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MICROBIOLOGY
Lithobiotic cyanobacteria diversity of the Karelian Isthmus
Oksana Rodina12, Denis Davydov2, and Dmitry Vlasov3
1Department of Crystallography, Institute of Earth Sciences, Saint Petersburg State University, per. Dekabristov, 16, Saint Petersburg, 199155, Russian Federation 2Polar-Alpine Botanical Garden-Institute, ul. Fersmana, 18A, Apatity, 184209, Russian Federation
3Department of Botany, Faculty of Biology, Saint Petersburg State University, Universitetskaya nab., 7-9, Saint Petersburg, 199034, Russian Federation
Address correspondence and requests for materials to Oksana Rodina, o.rodina@ksc.ru
Abstract
Citation: Rodina, O., Davydov, D., and Vlasov, D. 2022. Lithobiotic cyanobacteria diversity of the Karelian Isthmus. Bio. Comm. 67(2): 97-112. https://doi. org/10.21638/spbu03.2022.203
Authors' information: Oksana Rodina, Research Engineer, orcid.org/0000-0002-6598-6953; Denis Davydov, PhD, Senior Researcher, orcid.org/0000-0002-0866-4747; Dmitry Vlasov, Dr. of Sci. in Biology, Professor, Leading Researcher, orcid. org/0000-0002-0455-1462
Manuscript Editor: Vladimir Onipchenko, Department of Geobotany, Faculty of Biology, Moscow Lomonosov State University, Moscow, Russia
Received: July 16, 2021;
Revised: October 20, 2021;
Accepted: January 13, 2022.
Copyright: © 2022 Rodina et al. This is an open-access article distributed under the terms of the License Agreement with Saint Petersburg State University, which permits to the authors unrestricted distribution, and self-archiving free of charge.
Funding: The sampling and identification of cyanobacteria were performed with Russian Science Foundation support project No. 1917-00141. The cultivating and estimate of taxa features were performed with Russian Science Foundation support project No. 21-14-00029. The storage of samples is provided by the herbarium at the PolarAlpine Botanical Garden-Institute (KPABG). The research was done using the large-scale research facilities of the herbarium at the Polar-Alpine Botanical Garden-Institute (KPABG; Kirovsk, Russia) reg. No. 499397.
Ethics statement: This paper does not contain any studies involving human participants or animals performed by any of the authors.
Competing interests: The authors have declared that no competing interests exist.
This work presents data obtained as a result of studying the composition of cyanobacteria in lithobiotic communities on various substrates (Ruskeala marble, rapakivi-granite, granite gneiss) in different light conditions on the territory of the Karelian Isthmus: Leningrad Oblast, Republic of Karelia, and South Finland. The species composition of cyanobacteria was revealed, and the species composition on certain types of substrates was analyzed. A total of 49 species of cyanobacteria were noted for the Republic of Karelia (13 of which were not previously recorded in this territory). The detailed taxonomic and environmental characteristics of species are given. Changes in the species diversity of cyanobacteria in connection with specific habitats are shown. The type of substrate, the degree of moisture, and illumination are noted as the main factors determining the diversity of cyanobacteria in lithobiotic communities. Keywords: biofilms, cyanobacteria, lithobiotic communities, Ruskeala marble, rapakivi-granite
Introduction
Cyanobacteria are one of the most diverse groups of bacteria. They are often primary colonizers of bare areas of rock (Whitton, 1992; Gromov, 1996) because their life processes require only water, carbon dioxide, inorganic substances, and light.
Due to their efficient adaptive capacity, cyanobacterial colonies form frequent biofilms in different terrestrial habitats (Gorbushina, 2007; Rossi and De Philip-pis, 2015; Davydov and Patova, 2018). Different cyanobacterial species often were noted on infertile substrates such as desert sand or volcanic ash, at the stone-soil interface, and in endolithic niches in all Earth biomes (Jaag, 1945; Weber, Wessels and Budel, 1996; Budel, 1999; Mur, Skulberg and Utkilen, 1999; Pentecost and Whitton, 2000; Golubic and Schneider, 2003).
The functioning processes of lithobiotic systems and the various environments that influence these processes have not been sufficiently studied. The conditions of existence on the surface of the stone are considered close to extreme. Being in a thin surface layer, microorganisms are exposed to sharp fluctuations in humidity, temperature, pH, and light. Most often these are slow-growing organisms that are resistant to harsh environmental conditions. (Gorbushina, Lia-likova, Vlasov and Khizhniak, 2002; Keshari and Adhikary, 2013).
The exopolysaccharide substance (EPS) of microbial origin contributes to the colonization of the mineral substrate by cyanobacteria and has an integrating function (Beech and Gaylarde, 1991). Cyanobacteria that actively produce EPS
28.0 32.0
Fig. 1. Map of sampling sites: 1-4 are granite quarries, Southern Finland; 5 — Monrepos Park; 6 — Vyborg tunnels; 7 — Ristijarvi Park; 8 — Owl Mountain; 9 — Ruskeala Park. Numbers of sample plots as in Table 1.
contribute to the retention of moisture and the accumulation of organic matter, creating conditions for the development and accumulation of saprotrophic bacteria and fungi (Crispim, Gaylarde and Gaylarde, 2003).
The lithobiotic organisms could be considered as several different groups: chasmoendoliths and cryptoen-doliths occupy preexisting fissures and structural cavities in the rocks, whereas euendoliths penetrate soluble carbonate and phosphate substrates (Golubic, Friedmann and Schneider, 1981).
If the biofilm is formed on a rock surface (at the boundary of the solid and air phase), it is called a sub-aerial biofilm (SAB). In natural conditions, such biofilms are hard, dry plates or biological soil crusts. On vertical surfaces, they form films in the form of colored smudges (Gorbushina and Broughton, 2009). Biofilms occupy a significant part of the earth's surface and play a significant role in the circulation of matter and energy, in the weathering of rocks and soil formation processes (Krasilnikov, 1949; Glazovskaya and Dobrovolskaya, 1984; Grbic et al., 2010; Sancho, Maestre and Budel, 2014; Davydov and Redkina, 2021).
In general, the analysis of the literature data indicates the important role of cyanobacteria in primary lithobiotic communities formed under a wide variety of environmental conditions.
The relevance of the topic is associated with the fundamental role of cyanobacteria in the colonization of the mineral substrate. At the same time, a comparative study of the composition and structure of cyanobacterial communities on different rocks under different environmental conditions may be of particular interest. Based on the results of such a study, it is possible to answer the question of what factors most affect the diversity of this group of prokaryotes inhabiting carbonate and silicate rocks. These issues still remain insufficiently studied. The biodiversity of cyanobacteria can be very high, including in poorly studied geographic regions (Nabout, Da Silva Rocha, Carneiro and Sant'Anna 2013; Gaysina, Bohunicka, Hazukova and Johansen, 2018). Since morphological features have traditionally been the main criterion for the classification and identification of cya-nobacteria, most of the studies conducted to date have relied almost entirely on morphology-based methods
Table 1. Description of localities studied
Substrate Place Location Description of localities Number of samples
Marble Ruskeala (9)* N 61°56'45" E 30°34'49" The Ruskeala quarry is located in the Northern Ladoga region (the Republic of Karelia, near Sortavala town). Currently, it is a monument of mining. We examined this area for the first time to determine the composition of cyano-bacteria in lithobiotic biofilms at open rock surfaces in 2016 (Kuznetsova et al., 2016). For this research, sampling was carried out in open areas of rock surfaces and in tunnels with artificial lighting and poorly lit areas. 40
Southern Finland (1-4)* Montferrand quarry (Quarry I) (3) N 60°34'12,4", E 027°43'50.1"; Quarry II (2) N 60°31'51,3" E 027°39'41.9"; Quarry III (1) N 60°32'6,1" E 027°39'49.4"; Quarry IV (4) N 60°44'24,8" E 028°0'33.8". For the study of granites, four rapakivi-granite quarries were selected on the territory of Finland, where a stone was extracted for the construction of famous architectural structures in St Petersburg. Montferrand Quarry is one of the famous quarries. Granite mining here was stopped in the 19th century. The quarry was preserved as a mining monument. Now the quarries are privately owned by the Finns, and they are open to visitors. The quarry is located in the forest near the settlement of Peterlahti (Virolahti) on the shore of the Gulf of Finland in southeastern Finland, almost on the border with Russia. Currently, the quarry is undergoing gradual natural overgrowth. In addition to the Montferrand Quarry, which was the main focus, there are other granite outcrops in the same area, which were also investigated. 24
Granite and Granite gneiss Ristijarvi (7)* N 61°46'48" E 30°44'6" About two billion years ago, this area was the mouth of the Kiryavolakhtinsky volcano (Ladoga volcano of the Lower Proterozoic era). Its length reached 60-80 km, width was 30-40 km, and height — 2.5 km. Over the past two billion years, under the influence of the sun, wind, and precipitation, the mighty Kiryavolakhtinsky volcano was largely destroyed. Its central part, which was in the zone of tectonic-magmatic uplift of the granite gneiss dome, was completely eroded, up to the granite base. It is here, in the center of the former volcano, that the modern Ristijarvi Lake is located. The lake is surrounded by high (up to 100-140 m) and steep rocks, composed of granites and granite gneisses, formed in the Archean. 6
Monrepos Park (5)* N 60°44'01" E 28°43'29" Rock landscape park on the shore of the Protective Bay of the Vyborg Bay, on Tverdysh Island in the northern part of Vyborg town, Leningrad Oblast, located on the territory of the State Historical, Architectural and Natural Museum-Reserve Monrepos Park. The territory of the park consists of two large sections, adjacent to the south and north to the historical core of the park. It is characterized by unique Ice Age stone ridges of rapakivi-granite (Karel. rapakivi — "rotten stone"), in some areas reaching a height of 20 m. 2
Vyborg tunnels (6)* N 60046'23" E 28040'13" Biofilms from poorly lit and heavily shaded (underground) areas of the rapakivi-granite rock surface were studied in the territory of three tunnels near the "Iron Forest" Museum ("Rautakorpi"), Vyborg District. These are tunnels cut out of the rock, from the time of the First World War, which served as Finnish artillery depots. Sampling was at a depth of 3 to 5.5 m from the entrance with a gradient decrease in illumination. 13
Owl Mountain (8)* N 61032'59.9" E 3Q011'56.9" Owl Mountain is a military-historical and geological museum, located in South Karelia in Lahdenpokhya town. The location of the object is unique — it is a huge man-made bunker inside a powerful granite rock with two entrances inside. On the vertical walls at the entrance to the bunker, biofilms with the participation of cyanobacteria are actively formed. The substrate is a gray granite gneiss (close to the rapakivi-granite). 8
*The numbers in brackets correspond to the points in Fig. 1.
(Alvarenga, Rigonato, Branco and Fiore, 2015). Nienow (1996) indicates 70 cyanobacteria genera involved in the formation of subaerial communities. Of these, the order Chroococcales is the leader in the number of spe-cies—34 genera (49 %) (Nienow, 1996). In this order, the genus Gloeocapsa is represented by the greatest variety of species for the carbonate substrates of caves in Bulgaria (a total of 59 cyanobacteria species were noted) (Draganov and Dimitrova-Burin, 1977). For speleoob-jects, the intensity of illumination is also a key factor in the algae distribution. For example, green algae predominate in grottoes, as ecologically more comfortable habitats, while the algoflora of caves was characterized by lower species diversity and a significantly higher proportion of cyanobacteria (40-70 % of the species composition) (Vinogradova and Mikhailyuk, 2009). In the Left-bank cave in Leningrad Oblast, nine cyanobacteria species (31 % of the species composition of algae) were identified (Abdullin, 2012). The flora of lithobiotic communities on the territory of the Karelian Isthmus remains poorly studied. For the Republic of Karelia and Leningrad Oblast, the flora of aquatic habitats is mainly known.
The aim of our work was to identify the species composition of cyanobacteria on rock surfaces (Ruskeala marble and rapakivi-granite) under various environments (rock outcrops, quarries, and tunnels).
Materials and methods
The investigated area is situated on the northeastern part of the Baltic region. We studied cyanobacterial diversity of the Karelian Isthmus: Leningrad Oblast, Karelia, and South Finland. All samples were collected by the senior author in 2015-2018 (Fig. 1, Table 1).
Samples were taken at the sites of color and surface substrate changes. Wet samples were collected in sterile containers and tubes with a volume of up to 120 ml. Dry samples were collected in sterile containers and Kraft envelopes. Samples were taken with a sterile scalpel, because biofilms dominated by cyanobacteria are most often easily separated from the substrate. Where possible, biofilms were taken together with small pieces of substrate. The storage of samples is provided by the herbarium of the Polar-Alpine Botanical Garden-Institute (KPABG).
Characteristics of the studied substrates
In the study area, there are rocks of different geological origin. Most of the territory of study is located within felsic rocks. In contrast, the Ruskeala marble quarry site belongs to carbonate rocks. It includes not only calcite but also dolomite, unlike numerous analogs. The color of Ruskeala marble varies from dark gray and black to
snow-white, sometimes with greenish stripes and nests up to several centimeters wide; the structure is finegrained. Ruskeala marble is divided into 3 groups: cal-cite, dolomite, and calcite-dolomite (Bulakh, 1999).
The physical and mechanical properties of Ruskeala marbles are the following: density 2750-2820 kg/m3; water absorption 0.1-0.2 %; compressive strength from 200 MPa to 80 MPa (Borisov, 2001).
A greater variety of substrata were found among felsic rocks.
Rapakivi-granite (Finnish "rapakivi", of "rapa" (meaning mud or sand) and "kivi" (meaning rock or stone)) refers to double-feldspar granites of high alkalinity with a characteristic structure due to the presence of large ovoids of potassium feldspar, usually surrounded by oligoclase borders. This structure causes the relatively rapid destruction of the rock to which it owes its name, which in Finnish means "rotten stone". The color of rapakivi is gray and pink. Dark-colored minerals are represented by biotite and high-ferruginous hornblende; accessory minerals are titanomagnetite, olivine, fluorite, apatite, and zircon. In composition, rapakivi belongs to alkaline granites or granosienites with a high content of Fe. Rapakivi density is about 2650 kg/m3, porosity is 0.3 %, water absorption is 0.15-1.30 %, and compression resistance is 100-200 MPa.
Granite gneiss is full-crystalline banded or shale rock, and its composition is similar to granite. Granite gneiss structure occupies an intermediate position between granite and gneiss. The texture is due to the subparallel arrangement of tabular and prismatic crystals (mica, hornblende, feldspar) and elongated inclusions, as well as the accumulation of individual minerals in alternating bands or interlayers (the so-called gneiss-like texture). Most researchers consider granite gneisses as granites that crystallized in the deep zones of the earth's crust during the cooling of the magmatic melt under conditions of directional pressure or during the movement of magma, resulting in a parallel orientation of minerals. The bodies of such granite gneisses have secant contacts with the host rocks.
Laboratory research methods
Species identification was carried out using light microscopy (Leica DM 1000 microscope) by direct microscopy of samples. Cultural methods were also used. To obtain accumulative cultures, fragments (biofilm pieces or fragments of the substrate with biofilms on their surface) of all samples were placed in distilled water, in the liquid Gromov-6 medium, as well as in the medium Z8 and BG-11 (Kotai 1972; Rippka, 1988; Waterbury, 2006). Isolation of pure cultures was carried out from accumulative cultures by successive inoculations onto BG-11 agar medium. The cultures
were stored on BG-11 agar medium and Z8 liquid medium.
Part of the material was examined under a scanning electron microscope in the magnification range from 100x to 10,000x. SEM studies were performed on a TM 3000 electron microscope (HITACHI, Japan, 2010) with an OXFORD energy-dispersive microanalysis device at the SPbU Microscopy and Microanalysis Resource Center.
The species composition of cyanobacteria was determined by morphological characteristics, using identification guides (Komarek and Anagnostidis, 1998, 2005; Komarek, 2013). Data on the geographical distribution of the species and their ecological characteristics are provided in accordance with CRIS database (Melechin, Davydov, Shalygin and Borovichev, 2013; Melekhin et al., 2019). The names of geographical elements are cited according to Konstantinova (2000). The geographical elements are distinguished by Davydov (2010b).
The principal component method was performed using the Statistica 8.0 software. Samples were selected as objects (n = 76), and the presence of species taxa in the sample (90 species in total) was selected as features.
Flora similarity was determined by the ExcelToR software (Novakovskiy, 2016) based on the Sorensen index: KS = 2a/(2a + b + c), where a is the number of species common to both sets, b — the number of species unique to the first set, and c — the number of species unique to the set.
Results and discussion
Taxonomic analysis of cyanobacteria diversity in lithobiotic communities
In this research, 90 species taxa of cyanobacteria belonging to 4 orders, 17 families, and 31 genera were identified by morphological features. A complete taxonomic list of identified cyanobacteria is given in Table 2.
The order Nostocales is represented by the largest number of families (6). The smallest number of genera (4) is represented by the order Oscillatoriales; the other orders contain 9 genera each. A total of 30 taxa at the species level were identified in Chroococcales; in Synechococcales — 33 taxa. Of the 17 families, the most widely represented is Leptolyngbyaceae (16 species). Leptolyngbya is the genus with the most species diversity and occurrence frequency (Fig. 2), which includes 13 species (14 % of the identified diversity). The taxonomic analysis of the identified cyanobacteria of the lithobiotic communities is given in Table 3.
The presented results are corresponding with some published data. For example, species of the genera Gloeo-capsa, Gloeothece, Chamaesiphon, Calothrix, Tolypothrix, and Scytonema are especially characteristic of irrigated
Fig. 2. The number of species taxa for the most widely represented genera of cyanobacteria.
rocks (Wasser, Kondratyeva and Masyuk, 1989), most of which were noted in our studies.
However, when compared with a specific region (Murmansk Oblast), the most studied in relation to terrestrial cyanobacterial communities, we noted some differences, primarily related to the diversity of species within genera. Thus, the genera Gloeocapsa (7 species), Nostoc (7), Phormidium (7), Leptolyngbya (6), Chroococ-cus (5), and Calothrix (5) are widely represented in this territory (Davydov, 2010a). In the present study, the genus Leptolyngbya (13 species) is in the first place. The genera Nostoc and Calothrix contained only two species of taxa each. Such differences are most likely associated with the characteristics of the studied biotopes. In this study, only lithobiotic (epilitic) communities were considered, while in Murmansk Oblast, soil habitats were also studied. As an example, the genus Nostoc, which is more typical for soils, falls out of our general list.
We found 38 species on marble and 76 on granite. A comparison of the species lists of cyanobacteria found on carbonate and silicate rocks using the Sorensen index of species similarity shows a medium degree of similarity — 53 %.
The distribution of cyanobacteria species by geographical elements (Davydov, 2010b) shows that among species with a known geographical distribution (49 species), there is a predominance of cosmopolitan species (Fig. 3). Stony substrates can be classified as difficult for living organisms. These substrates are usually colonized by either highly specialized species or species with a wide range of resistance to environmental factors. Only 11 % of identified species are associated with mountain habitats (montane, arctomontane, arctoborealmontane (Davydov, 2010b)).
Table 2. Cyanobacteria species composition on rock surfaces from Ruskeala marble and granite of Leningrad Oblast, Karelia and South Finland
Taxon Ruskeala (marble) Common For Granite Vyborg tunnels Quarry I Quarry II Quarry III Quarry IV Ristijarvi Owl mountain Monrepos Park
Anabaena sp. 1
Aphanocapsa fusco-lutea Hansg. 1 1
Aphanocapsa parietina Näg. 1 1 1
Aphanocapsa sp.1 1 1 1 1
Aphanocapsa sp.2 1 1
Aphanocapsa sp.3 1 1
Aphanocapsa holsatica (Lemm.) G. Cronb. et Komärek 1 1
Aphanocapsa muscicola (Menegh.) Wille 1 1 1
Aphanothece castagnei (Breb.) Rabenh. 1 1
Aphanothece saxicola Näg. 1 1
Calothrixparietina Thür, ex Born, et Flah. 1 1 1 1 1 1 1
Calothrix sp. 1 1 1
Chalicogloea sp. 1 1 1 1
Chondrocystis dermochroa (Näg.) Komärek et Anagn. 1 1
Chroococcus cohaerens (Breb.) Näg. 1
Chroococcus minor {Kütz.) Näg. 1 1 1 1
Chroococcus minutus (Kütz.) Näg. 1 1 1
Chroococcus pallidus (Näg.) Näg. 1 1 1
Chroococcus sp. 1
Chroococcus spelaeus Erceg. 1 1 1
Chroococcus turgidus (Kütz.) Näg. 1
Chroococcus varius A. Braun 1 1 1
Cyanobium gaarderi (Älvik) Komärek et al. 1 1
Cyanothece aeruginosa (Näg.) Komärek 1 1 1
Eucapsis sp. 1
- s
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Leptolyngbya lagerheimeii (Gom.) Anagn. et Komárek Leptolyngbyagracillima (Hansg.) Anagn. et Komárek Leptolyngbya foveolarum (Rabench. ex Gom.) Anagn. et Komárek Leptolyngbya cebennensis (Gom.t) 1. Umezaki et M. Watanabe Leptolyngbya amplivaginata (Van Goor) Anagn. et Komárek Johannesbaptistia schizodichotoma (J.J. Copeland) Komárek et Anagn. Johannesbaptistia pellucida (Dickie) W. R. Taylor et Drouet Jaaginema pseudogeminatum (G. Schmid) Anagn. et Komárek Jaaginemageminatum (Menegh. ex Gom.) Anagn. et Komárek Hapalosiphon pumilus Kirchn. ex Born, et Flah. Gloeothece sp. Gloeothece rupestris (Lyngb.) Born. Gloeothece palea (Küzt.) Rabenh. Gloeocapsopsis crepidinum (Thur.) Geitl. ex Komárek Gloeocapsopsis sp. 2 Gloeocapsopsis sp. 1 Gloeocapsopsis magma (Bréb.) Komárek et Anagn. Gloeocapsa atrata Kütz. Gloeocapsa violascea (Corda) Rabenh. Gloeocapsa sp. Gloeocapsa rupestris Kütz. Gloeocapsa punctata Nag. Gloeocapsa kuetzingiana Nag. Gloeocapsa compacta Kütz. Gloeocapsa alpina (Nág.) Brand Taxon
- - - - - - - - - - - - - - Ruskeala (marble)
Common For Granite
- - - Vyborg tunnels
- - - - - Quarry I
- - - - - - - - Quarry II
- - - - - - Quarry III
- - - - - - - Quarry IV
- - Ristijarvi
- - - - Owl mountain
Monrepos Park
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Phormidium sp. Phormidium papyraceum (C. Ag.) Gom. Phormidium inundatum Kutzi. ex Gom.t Phormidium interruptum Kutz. ex Forti Phormidiumgranuiatum (N. L. Gardner) Anagn. Phormidium corium Gom. ex Gom. Phormidium breve (Kutz. ex Gom.) Anagn. et Komarek Phormidesmis moiie (Gom.) Turicchia et al. Petaionema incrustans Komarek Nostoc microscopicum Carm. ex Born, et Flah. Nostoc commune Vauch. ex Born, et Flah. Microcystis puiverea (H. C. Wood) Forti Microcoieus vaginatus Gom. ex Gom. Microcoieus paludosus Gom. Microchaete fe/iera Thur. ex Born, et Flah. Lyngbya sp. Lyngbya martensiana Menegh. ex Gom. Leptolyngbya tenuis (Gom.) Anagn. et Komarek Leptolyngbya angustissima (W. West et G. S. West) Anagn. et Komarek Leptolyngbya valderiana (Gom.) Anagn. et Komarek Leptolyngbya sp. 3 Leptolyngbya sp. 2 Leptolyngbya sp. 1 Leptolyngbya sieminskae D. Richter et Matula Leptolyngbya perelegans (Lemm.) Anagn. et Komarek Taxon
- - - - - - - - Ruskeala (marble)
- - - Common For Granite
- - Vyborg tunnels
- Quarry I
- - - - Quarry II
- - Quarry III
- - - - - - - - - Quarry IV
- - - Ristijarvi
- - - - Owl mountain
- - - Monrepos Park
£0Z'ZZ0Z'£0nqds/8£9LZ'0L/SJO-!op//:sdjjL| | ZZOZ '3unf-|udv 'Z snssi 7.9'|oa 'SNOIlVOINniAIIAlOO "lVOIDOIOia POl
Taxon Ruskeala (marble) Common For Granite Vyborg tunnels Quarry I Quarry II Quarry III Quarry IV Ristijarvi Owl mountain Monrepos Park
Phormidium tergestinum [Kütz.] Anagn. et Komárek 1
Phormidium puteale (Mont. ex Gom.) Anagn. et Komárek 1
Planktolyngbya bipunctata (Lemm.) Anagn. et Komárek 1 1
Planktolyngbya limnetica (Lemm.) Kom.-Legn. et G. Cronberg 1 1
Scytonema hofmanii C. Ag. ex Born, et Flah. 1 1
Stigonema hormoides Kütz. ex Born, et Flah. 1 1
Stigonema mesentericum Geltler 1 1
Stigonema ocellatum Thur. ex Born, et Flah. 1 1
Synechococcus sciophiius Skuja 1 1
Synechocystis aquatilis Sauv. 1 1 1 1
Synechocystis parvula Perflllev 1 1
Synechocystis pevaiekii Ercegovic 1 1
Synechocystis salina Wlslouch 1 1
Synechocystis minuscula Woronlchln 1 1 1 1
Tolypothrix tenuis Kütz. ex Born, et Flah. f. terrestrisJ. B. Petersen 1 1
Total 38 76 12 10 19 9 26 14 15 5
o Ln
MICROBIOLOGY
Table 3. Taxonomic structure of identified cyanobacteria of lithobiotic communities
ORDER NUMBER OF FAMILIES FAMILY NUMBER OF GENERA GENUS NUMBER OF SPECIES TAXA
Chroococcales 4 Aphanothecaceae 2 Aphanothece 2
Gloeothece 3
Chroococcaceae 4 Chalicogloea 1
Chondrocystis 1
Chroococcus 8
Gloecapsopsis 4
Cyanothrichaceae 1 Johannesbaptistia 2
Microcystaceae 2 Gloeocapsa 8
Microcystis 1
Nostocales 6 Hapalosiphonaceae 1 Hapalosiphon 1
Nostocaceae 2 Anabaena 1
Nostoc 2
Rivulariaceae 2 Calothrix 2
Microchaete 1
Scytonemataceae 2 Petalonema 1
Scytonema 1
Stigonemataceae 1 Stigonema 3
Tolypothrichaceae 1 Tolypothrix 1
Oscillatoriales 4 Cyanothecaceae 1 Cyanothece 1
Leptolyngbyaceae 3 Leptolyngbya 13
Phormidesmis 1
Planktolyngbya 2
Microcoleaceae 1 Microcoleus 2
Oscillatoriaceae 2 Lyngbya 2
Phormidium 9
Synechococcales 3 Merismopediaceae 3 Aphanocapsa 7
Eucapsis 1
Synechocystis 5
Pseudanabaenaceae 1 Jaaginema 2
Synechococcaceae 2 Cyanobium 1
Synechococcus 1
cosmopolits unclear distribution species a reto bo rea I boreal
a reto montane arctoborealmontane montane
Fig. 3. Distribution of identified cyanobacteria species by geographical elements.
The assessment of the distribution of the found species by type of habitat also shows the predominance of cosmopolitan species (37 % of the total number). It is interesting to note that one species we found, Gloeocapsop-sis crepidinum, belongs to the amphioceanic type (found in terrestrial habitats along the sea coast, the supralithoral zone, or along the shores of brackish reservoirs); at the same time, we found it on granites irrigated with fresh water. This species was previously recorded in Russia on Kamchatka, on Medny Island (arch. Commander Islands, Kamchatka) and in Murmansk Oblast (Davydov, 2010a). In this research, the presence of 49 species of cyanobacteria was noted for the Republic of Karelia (13 of which were not previously recorded in this territory).
A floristic comparison of the studied territories with each other according to the identified species of cyanobacteria was carried out. In addition, for comparison, we selected territories where the composition of lithobiotic cyanobacteria in mountain conditions had been studied: the flora of ravines in the Vatsuoi River valley (Sal'nye Tundra
ridge), Kerkchorr (Chunatundra ridge) (Shalygin, 2012), and the Aikuaivenchorr ravine (Davydov, 2018). The number of species in the studied territories is comparable: for Aikuivenchorr flora 36 species were found; the Vatsuoi Valley flora includes 23 taxa; and for Kerkchorr ravine, 26 species were noted. With other territories, the comparison can only be made conditionally, since the substrates and environmental conditions are very different. The similarity coefficient for the studied territories is low. The highest S0rensen index of 0.48 was obtained when comparing Ris-tijarvi and Owl Mountain (Fig. 4). The flora are clustered according to the geographical principle. The territories are close to each other geographically and have a similar substrate in general. Since there was only one collection area for Ruskeala marble, it is not possible to trace the clustering by substrate when comparing the studied territories.
Component analysis (the principal component method) was used to identify the possible association of cyanobacteria with certain types of habitats. The analysis results show that the entire sampling is practically homogeneous. The first two factors together reflect only a little less than 12 % of the explained variability. The first factor is most likely associated with the type of substrate (granite, marble); the second—with the degree of saturation. On the graph (Fig. 5) most species are grouped in the central part of the graph (located in the area of zero coordinates), which is due to their rare and rather random finds. There is also a division of species in three directions. In the first sector, the species common to marble and granite are grouped. Sector II is occupied by species typical of granites. In the third sector, the species of marble are grouped.
On the surface of rocks, both monospecific and multi-species communities can form. For example, a rich biofilm with Stigonema ocellatum dominance was found on granite in the Montferrand Quarry at the site of natural water seepage (Fig. 6). This species forms a biofilm of tightly intertwined threads, which is clearly visible on the SEM image. As you can see in the figure, the biofilm from green to black is represented by Stigo-nema ocellatum, and in the wetter part of the biofilm, you can see a change in color to brown-red, which is associated with the replacement of the dominant species by Gloeocapsopsis magma. In addition, other cyanobacteria also appear: Lyngbya sp., Leptolyngbya foveolarum. For this case, the degree of moisture can be noted as the leading factor affecting the biofilm composition.
Lighting also plays an important role in the formation of biofilms. For comparison, we studied communities from the surface of Ruskeala marble and rapakivi-granite in open areas and in tunnels (under artificial lighting and in shaded areas).
For biofilms on open rock surfaces under natural light on the surface of marble, the list of species included 21 taxa. For open areas of granite quarries, 63 species of cyanobacteria were identified.
Index:1 brayr. Binary:' TRUE'. Agglomerative method:' average'
0.12
0.11
0.2
0.24
0.27
0.34
0.38
0.28
0.32
0.4S
Ker
Vat
OMt
Ris
Vyb
Ayk
QIV
Rus
Oil
QUI
Ql
0
0.2
0.4
0.6
OS
Fig. 4. Graph of similarity of cyanobacteria flora on rock surfaces from Ruskeala marble (Rus) and granite of Leningrad Oblast (Vyborg tunnels — Vyb), Karelia (Ristijarvi Park — Ris, Owl Mountain — OMt, South Finland quarries — Q l-IV and in the ravines of Aykuivenchorr — Ayk (Davydov, 2018), Vatsuoi — Vat, Kerkchorr — Ker (Shalygin, 2012). S0rensen's similarity coefficient, clustering by average method.
Projection of the variables on the factor-plane ( 1 x 2)
Leptolyngbya foveolamm
..Gloeocapsa atrata
1 0
Active
Gloeocapsa violascea ya sp. 1 :: ' Chroecoccus cohaerens
o
0,0 0,5
Factor 1 : 6,35%
Aphanocapsa holsatica:
Planktolyngbya limnetica|
Phormidium interruptum
_ J_
Phormidium papyraceum
Leptolyngbya angustissima
Phormidium coriuml
Fig. 5. Ordination of features (occurrence of species in samples) in the space of the first and second factors (I, II, III — sector number).
MICROBIOLOGY
Fig. 6. Biofilm with a change of dominant species on the rapakivi-granite surface in the Montferrand Quarry (a — Gloeocapsopsis magma, b — Stigonema ocellatum).
Conclusion
The conducted studies have shown that the cyanobacte-ria composition on rock outcrops (marble and granite) in Leningrad Oblast, Karelia and Southern Finland is characterized by significant diversity. The obtained data expand the information of cyanobacteria in lithobiotic communities, but further research in this direction is needed. The diversity of cyanobacteria on rapakivi-granite was two times higher than on Ruskeala marble, which is probably due to the more complex mineral composition of rapakivi-granite. Daylight, along with the degree of moisture, plays an important role in the species distribution on the rocky substrate's surface (the cyanobacteria diversity decreases significantly with a decrease in the illumination of the habitat, which is especially noticeable for tunnels). The revealed differences probably affect the composition of the metabolites of the lithobiotic biofilms. The role of cyanobacteria in geochemical processes and the destruction of stone requires in-depth study, and our work can be considered as a stage of this research.
Acknowledgments
The authors are grateful to Elena Gennadievna Panova for help in sampling organization and Valentina Nikolaevna Nikitina1 for help in cyanobacteria identification.
Our studies have shown that the species composition in the tunnels is similar on different substrates and, at the same time, significantly poorer in comparison with open rock surfaces areas. In total, seven cyano-bacteria species were detected on granites in conditions of limited light: Chroococcus sp. 1, Gloeocapsa kuetz-ingiana, Gloeocapsopsis magma, Gloeocapsa violascea, Leptolyngbya sp., Aphanocapsa sp., and Aphanocapsa cf. fusco-lutea. For comparison, six species of cyanobacteria were detected in one combined sample on open granite-rapakivi surface area near the tunnels (under natural light): Gloeocapsopsis magma, Nostoc commune, Calo-thrix parietina, Scytonema hofmanii, Aphanocapsa sp. 1, and Aphanocapsa cf. fusco-lutea.
Only six cyanobacteria species were identified in the marble tunnels: Aphanocapsa sp., Chroococcus sp. 1, Chroococcus sp. 2, Gloeocapsa atrata, Leptolyngbya gracillima, and Leptolyngbya sp.
The results show that biofilm diversity is significantly reduced in the absence of bright daylight. Light is the limiting factor in the distribution of phototrophs on rocky surfaces. This is typical not only for epilitic species in caves and tunnels, but, for example, for the desert regions of Antarctica, where the distribution of endolithic species also depends on the daylight penetration deep into the substrate (Nienow, McKay and Friedmann, 1988).
References
Abdullin, Sh.R.2012. Cyanobacteria and algae of Levober-ezhnaya cave (Leningrad region). Botanicheskii zhurnal 97(8):1040-1051. (In Russian) Alvarenga, D. O., Rigonato, J., Branco, L. H.Z., and Fiore, M. F. 2015. Cyanobacteria in mangrove ecosystems. Biodiversity and Conservation 24(4):799-817. https://doi. org/10.1007/s10531-015-0871-2 Beech, I. B. and Gaylarde, C. C. 1991. Microbial polysaccharides and corrosion. International Biodeterioration 27(2):95-107. https://doi.org/10.1016/0265-3036(91)90002-9 Borisov, I.V.2001. Ruskeala marble. Abstract. Fondy RMSP.(In Russian)
Budel, B. 1999. Ecology and diversity of rock-inhabiting cyanobacteria in tropical regions. European Journal of Phy-cology 34(4):361-370. https://doi.org/10.1080/09670269 910001736422
Bulakh, A.G. 1999. Saint-Petersburg stone decoration.
Sudarynia Publ., St Petersburg. (In Russian) Crispim, C.A., Gaylarde, P.M., and Gaylarde, C.C.2003. Algal and cyanobacterial biofilms on calcareous historic buildings. Current Microbiology 46(2):79-82. https://doi. org/10.1007/s00284-002-3815-5 Davydov, D.A. and Redkina, V.V.2021. Algae and cyano-prokaryotes on naturally overgrowing ash dumps of the Apatity thermal power station (Murmansk Region). Transactions of Karelian Research Centre of RAS 1:51-68. https://doi.org/10.17076/bg1270 Davydov, D. and Patova, E.2018. The diversity of Cyanopro-karyota from freshwater and terrestrial habitats in the Eurasian Arctic and Hypoarctic. Hydrobiologia 811(1):119-137. https://doi.org/10.1007/s10750-017-3400-3 Davydov, D. A. 2010a. Cyanoprokaryotes and their role in the process of nitrogen fixation in terrestrial ecosystems of the Murmansk region. GEOS Publ., Moscow. 178 pp. (In Russian)
Davydov, D. A. 2010b. Features of the geographical distribution and analysis of cyanoprokaryotes (Cyanoprokaryota / Cyanobacteria) on the example of the biota of the Murmansk region. Bulletin MoIP 115(4):43-54. (In Russian) Davydov, D.A.2018. Finds of new species of cyanoprokaryotes in the Aikuayvenchorr gorge (Khibiny, Murmansk region). Trudy Karel'skogo nauchnogo centra RAN 8:132140. https://doi.org/10.17076/bg734 (In Russian) Draganov, S. and Dimitrova-Burin, E. D. 1977. Speleoalgologi-cal research in Bulgaria. Proceedings of the 6th International Congress of Speleology, Olomouc 5:11-17. Gaysina, L.A., Bohunicka, M., Hazukova, V., and Johansen, J. R. 2018. Biodiversity of terrestrial cyanobacteria of the South Ural Region. Cryptogamie, Algologie 39(2):167-198. https://doi.org/10.7872/crya/v39.iss2.2018.167 Glazovskaya, M. A. and Dobrovolskaya, N.G.1984. Geochemi-cal function of microorganisms. Izdatel'stvo MGU Publ., Moscow. 152 pp. (In Russian) Golubic, S. and Schneider, J.2003. Microbial endoliths as internal biofilms; pp. 249-263 in Krumbein, W.E., Pater-son, D.M., and Zavarzin, G.A. (eds), Fossil and recent biofilms. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-0193-8_16 Golubic, S., Friedmann, I., and Schneider, J. 1981. The lithobi-ontic ecological niche, with special reference to microorganisms. Journal of Sedimentary Research 51(2):475-478. https://doi.org/10.1306/212F7CB6-2B24-11D7-8648000102C1865D Gorbushina, A. A. 2007. Life on the rocks. Environmental Microbiology 9(7):1613-1631. https://doi.org/10.1111Zj.1462-2920.2007.01301.x
Gorbushina, A.A. and Broughton, W.J.2009. Microbiology of the atmosphere-rock interface: how biological interactions and physical stresses regulate a sophisticated microbial system. Annual Reviews of Microbiology 63(1):431-450. https://doi.org/10.1146/annurev.mi-cro.091208.073349 Gorbushina, A.A., Lialikova, N. N., Vlasov, D.Yu., and Khizhniak, T.V.2002. Microbial communities on the monuments of Moscow and St. Petersburg: biodiversity and trophic relations. Mikrobiologiia 71(3):409-417. (In Russian) Grbic, M.L., GrbicVukojevic, J., Simic, G.S., Krizmanic, J., and Stupar, M.2010. Biofilm forming cyanobacteria, algae and fungi on two historic monuments in Belgrade, Serbia. Archeological Biological Science 62(3):625-631. https://doi.org/10.2298/ABS1003625L Gromov, B.V.1996. Cyanobacteria in the biosphere. So-
rosovskij obrazovatel'nyj zhurnal 9:33-39. (In Russian) Jaag, O.1945. Untersuchungen tiber die Vegetation und Biologie der Algen des nackten Gesteins in den Alpen, im Jura und im schweizerischen Mittelland. Beitr. Kryptoga-mentl. Schweiz 9. 560 pp. Keshari, N. and Adhikary, S. P. 2013. Characterization of cyanobacteria isolated from biofilms onstone monuments at Santiniketan, India. Biofouling: The Journal of Bioadhe-sion and Biofilm Research 29(5):525-536. https://doi.org/ 10.1080/08927014.2013.794224 Komarek, J. 2013. Cyanoprokaryota. Teil 3: Heterocytous genera. Süsswasserflora von Mitteleuropa. Bd 19/3. Springer Spektrum, Berlin, Heidelberg. 1133 pp. Komarek, J. and Anagnostidis, K.1998. Cyanoprokaryota. 1.
Teil. Part: Chroococcales. Spektrum, Berlin. 548 pp. Komarek, J. and Anagnostidis, K.2005. Cyanoprokaryota. 2.
Teil. Part: Oscillatoriales. Spektrum, Berlin. 759 pp. Konstantinova, N.A.2000. Distribution patterns of the North Holarctic hepatics. Arctoa 9:29-94. https://doi. org/10.15298/arctoa.09.06 (In Russian) Kotai, J.1972. Instructions for preparation of modified nutrient solution Z8 for algae. Norwegian Institute for Water Research, Blindern, Oslo 11/69, 5 pp. Krasilnikov, N.A.1949. Role of microorganisms in weathering of rocks. I. Microflora of surface rocks. Mikrobiologiia 18(4):318-323. (In Russian) Melechin, A.V., Davydov, D.A., Shalygin, S.S., and Borovi-chev, E.A. 2013. Open information system on biodiversity cyanoprokaryotes and lichens CRIS (Cryptogamic russian Information System). Bulletin MoIP 118:51-56. (In Russian) Melekhin, A. V., Davydov, D. A., Borovichev, E.A., Shalygin, S. S., and Konstantinova, N.A.2019. CRIS — service for input, storage and analysis of the biodiversity data of the cryptogams. Folia Cryptogamica Estonica 56:99-108. https:// doi.org/10.12697/fce.2019.56.10 Mur, L.R., Skulberg, O.M., and Utkilen, H.1999. Cyanobacteria in the environment; Chapter 2 in Chorus, I. and Bartram, J.(eds), Toxic cyanobacteria in water: a guide to their public health consequences, monitoring and management.
Nabout, J.C., Da Silva Rocha, B., Carneiro, F.M., and Sant'Anna, C. L. 2013. How many species of Cyanobacteria are there? Using a discovery curve to predict the species number. Biodiversity and Conservation 22(12):2907-2918. https://doi.org/10.1007/s10531-013-0561-x Nienow, J. A. 1996. Ecology of subaerial algae. Nova Hedwigia 1 12:537-552.
Nienow, J. A., McKay, C. P., and Friedmann, E.I.1988. The cryp-toendolithic microbial environment in the Ross Desert of Antarctica: Light in the photosynthetically active region. Microbial Ecology 16(3):271-289. https://doi.org/10.1007/ BF02011700
Novakovskiy, A.B.2016. Interaction between Excel and statistical package R for ecological dataanalysis. Vestnik IB Komi NC Uro RAN 3:26-33. (In Russian) Pentecost, A. and Whitton, B. A. 2000. The ecology of cyano-bacteria — their diversity in time and space; pp. 257279 in Whitton, B.A. and Potts, M.(eds), Limestones. Kluwer Academic Publishers. Rippka, R.1988. Isolation and purification of cyanobac-teria. Methods of Enzymology 167:28-67. https://doi. org/10.1016/0076-6879(88)67005-4 Rossi, F. and De Philippis, R.2015. Role of cyanobacteri-al exopolysaccharides in phototrophic biofilms and in complex microbial mats. Life 5:1218-1238. https://doi. org/10.3390/life5021218 Sancho, L. G., Maestre, F.T., and Budel, B.2014. Biological soil crusts in a changing world: introduction to the special issue. Biodiversity and Conservation 23(7):1611-1617. https://doi.org/10.1007/s10531-014-0727-1 Shalygin, S.S.2012. Groupings of epilithic and epiphytic cy-anoprokaryotes of the Lapland nature reserve. Avtore-ferat dissertacii kand. biol. nauk, Ufa. 17 pp. (In Russian)
Vinogradova, O. N. and Mikhailyuk, T. 1.2009. Algoflora of caves and grottoes of the National Natural Park "Podolskie Tov-try" (Ukraine). Al'gologiia 19(2):155-171. (In Russian) Wasser, S. P., Kondratyeva, N.V., and Masyuk, N. P.1989. Algae. Directory. Naukova Dumka Publ., Kiev. 608 pp. (In Russian)
Waterbury, J. B. 2006. The Cyanobacteria — isolation, purification and identification of major groups of Cyanobacteria; pp. 1053-1073 in Dworkin, M., Falkow, S., Rosenberg, E., Schleifer, K.-H., and Stackebrandt, E.H.(eds), The Pro-karyotes. Springer, New York. https://doi.org/10.1007/0-387-30744-3_38 Weber, B., Wessels, D.C.J., and Budel, B.1996. Biology and ecology of cryptoendolithic cyanobacteria of a sandstone outcrop in the Northern Province, South Africa. Algological Studies 83:565-579. https://doi.org/10.1127/ algol_stud/83/1996/565 Whitton, B.A. 1992. Diversity, ecology and taxonomy of the cyanobacteria; pp. 1-51 in Mann, N.H. and Carr, N.G. (eds), Photosynthetic prokaryotes. Plenum Press, New York. https://doi.org/10.1007/978-1-4757-1332-9_1
