Vertical distribution of the snow alga Chlamydomonas nivalis (Chlorophyta,Volvocales)

Summary Chlamydomonas nivalis commonly forms large blooms, visible as a red coloration, in the snow during summer. Fewer algae are seen in the top layer of snow during days of intense sunlight than on cloudy days. The present experiments were done to investigate this change of vertical distribution of algae. Apparently the algae are able to associate with the watersurface surrounding snow crystals. Due to this association they avoid being washed away by the water from melting snow. Intense sunlight, however, decreases the degree to which the algae associate with the water-surface, and thereby increasing the number of algae being removed from the top layer of snow by the melting water. If the weather becomes cloudy again, the algae do not move upwards, but stay attached to the water-surfaces. Thus when the snow above melts, they will reappear in the top layer.     

EVIDENCE A PHOTOPROTECTIVE FOR SECONDARY CAROTENOIDS OF SNOW ALGAE1

Snow algae occupy a unique habitat in high altitude and polar environments. These algae are often subject to extremes in nutrient availability, acidity, solar irradiance, desiccation, and ambient temperature. This report documents the accumulation of secondary carotenoids by snow algae in response to the availability of nitrogenous nutrients. Unusually large accumulations of astaxanthin esters in extra-chloroplastic lipid globules produce the characteristics red pigmentation typical of some snow algae (e.g. Chlamydomonas nivalis (Bauer) Wille). Consequently these compounds greatly reduce the amount of light available for absorption by the light-harvesting pigment-protein complexes, thus potentially limiting photoinhibition and photodamage caused by intense solar radiation. The esterification of astaxnthin with fatty acids represents a possible mechanism by which this chromophore can be concentrated within cytoplasmic globules to maximize its photoprotective efficiency.     

A snow microflora in the Cairngorm Mountains,Scotland

An association of snow algae and fungi from a corrie in the Cairngorms is described and illustrated. This is the first detailed account of a snow microflora from Britain, though Chlamydomonas nivalis was collected twice last century.     

FINE STRUCTURE OF THE SNOW ALGA (CHLAMYDOMONAS NIVALIS) AND ASSOCIATED BACTERIA1

Chlamydomonas nivalis (Bau.) Wille is present in red snow as large spherical resting cells. Fine structural studies reveal an abundance of clear granules in the cytoplasm and occasional starch grains in the chloroplast. Individual cells display a thick cell wall with a smooth outer surface. Cells may be surrounded by a loose fibrous network in which encapsulated bacteria are seen. The bacteria have a characteristic Gram-negative cell wall and constrictive mode of division. The algal-bacterial association appears to be characteristic of red snow populations.     

Red-coloured snow algae in svalbard — Some environmental factors determining the distribution of Chlamydomonas nivalis (Chlorophyta volvocales)

Summary Studies of colonization of snow slopes by the snow algae Chlamydomonas nivalis (Chlorophyta volvocales) show that various environmental factors can be correlated with regions of algal colonization. All colonized sites discovered faced west, and when transects were made across the slopes it was found that areas of maximum colonization showed minima in temperature and pH and maxima of conductivity and chloride ion concentration.Svalbard Expedition (1980) Perse School, Cambridge     

PHOTOSYNTHESIS IN THE SNOW: THE ALGA CHLAMYDOMONAS NIVALIS (CHLOROPHYCEAE)1

Red blooms of snow algae consisting almost exclusively of large spherical red cells of Chlamydomonas nivalis (Bauer) Wille are widespread during the summer in the Beartooth Mountains in Montana and Wyoming. Field studies designed to examine the effects of temperature, light, and water potential on algal activity were performed with natural populations using photosynthetic ¹⁴C-HCO₃- or ¹⁴CO₂ incorporation as a measure of activity. The algae photo-synthesized optimally at 5.4 × 10⁴ lx, but were not inhibited by increased light intensity up to 8.6 × 10⁴ lx, the maximum observed in the field. Photosynthesis was sensitive to a reduction in water potential, and since low water potentials develop in snow at temperatures below 0 C, it is unlikely that significant algal activity occurs at the sub-0 temperatures which occur throughout winter. Photosynthesis was much lower following melting of the snow, but this was probably due to decreased diffusion of CO₂. The optimal temperatures varied considerably among the different algal populations. Most samples photo-synthesized optimally at 10 or 20 C but retained substantial activity at temperatures as low as 0 or -3 C. Exceptional samples photosynthesized optimally at 0 or -3 C. It is proposed that the varied temperature responses reflect the presence of different temperature strains. Taken together, the data suggest that development of the snow algae can occur only during the summer months.     

LC–MS/APCI identification of glucoside esters and diesters of astaxanthin from the snow alga Chlamydomonas nivalis including their optical stereoisomers

HPLC methods (LC–MS/APCI and chiral HPLC) were used for the identification of astaxanthin derivatives from the red snow alga Chlamydomonas nivalis collected in Austrian Alps, Slovak High Tatra Mountains and Bulgarian Pirin. We observed a striking difference in the composition of astaxanthin optical isomers in C. nivalis collected in geographically distinct regions. Furthermore, algae from the Pirin Mountains differed in the dominance of astaxanthin diglucoside diesters, suggesting an alternative strategy to enhance cell viability at low temperatures.     

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.     

CHLAINOMONAS KOLII(HARDY ET CURL) COMB. NOV. (CHLOROPHYTA,VOLVOCALES), A REVISION OF THE SNOW ALGA,TRACHELOMONAS KOLII HARDY ET CURL (EUGLENOPHYTA,EUGLENALES)1,2

The snow alga Trachelomonas kolii is transferred from the Euglenophyta (Euglenales) to the Chlorophyta (Volvocales) as Chlainomonas kolii comb. nov. As a result of critical examination of both living and type material, this species was found to have 4 flagella per vegetative cell, true starch, and 1 axial plastid per cell with several peripheral lobes. Vegetative cells of Trachelomonas kolii were described originally as having 1 flagellum, lacking true starch but having paramylum, and as having several parietal plastids per cell. The reticulate markings on the outer envelope of vegetative cells were found to be different from those in the original illustrations. Vegetative cells and resting spores of Chlainomonas kolii and Chlainomonas rubra are compared. The similarities of resting spores of Chlainomonas kolii and Chlamydomonas nivalis are discussed. These are the first records of Chlainomonas kolii from snow in Washington State.     

OBSERVATIONS ON SNOW ALGAE IN CALIFORNIA1,2

Algae that impart a red color to snowfields are rather common in California. Red snow occurs mainly in the Sierra Nevada at altitudes of 10,000–12,000 ft (3050–3600 in) and can occur at high altitudes where snow persists in other parts of the state. The distribution in the Sierra was similar in 1969 and 1970, contrasting snowfall years. Colored snow was found from May to October in old, wet snow-fields. The predominant color was red and occurred as surface patches in depressions in the snow. The color could extend as deep as 30 cm below the snow surface. Algae in the snowfields of the Tioga Pass area (Sierra Nevada) were large, red, spherical cells of Chlamydomonas nivalis. No other algae were seen. Their distribution, as measured by cell numbers and chlorophyll a, was patchy. Algal cells and chlorophyll a were mainly distributed at or near the snow surface but extended down to a depth of 10 cm. Light intensity was greatly attenuated by snow, but enough light for photosynthesis was found at 50 cm below the surface. Nutrient content of one snow sample was very low. The populations were very actively photosynthetic and took up as much as 65% of added ¹⁴CO₂ in only 3 hr. It was tentatively concluded that CO₂ limits in situ photosynthesis. Photosynthesis was inhibited by melting snow samples. Rough calculations of the growth rate suggested in situ generation times of only a few days for these algae.     

Snow algae from northwest Svalbard: their identification, distribution, pigment and nutrient content

We mapped coloured snow during the summers of 1995 and 1996 at about 60 localities in the coastal region of northwest Spitsbergen. The colour was mainly induced by snow algae (Chlamydomonas spp. and Chloromonas spp.). In the late summer of 1996, snow algal fields of several hundred meters in size were observed along the west and north coasts. They had no preferred geographical orientation. We studied the abundance of primary pigments and secondary carotenoids from different developmental stages of the snow algae of Chlamydomonas spp. under natural conditions. Extensive accumulation of astaxanthin and its esters accompanied the transition from green biflagellated cells to orange spores, hypnozygotes and dark-red cysts. The photoprotective effect of the secondary carotenoids is enhanced by concentration in cytoplasmic lipid droplets around the nucleus and chloroplast. The nutrient content of melt-water and snow algae had no direct correlation with the content of secondary carotenoids. Relatively high Fe, Ca, P, K and Al contents of snow algae were found, suggesting a good supply of these mineral elements. Received: 20 May 1997 / Accepted: 18 March 1998     

Detection and Quantification of Snow Algae with an Airborne Imaging Spectrometer

We describe spectral reflectance measurements of snow containing the snow alga Chlamydomonas nivalis and a model to retrieve snow algal concentrations from airborne imaging spectrometer data. Because cells of C. nivalis absorb at specific wavelengths in regions indicative of carotenoids (astaxanthin esters, lutein, β-carotene) and chlorophylls a and b, the spectral signature of snow containing C. nivalis is distinct from that of snow without algae. The spectral reflectance of snow containing C. nivalis is separable from that of snow without algae due to carotenoid absorption in the wavelength range from 0.4 to 0.58 μm and chlorophyll a and b absorption in the wavelength range from 0.6 to 0.7 μm. The integral of the scaled chlorophyll a and b absorption feature (I_0.68) varies with algal concentration (C_a). Using the relationship C_a = 81019.2 I_0.68 + 845.2, we inverted Airborne Visible Infrared Imaging Spectrometer reflectance data collected in the Tioga Pass region of the Sierra Nevada in California to determine algal concentration. For the 5.5-km² region imaged, the mean algal concentration was 1,306 cells ml⁻¹, the standard deviation was 1,740 cells ml⁻¹, and the coefficient of variation was 1.33. The retrieved spatial distribution was consistent with observations made in the field. From the spatial estimates of algal concentration, we calculated a total imaged algal biomass of 16.55 kg for the 0.495-km² snow-covered area, which gave an areal biomass concentration of 0.033 g/m².     

An update on carotenoid biosynthesis in algae: phylogenetic evidence for the existence of two classes of phytoene synthase

Carotenoids play crucial roles in structure and function of the photosynthetic apparatus of bacteria, algae, and higher plants. The entry-step reaction to carotenoid biosynthesis is catalyzed by the phytoene synthase (PSY), which is structurally and functionally related in all organisms. A comparative genomic analysis regarding the PSY revealed that the green algae Ostreococcus and Micromonas possess two orthologous copies of the PSY genes, indicating an ancient gene duplication event that produced two classes of PSY in algae. However, some other green algae (Chlamydomonas reinhardtii, Chlorella vulgaris, and Volvox carteri), red algae (Cyanidioschyzon merolae), diatoms (Thalassiosira pseudonana and Phaeodactylum tricornutum), and higher plants retained only one class of the PSY gene whereas the other gene copy was lost in these species. Further, similar to the situation in higher plants recent gene duplications of PSY have occurred for example in the green alga Dunaliella salina/bardawil. As members of the PSY gene families in some higher plants are differentially regulated during development or stress, the discovery of two classes of PSY gene families in some algae suggests that carotenoid biosynthesis in these algae is differentially regulated in response to development and environmental stress as well.     

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.     

ALGAL TRANSGENICS IN THE GENOMIC ERA1

The last few years have witnessed significant advances in the field of algal genomics. Complete genome sequences from the red alga Cyanidioschyzon merolae and the diatom Thalassiosira pseudonana have been published, the genomes for two more algae (Chlamydomonas reinhardtii and Ostreococcus tauri) are nearing completion, and several others are in progress or at the planning stage. In addition, large‐scale cDNA sequencing projects are being carried out for numerous algal species. This wealth of genome data is serving as a powerful catalyst for the development and application of recombinant techniques for these species. The data provide a rich resource of DNA elements such as promoters that can be used for transgene expression as well as an inventory of genes that are possible targets for genetic engineering programs aimed at manipulating algal metabolism. It is not surprising therefore that significant progress in the genetic engineering of eukaryotic algae is being made. Nuclear transformation of various microalgal species is now routine, and progress is being made on the transformation of macroalgae. Chloroplast transformation has been achieved for green, red, and euglenoid algae, and further success in organelle transformation is likely as the number of sequenced plastid, mitochondrial, and nucleomorph genomes continues to grow. Importantly, the commercial application of algal transgenics is beginning to be realized, and algal biotechnology companies are being established. Recent work has shown that recombinant proteins of therapeutic value can be produced in microalgal species, and it is now realistic to envisage the genetic engineering of commercially important species to improve production of valuable algal products. In this article we review the recent progress in algal transgenics and consider possible future developments now that phycology has entered the genomic era.     

A COMBINED 18S rDNA AND rbcL PHYLOGENETIC ANALYSIS OF CHLOROMONAS AND CHLAMYDOMONAS (CHLOROPHYCEAE,VOLVOCALES) EMPHASIZING SNOW AND OTHER COLD‐TEMPERATURE HABITATS1

An extensive phylogenetic analysis of the biflagellate genera, Chlamydomonas Ehrenberg and Chloromonas Gobi emend. Wille, was undertaken using 18S rDNA and rbcL gene sequence analysis. Emphasis was placed on 21 cold‐tolerant taxa of which 10 are from snow. These taxa occurred in four distinct clades each in the 18S rDNA and rbcL phylogenies, and when taken together suggest at least five distinct origins in cold habitats. Most of these taxa occur in a single clade (A), and all snow species occurred in this clade. In the rbcL and combined rbcL–18S rDNA analyses, the snow taxa fell into three groups. Two groups occurred in subclade 1: Chlamydomonas augustae Skuja CU, Chlamydomonas augustae UTEX, and Chlamydomonas sp.‐A and Chloromonas clathrata Korshikov, Chloromonas rosae Ettl CU, and Chloromonas rosae v. psychrophila var. nov. The third snow group, subclade 2, included three species with unique cell divisions, Chloromonas brevispina (Fritsch) Hoham, Roemer et Mullet, Chloromonas pichinchae (Lagerheim) Wille, and Chloromonas sp.‐D, and the basal Chloromonas nivalis (Chodat) Hoham et Mullet with normal cell divisions. This suggests that the snow habitat has been colonized at least twice and possibly three times in the history of these biflagellates. In the 18S rDNA tree, one cold‐tolerant Chloromonas species fell outside clade A: Chloromonas subdivisa (Pascher et Jahoda) Gerloff et Ettl. In the rbcL tree, three cold‐tolerant Chloromonas species fell outside clade A: Chloromonas subdivisa, Chloromonas sp.‐ANT1, and Chloromonas sp.‐ANT3. These results support previous findings that pyrenoids have been gained and lost several times within this complex.     

10. Snow algae of the Windmill Islands region, Antarctica

A list of the 24 species of snow algae identified from the region, a resume of what is currently known about the major species, and avenues for further research are provided. New species discovered include 2 Desmotetra spp., one Chlorosarcina sp., 2 Chloromonas spp. and a Palmellopsis sp. Several of these are from genera whose members have previously been found only in the soil flora. Not only was it necessary to elucidate the life cycle of these species, but it was also essential to examine them ultrastructurally to determine their taxonomic positions.     

ALGAL BIOREMEDIATION OF THE BERKELEY PIT LAKE

Ongoing research is unraveling the intricacies of the microbial ecology of the Berkeley Pit Lake System, with ever increasing information becoming available regarding the diversity of Algae, Protistans, Fungi and Bacteria that inhabit this mine waste site. Defining the baseline community structure has been the first step not only toward understanding the interactions of the different groups of organisms, but also toward assessing any improvement in biodiversity within the biotic community. Now that this first step has begun, some of these extremophiles, specifically algae, that have been isolated from the Berkeley Pit Lake System are being used as a potential solution for bioremediation. The specific objectives of this research are fivefold: 1) To evaluate the bioremediative potential of our four most rapidly growing species: (Chromulina freiburgensis Dofl., Chlorella ellipsoidea Gerneck, Chlorella vulgaris Beyerinck and Chlamydomonas acidophilla Negoro) in Berkeley Pit Lake System Water with the additions of NaNO₃ and NaPO₄ by using an experimental matrix. This matrix will be used to estimate the minimum nutrient concentrations that would be necessary to achieve the maximum growth of algae and maximum bioremediation of the Berkeley Pit Lake System. 2) To determine which combination of nutrients will stimulate growth of the best bioremediator of our four isolated species in natural Berkeley Pit Lake System waters. In other words, what nutrient combination will give the best bioremediator a competitive edge over the other species. If time permits, different species may be grown in combination to determine if there are synergistic effects (protocooperation) between/among species. 3) To determine a temperature profile for these four species in order to determine their optimal growth temperature in Berkeley Pit Lake System water. 4) To continue to isolate organisms from the Berkeley Pit Lake System and determine their bioremediative potential. 5) Monitor algal and bacterial counts from a profile of Pit Lake System waters. The results to date will be presented for this conference.     

The Distribution and Phytogeographical Status of the Marine Algal Flora of Gambia

The littoral algae and their zonation on the short and hitherto phycologically virtually unexplored coast of Gambia (West Africa) were investigated during November 1975. Littoral zonation of benthic organisms was essentially similar to that found to the south in the Gulf of Guinea. Formerly only 4 species of algae had been reported specifically for Gambia but only 1 of these species was included in the 69 found in the present study. These new records, with the exception of just 2 species (Hypnea arbuscula and Ulva popenguinensis), are all found in the Gulf of Guinea and thus the previously known limit (Sierra Leone) of the truly tropical marine flora of West Africa is extented over 600 km to the north.     

Rosetta gen. nov. (Chlorophyta): Resolving the identity of red snow algal rosettes

Thick-walled rosette-like snow algae were long thought to be a life stage of various other species of snow algae. Rosette-like cells have not been cultured, but by manually isolating cells from 38 field samples in southern British Columbia, we assigned a variety of rosette morphologies to DNA sequence. Phylogenetic analysis of Rubisco large-subunit (rbcL) gene, ribosomal internal transcribed spacer 2 (ITS2) rRNA region, and 18S rRNA gene revealed that the rosette-like cells form a new clade within the phylogroup Chloromonadinia. Based on these data, we designate a new genus, Rosetta, which comprises five novel species: R. castellata, R. floranivea, R. stellaria, R. rubriterra, and R. papavera. In a survey of 762 snow samples from British Columbia, we observed R. floranivea exclusively on snow overlying high-elevation glaciers, whereas R. castellata was observed at lower elevations, near the tree line. The other three species were rarely observed. Spherical red cells enveloped in a thin translucent sac were conspecific with Rosetta, possibly a developmental stage. These results highlight the unexplored diversity among snow algae and emphasize the utility of single-cell isolation to advance the centuries-old problem of disentangling life stages and cryptic species.