ę ę Acta Mycologica DOI: 10.5586/am.1101 ORIGINAL RESEARCH PAPER Publication history Received: 2017-06-07 Accepted: 2017-12-12 Published: 2017-12-30 Handling editor Maria Rudawska, Institute of Dendrology, Polish Academy of Sciences, Poland Disciseda verrucosa (Agaricomycetes, Basidiomycota) – a species new to Poland Agnieszka Tomaszewska1*, Janusz Łuszczyński1, Gabriel Moreno2 Department of Botany, Institute of Biology, Jan Kochanowski University in Kielce, Świetokrzyska 15, 25-406 Kielce, Poland 2 Department of Life Sciences, University of Alcalá, 28871 Alcalá de Henares, Madrid, Spain 1 Authors’ contributions AT: mycological collections, identification of Disciseda verrucosa fruit bodies, writing the manuscript; JŁ: identification of Disciseda verrucosa fruit bodies, writing the manuscript; GM: verification of the correctness of the Disciseda verrucosa fruit bodies Funding Financial support was provided by the Polish Ministry of Science and Higher Education, grant No. 612431. Competing interests No competing interests have been declared. Copyright notice © The Author(s) 2017. This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. Citation Tomaszewska A, Łuszczyński J, Moreno G. Disciseda verrucosa (Agaricomycetes, Basidiomycota) – a species new to Poland. Acta Mycol. 2017;52(2):1101. https://doi. org/10.5586/am.1101 Digital signature This PDF has been certified using digital signature with a trusted timestamp to assure its origin and integrity. A verification trust dialog appears on the PDF document when it is opened in a compatible PDF reader. Certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. If the certificate is missing or invalid it is recommended to verify the article on the journal website. * Corresponding author. Email: sikorka105@wp.pl Abstract The genus Disciseda in Poland until now was represented by two species: Disciseda bovista and D. candida. During the mycological investigations on the macromycetes fungi in the xerothermic grasslands of the Nida Basin, a species new for Poland from this genus, namely Disciseda verrucosa, was recorded. The basidiocarps of this species were found in the village Gacki (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), in the village Wola Zagojska (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), and in the Krzyżanowice Reserve (50°27'13" N 20°33'36" E; ATPOL square Fe 14), in the Ponidzie region. The basidiocarps of D. verrucosa were collected in the xerothermic grass, within Festucetum pallentis and Sisymbrio-Stipetum capillatae associations in the neighborhood of Festuca pallens, Stipa capillata, and Thymus marschalianus. On the basis of collected basidiocarps, macro- and microscopic characteristics were described and compared with other European Disciseda species. An identification key to the species of Disciseda occurring in Poland is given. Keywords Agaricomycetes; xerothermic fungi; Nida Basin Introduction The genus Disciseda is in the whole world represented by 15 species [1]. In Europe, about five species were found [2,3], whereas in Poland, Discidea is represented by two species: D. bovista (Klotzsch) P. Henn. and D. candida (Schwein.) C. G. Lloyd [4]. Both species in Poland belong to a highly endangered fungi, and classified to the category endangered – E [5]. Due to a rather characteristic basidiocarps, fungi from genus Discidea are relatively easy to identify. The taxonomy of species from this genus is based on the knowledge of individual features of macro- and micromorphological structure. Such features are: the shape, size and color of the endoperidium, color of the exoperidium, degree of exoperidium incrustation by the substrate particles, the shape and size of peristome, and also the spores size and episporium ornamentation [2,5]. In terms of habitat these fungi are thermophilous, often found on the dry, warm, sunny xerothermic and psammophilous sites. The mycelium and basidiocarps develop often in the open places with a southern exposure [2,5,6]. The aim of this paper is to increase the knowledge about the richness and distribution of Disciseda species in Poland. Material and methods The basidiocarps of Disciseda verrucosa were collected during the mycological investigations carried out in the xerothermic sites of the Nida Basin (SE Poland). The Published by Polish Botanical Society 1 of 7 Tomaszewska et al. / Disciseda verrucosa – a species new to Poland investigations included the communities of xerothermic plants distributed in the areas legally protected, such as: the areas of Nature 2000 (the Nida refuge, PLH 260003), the nature reserves (the Skorocice and Krzyżanowice reserve), and the landscape parks (Nida, Szaniec, Kozubów). The field studies included the cyclic mycological observations carried out on 30 research plots and supplemented by searching for fungi using the route method. Field studies were conducted in the village Gacki (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), in the village Wola Zagojska (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), and in the Krzyżanowice Reserve (50°27'13" N, 20°33'36" E; ATPOL square Fe 14), in the Ponidzie region. During the survey, the number of basidiocarps of given species and the organoleptic features such as: the shape, size, color of endoperidium, and color of exoperidium were taken into consideration. The laboratory studies were carried out using the light microscope, scanning electron microscope (SEM), and the standard chemical reagents (10% KOH, JKJ). The investigations using the light microscope (LM) included the measurements of capillitium and spores, which the structure, size, and shape have significant meaning for taxonomical species identification of this genus. The preparations were made from each collected basidiocarps. In each preparation, 10 randomly selected spores were measured. All the microscopic structures were carried out using the immersion lens. The investigations using the scanning electron microscope (SEM) were carried out in the Independent Department of Environment Protection and Modelling, Jan Kochanowski University in Kielce. The electron micrographs were taken at the magnifications: ×3000, ×5000, ×10 000, and ×12 000. Description of the basidiocarps was made on the basis of own specimens. The taxonomical identification of collected fungi fruit bodies was made based on the following works: Moravec [6], Rudnicka-Jezierska [7], Pérez-Silva et al. [8], Bates et al. [9], and Lizárraga et al. [10]. The nomenclature of plants follows Mirek et al. [11]. The vouchers were deposited in the Fungarium (KTC), Faculty of the Mathematics and Science, Jan Kochanowski University in Kielce, Poland Results Disciseda verrucosa G. Cunn., Trans. & Proc. New Zealand Inst. 57: 205 (1926) Syn. Disciseda arida Velenovský, Novit. Mycol. p. 169, 1939. – Moravec, Čas. čes. Houbařu 29: 15, 1952. Description. Disciseda verrucosa produces the basidiocarps typical for the genus Disciseda. The basidiocarps globose to discoid, slightly flattened 10–15(–20) mm in diameter. Exoperidium whitish, strongly encrusted with the particles of substratum. Endoperidium smooth, gray-brown, rigid, leathery and permanent (Fig. 1). The endoperidium dehiscing by an mammose laciniate and toothed stoma about 1 mm diam. Gleba brown to dark brown. Subgleba absent. Spores globose 7.0–(–8.0)–8.5 µm (measurement with warts), with short colorless sterigmata. Under LM, the spores have bright brown color and are from distinctly verrucose to delicately spiny. Under SEM, the ornamentation of these structures appears formed by thick verrucae 1.6 µm long (Fig. 1b–d). Capillitium septate, threads thick-walled, (up to 0.8 µm thick). Capillitium wavy, fragile (2.5–5.5 µm thick), with very small pores, the walls brightly brown (Fig. 1). Specimens examined. POLAND. 1 – Nida Basin, Gacki village, 45 km south from Kielce (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), two basidiocarps in xerothermic plant associations of Festucetum pallentis, September 2015; 2 – Nida Basin, Wola Zagojska village, 45 km south from Kielce (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), two basidiocarps in xerothermic plan associations of Festucetum pallentis, October 2015; 3 – Nida Basin, Krzyżanowice Reserve (50°27'13" N, 20°33'36" E; ATPOL square Fe 14), in the vicinity of the Pińczów town in the Nida Basin, 48 km south from Kielce, one basidiocarp in xerothermic plant associations of Sisymbrio-Stipetum capillateae, September 2016. © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1101 2 of 7 Tomaszewska et al. / Disciseda verrucosa – a species new to Poland a b c d e f Fig. 1 Disciseda verrucosa – mature basidiocarps (photo by A. Tomaszewska) (a). Disciseda verrucosa – spores (SEM; scale bar: 2 µm; photo by G. Moreno) (b). Disciseda verrucosa – spores (SEM; scale bar: 5 µm) (c). Disciseda verrucosa – spores (SEM; scale bar: 5 µm) (d). Disciseda verrucosa – capillitium with spores (LM; scale bar: 5 µm) (e). Disciseda verrucosa – capillitium (LM; scale bar: 5 µm) (f). Five basidiocarps of Disciseda verrucosa were collected during the mycological investigations (Tab. 1). The basidiocarps of D. verrucosa were recorded in the village Gacki (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), in the village Wola Zagojska (50°26'41" N, 20°36'34" E; ATPOL square Fe 24), and in the Krzyżanowice Reserve (50°27'13" N, 20°33'36" E; ATPOL square Fe 14), in the vicinity of the Pińczów town in the Nida Basin, 48 km south from Kielce (Fig. 2). The specimens were collected in the xerothermic grass, in the patches of Festucetum pallentis and Sisymbrio-Stipetum capillatae © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1101 3 of 7 Comparison of structure features of Disciseda species recorded in Poland (according to: (1) Moravec [6]; (2) Rudnicka-Jezierska [7]; (3) Bates et al. [9]; (4) Lizárraga et al. [10]; (6) Dörfelt, Nowak [3]). Published by Polish Botanical Society Acta Mycol 52(2):1101 No. Structure features Disciseda bovista D. candida D. nigra D. hyaalothrix D. verrucosa 1. Exoperidium White, whitish yellow, mature brown pale grey Dirty whitish yellowish, mature from brownish to earth colored Dark brownish White, whitish yellow. White, mature grey-orange 2. Endoperidium Rigid, pergameneous, color of hazel nut Strong, leathery, pergameneous, brown-grey Thin, papyraceous, blackish brown to black, very fine-grained Thin, papyraceous, brownish gray to dark brown, Permanent, leathery, color of “mouse”-grey brown 3. Peristome Frayed Frayed, fimbrillate Irregularly round, sometimes blown radially Frayed Slightly frayed at edges 4. Gleba Red brown Brightly brown, rusty brown Blackish brown Greyish brown or somewhat purplish Brown-orange, brightly brown 5. Spores (1) (5–)6.5–7.8(–8.6) µm; (2) 5–6(–7.1) µm; (4)4–7 µm in diameter. Distinctly strongly verrucose (verrucae 1–1.5 µm). (1) 3.0–4.2 µm; (2) 3.8–4.5–5 µm; (3) 4.0–5.6(–6.4) × 4.0– 5.6(–6.4) µm; (4) 4–5 µm in diameter. Smooth or puncticulate, very fine verrucose. (6) 7.5–8.5 µm in diameter with ornamentation. Strongly verrucose, verrucae 1.8 µm. (4) 8–11 μm in diameter, with spines. (1) 6. Sterigmata Present Present Very often the lack, if present, up to 1.5 µm long Present Short sterigma 0.8 µm long 7. Capillitium Wavy, fragile, 2.7–3.5 µm thick, without the pores Wavy, fragile, 2.5 µm thick, without the pores Wavy, fragile, 3.5–4.5 µm thick, without the pores Little branched, fragmentable, 3–5 μm diam., without pores Wavy, fragile, 2.4–5.6 µm thick, with small pores 8.5–11.5 µm; (3) (8.0–)8.8– 12.0(–12.8) × (8.0–)8.8– 12.0(–12.8) µm; (4) 7–8(–8.5) µm in diameter. Strongly verrucose, verrucae 1.6 µm. Tomaszewska et al. / Disciseda verrucosa – a species new to Poland © The Author(s) 2017 Tab. 1 4 of 7 Tomaszewska et al. / Disciseda verrucosa – a species new to Poland Fig. 2 Distribution of Disciseda verrucosa in Poland. associations. The specimens were recorded on the slope with a southern exposure and 45° inclination angle. The phytocoenosis of Festucetum pallentis (location: Gacki village) of the investigated xerothermic plant association is as follow: cover of herb layer – 60%, size of the area – 80 m2, Allium montanum 1.2, Anthericum ramosum +.2, Anthyllis vulneraria +, Artemisia campestris 1.2, Calamintha acinos +, Carex humilis 1.2, Festuca pallens 2.2, Festuca sulcata +.2, Galium verum +.2, Gypsophila fastigiata 2.2, Koeleria glauca +.2, Potentilla arenaria 2.2, Sedum sexangulare 2.2, Silene otites +, Stipa capillata +.2, Thymus marschalianus 2.2. The phytocoenosis of Sisymbrio-Stipetum capillatae (location: Krzyżanowice Reserve) of the investigated xerothermic plant association is as follow: cover of herb layer – 80%, size of the area – 100 m2, Stipa capillata 4.4, Artemisia campestris 1.2, Asperula cynanchica +.2, Astragalus dannicus +.2, Campanula sibirica 1.1, Carex humilis 2.2, Centaurea stoebe +.2, Euphorbia cyparissias 1.2, Festuca rupicola 1.2, Thymus marschallianus 2.2. Key for species identification of the genus Disciseda recorded in Poland 1 Spores delicately verrucose or smooth Mature basidiocarps discoid. Endoperidium strong, matt, color from brown-grey to pale grey. Spores globose, 3.8–4.5 µm. Capillitium brightly yellow, hyaline, thick-walled, without the pores. Disciseda candida (Schwein.) Lloyd 1* Spores thickly verrucose 2 2 Spores 8.0–12.0(–12,8) × (8.0–)8.8–12.0(–12.8) µm in diameter Basidiocarps globose, discoid, slightly flattened. Endoperidium grey-brown, rigid. Spores globose, distinctly thickly verrucose, with verrucae up to 1.6 µm long. Capillitium brightly yellow, hyaline, thick-walled, with small pores. Disciseda verrucosa G. Cunn., Trans. & Proc. New Zealand Inst. 57: 205 (1926) 2* Spores with diameter 5–6.5–7.8(–8.6) µm Fruitbodies globose, rarely flattened. Endoperidium with hazel nut color, pergameneous like, spores with verrucae 1–1.5 µm long. Capillitium brightly yellow, hyaline, rather thick-walled, without the pores. Disciseda bovista (Klotzsch) Henn. Discussion During the taxonomical identification of collected Disciseda verrucosa basidiocarps, a large similarity of this species to D. bovista was observed. Some authors [8] also point out the possibility of misidentifying D. verrucosa with other species of this genus, in particular with D. bovista. The feature distinguishing D. verrucosa from D. bovista is, first of all, the size of spores. The differences between these species are also due to a different ornamentation of these structures. Strongly verrucose of episporium is a characteristic feature for D. verrucosa. Examination of the morphology of the spore ornamentation of D. verrucosa shows no considerable differences in spore structures of D. verrucosa and D. hyalothrix. However, the examined spores of D. verrucosa were smaller than D. © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1101 5 of 7 Tomaszewska et al. / Disciseda verrucosa – a species new to Poland hyalothrix and had significantly smaller warts on the episporium surface [10]. Disciseda verrucosa and D. nigra have very similar spores in terms of size and ornamentation but both species differ very clearly in the color of the basidiocarps and gleba [3]. The list of macro- and micromorphological features of Disciseda species recorded in Poland is presented in Tab. 1. Disciseda verrucosa in Europe was up to now recorded in the Czech Republic and Slovakia [6], Spain [10], and recently also in Germany [12]. Outside of Europe, the species is known from: North America [8–10,13–18], Australia, New Zealand [19,20], South Africa [9,21], and South America [22–24]. Disciseda verrucosa is a thermophilous species. This species produces the mycelium and basidiocarps mostly in the open, sandy areas [6,9], but it was also found in the tropical deciduous forests under the plantings: Ceiba sp., Cercidium microphyllum, Olneya sp., Juniperus sp., Pinus edulis, and Prosopis sp. [8,9]. The following conclusions can be drawn from our studies: ■ Xerothermic plant communities developing in the area of the Nida Basin are characterized by an extraordinary high richness of gasteroid fungi species from warmer climates. An example described in this work is Disciseda verrucose, a species new to the Polish mycobiota. ■ In the context of the obtained results, we suggest the need of the revision of the national herbarium collections of fungi of the genus Disciseda, including molecular taxonomic studies. ■ The authors are expecting that further field studies will help to find the new localities of the others European species of Disciseda in Poland. References 1. Kirk PM, Cannon PF, David JC, Stalpers JA. Ainsworth and Bisby’s dictionary of the Fungi. 10th ed. Cambridge: University Press; 2008. https://doi.org/10.1079/9780851998268.0000 2. Sarasini M. Gasteromiceti epigei. Trento: Associazione Micologica Bresadola – Via A. Volta; 2005. 3. Dörfelt H, Nowak H. Disciseda nigra – ein verkannter Gasteromycetes. Feddes Repert. 2002;113:24–29. https://doi.org/10.1002/1522-239X(200205)113:1/2%3C24::AID-FEDR24%3E3.0.CO;2-S 4. Wojewoda W. Checklist of Polish larger Basidiomycetes. Cracow: W. Szafer Institute of Botany, Polish Academy of Sciences; 2003. (Biodiversity of Poland; vol 7). 5. Wojewoda W, Ławrynowicz M. Red list of the macrofungi in Poland. In: Mirek Z, Zarzycki K, Wojewoda W, Szeląg Z, editors. Red list of plants and fungi in Poland. Cracow: W. Szafer Institute of Botany, Polish Academy of Sciences; 2006. p. 53–70. 6. Moravec Z. 1. rod Disciseda CZERN. – Žaludice. In: Pilát A, editor. Flora ČSR, Gasteromycetes, Houby – břichatky. Praha: Československé Akademie Věd; 1958. p. 377–386. 7. Rudnicka-Jezierska W. Grzyby (Mycota). T. 23. Podstawczaki (Basidiomycetes), purchawkowe (Lycoperdales), tęgoskórowe (Sclerodermatales), pałeczkowe (Tulostomatales), gniazdnicowe (Nidulariales), sromotnikowe (Phallales), osiakowe (Podaxales). Kraków: Instytut Botaniki PAN; 1991. (Flora Polska. Rośliny Zarodnikowe Polski i Ziem Ościennych). 8. Pérez-Silva E, Esqueda M, Herrera T, Moreno G, Altés A. Disciseda verrucosa (Gasteromycetes) in Mexico. Mycotaxon 2000;76:337–341. 9. Bates ST, Roberson RW, Desjardin DE. Arizona gasteroid fungi I: Lycoperdaceae (Agaricales, Basidiomycota). Fungal Divers. 2009;37:153–207. 10. Lizárraga M, Esqueda M, Gutiérrez A, Piña C, Barredo-Pool F. El género Disciseda (Agaricales, Agaricaceae) en la Planicie Central del Desierto Chihuahuense, México. Rev Mex Micol. 2010;32:41–47. 11. Mirek Z, Piękoś-Mirkowa H, Zając A, Zając M. Flowering plants and pteridophytes of Poland a checklist. Cracow: W. Szafer Institute of Botany, Polish Academy of Sciences; 2002. (Biodiversity of Poland; vol 1). © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1101 6 of 7 Tomaszewska et al. / Disciseda verrucosa – a species new to Poland 12. Specht P, Schubert H. Seltene Gasteromyceten in Deutschland I – Disciseda verrucosa. Z Mykol. 2012;78(2):169–178. 13. Moreno G, Altés A, Ochoa C. Notes on some type materials of Disciseda (Lycoperdaceae). Persoonia. 2003;18:215–223. 14. Morales MI, Kimbrough JW. The Lycoperdaceae of North Central Florida. I. The genera Calvatia and Disciseda. Rev Biol Trop. 1978;26:227–236. 15. Gilbertson RL, Desjardin DE, Rogers JD, Hemmes DE. Fungi from the Mamane-Naio vegetation zone of Hawai’i. Fungal Divers. 2001;6:35–69. 16. Bates ST. A preliminary checklist of Arizona macrofungi. Canotia. 2006;2:47–78. 17. Esqueda M, Coronado M, Sanchez A, Perez-Silva E, Herrera T. Macromycetes of pinacate and great altar desert biosphere reserve, Sonora, Mexico. Mycotaxon. 2006;95:81–90. 18. Moreno G, Lizárraga M, Esqueda M, Coronado ML. Contribution to the study of gasteroid and secotioid of Chihuahua, Mexico. Mycotaxon. 2010;112:291–315. https://doi.org/10.5248/112.291 19. Moreno G, Esqueda M, Perez-Silva E, Herrera T, Altes A. Some interesting gasteroid and secotioid fungi from Sonora, Mexico. Persoonia. 2007;19:265–280. 20. Cunningham GH. The Gasteromycetes of Australia and New Zealand. Dunedin: J. McInhoe; 1942. 21. Grgurinovic AC. Larger fungi of South Australia. Adelaide: Botanic Gardens of Adelaide and State Herbarium and Flora and Fauna of South Australia Handbooks Committee; 1997. 22. Bottomley AM. Gasteromycetes of South Africa. Bothalia. 1948;4:473–810. https://doi.org/10.4102/abc.v4i3.1859 23. Kuhar F, Castiglia V, Zamora JC, Papinutti L. New records and notes on gasteroid fungi of arid regions in Argentina. Sydowia. 2012;64:233–244. 24. Silva BDB, Baseia IG. New records of Disciseda (Agaricales, Fungi) in the semiarid regions of Northeast Brazil. J Torrey Bot Soc. 2014;141(4):353–362. https://doi.org/10.3159/TORREY-D-14-00018.1 © The Author(s) 2017 Published by Polish Botanical Society Acta Mycol 52(2):1101 7 of 7