The Intracellular Localization of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase in Chlamydomonas reinhardtii1

The pyrenoid is a proteinaceous structure found in the chloroplast of most unicellular algae. Various studies indicate that ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is present in the pyrenoid, although the fraction of Rubisco localized there remains controversial. Estimates of the amount of Rubisco in the pyrenoid of Chlamydomonas reinhardtii range from 5% to nearly 100%. Using immunolocalization, the amount of Rubisco localized to the pyrenoid or to the chloroplast stroma was estimated for C. reinhardtii cells grown under different conditions. It was observed that the amount of Rubisco in the pyrenoid varied with growth condition; about 40% was in the pyrenoid when the cells were grown under elevated CO2 and about 90% with ambient CO2. In addition, it is likely that pyrenoidal Rubisco is active in CO2 fixation because in vitro activity measurements showed that most of the Rubisco must be active to account for CO2-fixation rates observed in whole cells. These results are consistent with the idea that the pyrenoid is the site of CO2 fixation in C. reinhardtii and other unicellular algae containing CO2-concentrating mechanisms.

The selection of a model microalgal species as biomaterial for a novel aquatic phytotoxicity assay

A phytotoxicity assay based on the ToxY-PAM dual-channel yield analyser has been developed and successfully incorporated into field assessments for the detection of phytotoxicants in water. As a means of further exploring the scope of the assay application and of selecting a model biomaterial to complement the instrument design, nine algal species were exposed to four chemical substances deemed of priority for water quality monitoring purposes (chlorpyrifos, copper, diuron and nonylphenol ethoxylate). Inter-species differences in sensitivity to the four toxicants varied by a factor of 1.9-100. Measurements of photosystem-II quantum yield using these nine single-celled microalgae as biomaterial corroborated previous studies which have shown that the ToxY-PAM dual-channel yield analyser is a highly sensitive method for the detection of PS-II impacting herbicides. Besides Phaeodactylum tricornutum, the previously applied biomaterial, three other species consistently performed well (Nitzschia closterium, Chlorella vulgaris and Dunaliella tertiolecta) and will be used in further test optimisation experiments. In addition to sensitivity, response time was evaluated and revealed a high degree of variation between species and toxicants. While most species displayed relatively weak and slow responses to copper, C. vulgaris demonstrated an IC10 of 51 μ g L-1, with maximum response measured within 25 minutes and inhibition being accompanied by a large decrease in fluorescence yield. The potential for this C vulgaris-based bioassay to be used for the detection of copper is discussed. There was no evidence that the standard ToxY-PAM protocol, using these unicellular algae species, could be used for the detection of chlorpyrifos or nonylphenol ethoxylate at environmentally relevant levels. © 2005 Elsevier B.V. All rights reserved.

Étude du génome chloroplastique des algues vertes de la classe Chlorophyceae

Les algues unicellulaires de la classe Chlorophyceae sont particulièrement étudiées pour leur potentiel économique dans la production de biocarburant. La première taxonomie de cette classe a été faite avec l’avènement de la microscopie électronique et par la suite avec des phylogénies moléculaires. Cette lignée se divise en deux groupes : OCC (Oedogoniales + Chaetophorales + Chaetopeltidales) et CS (Chlamydomonadales + Sphaeropleales). Il existe de profondes incertitudes sur les positions phylogénétiques des organismes à la base du groupe CS. Afin de renforcer la phylogénie de ces organismes, les génomes chloroplastiques de cinq algues basales ont été séquencés à l’aide de la technologie de nouvelle génération 454 et assemblés de novo. Une analyse phylogénétique de 69 séquences de protéines a permis de montrer que trois des cinq organismes classés dans l’ordre Chlamydomonadales par la littérature actuelle sont en fait basaux dans l’ordre Sphaeropleales. Ce reclassement phylogénétique implique de nouvelles hypothèses sur l’évolution des corps flagellaires.

Immunodetection of planktonic algae

Interest in the identification and characterisation of cyanobacteria and dinoflagellates in aquatic environments is increasing rapidly due to the perceived roles of these organisms in primary production and nuisance aspects in terms of water treatment and public health. Techniques for the identification and quantification of these organisms currently are limited, and the application of molecular approaches provides fundamental taxonomic information and techniques of practical value. Antigenic properties of algal cells may be useful taxonomic markers. Immunodetection techniques utilise the specificity of the antibody/antigen association as a probe for recognising and distinguishing between microorganisms according to their cell- surface chemistry. Immunofluorescent detection of unicellular cyanobacteria and dinoflagellates has been studied with success in marine and freshwater ecosystems and a range of techniques and results are presented and discussed. The most recent advances in the study of planktonic algae have come with the application of continuous flow cytometric methods (CFC). Flow cytometry makes use of the autofluorescence properties of the algal cells, which alone can be used to demonstrate their presence and permit their quantification in natural water samples. When used in conjunction with immunolabelling techniques, the potential of CFC analysis is broadened to study the serological/strain composition of plankters in natural populations. Changes in algal strains represented within and between waters over periods of time are reported and discussed, along with the ecological issues thus raised.

Effect of two green algae species (Chlorella sp. & Scenedesmus obliquus) enriched with B group vitamins on food chain of Rutilus frisii kutum (Kamensky, 1901) fry

The Effect of two freshwater green algae species Chlorella sp. & Scenedesmus obliquus enriched (from the beginning of culture and after 96 hours) with different dosages of B group vitamins (0, 0.5, 1, and 2 ml of enriching solution per each liter of algae medium) on fecundity of Daphnia magna and growth of Rutilus frisii kutum fry were investigated in a research from spring, 2008 to autumn, 2009. First, each of the green algae species were cultured purely and massively in the Zander (Z-8+N) medium and then the nutritional value (the amount of protein, lipid, and carbohydrate) of enriched algae were meausered. In this study, enriching of Chlorella sp. & S. obliquus with a suitable mix of B group vitamins significantly improved their nutritive value. So the highest amount of nutritional value of Chlorella sp. was obtained because of enriching with dosage 0.5 ml.l-1 (366.654Kcal) and for Scenedesmus obliquus with dosage of 1 ml.l-1 (376.95Kcal). The acquired amount from control group showed an increase of respectively 42% and 11%. According to the results, increased dosages of enriching solution caused Daphnia fecundity to increase (at both stages : enrichment from the beginning of culture and after 96 hours). So the highest average of D. magna reproduction rate was obtained through being fed with Chlorella sp. and S. obliquus enriched with dosage of 2 ml enriching solution per liter of algae medium. The average fecundity of D. magna fed with Chlorella sp. enriched with dosage of 2 ml.l-1 enriching solution from the beginning of culture and after 96 hours was obtained respectively 2.128 ± 0.375 and 2.1 ± 0.69 and the average fecundity of D. magna fed with S. obliquus enriched with dosage of 2 ml enriching solution from the beginning of culture and after 96 hours was obtained respectively 2.128 ± 0.375 and 2.1 ± 0.69 which showed respectively an increase of 61 ٪, 91٪, 77 ٪, and 83٪ in proportion to the acquired amount from control group. When enriching solution was added to either algae culture medium from the beginning of culture, showed statistically significant differences (P<0.05) between dosages of 0 and 2 ml.l-1, 1 and 2 ml.l-1, and 0.5 and 2 ml enriching solution per each liter of Chlorella sp. culture medium and between dosages of 0 and 1 ml.l-1, and 0 and 2 ml enriching solution per each liter of S. obliquus culture medium. The highest average of body weight gain percentage and specific growth rate of kutum fry was obtained respectively 21.19%, 26.63%, 1.92, and 2.34 from the beginning of culture and after 96 hours with dosage of 1 ml B group vitamins per each liter of Chlorella sp. culture medium, which showed respectively an increase of 50%, 70%, 46%, and 62% in proportion to the acquired amount from control group. In the cases which Chlorella sp. were grown in the medium containing vitamin, from point of view of the average percentage of weight and specific growth rate of kutum fry significant differences were observed on the basis of the result of One-way ANOVA between dosages of 0 and 1, 1 and 2 , 0.5 and 1 ml B group vitamins per each liter. The highest average of body weight gain percentage and specific growth rate of kutum fry was obtained respectively 32.02%, 29.42%, 2.78, and 2.34 from the beginning of culture and after 96 hours with dosage of 2 ml B group vitamins per each liter of S. obliquus culture medium, which showed respectively an increase of 32%, 19%, 28%, and 17% in proportion to the acquired amount from control group. In the cases which S. obliquus were grown in the medium containing vitamin, from point of view of the average percentage of weight and specific growth rate of kutum fry significant differences were observed on the basis of the result of One-way ANOVA between dosages of 0 and 1, 0 and 2. According to the results of the present research we can say that considerable enhancement in the quality of the food of D. magna can be made by manipulation of the nutritional value of fresh water unicellular green algae with suitable mixture of B group vitamins, so that both the fecundity of D. magna will increase and the nutritional requirements of the kutum fry will be filled in this way.

Transient expression of lacZ in bombarded unicellular green alga Haematococcus pluvialis

This paper reports for the first time the transient expression of a reporter gene, LacZ, in the unicellular green alga Haematococcus pluvialis. By employing the micro-particle bombardment method, motile cells in the exponential phase showed transient expression of lacZ. This was detected in bombarded motile cells under the rupture-disc pressures of 3103 KPa and 4137 KPa. Transient expression of LacZ gene could not be observed in non-motile cells of this alga under the same transformation condition. No LacZ background was found in either the motile cells or the non-motile cells. The study suggests a promising potential of the SV40 promoter and the lacZ reporter gene in genetic engineering of unicellular green algae.

Feeding on micro-algae in the invasive Ponto-Caspian amphipod Dikerogammarus villosus (Sowinsky, 1894)

Feeding on micro-algae is shown in the invasive Ponto-Caspian amphipod Dikerogammarus villosus. Compared with controls, males, females and juveniles of this species significantly reduced the concentration in suspension of unicellular micro-algae. Juveniles had higher concentrations of algae in the cardiac gut than adults. The presence of these algae in the mid- and hindgut was also recorded. This feeding behaviour was filmed and the mechanisms involved are described and discussed. We comment on the use of the Functional Feeding Group (FFG) concept to classify feeding in amphipods. The role of being a feeding-generalist in aiding the invasion process is also discussed.

Extracellular matrix of the charophycean green algae

A comprehensive knowledge of cell wallstructure and function throughout the plant kingdom is essential to understanding cell wall evolution. The fundamental understanding of the charophycean green algal cell wall is broadening. The similarities and differences that exist between land plant and algal cell walls provide opportunities to understand plant evolution. A variety of polymers previously associated with higher plants were discovered in the charophycean green algae (CGA), including homogalacturonans, cross-linking glycans, arabinogalactan protein, β-glucans, and cellulose. The cellulose content of CGA cell walls ranged from 6% to 43%, with the higher valuescomparable to that found in the primary cell wall of land plants (20-30%). (1,3)β-glucans were found in the unicellular Chlorokybus atmophyticus, Penium margaritaceum, and Cosmarium turpini, the unbranched filamentous Klebsormidium flaccidum, and the multicellular Chara corallina. The discovery of homogalacturonan in Penium margaritaceum representsthe first confirmation of land plant-type pectinsin desmids and the second rigorous characterization of a pectin polymer from the charophycean algae. Homogalacturonan was also indicated from the basal species Chlorokybus atmophyticus and Klebsormidium flaccidum. There is evidence of branched pectins in Cosmarium turpini and linkage analysis suggests the presence of type I rhamnogalacturonan (RGI). Cross-linking β-glucans are associated with cellulose microfibrils during land plant cell growth, and were found in the cell wall of CGA. The evidence of mixed-linkage glucan (MLG) in the 11 charophytesis both suprising and significant given that MLG was once thought to be specific to some grasses. The organization and structure of Cosmarium turpini and Chara corallina MLG was found to be similar to that of Equisetumspp., whereas the basal species of the CGA, Chlorokybus atmophyticus and Klebsormidium flaccidum, have unique organization of alternating of 3- and 4-linkages. The significance of this result on the evolution of the MLG synthetic pathway has yet to be determined. The extracellular matrix (ECM) of Chlorokybus atmophyticus, Klebsormidium flaccidum, and Spirogyra spp. exhibits significant biochemical diversity, ranging from distinct “land plant” polymers to polysaccharides unique to these algae. The neutral sugar composition of Chlorokybus atmophyticus hot water extract and Spirogyra extracellular polymeric substance (EPS), combined with antibody labeling results, revealed the distinct possibility of an arabinogalactan protein in these organisms. Polysaccharide analysis of Zygnematales (desmid) EPS, indicated a probable range of different EPS backbones and substitution patterns upon the core portions of the molecules. Desmid EPS is predominately composed of a complex matrix of branched, uronic acid containing polysaccharides with ester sulfate substitutions and, as such, has an almost infinite capacity for various hydrogen bonding, hydrophobic interaction and ionic cross-bridging motifs, which characterize their unique function in biofilms. My observations support the hypothesis that members of the CGA represent the phylogenetic line that gave rise to vascular plants and that the primary cell wall of vascular plants many have evolved directly from structures typical of the cell wall of filamentous green algae found in the charophycean green algae.

(Table 1) Abundance of unicellular eukaryotes in ice cores collected during the July 2008 CCGS Amundsen cruise in Darnley Bay

Influence of methanogenic populations in Holocene lacustrine sediments revealed by clone libraries and fatty acid biogeochemistry.Biological characteristics of ice-associated algal communities were studied in Darnley Bay (western Canadian Arctic) during a 2-week period in July 2008 when the landfast ice cover had reached an advanced stage of melt. We found two distinct and separate algal communities: (1) an interior ice community confined to brine channel networks beneath white ice covers; and (2) an ice melt water community in the brackish waters of both surface melt ponds and the layer immediately below the ice cover. Both communities reached maximum chlorophyll a concentrations of about 2.5 mg/m**3, but with diatoms dominating the interior ice while flagellates dominated the melt water community. The microflora of each community was diverse, containing both unique and shared algal species, the latter suggesting an initial seeding of the ice melt water by the bottom ice community. Absorption characteristics of the algae indicated the presence of mycosporine-like amino acids (MAAs) and carotenoid pigments as a photoprotective strategy against being confined to high-light near-surface layers. Although likely not contributing substantially to total annual primary production, these ice-associated communities may play an important ecological role in the Arctic marine ecosystem, supplying an accessible and stable food source to higher trophic levels during the period of ice melt.

The effect of a reduction of nitrogen and phosphorus to eutrophic and mesotrophic conditions towards silver nanoparticle (AgNP) on freshwater green algae species

We investigated the sensitivity of algae towards silver nanoparticles with OECD test medium and lower nutrient concentrations under standard test conditions to improve comparability and to exclude any other confounding factor aside nutrient levels. Two unicellular freshwater microalgae Desmodesmus subspicatus and Raphidocelis subcapitata were chosen due to their status as standard test organisms for the algae growth inhibition test and the response to changes in nutrient supply was compared. The original medium was used as the reference (standard). For the other four media, the amount of either nitrogen or phosphorus in the medium was lowered from half (50%) to one-fourth (25 %) of that of the OECD guideline, resulting in the following media: 50% N, 25% N, 50% P and 25% P medium. As test substance, the OECD reference material NM-300K was used. For this reason, the characterization of AgNP was done using DLS and Absorption spectra (UV/vis). Actual silver concentrations and ionic silver concentrations were measured at the highest test concentration used (100 µg Ag L-1) in R. subcapitata treatments only to reduce the number of samples. All tests were run according to the OECD guideline 201 with sterilized 50 mL cell culture flask. Each medium was tested using the test conditions for culturing with 3 replicates. Test concentrations for both algae species were 0, 25, 50 and 100 µg Ag L-1 for OECD, 50% P and 25% P while for both N reductions, the silver concentrations were 0, 10, 25 and 100 µg Ag L-1. Samples for determining the algal density were taken at every 24 h.

Influence of fatty acid structure on fuel properties of algae derived biodiesel

Physical and chemical properties of biofuel are influenced by structural features of fatty acid such as chain length, degree of unsaturation and branching of the chain. A simple and reliable calculation method to estimate fuel property is therefore needed to avoid experimental testing which is difficult, costly and time consuming. Typically in commercial biodiesel production such testing is done for every batch of fuel produced. In this study 9 different algae species were selected that were likely to be suitable for subtropical climates. The fatty acid methyl esters (FAMEs) of all algae species were analysed and the fuel properties like cetane number (CN), cold filter plugging point (CFPP), kinematic viscosity (KV), density and higher heating value (HHV) were determined. The relation of each fatty acid with particular fuel property is analysed using multivariate and multi-criteria decision method (MCDM) software. They showed that some fatty acids have major influences on the fuel properties whereas others have minimal influence. Based on the fuel properties and amounts of lipid content rank order is drawn by PROMETHEE-GAIA which helped to select the best algae species for biodiesel production in subtropical climates. Three species had fatty acid profiles that gave the best fuel properties although only one of these (Nannochloropsis oculata) is considered the best choice because of its higher lipid content.

Dewatering of microalgal cultures : a major bottleneck to algae-based fuels

Microalgae dewatering is a major obstruction to industrial-scale processing of microalgae for biofuel prodn. The dil. nature of harvested microalgal cultures creates a huge operational cost during dewatering, thereby, rendering algae-based fuels less economically attractive. Currently there is no superior method of dewatering microalgae. A technique that may result in a greater algal biomass may have drawbacks such as a high capital cost or high energy consumption. The choice of which harvesting technique to apply will depend on the species of microalgae and the final product desired. Algal properties such as a large cell size and the capability of the microalgae to autoflocculate can simplify the dewatering process. This article reviews and addresses the various technologies currently used for dewatering microalgal cultures along with a comparative study of the performances of the different technologies.

Spirulina (Spirulina platensis) algae supplementation increases microbial protein production and feed intake and decreases retention time of digesta in the rumen of cattle

Cattle consuming pastures low in protein have low liveweight gain due to low rumen degradable protein (RDP) supply and thus low microbial crude protein (MCP) production and efficiency of MCP production [EMCP, g MCP/kg digestible organic matter (DOM)]. Nitrogen supplements can increase MCP production and EMCP of cattle grazing low protein pastures. The objective of this study was to compare the effects of supplementation with a non-protein-N source (NPN), in this case urea and ammonium sulfate (US), with a single-cell algal protein source (Spirulina platensis), on intake, microbial protein supply and digestibility in cattle. Nine cannulated Bos indicus steers [initial liveweight 250.1 ± 10.86 (s.d.) kg] were fed Mitchell grass hay (Astrebla spp; 6.1 g N, 746 g NDF/kg DM) ad libitum and were supplied with increasing amounts of US (0, 6, 13, 19 and 33 g US DM/kg hay DM) or Spirulina 0, 0.5, 1.4, 2.5 and 6.1 g Spirulina DM/kg W.day in an incomplete Latin square design. The response of MCP production and EMCP to increasing amounts of the two supplements was different, with a greater response to Spirulina evident. The MCP production was predicted to peak at 140 and 568 g MCP/day (0.64 and 2.02 g MCP/kg W.day) for the US and Spirulina supplements, respectively. The highest measured EMCP were 92 and 166 g MCP/kg DOM for the US and Spirulina treatments at 170 and 290 g RDP/kg DOM, respectively, or a Spirulina intake of 5.7 g DM/kg W.day. Increasing RDP intake from US and Spirulina resulted in an increase in Mitchell grass hay intake and rumen NH3-N concentration and reduced the retention time of liquid and particulate markers and digesta DM, NDF and lignin in the rumen with greater changes due to Spirulina. Total DM intake peaked at a Spirulina supplement level of 4.6 g Spirulina DM/kg W.day with a 2.3-fold higher DOM intake than Control steers. Rumen NH3-N concentrations reached 128 and 264 mg NH3-N/L for the US and Spirulina treatments with a significant increase in the concentration of branched-chain fatty acids for the Spirulina treatment. The minimum retention time of liquid (Cr-EDTA; 23 and 13 h) and particulate (Yb; 34 and 22 h) markers in the rumen were significantly lower for Spirulina compared with US and lower than unsupplemented animals at 24 and 34 h for Cr-EDTA and Yb, respectively. Spirulina could be provided safely at much higher N intakes than NPN supplements. The results suggest that, at an equivalent RDP supply, Spirulina provided greater increases than US in MCP production, EMCP and feed intake of Bos indicus cattle consuming low protein forage and could also be fed safely at higher levels of N intake.

Harmful algae in the planktonic food web of the Baltic Sea

This study deals with algal species occurring commonly in the Baltic Sea: haptophyte Prymnesium parvum, dinoflagellates Dinophysis acuminata, D. norvegica and D. rotundata, and cyanobacterium Nodularia spumigena. The hypotheses are connected to the toxicity of the species, to the factors determining toxicity, to the consequences of toxicity and to the transfer of toxins in the aquatic food web. Since the Baltic Sea is severely eutrophicated, the fast-growing haptophytes have potential in causing toxic blooms. In our studies, the toxicity (as haemolytic activity) of the haptophyte P. parvum was highest under phosphorus-limited conditions, but the cells were toxic also under nitrogen limitation and under nutrient-balanced growth conditions. The cellular nutrient ratios were tightly related to the toxicity. The stoichiometric flexibility for cellular phosphorus quota was higher than for nitrogen, and nitrogen limitation led to decreased biomass. Negative allelopathic effects on another algae (Rhodomonas salina) could be observed already at low P. parvum cell densities, whereas immediate lysis of R. salina cells occurred at P. parvum cell densities corresponding to natural blooms. Release of dissolved organic carbon from the R. salina cells was measured within 30 minutes, and an increase in bacterial number and biomass was measured within 23 h. Because of the allelopathic effect, formation of a P. parvum bloom may accelerate after a critical cell density is reached and the competing species are eliminated. A P. parvum bloom indirectly stimulates bacterial growth, and alters the functioning of the planktonic food web by increasing the carbon transfer through the microbial loop. Our results were the first reports on DSP toxins in Dinophysis cells in the Gulf of Finland and on PTX-2 in the Baltic Sea. Cellular toxin contents in Dinophysis spp. ranged from 0.2 to 149 pg DTX-1 cell-1 and from 1.6 to 19.9 pg PTX-2 cell-1 in the Gulf of Finland. D. norvegica was found mainly around the thermocline (max. 200 cells L-1), whereas D. acuminata was found in the whole mixed layer (max. 7 280 cells L-1). Toxins in the sediment trap corresponded to 1 % of DTX-1 and 0.01 % PTX-2 of the DSP pool in the suspended matter. This indicates that the majority of the DSP toxins does not enter the benthic community, but is either decomposed in the water column, or transferred to higher trophic levels in the planktonic food chain. We found that nodularin, produced by Nodularia spumigena, was transferred to the copepod Eurytemora affinis through three pathways: by grazing on filaments of small Nodularia, directly from the dissolved pool, and through the microbial food web by copepods grazing on ciliates, dinoflagellates and heterotrophic nanoflagellates. The estimated proportion of the microbial food web in nodularin transfer was 22-45 % and 71-76 % in our two experiments, respectively. This highlights the potential role of the microbial food web in the transfer of toxins in the planktonic food web.

A systematic-ecological approach to Baltic Sea ice studies of algae and protists

The seasonal occurrence of sea ice that annually covers almost half the Baltic Sea area provides a unique habitat for halo- and cold temperature-tolerant extremophiles. Baltic Sea ice biology has more than 100 years of tradition that began with the floristic observation of species by the early pioneers using light microscopic techniques that were the only thing available at the time. Since the discovery of life within sea ice, more technologies have become available for taxonomy. Electron microscopy and genetic evidence have been used to identify sea ice biota revealing increased numbers of taxa. Meanwhile ecologists have used light microscopic cell enumeration in addition to the chemical and physical properties of sea ice in attempts to explain the food web structure of sea ice and its functions. Thus, during the Baltic winter, the sea ice hosts more abundant and diverse microbial communities than the water column beneath it. These communities are typically dominated by autotrophic diatoms together with a diverse assortment of dinoflagellates, auto- and heterotrophic flagellates, ciliates, metazoan rotifers and bacteria, which are mostly responsible for the recycling of nutrients. This thesis comprises ecological and systematic studies. In addition to the results of the previous studies carried out on landfast ice, the data presented here provide new insight into the spatial distribution of pelagial sea ice, which has remained largely unexplored. The studies reveal spatial heterogeneity in the pelagial sea ice of the Gulf of Bothnia. There were mismatches in chlorophyll-a concentrations and in photosynthetic efficiencies of the communities studied. The temporal succession was followed and experimental studies performed investigating the community responses towards increased or decreased light in landfast ice in the Gulf of Finland. The systematic studies carried out with established dinoflagellate cultures revealed a new resting cyst belonging to common sea ice dinoflagellate, Scrippsiella hangoei (Schiller) Larsen 1995. The cyst can be used to explain the overwintering of this species during prolonged periods of darkness. The dissimilarities and similarities in the material isolated from the sea ice called for description of a new subspecies Heterocapsa arctica ssp. frigida. The cells obtained in the cultured material were unlike those of the previously described species, necessitating description of ssp. frigida. As a result of its own unique habitus, the subspecies had been noted by Finnish taxonomists during the past three decades and thus its annual occurrence and geographical distribution in the Baltic Sea. This illustrates how combining ecology and systematics increases our understanding of organisms.