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Abstracts of Poster Presentations
The abstracts are in alphabetical order by lead author.
1. Achilles-Day U(1), Pröschold T(1), Harvey R(2) & Day JG(1)
Elucidating the phylogenetic position of Euplotes daidaleos as a pilot-study for resolving the taxonomy of CCAP's protistan strains
The Culture Collection of Algae and Protozoa (CCAP) (http://www.ccap.ac.uk) is the largest and most genotypically diverse microbial Biological Resource Centre (BRC) in Europe. The collection includes a wide variety of protists, many of which have never been examined using modern molecular methods. One of the objectives of CCAP over the next four and a half years is to resolve taxonomic uncertainties and where possible resolve the phylogeny of the collections’ holdings where possible. This pilot-study focuses on the freshwater ciliate Euplotes daidaleos (Diller,WF & Kounaris, D, 1966), where we have used Small Subunit rRNA Gene Sequence (SSrRNA) data from E. daidaleos CCAP 1624/15 and data available from GenBank on other Euplotes species to elucidate its taxonomic position.
(1)CCAP, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Oban, PA37 1QA, UK
(2)Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Oban, PA37 1QA, UK
2. Amorim A & Veloso V
The Algae Culture Collection at the University of Lisbon - ALISU
The Algae Culture Collection at the University of Lisbon (ALISU) was started in 1997 and is dedicated to marine phytoplankton with emphasis on Harmful Algae (HABs). In addition, several freshwater and estuarine microphytobenthos species are kept in culture for teaching or research purposes. The main objectives of the collection are to contribute for research activities on the biology, ecology and taxonomy of marine and estuarine microalgae, and support undergraduate, graduate, MSc and PhD research projects at the University of Lisbon. The collection is based on strains isolated from Portuguese coastal waters and aims at the ex situ conservation of regional representatives of all HAB species. Currently the collection hosts species assigned to Cyanobacteria, Bacillariophyceae, Chlorophyceae, Cryptophyceae, Dinophyceae, Haptophyceae and Raphidophyceae. Some strains are still in the process of being described or studied.
Faculty of Sciences University of Lisbon, Centre of Oceanography, 1749-016 Lisboa, Portugal
3. Bejoy T(1), Green DH(2) & Bolch CJS(1)
Influence of marine bacteria on growth of the toxic dinoflagellate Gymnodinium catenatum
Cultures of the dinoflagellate Gymnodinium catenatum were established with bacterial communities composed one to three bacterial types, to examine two hypotheses: a) that G. catenatum has an obligate requirement for marine bacteria, and b) that the composition of the bacterial community affects the growth of G. catenatum. Cultures of G. catenatum showed a 10-fold decline in cell density after treatment with antibiotics to remove bacteria. In contrast, control cultures established with antibiotic-resistant bacteria continued to grow showing that removal of the bacteria, not the antibiotic, caused the decline of the dinoflagellate. Dinoflagellate growth rates in the presence of strains of Alcanivorax or Marinobacter exhibited significantly higher growth rates than cultures grown with Roseobacter. Cultures grown with communities composed of all 3 bacterial types showed similar growth rate and batch culture dynamics to controls containing “natural” communities from non-axenic G. catenatum cultures. Our data demonstrate that 1) the presence of a bacterial community is essential for growth of G. catenatum, 2) that the community composition affects dinoflagellate growth rate and dynamics, 3) that the experimental model can be used to mimic responses to complex natural bacterial communities, and 4) that microbial interactions can play a major role in the development, maintenance and decline of natural phytoplankton populations.
(1)School of Aquaculture, University of Tasmania, Launceston, Tasmania, 7250, Australia
(2)Scottish Association of Marine Science, Dunstaffnage Marine Laboratory, Oban, Argyll, PA37 1QA, UK
4. Blackburn SI(1),(2),(3),(4), Jameson I(1),(4), Johnston C(1), Frampton D(1),(4), Nichols CM(1),(5), Mansour MP(2),(4), Nichols PD(2),(4), Robert S(2),(4) & Volkman JK(1),(3),(4)
Bioapplications of Microalgae
Microalgae are microscopic plants inhabiting the world’s oceans and other aquatic environments. Crucial for the health of the planet, microalgae are responsible for half global primary productivity, converting solar energy to organic energy and fixing carbon dioxide as part of the process. The Collection of Living Microalgae (http://www.cmar.csiro.au/microalgae/) (CCLM) is a living bank of microalgal biodiversity isolated from the tropics to Antarctica. Our research has demonstrated biodiversity at the strain level, with Australian microalgae having unique chemical, molecular and physiological characteristics. This biodiversity is displayed in a range of bioactive compounds that have medical, human health, aqua-feed, and energy applications. Australian strains from the CCLM are being explored for the adhesive potential of their exopolysaccharides for biomedical applications. CCLM have been the subject of considerable lipid characterisation over many years. This is now being used in intelligent interrogation of strains for lipid biosynthesis and production. Application areas being targeted are biodiesel from microalgae for energy applications and omega-3 long chain polyunsaturated fatty acid biosynthesis for human health applications.
(1)Wealth from Oceans National Research Flagship
(2)Food Futures National Research Flagship
(3)Energy Transformed National Research Flagship
(4)CSIRO Marine and Atmospheric Research, Hobart, 7001, Australia
(5)CSIRO Molecular Health Technologies, Melbourne, Australia
5. Bock C(1), Krienitz L(1), Wei L(1) & Pröschold T(2)
The Dictyosphaerium-morphotype within the Chlorellaceae
The colonial green alga Dictyosphaerium is characterized by the formation of autospores, which remain attached to the remnants of their mother cell wall and are surrounded by a mucilage envelope. Our phylogenetic analyses based on the concatenated SSU and ITS rDNA data set, revealed a relationship of Dictyosphaerium to the Chlorellaceae. This study confirmed the recent division of Chlorellaceae into two sister groups: the Chlorella-clade and the Parachlorella-clade and showed that Chlorella-related coccoid green algae evolved in both clades within the Chlorellaceae. Beside the common spherical phenotype, several Taxa with considerably different morphology cluster within the Chlorella-clade, e.g. Actinastrum Lagerheim (ellipsoidal, coenobial), Didymogenes Schmidle (ellipsoidal, coenobial with or without spines), Meyerella Fawley et K. Fawley (solitary without pyrenoids) and Micractinium Fresenius (colonial, with bristles). However, three strains formerly classified as Dictyosphaerium cluster within the Chlorella-clade. The cells exhibit the same morphology and ultrastructure like typical Chlorella species, but differ in the possession of mucilaginous envelopes. In addition, strains with the Dictyosphaerium-morphotype cluster in the Parachlorella-clade. Further investigations need to be carried out to resolve the relationship of the genus Dictyosphaerium within the Chlorellaceae and whether or not the possession of mucilage is a phenotypical adaptation to environmental factors.
(1)Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Alte Fischerhütte 2, D-16775 Stechlin, Germany
(2)Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Oban, UK
6. Borisova EV(1), Tsarenko PM(1), Rad-Menendez C(2) & Pröschold T(2)
Strains of the Dunaliella species in microalgae culture collection of Ukraine (IBASU-A)
The collection of microalgae culture of Ukraine (IBASU-A) maintains more than 100 strains of Dunaliella Teodoresco. Most of the strains were isolated by N. P. Massjuk, one of founders of the collection, for morphological, systematic, ecological and biogeographical studies in 1960-1970s. Several strains were also used in biotechnology of mass culturing for the production of beta-carotene and high protein feed. In 1980s ten species (2)5 strains) were subjected to microbiological analysis to study their bacterial composition. Since the early 1990s some strains have been used in experiments to study the photo-movement of flagellates. Now IBASU-A has the 14 authentic strains ("type cultures") of Dunaliella, which are still of interesting for taxonomic and phylogenetic studies. The world wide distributed species such as D. salina Teodoresco and D. viridis Teodoresco are presented in collection by the most strains isolated from European and Asian territories (Israel, Russian Federation: Western Siberia, Turkmenistan, Ukraine). D. minuta Lerche with the Eurasian-Australian area of distribution are also presented by the European (Russian Federation, Ukraine) and Asian (Kyrghyzstan, Turkmenistan) strains. D. asymmetrica Massjuk and D. terricola Massjuk having rather restricted Eurasian areas of distribution are presented by the Asian strains from Azerbaijan, Turkmenistan and Uzbekistan. In our preliminary studies we sequenced now all strains of both species and compared their morphology with the original descriptions. The phylogenetic analyses of the concatenated data set of SSU and ITS rDNA sequences resulted the monophyly of the strains designated as D. asymmetrica; however, the strains designated as D. terricola split into two groups. The morphology of only two strains (IBASU-A 130 and IBASU-A 131) fits with the original description. The molecular and morphological characterization of the remaining strains in the collection is still in progress.
(1)M.G. Kholodny Institute of Botany National Academy of Science of Ukraine, Kyiv, Ukraine
(2)Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Oban, UK
7. Bresnan E(1), Graham J(1), Collins C(1), McGunnigal C(2), Mayor DJ(3), Brown L(1) & Turrell E(1)
Phytoplankton cultures: an essential tool for investigating the identity and ecology of Phaeocystis in Scottish waters
The haptophyte genus Phaeocystis is currently being used as an indicator of marine eutrophication as part of the Water Framework Directive and the OSPAR assessment criteria. While recognised as a problem genus in the Southern North Sea, to date there has been little work performed on the identity or ecology of Phaeocystis in Scottish waters. Phaeocystis is difficult to identify to species level using light microscopy. In many instances, the use of preservatives and sample homogenisation prior to analysis, ruptures the characteristic colonies of cells. During 2006, six cultures of Phaeocystis from around the Scottish coast were established using single cell isolation. Molecular analysis, targeting the 18S rDNA confirmed the identity of these cultures as P. globosa. ITS rDNA sequence analysis performed on these cultures revealed variation between Scottish P. globosa and isolates from other geographic regions. Culture experiments examining the response of Scottish isolates to different environmental regimes have commenced. These P. globosa isolates will be lodged with the CCAP once investigations into their morphological identity have been completed.
(1)Fisheries Research Services Marine Laboratory (FRS), 375 Victoria Road, Aberdeen, AB11 9DB, UK
(2)School of Biological Sciences, University of Aberdeen, Zoology Building, Tillydrone Av., Aberdeen, AB24 2TZ, UK
(3)School of Biological Sciences, University of Aberdeen, Oceanlab, Newburgh, Aberdeenshire, AB41 6AA, UK
8. Brown L, Bresnan E, Graham J, Collins C, Lacaze J-P & Turrell EA
Diversity of Alexandrium (Dinophyceae) in Scottish waters
The shellfish industry in Scotland has experienced repeated annual enforced closures of shellfish harvesting areas due to concentrations of paralytic shellfish toxins exceeding the closure limit of 80 µg STX eq.100 g-1. To investigate the diversity and toxicity of Alexandrium species in Scottish waters, phytoplankton cultures for morphological, molecular and toxin analyses were established from sediment and water samples taken from around the Scottish coast. Twenty one Alexandrium cultures were established and four species of Alexandrium identified: A. tamarense (Group I and Group III), A. minutum, A. ostenfeldii and A. tamutum. Identification to species level using thecal plate morphology was confirmed by LSU analysis. Toxin analysis using LC/MS/MS confirmed the production of paralytic shellfish toxins by A. tamarense (Group I) and A. ostenfeldii. Spirolides were also detected in A. ostenfeldii cultures. No toxins were detected in the A minutum, A. tamutum or A. tamarense (Group III) cultures established. This study highlights the diversity of Alexandrium in Scottish waters and also identifies the presence of a number of non-toxic species. All cultures established during this study will be lodged with the CCAP on completion.
Fisheries Research Services Marine Laboratory (FRS), 375 Victoria Road, Aberdeen, AB11 9DB, UK
9. Darienko T(1) & Pröschold T(2)
Molecular phylogeny and systematics of terrestrial Ulvophyceae (Preliminary results)
Successful invasion of the land by marine organisms, or of the sea by freshwater organisms, is generally considered as a relatively rare event in evolution. Among the coccoid green algae, once placed together in the order Chlorococcales, there are a number of marine forms amongst the overwhelming freshwater majority. A similar situation occurs among the filamentous green algae, which are included in the orders Ulotrichales or Chaetophorales. What does this represent? Is this disparity in number a chance event, or is it controlled by biogeographic, systematic or molecular effects? Most recent phylogenetic analyses of SSU rRNA sequences have shown that coccoid and filamentous green algae are distributed among all classes of Chlorophyta. One of these classes, the Ulvophyceae, mostly contains marine seaweeds like Ulva, or marine filamentous microalgae, which grow as epiphytes on other seaweeds. However, new studies showed that there are freshwater and terrestrial species (including symbionts in lichens) among the Ulvophyceae, but very little is known about these species. Most marine coccoid and filamentous green algal species have traditionally been placed within genera dominated by species from freshwater or soil habitats. Molecular data have confirmed that these marine species are members of the Ulvophyceae: In contrast, most of the freshwater and terrestrial species belong to the Chlorophyceae or Trebouxiophyceae. The aim of this study is to investigate species of the genera Dilabifilum, Planophila, Trichosarcina, Pseudendoclonium, Pseudendocloniopsis, Hazenia, Ulothrix and Gloeotilopsis, which were mostly isolated from freshwater, or terrestrial habitats (some are symbionts in lichens) and were considered to be ulvophytes. Ultrastructural studies on some species have shown that the zoospores form the flagellar apparatus (counterclockwise basal body orientation), which is typical for the Ulvophyceae. In addition to ultrastructural features, the presence of a "Codiolum"-stage is characteristic for this algal class. Using a polyphasic approach we compared phylogenetic analyses of SSU and ITS rDNA sequences with morphology and life cycle of these species. Our preliminary phylogenetic analyses of SSU rDNA sequences have revealed that the terrestrial species studied, so far, belong to three independent lineages of the Ulvophyceae called here: Chlorocystis-, Pirula, and Trichosarcina-clades. Strains designated as Dilabifilum sp. are members of different subclades of the Pirula-clade (two strains labelled as Dilabifilum sp. are species of Aphanochaete, a genus of the Chaetophora-clade in the Chlorophyceae). The Tricho-sarcina-clade contains the majority of terrestrial ulvophytes studied (the only marine species belong to the Gomontia-subclade). The Trichosarcina-clade is known in the literature as the Ulothrix-clade or Ulotrichales; however, our morphological studies have revealed that the only species studied, Ulothrix zonata, does not belong to the genus Ulothrix and has to be transferred to a new genus. In our investigations, the strains demonstrated a high degree of phenotypic plasticity depending on the culture conditions (marine or freshwater, liquid or solid media, different temperatures), which could lead to different identifications of the same organism. Based on these results the morphological species concept of these genera will be proven and if necessary, a taxonomic revision will be proposed.
(1)M.G. Kholodny Institute of Botany National Academy of Science of Ukraine, Kyiv, Ukraine
(2)Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Oban, Scotland
10. Day JG(1), Achilles-Day UEM(1), Martinez-Montero ME(2), Müller H(3), Harding K(4) & Benson EE(4)
The use of vitrification for the long-term conservation of protists
Colligative cryopreservation of protists depends on the application of penetrating cryoprotectants (commonly, DMSO or methanol) and slow cooling (e.g. -1oC min-1) to an intermediate sub-zero temperature (usually the temperature of homogeneous ice nucleation -40oC) prior to plunging into liquid nitrogen (-196oC). This approach has been extremely successful in preserving the cells of small, morphologically simple Taxa. Although, this approach is generally applicable, to many organisms some remain freeze-recalcitrant they include most euglenoids, ciliates and many diatoms. These organisms may be preserved by vitrification, an alternative cryopreservation strategy, which is the conversion of an aqueous system to an amorphous, non-crystalline solid, termed the ‘glassy state’. This is achieved by increasing cell viscosity to a critical point termed the glass transition (Tg) temperature. This obviates damage due to ice crystallisation as can occur during slow cooling. A variety of strategies have been applied to achieve the viscosity required for the vitrification of protistan cultures including the use of high concentration cocktails of cryoprotectants, alginate encapsulation and air-drying of cyst suspensions in soil. The successful application of the above approaches to marine diatoms, Euglena gracilis, and Meseres corlissi respectively will be discussed.
(1)Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunbeg, Argyll, UK
(2)Bioplantas Center, University of Ciego de Avila, CUBA
(3)Private Laboratory, Jacob-Burckhardt-Str.18, Konstanz, Germany
(4)Damar, Research Scientists, Conservation, Environmental Science & Biotechnology, Drum Road, Cupar, Fife, KY15 5RJ UK
11. Fraga S, Bravo I, Fernández-Villamarín A, Ramilo I & Rial P
CCVIEO, the Culture Collection of harmful marine microalgae of the Instituto Español de Oceanografía in Vigo, Spain
In order to identify the causative organisms of shellfish toxicity in Galicia, the isolation and culture of suspected organisms started in the 80's. Alexandrium tamarense was isolated and unexpectedly, resulted as non-toxic, while another dinoflagellate, Gymnodinium catenatum was found to be toxic and identified as the main causative organism of PSP in the area. Since then, many toxic strains have been isolated like the benthic dinoflagellate Prorocentrum lima, an okadaic acid producer, the diatom Pseudo-nitzschia australis, a domoic acid producer, or the fish killer raphidophycean Heterosigma akashiwo. While at the beginning, the collection held only local strains, posteriorly, strains from other collections were incorporated for comparative studies. Strains were isolated as well from water samples taken from other areas of Spain or from other countries, like toxic strains of Ostreopsis spp. from the Mediterranean and Brazil, or Alexandrium cantenella, Alexandrium minutum and Protoceratium reticulatum also from the Mediterranean. Non toxic strains, but taxonomically related with toxic ones, or red tide producers are also in the collection, like Alexandrium affine, Prorocentrum spp. or Akashiwo sanguinea. In addition to toxins studies, many of these strains are currently being used also for cell cycle studies and pigment analysis as an aid to taxonomy.
Instituto Español de Oceanografía, Subida á Radio Faro, 50, E-36390 Vigo, Spain
12. Graham J, Collins C, Brown L, Bresnan E, Lacaze J-P & Turrell EA
Alexandrium tamarense (Dinophyceae) in Scottish waters
Contamination of shellfish with paralytic shellfish poisoning (PSP) toxins produced by Alexandrium species, poses a potential threat to the sustainability of the Scottish aquaculture industry. Routine light microscopy (LM) analysis of water samples from around the Scottish coast has previously identified Alexandrium Halim (Dinoflagellata) as a regular part of the spring and summer phytoplankton communities in Scottish coastal waters. In this study, Alexandrium tamarense isolated from sediment and water samples were established in laboratory culture. Species identification of these isolates was confirmed using thecal plate dissections and by molecular characterisation based on their LSU rDNA sequence. Molecular characterisation and phylogenetic analysis showed the presence of two ribotypes of A. tamarense: Group I (North American ribotype) and Group III (Western European ribotype). Assessment of PSP toxin production using hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-MS/MS) showed that A. tamarense Group I produced a complex array of toxins (ca. 2,000 fg STX equivalents cell-1) with the major toxins being C2, neosaxitoxin (NEO), saxitoxin (STX), gonyautoxin-4 (GTX-4) and GTX-3 while A. tamarense Group III did not produce toxins. Historically, it was considered that all Alexandrium species found in Scottish waters produce potent PSP toxins. This study has highlighted the presence of both PSP toxin producing and benign species of A. tamarense. Use of methodology to differentiate between the ribotypes in future monitoring programmes should provide a better picture of the composition of benign and toxic A. tamarense in Scottish waters and help to understand environmental and seasonal conditions influencing their occurrence. The cultures generated in this study will be placed with the CCAP. By placing these species in the public domain, the potential for research on phytoplankton from Scottish coastal waters by the international community is increased.
Fisheries Research Services Marine Laboratory (FRS), 375 Victoria Road, Aberdeen, AB11 9DB, UK
13. Haworth EY, Monaghan E, Devlin G & Durrell L
Fritsch and his Collection of Algal Illustrations
When Felix Eugen Fritsch was Professor of Botany of Queen Mary College, London he began filing any algal illustrations that he received by species, for comparison. At his death this collection came to Dr J.W.G. Lund at the Freshwater Biological Association, Windermere. Together with comprehensive citations, it now contains several million published figures and taxonomic entries on fresh- and brackish-water algae from worldwide distributions.
Freshwater Biological Association, Ferry Landing, Far Sawrey, Ambleside, Cumbria, LA23 1JA, UK
14. Jameson I(1),(2), Blackburn SI(1),(2),(3), Johnston C(2), Albinsson ME(1),(2),(3), Skerratt J(1),(2), Hassler C(1),(2), Robert S(1),(2), Frampton DMF(1),(2), Bolch CJ(4), Wynne J(2), Holmes B(2) & Volkman JK(1),(2),(3)
Environmental applications of the CSIRO Collection of Living Microalgae
The CSIRO Collection of Living Microalgae (http://www.cmar.csiro.au/microalgae/) maintains over 800 strains from 140 genera representing the majority of marine and some freshwater microalgal classes. As part of the Collection, the CSIRO Microalgae Supply Service provides microalgal strains to industry, government research organisations and universities in 50 countries for both research and industry applications. The Collection underpins research within CSIRO and in partnership with collaborators spanning environmental issues, aquaculture and biotechnology. Studies of cultured strains under controlled conditions provide key information on phytoplankton dynamics and harmful algal blooms (HABs) as part of system-wide environmental research for the management of estuarine and coastal aquaculture activities in south-east Tasmania and for understanding the Southern Ocean ecosystem. Aquafin Cooperative Research Centre research includes the interactions of the toxic HAB dinoflagellate Gymnodinium catenatum with bacteria and their influences on algal growth / bloom dynamics and expression of key functional genes, as well as investigations into mortality of G. catenatum and other phytoplankton. Climate research uses cultured Southern Ocean microalgal strains to investigate iron bioavailability and limitation and their effects on phytoplankton dynamics. Outcomes of controlled culture studies using Collection strains include better parameterisation of biogeochemical models and inputs for biological sensor development to better predict phytoplankton dynamics and manage harmful algal blooms.
(1)CSIRO Wealth from Oceans National Research Flagship, Australia
(2)CSIRO Marine and Atmospheric Research, GPO Box 1538, Hobart, Tasmania, 7004, Australia
(3)Aquafin Cooperative Research Centre, Australia
(4)University of Tasmania, School of Aquaculture
15. Jodlowska S, Pniewski F & Latala A
Ecophysiological and morphological study of two diatoms from the Baltic Sea
The experiments were carried out on two diatom strains: planktonic, Skeletonema marinoi (BA-98), and benthic, Fistulifera saprofila (BA-55). The strains were isolated from the Gulf of Gdansk (the Southern Baltic) and are maintained as unialgal cultures in Culture Collection of Baltic Algae (CCBA). The objective of this work was to determine the ecophysiological and morphological properties of the strains. The influence of irradiance and temperature, and their interaction on the growth, pigment concentration and the performance of photosynthesis was investigated in 40-160 μmol photons·m-2·s-1 and 15-25 °C for S. marinoi, and 10-290 μmol photons·m-2·s-1 and 15-30 °C for F. saprofila strains. The experiments showed that both irradiance and temperature are important factors contributing to the variation in diatom growth but with certain differences between two strains. The concentrations of chlorophyll a were significantly higher at low irradiances at each strains, while the carotenoids/chlorophyll a ratio increased with an increase in irradiance. On the basis of P-E curves two mechanisms of photoacclimation were recognized for benthic strain: a change in the number and in the size of PSU. Both Fv/Fm and ΦPSII was higher for cultures grown at low irradiance than at higher one. The morphology was studied using Hitachi S-4700 SEM.
Institute of Oceanography, University of Gdansk, Av. Pilsudskiego 46, 81-378 Gdynia, Poland
16. Kasai F(1), Ishimoto M(1), Erata M(1), Sakayama H(2), Nozaki H(2), Miyaji K(3), Kato S(3), Sano S(4), Morishima H(5), Omori Y(6), Higuchi S(7), Kume O(8), Hirabayashi S(1) & Watanabe MM(9)
Loss of Charales algae from ponds and lakes in Japan and their ex situ conservation
Charales algae are ecologically, as well as evolutionarily, important organisms, which inhabit a lower layer of the littoral zone, suppressing nutrient retrieval from bottom mud and providing nurseries to aquatic organisms. In recent years, those Charales algae have declined in lakes and ponds of Japan. Based on the survey of lakes all over Japan conducted during 1992-1998, the first list of endangered Charales algae was published in 2000 with other groups of algae. After that we have followed up the survey of Charales, specially focusing on the other type of habitat, relatively small and shallow waters such as agricultural reservoirs, ponds and paddy fields, because a large number of species have been reported from such habitats. Based on the follow-up survey, the Red List of Japan (a list of endangered species) was revised and lists 58 Taxa of Charales, as Extinct, Extinct in the Wild, Critically Endangered and Vulnerable species in 2007. The NIES-Collection started maintaining such Charales strains to provide research materials for conservation studies from the mid 1990s. And now, 55 strains, including 10 uni-charalean strains, are maintained.
(1)National Institute for Environmental Studies, Tsukuba, Japan
(2)University of Tokyo, Japan
(3)Toho University, Japan
(4)Funabashi Sibayama High School, Japan
(5)Funabashi Kowagama High School, Japan
(6)Yokosuka City Museum, Japan
(7)Nagano Env. Cons. Res. Institute, Japan
(8)Kagawa Prefecture, Japan
(9)University of Tsukuba, Japan
17. Kynclová A, Škaloud P, Rezácová-Škaloudová M
Molecular diversity and species concept in Synura petersenii (Synurophyceae, Heterokontophyta)
Synura petersenii cells are covered with characteristic siliceous scales with a great deal of intraspecific variation. Besides S. petersenii f. petersenii there is a wide range of other more or less divergent morphotypes. Some of these are unnamed, others have been given status of forma, or previously also of variety. Therefore, molecular investigation of this taxon is necessary to clarify a species concept in S. petersenii. This study is based on the comparative molecular and morphological investigation of S. petersenii cultured strains. Altogether, 25 strains were investigated. The strains have been collected from different localities in the Czech Republic, two others are from USA. ITS sequence analysis has unambiguously distinguished six clearly separated clades within S. petersenii species complex. Ten strains were included in the clade with a wide distribution. The acquired molecular data were then compared with morphology of the siliceous scales. Geometric morphometric methods were used to evaluate scale morphology data of several strains. In our opinion, the different clades represent separated species, clearly defined by 1) unique ITS sequences, 2) specific combination of (hemi-) CBCs in ITS secondary structure, and 3) combinations of several morphological characteristics.
Charles University in Prague, Faculty of Science, Department of Botany, Benátská 2, 12801, Praha 2, Czech Republic
18. Le Gall F, Rigaut-Jalabert F, Marie D, Garczarek L, Viprey M, Gobet A & Vaulot D
Picoplankton diversity in the South-East Pacific Ocean from cultures
In late 2004, the BIOSOPE cruise sailed between the equatorial influenced waters off Marquesas Islands and the nutrient enriched waters of the Chilean upwelling through the extremely oligotrophic Southeast Pacific gyre. We undertook a vigorous effort to isolate novel photosynthetic picoplanktonic eukaryotes following two strategies: enrichment of samples with culture medium and sorting of specific populations by flow cytometry. After purification, the 212 strains obtained were characterized morphologically and genetically (1)8 S rRNA). Among the characterized strains, the largest number belonged to stramenopiles with in particular 38 related to species Pelagomonas calceolata. Strains belonging to the recently described genera Bolidomonas and Florenciella were re-isolated for the first time since their description. Two other abundant groups were Prasinophyceae, and Prymnesiophyceae, especially Phaeocystis and Emiliania. A few heterotrophic flagellates, all belonging to known genera, and some unicellular Synechococcus cyanobacteria strains were isolated. Very few novel taxa have been obtained. One set of strains is related to Prasinoderma coloniale but their sequences are sufficiently different from the latter to probably belong to a new genus or species. The sequences of two other strains (unfortunately lost thereafter) were affiliated to stramenopile environmental sequences and probably corresponded to a novel algal class.
Station Biologique de Roscoff, UMR 7144, CNRS et Université Pierre et Marie Curie, Place G. Tessier, 29682, Roscoff, France
19. Oivanen P, Saari L, Wahlsten M & Sivonen K
Cryopreservation of cyanobacteria in UHCC (University of Helsinki Cyanobacteria Culture Collection)
University of Helsinki Cyanobacteria Culture Collection (UHCC) is a collection built up during the research work of the "Cyanobacteria research group" at the department of Applied Chemistry and Microbiology in the University of Helsinki from the late 1980's until today. Nearly 1000 strains are deposited in the collection, of which about 200 are axenic. This culture collection contains a number of planktonic, benthic and symbiotic strains which produce toxin and other bioactive compounds. All strains are maintained by serial transfer. However, this is not an optimal preservation method, due to the potential for genetic drift and human handling errors. To reduce the risk of genetic change and to reduce the maintenance work cryopreservation was initiated. A total of 30 strains were frozen and 15 strains were thawed out. Seven strains of genera Microcystis, Nostoc, Oscillatoria and Snowella survived freezing and thawing. Eight strains of genera Anabaena, Aphanizomenon, Nodularia and Planktothrix did not. The recovery was very slow and it took at least one month of incubation on agar plate, until growth could be observed. Future attempts will include additional cryopreservation methods and their optimization to improve the preservation of the culture collection.
Department of Applied Chemistry and Microbiology, PO Box 56, University of Helsinki, FIN-00014, Finland
20. Pazoutová M & Škaloud P
Phylogeny and the generic concept of capsal green algae, family Radiococcaceae
The family Radiococcaceae, defined broadly as coccoid green algae with mucilaginous cover reproducing only by autospores, is one of the most taxonomically problematic groups among green algae. Radiococcaceae are common organisms of freshwater as well as terrestrial habitats worldwide and they have been studied for more than 100 years, yet their taxonomy remains unclear. The generic concept differs from author to author and some of the traditional morphological traits, like the presence of mucilage itself, proved to be unreliable. The first molecular study on members of the family (Wolf et al. 2003) showed the polyphyly of the group, with species divided into two different classes of green algae, Chlorophyceae and Trebouxiophyceae. We examined the phylogenetic position of 25 strains of Radiococcaceae from several culture collections representing different traditional species with different morphology. In accordance with the previous study, the strains were placed in two classes, 10 strains in Chlorophyceae and 15 in Trebouxiophyceae. Moreover, we distinguished 7 distinct clades in the former and 5 in the latter class. These molecular data are well supported by some morphological as well as ecological characteristics. We discuss possible revisions of the generic concept within this traditionally recognized group.
Department of Botany, Charles University Prague, Czech Republic
21. Pingle SD
Culture studies on Chaetonema irregulare Nowakowski
Chaetonema irregulare Nowakowski alga came up in an enrichment culture started with soil samples from Pashan Lake (located 4km away from University of Pune campus). The alga is first time reported from India; hence it was isolated and cultured for further detail studies. Alga came up as heterotrichous free growth with irregularly branched creeping filaments, erect system was developed from creeping filaments. A small number of unicellular hyaline hairs (setae) with slightly swollen bases were observed to arise on terminal or intercalary cells of the erect filaments as well as the creeping filaments. Detailed study of structure and reproduction were made using following media: BBM Agar and Liquid Medias, Biphasic medium, Modified BBM, Chu 10 and De’s liquid media. Off all these the biphasic, Chu 10, and De’s media were found just suitable for normal morphological growth and reproduction of the alga. The normal BBM medium did not showed the development of setae where as in the absence of NaNO3, K2HPO4 and KH2PO4 development of setae was observed. Both sexual and asexual reproductions were observed in the culture media: Biphasic medium, Modified BBM, Chu 10 and De’s liquid media.
Padmashri Vikhe Patil College of Arts Science and Commerce Pravaranagar, Maharashtra, 413713, India
22. Pniewski F & Latala A
Growth response of Baltic microalgae from CCBA collection to salinity changes
The Baltic Sea is biggest brackish sea in the world. Its modern history goes back merely 13.000 years. Biological diversity of the sea is relatively poor since the short history did not allow to create a large number of species. In spite of this, its flora and fauna are unique and consist of freshwater, marine as well as brackish water species. Culture Collection of Baltic Algae (CCBA) has existed since 1980s. At present the collection contains more than hundred strains which cover 4 classes of algae including Cyanophyceae, Dinophyceae, Bacillariophyceae and Chlorophyceae. Microalgae are maintained in liquid f/2 and BG-11 media prepared from the Baltic water. The following culture conditions are applied: salinity 6.9 PSU, temperature 20 ºC, PAR intensity 40 µmol photons·m-2·s-1 and 16:8 L:D photoperiod. The best growth of most green algal strains was observed in freshwater (Monoraphidium and Scenedesmus genera). Although, some of them grew best in low salinities (e.g. Chlorella vulgaris, Oocystis submarina). All tested diatoms showed good growth in a wide range of salinities with a optimum at intermediate salinities (e.g. Amphora coffeaeformis, Bacillaria paxillifer, Haslea spicula). Similar tendency was observed in blue-green alga Nodularia spumigena, while Phormidium amphibium occurred to be a marine species.
Laboratory of Marine Plant Ecophysiology, Institute of Oceanography, University of Gdansk, Pilsudskiego 46, 81-378 Gdynia, Poland
23. Pribyl P
Search through CCALA for lipid-rich algal strains. The significance of Culture Collections.
Under specific conditions, some microalgae can accumulate substantial amounts of neutral lipids, predominantly triacylglycerols (TAGs), intracellularly. Biodiesel derived from algal TAGs seems to be the only one option of renewable transport fuels that is capable of meeting the global demands. The research into algal lipids is going to be in great demand in this context. Microalgae appear to be the substantial source not only of TAGs but also of highly valued polyunsaturated fatty acids applicable as nutritional supplements. Specialised algal collections should efficiently serve as background for this research. It is likely that many promising (although not revealed) algal strains are kept in many CC worldwide. This is the reason for establishment of COllection of BIotechnological Exploitable Microalgae (COBIEM) as the subcollection of CCALA for further investigations. The algal strains rich in neutral lipids will comprise the ground of COBIEM, therefore it is of high importance to find oleaginous algal strains. Part of CCALA (mainly Xanthophyceae, Eustigmatophyceae and Chlorophyceae) was scanned for neutral lipids recently: rapid estimation of lipid content of algae was done using lipophilic dye Nile red, which exhibit characteristic fluorescence in a nonpolar environment, primarily storage lipid droplets. Here, I present preliminary results of this long-term effort.
CCALA, Algological Centre, Institute of Botany, Academy of Sciences of the Czech Republic, v.v.i., Dukelska 135, CZ 37982 Trebon, Czech Republic
24. Pröschold T(1), Achilles-Day UEM(1), Day JG(1) & Krienitz L(2)
The systematics of "Zoochlorella" revisited employing polyphasic approaches
For more than hundred years, symbiosis of green algae with protozoa and invertebrates has been studied and is still for general interest in modern biology. Endosymbiotic green algae are widely distributed in different ciliates (e.g. Paramecium, Stentor), heliozoa (e.g. Acanthocystis) and invertebrates (e.g. Hydra, Spongilla) and were traditionally classified as members of the genera Chlorella or Zoochlorella. We studied ten strains isolated from different hosts and from different geographical origins using polyphasic approaches (SSU and ITS rDNA sequences including their secondary structures, morphology, and virus sensitivity). Phylogenetic analyses have revealed the polyphyletic origin of endosymbiotic green algae. The strains examined belong to four independent lineages within the Trebouxiophyceae (Chlorophyta). Based on our results, we propose a taxonomic revision of endosymbiontic "Chlorella"-like green algae. We transfer some of them to other genera and propose two new species.
(1)Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Dunbeg by Oban, UK
(2)Institute for Freshwater Ecology and Inland Fisheries, Alte Fischerhütte 2, Stechlin-Neuglobsow, Germany
25. Skaloud P, Neustupa J, Veselá J, Eliáš M & Škaloudová M
Algal culture collection in Prague - CAUP: a platform for education and taxonomic research
The Culture Collection of Algae at the Department of Botany, Faculty of Natural Science, Charles University in Prague, was established by B. Fott in 1961. The culture collection originated by merging a large number of green algal and heterokont cultures isolated by B. Fott with part of the former E.G. Pringsheim’s algal culture collection. Presently, CAUP represents rather small algal collection, holding 194 algal strains. The leading idea of our culture collection is not to conserve a very broad range of organisms, but to serve as a platform for education and taxonomic research in our department. Plans for the near future include the introduction of cryopreservation techniques to preserve a large number of strains isolated during various taxonomic studies, and to continue in detailed image documentation of maintained strains. In addition, we present several recent taxonomical studies connected with the CAUP culture collection: Taxonomy of the genus Dictyochloropsis (Trebouxiophyceae); striking phylogenetic position of Kentrosphaera (Ulvophyceae); morphological and molecular diversity of Frustulia saxonica (Bacillariophyceae); morphological variation and phylogeny of Micrasterias (Zygnematophyceae); phylogeny of Eustigmatophyceae; and morphology and phylogeny of Mallomonas kalinae (Synurophyceae).
Charles University in Prague, Faculty of Science, Department of Botany, Benátská 2, 12801, Praha 2, Czech Republic
26. Skaloud P(1) & Peksa O(1),(2)
Phylogeny, morphology and taxonomic revision of the symbiotic alga Asterochloris (Trebouxiophyceae, Chlorophyta)
The genus Asterochloris is one of the most numerous lichen photobionts. We present a revision of the genus, based on the comparative morphological and molecular investigation of 34 cultured strains. For all strains, we determined nuclear-encoded ITS rDNA and partial actin I sequences. In addition, we studied morphology and reproduction by conventional light and confocal microscopy. We recognize and designate 13 monophyletic lineages, 7 of which represent currently described species (Asterochloris phycobiontica, Trebouxia erici, T. excentrica, T. glomerata, T. irregularis, T. italiana and T. magna). Concurrently with proposing new combinations for former Trebouxia species, we describe 6 species new to science (A. echinata, A. friedlii, A. gaertneri, A. leprariae, A. lobophora and A. woessiae); furthermore, T. pyriformis is reduced to a synonym of A. glomerata. According to the polyphasic approach, all species can be clearly defined by unique actin and ITS sequences, combination of selected morphological characters (chloroplast morphology, cell shape, etc.), and hemi-CBCs in ITS secondary structures. Asexual reproduction takes place by autospores, aplanospores and zoospores, which often bear unique posterior extensions. Isogamous sexual reproduction was scarcely observed.
(1)Charles University in Prague, Faculty of Science, Department of Botany, Benátská 2, 12801, Praha 2, Czech Republic
(2)The West Bohemian Museum in Pilsen, Kopeckého sady 2, 30100, Plzen, Czech Republic
27. Tanabe Y(1), Erata M(2), Mori F(2), Yumoto K(2), Sato M(2), Ishimoto M(2), Kawachi M(1), Watanabe MM(3) & Kasai F(1)
NIES collection of Microcystis aeruginosa: including genotypic data
Microcystis aeruginosa is a toxic bloom-forming cyanobacterium often associated with eutrophication of freshwater environments worldwide. An increasing public health risk and environmental problems caused by the bloom and cyanotoxin (microcystins) produced by M. aeruginosa underscore the needs of the systematic and comprehensive collection of this species to serve as research material for researchers working on M. aeruginosa. NIES-Collection currently lists more than a hundred of strains of M. aeruginosa: these include both toxic and nontoxic strains, those representing all five known morphospecies, and type species NIES-843T of which the whole genome sequence information is available. We have recently further improved the quality of our collection by adding multilocus genotypic information (Multilocus Sequence Typing, MLST) to each strain, thereby researchers are able to select strains in response to the needs of their investigation.
(1)National Institute for Environmental Studies, Tsukuba, Japan
(2)Global Environmental Forum, Japan
(3)University of Tsukuba, Japan
28. Vaulot D, Gourvil P, Le Gall F, Rigaut-Jalabert F, Marie D & Probert I
The Roscoff Culture Collection (RCC)
The Roscoff Culture Collection (RCC) currently holds over 1300 strains with emphasis on marine cyanobacteria (mainly Prochlorococcus and Synechococcus) and microalgae (mostly picoplankton from several classes, but also >350 strains of Haptophyta, mainly coccolithophores, recently transferred from the Algobank-Caen Collection). Most strains were isolated from the English Channel, the Mediterranean Sea, the Equatorial Pacific, the Tropical Atlantic, the Red Sea and the North Sea. The main strategic focus of the collection remains the isolation, maintenance, characterization and distribution of strains used for fundamental research. In this context, multiple strains of several Taxa (both prokaryote and eukaryote) that exhibit cryptic speciation and/or ecotype variability are maintained, as well as novel isolates from several algal classes. The collection includes the majority of marine cyanobacterial and microalgal strains for which the full genome sequencing is complete or in progress. A large fraction of strains are characterized based on morphological/ultrastructural features, SSU/LSU rDNA sequences and pigment analyses. Images (LM and EM) and taxonomic information on culture strains has been integrated into the Plankton*Net web resource (http://planktonnet.sb-roscoff.fr/index.php). The RCC website (http://www.sb-roscoff.fr/Phyto/RCC) has recently been redesigned to improve accessibility.
Station Biologique de Roscoff, UMR 7144, CNRS et Université Pierre et Marie Curie, Place G. Tessier, 29682, Roscoff, France
Culture Collection of Algae and Protozoa (CCAP)
Dunstaffnage Marine Laboratory • Dunbeg • Oban • PA37 1QA • UK
Tel: +44 (0)1631 559000 • Fax: +44 (0)1631 559001 • Email: ccap@sams.ac.uk • Web: www.ccap.ac.uk
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