient variability within the locus (18S) or the scarcity of reference sequences (ITS2). Our evaluation reveals that HTS output needs to be thoroughly checked when the studied habitats or organisms are poorly represented in publicly available databases. We recommend an optimized workflow for an improved biodiversity evaluation of not only snow algal communities, but generally 'exotic' ecosystems where similar problems arise. A consistent sampling strategy, two- molecular marker approach, light microscopy-based guidance, generation of appropriate reference sequences and final manual verification of all taxonomic assignments are highly recommended.

• Klíčová slova
• 18S rDNA, ITS2 rDNA, Illumina, OTU clustering, Sanger, high-throughput sequencing, oligotyping, red snow, secondary structure, snow algae,
• Publikační typ
• časopisecké články 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