Unexpected high species diversity among European stalked puffballs - a contribution to the phylogeny and taxonomy of the genus Tulostoma (Agaricales) Author Jeppson, Mikael Author Altes, Alberto Author Moreno, Gabriel Author Nilsson, R. Henrik Author Yolanda Loarce, Author Bustos, Alfredo de Author Larsson, Ellen text MycoKeys 2017 21 33 88 http://dx.doi.org/10.3897/mycokeys.21.12176 journal article http://dx.doi.org/10.3897/mycokeys.21.12176 1314-4049-21-33 Tulostoma calcareum Jeppson, Altes , G. Moreno & E. Larss. sp. nov. Figure 3 Holotype. Sweden, Soedermanland , Moerkoe , Egelsvik, Kalkberget, on calcareous soil under Artemisia campestris in rocky slope, 5 Nov. 2004, M. Jeppson 6965 (GB!, isotype AH). Etymology. The name refers to its habitat requirement, on calcareous sandy soil or among calcareous rocks and cliffs. Description. Spore-sac subglobose, often somewhat depressed, 5-12 mm. Exoperidium hyphal-verrucose, deciduous, sometimes persisting as whitish verrucae scattered on the endoperidial surface. Endoperidium brownish-ochraceous, initially rather dark coloured, with age fading to greyish white. Mouth circular, shortly tubular (Figure 3f), surrounded by a greyish or brownish peristome. Socket separated from the stem, inconspicuous. Stem slender, 20-50 x 2-3 mm, initially orange brown, with age warm reddish brown, longitudinally furrowed, smooth to fissured and appressed scaly, with a basal mycelial bulb (Figure 3a,e). Gleba ochraceous to ferrugineous brown. Capillitium 4-10 µm with medium-thick walls, sometimes abundantly ramified. Septa not or very slightly widened (Figure 3b). Spores subglobose, 4.0-6.0 µm (av. 4.7-5.0 µm ), verrucose-echinate. SEM-photos show spines in groups with connected tips, sometimes coalescing to form ridges and crests (Figure 3 g-i ). Habitat and distribution. Occurs in dry, exposed to semi-shaded situations in calcareous, sandy habitats and on calcareous rocks and cliffs. It is currently on record from Hungary, Norway, Spain, and Sweden. Notes. Tulostoma calcareum was commonly misinterpreted as T. squamosum (cfr. Nitare 1997 ), under which name it is red-listed as Critically endangered (CR) in Sweden ( Artdatabanken 2015 ). It differs from T. squamosum by having a hyphal to slightly verrucose exoperidium, by lacking sphaerocysts, and by never forming the reticulate pattern typical of T. squamosum . The stipe is also less squamulose. Spores and capillitial characters are almost identical. It differs from T. melanocyclum , with which it can easily be confused, by having a more robust stature, more orange-reddish brown colours of the stipe, and slightly smaller spores (av. 4.7-5.0 µm vs 5.2-5.4 µm in T. melanocyclum ). SEM-photos of T. melanocyclum (Figure 2i) show similar spore ornamentation but with spines with less tendency to coalesce and form ridges. In macro-morphology T. calcareum is fairly similar to T. ferrugineum D.M. Oliver & Hosford, a species described from northwestern USA, whose type material no one has been able to locate ( Wright 1987 ). However, according to its original description, it differs from T. calcareum by having distinctly widened capillitial septa. Specimens examined. HUNGARY, Budapest: Pilisszanto , on dolomite grassland, 31 Dec. 2013, P. Finy 4, (GB)*. NORWAY, Oppland: Vaga , 15 Jul. 2010, A. Breili S0133 (GB)*. SPAIN, Girona: Lloret de Mar, Platja de Fenals, sandy grassland with scattered pines, near hotel buildings, Oct. 2010, J-O. Aarnaes 2010-1 (GB)*. SWEDEN, Soedermanland : Moerkoe , Egelsvik, Kalkberget, on calcareous soil under Artemisia campestris in rocky slope, 5 Nov. 2004, M. Jeppson 6966 (GB). Oeland : Boeda , Ramsnaes , Hagudden, torrmark mellan fritidshus och strandvallen, 5 Apr. 2007, E. Hultqvist, T. Knutsson, U. Andersson, T. Gunnarsson, M. Jeppson 9077 (GB); Gardby , sandgraeshed , 6 Mar. 1999, K. Bringer (GB); Gardby , V. om hembygdsgarden , sand steppe vegetation, 25 Sept. 2004, J. Jeppson, M. Jeppson 7141 (GB)*; Hoegsrum , Karums alvar, dry calcareous grassland around Juniperus bush, 11 Nov. 2006, Oelands Botaniska Foerening , M. Jeppson 8065 (GB)*; Hulterstad, Alby, pa Ancylusvallen vid infarten till skolan, vaeg-/traedgardskant i graesmatta med mossor pa sandig jord, 18 Apr. 1997, H. Lundqvist, T. Knutsson 97-034 (GB); Hulterstad, Alby skola, i graesmatta och rabatt, 15 Oct. 2002, S. Jeppson, J. Jeppson, M. Jeppson 6513 (GB); Hulterstad, Triberga, sand steppe vegetation, 15 Nov. 2003, J. Jeppson, M. Jeppson 6375 (GB)*; Ibidem, 15 Nov. 2003, M. Jeppson 6438 (GB)*; Norra Moeckleby , Doerby , torraeng i kanten av den forna jaernvaegsbanken , 4 Apr. 2009, E. Hultqvist, T. Knutsson, U. Andersson, T. Gunnarsson, M. Jeppson 9084 (GB); Norra Moeckleby , Bettorps vaegskael , sandstaeppsvegetation , 21 Jul. 2006, T. Knutsson, U. Andersson, T. Gunnarsson, M. Jeppson 8777 (GB); Resmo, "Resmokorridoren" , 5 May 2005, U. Andersson, T. Gunnarsson (GB); Raepplinge , Borgholms slottsalvar, kalkhaellmark i aengshavretorraeng , 11 Nov. 2001, J. Jeppson, M. Jeppson 5743 (GB); Ibidem, M. Jeppson 5744 (GB); Sandby, Sandby kyrkvaegskael , sandy roadside, 24 Sept. 2004, J. Jeppson, M. Jeppson 7121 (GB). Figure 3. T. calcareum , holotype (M. Jeppson 6965, GB): a, e basidiocarps b capillitium c, g-i spores f detail of spore-sac showing the tubular mouth d basidiocarps (M. Jeppson 6513, GB). Scale bars: a, d-e = 10 mm; b-c =10 µm ; f = 5 mm; g-i = 1 µm .