Novel collections of Zeus olympius and Cosmospora ganymede (Ascomycota) from Bulgaria and Greece Dimitar Y. STOYKOV Summary: First Bulgarian records of Zeus olympius and Cosmospora ganymede, two unusual ascomycetes, Boris ASSYOV described and so far known only from a single locality in Greece, are reported. Zeus olympius is related to Rossen ALEXOV Bosnian pine (Pinus heldreichii) and Cosmospora ganymede is associated with dead ascomata of the former. Krasimir GRAZDILOV Newly collected specimens of both species from the locus classicus on the Mt Olympus, and of Z. olympius from Mt Pindus were also examined. Description of the Bulgarian and Greek materials is provided, including data from in vivo study, supplemented with illustrations of macroscopic and microscopic features, including Ascomycete.org, 6 (4) : 73-80. the first colour photographs of C. ganymede. Germination of ascospores of Z. olympius was observed for the Octobre 2014 first time. The new collections significantly extend the geographic area of those apparently endemic asco- Mise en ligne le 17/10/2014 mycetes. We also provide the first record of Z. olympius on artificially plantedP. heldreichii outside its natural range. Keywords: Balkan Ascomycota, endemic fungi, Nectriaceae, Pinus heldreichii, Rhytismataceae. Résumé : premiers signalements bulgares de Zeus olympius et Cosmospora ganymede, deux ascomycètes peu courants, décrits et connus jusqu'à présent que d'une seule localité en Grèce, sont rapportés. Zeus olym- pius est lié au pin de Bosnie (Pinus heldreichii) et Cosmospora ganymede est associé aux ascocarpes morts du premier. De nouveaux spécimens des deux espèces recueillis du locus classicus sur le mont Olympe et de Z. olympius du mont Pinde ont également été examinés. Des descriptions des spécimens bulgares et grecques sont fournies, y compris les données de l’étude in vivo. Celles-ci sont complétées par des illustrations des caractéristiques macroscopiques et microscopiques, y compris les premières photographies en couleur de C. ganymede. L’observation de la germination des ascospores de Z. olympius est reportée pour la première fois. Les nouvelles récoltes étendent sensiblement la répartition géographique de ces ascomycètes appa- remment endémiques. Nous fournissons également les premiers enregistrements de Z. olympius dans des plantations artificielles deP. heldreichii en dehors de son aire de répartition naturelle. Mots-clés : Ascomycota des Balkans, champignons endémiques, Nectriaceae, Pinus heldreichii, Rhytismata- ceae. Riassunto: Sono riportati primi ritrovamenti bulgari di Zeus olympius e Cosmospora ganymede, due ascomi- ceti insoliti, descritti e finora conosciuti solo da una singola località in Grecia. Zeus olympius è legato a pino bosniaco (Pinus heldreichii) e C. ganymede è associata ad ascomi morti del primo. Sono stati esaminati anche esemplari delle due specie provenienti da nuove raccolte dal locus classicus sul Monte Olimpo e di Z. olympius dal Monte Pindo. Sono fornite descrizioni del materiale bulgaro e greco, inclusi i dati provenienti da studi in vivo, integrate con le illustrazioni di caratteri macroscopici e microscopici, tra cui le prime fotografie a colori di C. ganymede. Le nuove raccolte estendono significativamente la distribuzione geografica di tali ascomiceti apparentemente endemici. La germinazione di ascospore di Z. olympius è stata osservata per la prima volta. Si registra per Z. olympius, la prima raccolta avvenuta su piante da impianto artificiale di P. heldreichii, fuori dal suo areale naturale. Parole chiave: Ascomycota dei Balcani, funghi endemici, Nectriaceae, Pinus heldreichii, Rhytismataceae. Introduction cotton blue and lactophenol with cotton blue (heating applied with the last two reagents). The amyloid reaction of the microscopic structures was tested in IKI (BARAL, 1987) and Melzer’s reagent, upon Zeus olympius Minter & Diamandis is an unusual ascomycete, ap- the recipe in KIRK et al. (2008), with and without KOH pretreatment. parently restricted to its host, Bosnian pine (Pinus heldreichii Christ.; The microscopic study was conducted with the aid of Olympus BX- Fig. 1), and Cosmospora ganymede (Minter & Diamandis) Rossman 41 and AmScope T360B microscopes (oil immersion objectives & Samuels is yet another peculiar species, which seems associated 100×), equipped respectively with Olympus E330 and Amscope with old ascomata of Z. olympius. Until recently they were only MU900 digital cameras. Measurements were performed on micro- known from the locus classicus on the Mt Olympus in Greece (MINTER photographs with Piximetre v. 5.2 software. Spores were obtained et al., 1987), and considered as a putative endemism of this area. Ho- from fresh mature ascomata and were always measured in water. wever, intensive search revealed their presence in some localities in From each ascoma 25 or 30 spores were measured, depending on Bulgaria and also allowed the study of newly collected specimens the availability. Spore measurements are reported in the following from Mt Olympus and Mt Pindus. Detailed description and photo- format: (minimum–)mean±standard deviation(–maximum), Q = graphs of the new collections of the two species are presented he- length/width ratio (n = sample size), except for the descriptions, rein, expanding the data about their morphology. where they are given in the following manner: minimum–(meanmi- nimum–meanmaximum)–maximum. For the remaining microscopic Materials and methods structures, only minimum and maximum values are reported. Color photographs were taken from fresh collections, while these Taxonomy were later dried and preserved in the Mycological Collection of the Institute of Biodiversity and Ecosystem Research (SOMF), Sofia. The Zeus olympius Minter & Diamandis, Trans. Brit. Mycol. Soc., 88 (1): description of colours of ascomata follows the British Fungus Flora 55 (1987). Colour Chart (ANONYMOUS, 1969) with some approximation. Fungi were examined microscopically in living state in water (BARAL, 1992), Illustrations: MINTER et al. (1987: 56, habitus, ascomata and mi- and in dead state in 5% KOH, congo red in 10% ammonia, aqueous croscopic features, black and white drawing); DIAMANDIS (1992: 498, 73 Fig. 1 — Host tree and habitat of Zeus olympius a-b – leaves and cones of Pinus heldreichii; c – habitat; d – dead young trees with ascomata of Z. olympius developed on the main stem and lateral branches. Photos: B. Assyov. 74 ascomata, colour photograph); MINTER (1996: 183, reproduction of portions, and revealing the luteous, lemon chrome, yellowish the illustration from the protologue); DIAMANDIS & PERLEROU (2011: 30, orange to dirty orange coloured hymenium, in senescent ascomata ascomata, colour photograph); this paper (Figs. 2–3). color is fading to orange brown to brown, sometimes with oliva- ceous tinge. Asci develop from croziers or may occasionally have a Original diagnoses bifurcate base, present a thin wall and rounded apex, and measure GENERIC DIAGNOSIS. Hic fungus ascophorus, ad ordinem Rhytismatalium et fa- 105–195 × 10–17 μm (n=60). They are 8-spored, lacking a distinctive miliam Rhytismatacearum pertinens, corticola, erumpens, clypeo nigro asco- apical apparatus, mostly do not react with iodine solutions, but matum sane statu tectus, ascosporas habet ellipsoideas vaginis indutas some show striking dextrinoid reaction in IKI. Ascospores biseriate mirabilibus et ascos nil in iodo coerulescentes. Habet ascomata Therryae vel Coc- in very young asci, becoming uniseriate at maturity, hyaline, comyceti aliquid similia, et ascos multarum Rhytismatacearum generibus si- miles, sed ascosporas adspectu mentem revocantes Ceratophacidium. smooth, ellipsoidal, aseptate or very rarely presenting a single sep- SPECIES DIAGNOSIS. Haec species incolat Iovis montem, virgas Pini leucodermis tum (lactophenol), bearing a transparent mucous sheath somewhat et ramulos mortuos habitans. Habet ipsis e virgis erumpentia ascomata quae constricted at the equator and extending up to 20 μm beyond the corticem arboris reflectant ut aperiantur. Ascomata quodque diametrum circa apices of the ascospore. Spore content is with numerous small gut- 0.5–2 mm habet, et nigro a clypeo tegitur quod conpluribus in partibus statu tules in water mounts, KOH, congo red in ammonia and aqueous maturo fissum est ut discum videatur. Asci plus minusve cylindrici, circa 100– Cotton blue, and more or less uniform in reagents with lactophenol 120 × 10–12.5 μm magnitudinis respectu, nil in apicibus crassiores, sed saepius and chloral hydrate. Spores measure 10.8–(12.7–15.6)–18.1 × 5.0– aliquanto curvati, seriatim maturant et ascosporas octo quique ferunt, fissura (6.2–7.9)–9.2 μm (excluding sheath), Q = 1.5–(1.9–2.1)–2.6 (n = 345). apicali quique dehiscentes ut ipsae liberentur. Ascosporae nec colore nec septis sese gaudentes, tenuitunicatae sunt et unicum habent parietem. In quibusque Germination occurs through a single germinative tube at one of the adest vagina mucosa sane aliquid eleganter et suaviter in apicibus fastigiata. apices of the spore. Paraphyses measure up to 3.5 μm wide, and Sporae autem, sine vaginis circa 12–15 × 5–8 μm magnitudinis respectu me- are thin-walled, filiform, mostly unbranched, sometimes with anas- tiuntur. tomoses close to the base, rarely septate, slightly widened at the apex up to 7 μm, in water mounts with yellowish globular content Description of the new specimens and vacuoles. Hymenium up to 200 μm thick in cross section. It Ascomata up to 8 mm in diameter, appearing scattered on dead arises from