126 Proceedings of IV Balkan Botanical Congress • Sofia • 2006

Remarkable and newly recorded aeroterrestric cyanoprokaryotes and algae in Bulgaria

Maya P. Stoyneva1 & Georg Gärtner2

1 Department of Botany, Faculty of Biology, University of Sofia "St. Kliment Ohridski", 8, D. Tsankov Blvd., 1164 Sofia, Bulgaria, e-mail: mstoyneva@abv.bg;

2 Institute of Botany, University of Innsbruck, Sternwartestr. 15, A–6020 Innsbruck, Austria, e-mail: georg.gaertner@uibk.ac.at

Abstract. Studies on aerophytic and terrestric algae and Cyanoprokaryota from nature and after application of cultivation techniques are presented; 9 of the recorded taxa are firstly reported or firstly proved by cultures for the Bulgarian flora.

Key words: aeroterrestric algae, Bulgaria, cultures, Cyanoprokaryota

Introduction

The diversity and taxonomy of soil- and aerophytic al-gae and Cyanoprokaryota is still poorly known world-wide. In Southeast European countries including Bul-garia studies of edaphophytes and aerophytes are also scarce with some exceptions like investigations of cave algae (e.g., Petkoff 1939; Draganov & Dimitrova 1968; Draganov & Dimitrova-Burin 1980; Stoyneva & al. 2002) or the studies of soil algae and Cyanoprokaryota (e.g., Draganov 1964, 1968; Stoyneva 1991; Draganov & al. 1992). As taxonomic studies of aeroterrestric algae in most cases need special cultivation techniques under standard laboratory conditions, research in this field of algology is underrepresented if compared with aquatic research (Dindal 1990; Johansen & Shubert 2001).

During scientific cooperation projects between the algological working groups at the Botanical Institutes of Sofia and Innsbruck aerophytic algal material from different localities in Western Bulgaria was collect-ed and together with soil samples taken in cultures. Among the algae investigated 9 were firstly recorded in cultures or new for the Bulgarian flora.

Material and methodsSoil samples were taken from the soil surface and 2 cm beneath it from the Boris Garden in Sofia (B). Incubation

of soil material on agar-solidified media followed the method described in Ettl & Gärtner (1995). Water dilut-ed soil material was sprayed over the surface of agar Petri dishes. After 6 to 8 weeks of cultivation at room temper-ature, on a north-faced window, purification treatments on agar plates were made according to Ettl & Gärtner (1995) and Andersen & Kawachi (2005). Clonal cultures were used for taxonomic investigation. Some of the iso-lated strains are kept in agar tubes for a long-time collec-tion at the Botanical Institutes in Sofia and Innsbruck.

Aerophytic material was collected by scraping from rock surfaces of Erma Gorge in the vicinity of the town Trun (T) and from different sites in the cave Prokhod-na nearby Karloukovo village (K). Enrichment cultures were again made on agar plates, pure cultures were es-tablished by the same procedures as for soil algae.

Light microscopy was done with a Reichert Diapan and an Olympus SH2 microscope, photos taken with a ProGres C 10 digital camera system. Determinations follow the adequate references, as Geitler (1932), Ettl & Gärtner (1995), John & al. (2002), Wehr & Sheath (2003) and Komárek & Anagnostidis (1999, 2005).

Results and discussionBelow a commented list of isolated algae in alphabet-ical order is provided, where taxa newly recorded and proved in cultures for Bulgaria are marked with aster-

127Scientific Area B • Plant diversity – past and present

isks (*). Abbreviations of the localities follow the de-scriptions in Material and methods.

CyanoprokaryotaChroococcus tenax (Kirchn.) Hieron.

K, common in "Tintenstrichen" on walls of Prokhodna cave, ca. 5 metres inside of entrance. Cells 13–17 μm in diameter.*Chroococcus westii J.B. Petersen

K, nearby the entrance of Prokhodna cave, between juvenile stages of lichens (e.g., Dermatocarpon mini-atum (L.) Mann). Colonies usually of 2–4 cells, 14–15 μm, with granular content and conspicuous pink to violet colour. Envelopes distinctly striated.Gloeocapsa aeruginosa Kütz.

K, between mosses and young thalli of Dermato-carpon miniatum in the entrance of Prokhodna cave. A widespread aerophytic species on wet calcareous rocks and a main part of the "Tintenstrich" flora (Geitler 1932). It differs from G. sanguinea (C. Agardh ) Kütz. by colourless envelopes and smaller cells (Jaag 1945; Komárek & Anagnostidis 1999). Colonies in our ma-terial were 12–15 μm in diameter.Gloeocapsa cf. aeruginosa Kütz.

K, on moist walls near the entrance of Prokhodna cave. Colonies 12–15 μm, differs from the type species by its granular envelopes.Nostoc commune (Vaucher) Bornet et Flahault

T, tiny but macroscopically visible colonies in the surrounding of the jelly lichen Collema cristatum (L.) F. Weber ex F.H. Wigg. Cells short barrel-shaped, 4.5–6.5 μm wide, heterocysts spherical.Phormidium kuetzingianum (Kirchn.) Anagnostidis et Komárek

K, forming thin greyish layers on moist rocks to-gether with Gloeocapsa spp. along a "Tintenstrich" nearby the entrance area of the cave Prokhodna. Tri-chomes 3.5–4 μm wide, in thin sheaths.Phormidium sp.

K, forms greyish-violet bundles of filaments, in cul-ture without sheaths, resembles P. violaceum (Wallr. ex Gomont) Anagnostidis but differs in morphology of apical cell, without calyptra. Trichomes 5–7 μm wide, almost isodiametric.*Tolypothrix tenuis var. calcarata (Schmidle) B.A. Whitton

K, in Prokhodna cave together with crustaceous li-

chens (e.g., Caloplaca xantholyta (Nyl.) Jatta). The alga formed a greyish-blue to greenish-grey felt when moist. Cells 7–8 μm wide and 8–10 μm long, not constricted at cross walls, false branches single, subtended by 1-pored heterocysts. Sheath thin, close to the trichome and par-tially strongly calcified (Plate I, Figs 1, 2).Tolypothrix sp.

K, forming dark greyish crusts between moss pr-otonemata near the entrance of Prokhodna cave. Tri-chomes 9–11 μm wide and 4–6 μm long, with firm sheath. Resembles a form of T. byssoidea (Hassall) Kirchn.

Xanthophyceae

*Chloridella sp.B, cells in culture 7–9 μm wide, spherical, with thin

and smooth cell wall; chloroplasts 1 to 2, cup-shaped, without pyrenoid; a characteristic oil droplet be-tween the chloroplasts. Reproduction by 4 autospores. The isolated strain resembles C. neglecta (Pascher et Geitler) Pascher but differs in the number of chloro-plasts. The similar species, C. cystiformis Pascher has a thickened, brownish cell wall (Pascher 1939).*Heterococcus sp.

B. Heterococcus species are characteristic mem-bers of the soil algal flora; their determination is pos-sible on culture studies which contain juvenile, adult and reproductive stages. The isolated material shows branched filaments, cells 5–6 μm wide and up to 18 μm long. Reproduction with zoospores and aplanospores, but still not properly documented due to need of long-time cultivation (Plate I, Figs 3–5).

Ulvophyceae

Trentepohlia aurea (L.) Mart.T, on moist calcareous rocks between small thalli

of Nostoc commune (Vaucher) Bornet et Flahault and the lichen Gyalecta jenensis (Batsch) Zahlbr. Cells 10–25 μm wide, 20–35 μm long, apical cells with distinct pectose caps. Zoosporangia seldom observed in fresh material.

Chlorophyceae

*Chlorococcum ellipsoideum Deason et BoldB, cells ellipsoidic or irregular, 7–8 μm wide and

11 μm long, chloroplast parietal with some incisions, one pyrenoid with smooth continuous starch sheath. Resembles the type (refer to descriptions by Ettl & Gärtner 1988) – Plate II, Figs 7, 8.

128 Proceedings of IV Balkan Botanical Congress • Sofia • 2006

Plate I

Figs 1, 2. Tolypothrix tenuis var. calcarata from field material;Figs 3–5. Heterococcus sp. from culture;Fig. 6. Desmococcus olivaceus from culture. Scale bars = 10 μm (Figs 1, 3–6) and 50 μm (Fig. 2).

129Scientific Area B • Plant diversity – past and present

Plate II

Figs 7, 8. Chlorococcum ellipsoideum from culture;Figs 9, 10. Trebouxia arboricola from culture. Scale bars = 10 μm (Figs 9–10) and 15 μm (Figs 7, 8).

130 Proceedings of IV Balkan Botanical Congress • Sofia • 2006

Trebouxiophyceae

Apatococcus lobatus (Chodat) J.B. PetersenT, on calcareous rock (wall of a tunnel), forming

greenish layer. In agar culture this isolate forms short filaments of 3–4 cells, but no zoospores have been ob-served. Cells 6–14 μm in diameter.*Chlorella minutissima Fott et Nováková

K, between mats of Tolypothrix spp. and moss pro-tonemata on moist walls of the Prokhodna cave near-by the entrance. Cells 2.5–4 μm in diameter.*Chlorella trebouxioides Punčoch.

K, together with Cyanoprokaryota and layers of Apatococcus lobatus on moist rocks nearby the en-trance to Prokhodna cave. Resembles the original de-scription of Punčochářová (1994), known from soil and bark (Hanagata & al. 1997). Cells 5–15 μm long and 3–12 μm wide.*Desmococcus olivaceus (Persoon ex Acharius) J.R. Laundon

K, common on moist walls of Prokhodna cave, be-tween soredia of lichens and moss protonemata. Al-so found on a fragment of glass (!) on the ground at the same locality. This cosmopolitan aerophytic al-ga forms cuboidal cell packets, chloroplast parietal with lobed margin and a small sometimes badly vis-ible pyrenoid. Cells 6–10 μm in diameter. The repro-duction stages (aplanosporangia) with characteristic warty cell walls were observed (Plate I, Fig. 6).Trebouxia arboricola Puym.

T, found in free living but also lichenized stage on moist rocks. Single cells without lichenization were also found in K on rocks of Prokhodna cave among Desmococcus olivaceus layers. Seemingly wide-ly distributed in Bulgaria, earlier recorded from bark (Vodenicharov & al. 1971) and on monuments (Gärt-ner & Stoyneva 2003). Cells 10–15 μm in diameter (Plate II, Figs 9, 10).*Trebouxia cf. excentrica P.A. Archibald

K, only found in lichenized stage, perhaps from li-chen soredia, together with Chroococcus westii, Desmo-coccus olivaceus and juvenile Dermatocarpon min-iatum, at the entrance of Prokhodna cave. The very distinct deeply incised chloroplast fills not more than 2/3 of the cell lumen. Cells 10–14 μm in diameter.

For future research, nature conservation and edu-cation it would be of great interest to conserve all the newly isolated algal strains for long-term maintenance

in a culture collection under controlled environmen-tal conditions.

Acknowledgements. This study was carried out with the fi-nancial support of the Austrian Research Fund (MOEL project No. 180).

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