Novel insights in cryptic diversity of snow and glacier ice algae communities combining 18S rRNA gene and ITS2 amplicon sequencing.

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'.

The snow alga Chloromonas kaweckae sp. nov. (Volvocales, Chlorophyta) causes green surface blooms in the high tatras (Slovakia) and tolerates high irradiance.

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.

A DUF3494 ice-binding protein with a root cap domain in a streptophyte glacier ice alga.

Ice-binding proteins (IBPs) of the DUF3494 type have been found in many ice-associated unicellular photoautotrophs, including chlorophytes, haptophytes, diatoms and a cyanobacterium. Unrelated IBPs have been found in many land plants (streptophytes). Here we looked for IBPs in two streptophyte algae that grow only on glaciers, a group in which IBPs have not previously been examined. The two species, Ancylonema nordenskioeldii and Ancylonema. alaskanum, belong to the class Zygnematophyceae, whose members are the closest relatives to all land plants. We found that one of them, A. nordenskioeldii, expresses a DUF3494-type IBP that is similar to those of their chlorophyte ancestors and that has not previously been found in any streptophytes. The protein is unusual in having what appears to be a perfect array of TXT motifs that have been implicated in water or ice binding. The IBP strongly binds to ice and almost certainly has a role in mitigating the daily freeze-thaw cycles that the alga is exposed to during late summer. No IBP was found in the second species, A. alaskanum, which may rely more on glycerol production for its freeze-thaw tolerance. The IBP is also unusual in having a 280-residue domain with a ß sandwich structure (which we designate as the DPH domain) that is characteristic of root cap proteins of land plants, and that may have a role in forming IBP oligomers. We also examined existing transcriptome data obtained from land plants to better understand the tissue and temperature dependence of expression of this domain.

Coelastrella terrestris for Adonixanthin Production: Physiological Characterization and Evaluation of Secondary Carotenoid Productivity.

A novel strain of Coelastrella terrestris (Chlorophyta) was collected from red mucilage in a glacier foreland in Iceland. Its morphology showed characteristic single, ellipsoidal cells with apical wart-like wall thickenings. Physiological characterization revealed the presence of the rare keto-carotenoid adonixanthin, as well as high levels of unsaturated fatty acids of up to 85%. Initial screening experiments with different carbon sources for accelerated mixotrophic biomass growth were done. Consequently, a scale up to 1.25 L stirred photobioreactor cultivations yielded a maximum of 1.96 mg·L-1 adonixanthin in free and esterified forms. It could be shown that supplementing acetate to the medium increased the volumetric productivity after entering the nitrogen limitation phase compared to autotrophic control cultures. This study describes a promising way of biotechnological adonixanthin production using Coelastrella terrestris.

Investigating the Growth of Algae Under Low Atmospheric Pressures for Potential Food and Oxygen Production on Mars.

With long-term missions to Mars and beyond that would not allow resupply, a self-sustaining Bioregenerative Life Support System (BLSS) is essential. Algae are promising candidates for BLSS due to their completely edible biomass, fast growth rates and ease of handling. Extremophilic algae such as snow algae and halophilic algae may also be especially suited for a BLSS because of their ability to grow under extreme conditions. However, as indicated from over 50 prior space studies examining algal growth, little is known about the growth of algae at close to Mars-relevant pressures. Here, we explored the potential for five algae species to produce oxygen and food under low-pressure conditions relevant to Mars. These included Chloromonas brevispina, Kremastochrysopsis austriaca, Dunaliella salina, Chlorella vulgaris, and Spirulina plantensis. The cultures were grown in duplicate in a low-pressure growth chamber at 670 ± 20 mbar, 330 ± 20 mbar, 160 ± 20 mbar, and 80 ± 2.5 mbar pressures under continuous light exposure (62-70 µmol m-2 s-1). The atmosphere was evacuated and purged with CO2 after sampling each week. Growth experiments showed that D. salina, C. brevispina, and C. vulgaris were the best candidates to be used for BLSS at low pressure. The highest carrying capacities for each species under low pressure conditions were achieved by D. salina at 160 mbar (30.0 ± 4.6 × 105 cells/ml), followed by C. brevispina at 330 mbar (19.8 ± 0.9 × 105 cells/ml) and C. vulgaris at 160 mbar (13.0 ± 1.5 × 105 cells/ml). C. brevispina, D. salina, and C. vulgaris all also displayed substantial growth at the lowest tested pressure of 80 mbar reaching concentrations of 43.4 ± 2.5 × 104, 15.8 ± 1.3 × 104, and 57.1 ± 4.5 × 104 cells per ml, respectively. These results indicate that these species are promising candidates for the development of a Mars-based BLSS using low pressure (∼200-300 mbar) greenhouses and inflatable structures that have already been conceptualized and designed.

Unicellular versus Filamentous: The Glacial Alga Ancylonema alaskana comb. et stat. nov. and Its Ecophysiological Relatedness to Ancylonema nordenskioeldii (Zygnematophyceae, Streptophyta).

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.

Ecophysiological and ultrastructural characterisation of the circumpolar orange snow alga Sanguina aurantia compared to the cosmopolitan red snow alga Sanguina nivaloides (Chlorophyta).

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.

Tetraedron minimum, First Reported Member of Hydrodictyaceae to Accumulate Secondary Carotenoids.

We isolated a novel strain of the microalga Tetraedron minimum in Iceland from a terrestrial habitat. During long-term cultivation, a dish culture turned orange, indicating the presence of secondary pigments. Thus, we characterized T. minimum for growth and possible carotenoid production in different inorganic media. In a lab-scale photobioreactor, we confirmed that nitrogen starvation in combination with salt stress triggered a secondary carotenoid accumulation. The development of the pigment composition and the antioxidant capacity of the extracts was analyzed throughout the cultivations. The final secondary carotenoid composition was, on average, 61.1% astaxanthin and 38.9% adonixanthin. Moreover, the cells accumulated approx. 83.1% unsaturated fatty acids. This work presents the first report of the formation of secondary carotenoids within the family Hydrodictyaceae (Sphaeropleales, Chlorophyta).

Thorsmoerkia curvula gen. et spec. nov. (Trebouxiophyceae, Chlorophyta), a semi-terrestrial microalga from Iceland exhibits high levels of unsaturated fatty acids.

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.

Growth, fatty, and amino acid profiles of the soil alga Vischeria sp. E71.10 (Eustigmatophyceae) under different cultivation conditions.

In this study, a unicellular soil alga isolated from farmland in Germany was surveyed. The investigation of the hypervariable molecular markers ITS1 rDNA and ITS2 rDNA identified strain E71.10 as conspecific with Vischeria sp. SAG 51.91 (Eustigmatophyceae). The culture was tested for biomass generation and for the yield of fatty acids and amino acids. The survey included four different culture conditions (conventional, elevated CO2, nitrogen depletion, or sodium chloride stress) at room temperature. The best yield of dry biomass was achieved applying 1% CO2, whereas nitrogen-free medium resulted into least growth. The fatty acid content peaked in nitrogen-free medium at 59% per dry mass. Eicosapentaenoic acid was the most abundant fatty acid in all treatments (except for nitrogen free), accounting for 10.44 to 16.72 g/100 g dry mass. The highest content of amino acids (20%) was achieved under conventional conditions. The results show that abiotic factors strongly influence to which extent metabolites are intracellularly stored and they confirm also for this yet undescribed strain of Vischeria that Eustigmatophyceae are promising candidates for biotechnology.

Cysts of the Snow Alga Chloromonas krienitzii (Chlorophyceae) Show Increased Tolerance to Ultraviolet Radiation and Elevated Visible Light.

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.

Snow and Glacial Algae: A Review1.

Snow or glacial algae are found on all continents, and most species are in the Chlamydomonadales (Chlorophyta) and Zygnematales (Streptophyta). Other algal groups include euglenoids, cryptomonads, chrysophytes, dinoflagellates, and cyanobacteria. They may live under extreme conditions of temperatures near 0°C, high irradiance levels in open exposures, low irradiance levels under tree canopies or deep in snow, acidic pH, low conductivity, and desiccation after snow melt. These primary producers may color snow green, golden-brown, red, pink, orange, or purple-grey, and they are part of communities that include other eukaryotes, bacteria, archaea, viruses, and fungi. They are an important component of the global biosphere and carbon and water cycles. Life cycles in the Chlamydomonas-Chloromonas-Chlainomonas complex include migration of flagellates in liquid water and formation of resistant cysts, many of which were identified previously as other algae. Species differentiation has been updated through the use of metagenomics, lipidomics, high-throughput sequencing (HTS), multi-gene analysis, and ITS. Secondary metabolites (astaxanthin in snow algae and purpurogallin in glacial algae) protect chloroplasts and nuclei from damaging PAR and UV, and ice binding proteins (IBPs) and polyunsaturated fatty acids (PUFAs) reduce cell damage in subfreezing temperatures. Molecular phylogenies reveal that snow algae in the Chlamydomonas-Chloromonas complex have invaded the snow habitat at least twice, and some species are polyphyletic. Snow and glacial algae reduce albedo, accelerate the melt of snowpacks and glaciers, and are used to monitor climate change. Selected strains of these algae have potential for producing food or fuel products.

Two New Kremastochrysopsis species, K. austriaca sp. nov. and K. americana sp. nov. (Chrysophyceae)1.

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.

Ice-Binding Proteins in a Chrysophycean Snow Alga: Acquisition of an Essential Gene by Horizontal Gene Transfer.

All ice-associated algae examined so far have genes for ice-binding proteins (IBPs), which suggest that these proteins are essential for survival in icy habitats. The most common type of IBP, type 1 IBPs (also referred to as DUF3494 IBPs), is also found in ice-associated bacteria and fungi. Previous studies have suggested that algal IBP genes were acquired by horizontal transfer from other microorganisms (probably bacteria). However, it remains unclear whether this is also the case for algae distantly related to the ones examined so far and whether microorganisms other than bacteria could be the donors. Furthermore, there is only limited evidence that these proteins are expressed at low temperature. Here, we show that Kremastochrysopsis austriaca (Chrysophyceae), an Austrian snow alga that is not closely related to any of the ice-associated algae examined so far, also produces IBPs, although their activity was weak. Sequencing the algal genome and the transcriptomes of cells grown at 1 and 15°C revealed three isoforms of a type 1 IBP. In agreement with their putative function, the three isoforms were strongly upregulated by one to two orders of magnitude at 1°C compared to 15°C. In a phylogenetic tree, the K. austriaca IBPs were distant from other algal IBPs, with the closest matches being bacterial proteins. These results suggest that the K. austriaca IBPs were derived from a gene that was acquired from a bacterium unrelated to other IBP donor bacteria and confirm by their presence in yet another alga the essential role of algal IBPs.

Ecophysiology of Chloromonas hindakii sp. nov. (Chlorophyceae), Causing Orange Snow Blooms at Different Light Conditions.

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.

Evaluating High-Throughput Sequencing Data of Microalgae Living in Melting Snow: Improvements and Limitations1.

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.

Ecophysiological and morphological comparison of two populations of Chlainomonas sp. (Chlorophyta) causing red snow on ice-covered lakes in the High Tatras and Austrian Alps.

Based on analyses of multiple molecular markers (18S rDNA, ITS1, ITS2 rDNA, rbcL), an alga that causes red snow on the melting ice cover of a high-alpine lake in the High Tatras (Slovakia) was shown to be identical with Chlainomonas sp. growing in a similar habitat in the Tyrolean Alps (Austria). Both populations consisted mostly of smooth-walled quadriflagellates. They occurred in slush, and shared similar photosynthetic performances (photoinhibition above 1300 µmol photons m-2 s-1), very high levels of polyunsaturated fatty acids (PUFA, 64% and 74% respectively) and abundant astaxanthin accumulation, comparable to the red spores of Chlamydomonas nivalis (Bauer) Wille. Physiological differences between the Slovak and Austrian populations included higher levels of α-tocopherol and a 13Z-isomer of astaxanthin in the former. High accumulation of secondary pigments in the Slovak population probably reflected harsher environmental conditions, since the collection was made later in the growing season when cells were exposed to higher irradiance at the surface. Using a polyphasic approach, we compared Chlainomonas sp. with Chlamydomonas nivalis. The latter causes 'conventional' red snow, and shows high photophysiological plasticity, with high efficiency under low irradiance and no photoinhibition up to 2000 µmol photons m-2 s-1. Its PUFA content was significantly lower (50%). An annual cycle of lake-to-snow colonization by Chlainomonas sp. from slush layers deeper in the ice cover is proposed. Our results point to an ecologically highly specialized cryoflora species, whose global distribution is likely to be more widespread than previously assumed.

Chloromonas nivalis subsp. tatrae, subsp. nov. (Chlamydomonadales, Chlorophyta): re-examination of a snow alga from the High Tatra Mountains (Slovakia).

Melting snow fields populated by aplanozygotes of the genus Chloromonas (Chlamydomonadales, Chlorophyta) are found in polar and alpine habitats. In the High Tatra Mountains (Slovakia), cells causing blooms of brownish-red snow designated as Scotiella tatrae kol turned out to be genetically (18S, ITS1 and ITS2 rDNA, rbcL) very closely related to Chloromonas nivalis (Chodat) Hoham et Mullet from the Austrian Alps. Therefore, Sc. tatrae is transferred into the latter taxon and reduced to a subspecies as Cr. nivalis subsp. tatrae. Both exhibit a similar photosynthetic performance, thrive in similar habitats at open sites above timberline, but differ in astaxanthin accumulation and number of aplanozygote cell wall flanges. In a field sample of Cr. nivalis subsp. tatrae, polyunsaturated fatty acids formed nearly 50 % of total lipids, dominating in phospholipids and glycolipids. Cr. nivalis subsp. tatrae represents likely a variation of a common cryoflora species with distinct morphology.

Ecology, cytology and phylogeny of the snow alga Scotiella cryophila K-1 (Chlamydomonadales, Chlorophyta) from the Austrian Alps.

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.

Distribution and UV protection strategies of zooplankton in clear and glacier-fed alpine lakes.

Zooplankton, a group of aquatic animals important as trophic link in the food web, are exposed to high levels of UV radiation (UVR) in clear alpine lakes, while in turbid glacier-fed lakes they are more protected. To study the interplay between behavioral and physiological protection responses in zooplankton from those lakes, we sampled six lakes of different UVR transparency and glacial turbidity. Copepods were absent in the upper water layers of the clearest lake, while in glacier-fed lakes they were more evenly distributed in the water column. Across all lakes, the weighted copepod mean depth was strongly related to food resources (chlorophyll a and rotifers), whereas in the fishless lakes, glacial turbidity largely explained the vertical daytime distribution of these organisms. Up to ~11-times (mean 3.5) higher concentrations of photo-protective compounds (mycosporine-like amino acids, MAAs) were found in the copepods from the clear than from the glacier-fed lakes. In contrast to carotenoid concentrations and antioxidant capacities, MAA levels were strongly related to the lake transparency. Copepods from alpine lakes rely on a combination of behavioral and physiological strategies adapted to the change in environmental conditions taking place when lakes shift from glacially turbid to clear conditions, as glacier retreat proceeds.