Fottea 10(1): 141–144, 2010 141

First report of Makinoella tosaensis Ok a d a (Chlorophyta, Chlorococcales, Oocystaceae) outside East Asia

František Hi n d á k & Alica Hi n d á k o v á

Institute of Botany, Slovak Academy of Sciences, Dúbravská cesta 9, SK–84523 Bratislava, Slovakia; e–mail: [email protected], [email protected]

Abstract: The morphology of Makinoella tosaensis Okada 1949, a chlorococcalean coenobial alga described from Japan and since then observed only in Korea, has been studied under the LM from field material and in laboratory cultures. Its collection from a small fountain in the campus park of the Slovak Academy of Sciences in Bratislava, Slovakia, is the first record outside East Asia. The morphology of coenobia and cells in this material is in agreement with published data of the species.

Key words: Chlorococcales, coenobial green algae, morphology, taxonomy, Slovakia

Introduction & Fo t t 1983). Outside of Japan (cf. http:// protist.i.hosei.ac.jp/PDB/Images/chlorophyta/ In the algal flora of Slovakia project, special Makinoella/index.html), it was observed only in attention was paid to small artificial water bodies Korea (He g e w a l d et al. 1999). The strain SAG such as urban fountains. The diversity of algae 28.97, isolated by Hegewald from Korea, is kept was investigated in several fountains within the at the Algal Collection in Göttingen, Germany. city limits of Bratislava (Hi n d á k 1977, 1980 a, b, The ultrastructure of this culture was investigated 1984, 1988, 1990, 1996; Hi n d á k & Hi n d á k o v á in detail by Sc h n e p f & He g e w a l d (2000). This 1994, 1998; Hi n d á k o v á & Hi n d á k 1998), strain was also used for molecular analyses and including a small basin located at the campus compared with some other representatives of the of the Slovak Academy of Sciences (SAS) at Oocystaceae (He p p e r l e et al. 2000). Patrónka. From this locality a new tetrasporalean alga, Chlamydocapsa mucifera Hi n d á k 1980, was described, and an interesting green alga Material and methods Oocystis nephrocytioides (Hi n d á k 1977, 1980a) was observed which had originally been recorded Algal samples were collected from the water or by Fo t t and Ča d o from an artificial channel near periphyton growing on the concrete walls of a small Lake Ohrid, Macedonia, in 1966 (cf. Ko m á r e k & basin with fountain on the campus of the Slovak Academy of Sciences in Bratislava, Western Slovakia, Fo t t 1983). between July 2009 and January 2010 (Fig. 1). The In this contribution the morphology of basin is supplied by tap water, pH 7.2. Studies were cells and coenobia of the autosporic chlorococcal performed on living material. Light microscope alga Makinoella tosaensis found in this specific observations were carried out with a Leitz Diaplan man–made reservoir at the SAS campus is microscope, and photomicrographs were taken with presented. As the only representative species of a Wild Photoautomat MPS45. Preserved material the genus, it was described from Japan by Ok a d a in formaldehyde is stored at the Institute of Botany (1949) and studied by Ka m i y a (1956). The taxon SAS, Bratislava. Living material has been deposited was also reported by Ya m i g s h i & Ak i y a m a in as strain Hindák 2009/15 at the Culture Collection Photomicrographs of the Freshwater Algae (1984) of Autotrophic Organisms (CCALA) in Třeboň, the and mentioned in several books on green algae Czech Republic. but without adding any geographical data from other countries (e.g. Bo u r r e l l y 1966; Ko m á r e k 142 Hi n d á k & Hi n d á k o v á : Makinoella tosaensis outside Asia

Daughter cells remaining in the conspicuously enlarged mother cell wall for a long time, finally released by its gelatinisation. Occurrence: colonies were found quite frequently and abundantly in the water or on the concrete walls of the basin among other cyanophytes and algae [we use here the classification system by Hi n d á k & Hi n d á k o v á (1998), that has been adopted in the algal flora of Slovakia databank]. The colonies were accompanied by various species of cyanophytes (in the periphyton usually by Gloeocapsa 1 sp., Leptolyngbya, Pseudanabaena catenata, Fig. 1. Locality where Makinoella tosaensis Okada was Phormidium tenue, Calothrix stagnalis) or by collected: a basin with fountain in the campus area of the different types of algal flagellates (Gymnodinium Slovak Academy of Sciences at Patrónka, Bratislava. sp., Cryptomonas ovata, Rhodomonas rubra, Chlamydomonas debaryana, Euglena cf. caudata), chlorococcal algae (Chlorella vulgaris, Coelastrum Results and discussion reticulatum, Coenocystis ovalis, Dictyosphaerium tetrachotomum, Didymocystis inconspicua, Makinoella tosaensis Ok a d a 1949 (Figs 2–5) Golenkiniopsis solitaria, Kirchneriella aperta, K. obesa, Monoraphidium arcuatum, M. contortum, Coenobia free, usually 4–8–16–celled, rarely Oocystella solitaria, Pediastrum boryanum var. 2– or 32–celled or cells solitary, embedded by longicorne, Pseudodictyosphaerium minutum, wide, distinct, hyaline, slightly perpendicularly Quadrigula sp., Scenedesmus abundans, stratified mucilaginous envelopes, mostly clearly S. acuminatus, S. armatus, S communis, S. visible by radiating fibrils or radial arranged maximus, S. naegelii, S. obliquus, S. opoliensis, endogloeic rod–like bacteria. Four–celled S. serratus, Siderocystopsis fusca); ulotrichacean coenobia square, with cells in one plane, touching algae (Elakatothrix spirochroma, Geminella each other subpolarly, inner cells in young stages interrupta, Koliella longiseta, Oedogonium sp., and during autosporulation touching each other Cladophora glomerata) or zygnematophycean by their longitudinal sides. Subsequent cell algae (Mougeotia spp., Closterium incurvum, division in 4–celled coenobia perpendicular to Cosmarium leave). the plane of the coenobium, hence syncoenobia Makinoella tosaensis was originally becoming rectangular, oval or irregular. Cells in placed by its author in the family Oocystaceae, adult 4–celled coenobia or later in syncoenobia a classification that was followed by Bo u r r e l l y arranged rhomboidally, with inner space. Old (1966). On the basis of colony development 16–celled syncoenobia usually disintegrating into Ko m á r e k & Fo t t (1983) assigned it to the family solitary 4–celled coenobia or, very rarely, into Scenedesmaceae, but noted that according the solitary cells. Dimensions of 4–celled coenobia original diagnosis the alga could also be classified with mucilaginous envelopes 65–70 × 50–65 µm, in the Xanthophyceae, because of the absence of 16–celled coenobia 80–90 × 60–80 µm, 32–celled pyrenoids and presence of multiple chloroplasts. coenobia to 130 × 105 µm. The ultrastructural studies performed by Cells broad ellipsoidal to irregularly oval, He g e w a l d et al. (1999) and Sc h n e p f & He g e w a l d slightly asymmetrical, 12–18–(23) × 8–12–(15) (2000) proved the presence of many chloroplasts µm, during autosporulation to 28 × 18 µm. Cell with a distinct starch envelope associated with walls thick, smooth, without incrustations or the pyrenoids. The classification of Makinoella conspicuous polar thickenings. Chloroplasts tosaensis in the Oocystaceae received full support 8–16 or more, bright green, parietal, discoid to also from the molecular analyses by He p p e r l e et slightly asteroid, each with a large central starch– al. (2000). associated pyrenoid. The 4–celled flat coenobia, with two cells Reproduction by autocoenobium with forming an inner pair and two cells forming mostly 4, less frequently with 2 autospores. an outer pair, touching each other subpolarly, Fottea 10(1): 141–144, 2010 143

2 3

4 5

Figs 2–5. Coenobia of Makinoella tosaensis Okada from natural material: (2) coenobia at low magnification; (3, 4) autosporulation in 4-celled coenobia; (5) disintegration of a syncoenobium. Scale bar 20 mm. resemble colonies of the genus Tetrachlorella consider M. tosaensis to be an endemic species Ko r s h i k o v (Ko r s h i k o v 1953; Ko m á r e k & Fo t t for East Asia. Its finding in Central Europe shows 1983; Hi n d á k 1984). However, the internal that this species has probably a cosmopolitan space of coenobia in M. tosaensis is rectangular, distribution, occurring in temperate zone (Japan, rhomboidal or asymmetrical, with the exception Korea, Slovakia). Our material does not differ of young stages of cells or during autosporulation significantly from published descriptions of the when inner cells are connected or touched by their species. Under laboratory conditions, the alga longer sides. grows well both in liquid and on agar BBM media. Makinoella tosaensis shares the feature We were successful in isolating several strains, of the numerous discoid chloroplasts, each from which one strain, Hi n d á k 2009/15, was with a pyrenoid, with some representatives of selected to be deposited at the Culture Collection the genus Oocystis A. Braun, namely the group of Autotrophic Organisms (CCALA) in Třeboň, of O. solitaria Wi t t r . in Wi t t r . et No r d s t . (cf. the Czech Republic. Ko m á r e k & Fo t t 1983; Hi n d á k 1984). However, contrary to O. solitaria, cells in M. tosaensis are Acknowledgements never lemon shaped, because of the absence of The authors are obliged to Dr. H. Sluiman, Edinburgh, conspicuous polar thickenings. Only rarely very for valuable comments and correcting the English. This slight polar thickenings were seen in a culture, study is a part of APVV 0566–07 and VEGA projects Nos 2/7069/27 and 2/0130/10 supported by the Slovak strain Hi n d á k 2009/15. Academy of Sciences. The occurrence of the species in small water basins seems to be typical. In Japan M. tosaensis has been recorded from several places, References in similar small water bodies, mostly in park ponds, and in Korea from a small campus pond Bo u r r e l l y , P. (1966): Les algues d´eau douce. Initiation in the Sookmyung Women´s University in Seoul à la systématique. Tome I: Les algues vertes. – (He g e w a l d et al. 1999). Until recently we could 512 pp., Ed. N. Boubée, Paris. 144 Hi n d á k & Hi n d á k o v á : Makinoella tosaensis outside Asia

He g e w a l d , E., Sc h n e p f , E. & Je o n , S.L. (1999): Report cyanobacteria and algae of Slovakia], p. 12 on Makinoella tosaensis Okada (Chlorophyta, – 100. In: Ma r h o l d K. & Hi n d á k F. (eds), Oocystaceae), a new species to Korea. – Algae Zoznam nižších a vyšších rastlín Slovenska 14: 87-90. [Checklist of non–vascular and vascular plants He p p e r l e , D., He g e w a l d , E. & Kr i e n i t z , L. (2000): of Slovakia]. – Veda, Bratislava. Phylogenetic position of the Oocystaceae Hi n d á k o v á , A. & Hi n d á k , F. (1998): Green algae of (Chlorophyta). – J. Phycol. 36: 590–595. five city fountains in Bratislava (Slovakia). – Hi n d á k , F. (1977): Studies on the chlorococcal algae Biologia 53: 481–493. (Chlorophycea). – Biologické Práce 23/4. – 192 Ka m i y a , T. (1956): A report on the reproduction of pp., Veda, Bratislava. Makinoella tosaensis Okada. – J. Jap. Bot. 31: Hi n d á k , F. (1980a): Studies on the chlorococcal algae 257–261. (Chlorophyceae). II. – Biologické práce 26/6. – Ko m á r e k , J. & Fo t t , B. (1983): Chlorophyceae 196 pp., Veda, Bratislava. (Grünalgen), Ordnung: Chlorococcales. – In: Hi n d á k , F. (1980b): Three new species of green algae Huber–Pestalozzi, G. (ed.): Die Binnnengewässer (Chlorophyceae). – Preslia 52: 289–298. 16/7. – 1044 pp., E. Schweizerbart´sche Hi n d á k , F. (1984): Studies on the chlorococcal algae Verlagsbuchhandlung, Stuttgart. (Chlorophyceae). III. – Biologické práce 30/1. Ko r s h i k o v , O.A. (1953): Pidklas Protokokovi – 310 pp., Veda, Bratislava. (Protococcinae). – Vizn. Prisnovod. Vodorostei Hi n d á k , F. (1988): Studies on the chlorococcal algae Ukrainskoi RSR, Kiev, 5: 1–436. (Chlorophyceae). IV. – Biologické práce Ok a d a , Y. (1949): Makinoella tosaensis, a new genus 34/1–2. – 264 pp., Veda, VSAV, Bratislava. of he Oocystaceae. – J. Japan. Bot. 24: 166– Hi n d á k , F. (1990): Studies on the chlorococcal algae 168. (Chlorophyceae). V. – Biologické Práce 36. – Sc h n e p f , E. & He g e w a l d , E. (2000): The ultrastructure 228 pp., Veda, Bratislava. of Makinoella tosaensis Okada (Chlorophyta, Hi n d á k , F. (1996): Kľúč na určovanie nerozkonárených Oocystaceae). – Algological Studies 97: 79–91. vláknitých zelených rias (Ulotrichineae, Ya m a g s h i , T. & Ak i y a m a , M. (1984): Photomicrographs Ulotrichales, Chlorophyceae) [Key to of the Freshwater Algae 1: 58. unbranched filamentous green algae http://protist.i.hosei.ac.jp/PDB/Images/chlorophyta/ (Ulotrichineae, Ulotrichales, Chlorophyceae)]. Makinoella/index.html; Photomicrographs of – Bull. Slov. Bot. Spoločn., Bratislava, Supl. 1: the Freshwater Algae, 1984. 1–77. Hi n d á k , F. & Hi n d á k o v á , A. (1994): Some interesting observations on desmids (Conjugatophyceae). – Biologia 49: 525–533. © Czech Phycological Society Hi n d á k , F. & Hi n d á k o v á , A. (1998): Zoznam siníc Received December 23, 2009 a rias Slovenska [Checklist of cyanophytes/ Accepted January 15, 2010