Amanita griseofusca: A new species of Amanita in section Vaginatae from Malam Jabba, Swat, Pakistan Author Kiran, Munazza University of the Punjab, Department of Botany, Quaid-e-Azam Campus- 54590, Lahore, Pakistan & Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, USA Author Khan, Junaid University of Swat, Center for Plant Sciences and Biodiversity, Saidu Sharif- 19200, Swat, Pakistan Author Sher, Hassan University of Swat, Center for Plant Sciences and Biodiversity, Saidu Sharif- 19200, Swat, Pakistan Author Pfister, Donald H. Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, USA Author Khalid, Abdul Nasir University of the Punjab, Department of Botany, Quaid-e-Azam Campus- 54590, Lahore, Pakistan text Phytotaxa 2018 2018-08-09 364 2 181 192 http://dx.doi.org/10.11646/phytotaxa.364.2.5 journal article 10.11646/phytotaxa.364.2.5 1179-3163 13703878 Amanita griseofusca J. Khan & M. Kiran sp.nov . ( FIGURES. 3 & 4 ) MycoBank no : MB 825012 Etymology:—the epithet “ griseofusca ” Latin refers to greyish brown pileus color characterizing the present taxon Holotype :— PAKISTAN . Khyber Pakhtunkhwa , Swat District , Malam Jabba valley , 1950 m a.s.l. , under Quercus dilatata Royle in mixed coniferous forest, 16 August 2015 , Junaid Khan MJ1558 ( Holotype SWAT000137 ! , Genbank no. MH241057 for ITS & MH241058 for LSU ). Diagnosis: —greyish brown pileus with white to beige, membranous volval remnants present as one (large) to a few (small) warts, striations one third of the total pileus radius, close, cream lamellae with a pink tone, fibrillose stipe, with a white loose and saccate volva turning beige at maturity and broadly ellipsoidal to ellipsoidal inamyloid basidiospores. FIGURE 3: A–D: Morphology of Amanita griseofusca . A: young stage fruiting body of MJ1558 (SWAT000137!). B: Basidiomata of MJ1558 in natural habitat. C: View of the lamellae and stipe. D: Basidiomata of MJ1630 (LAH35366!). Scale bars: A = 8 cm; B = 16.5 cm; C = 10 cm; D = 20cm. Photographs by Junaid Khan. Description:— Basidiomata ( Fig. 3 ) medium-sized. Pileus 50‒80 mm diam., hemispherical to elliptical when young, mostly flat or broadly convex with a depressed center at maturity, greyish brown (10YR 5/2) with a darker (10YR 3/2) disc, striated to almost one third from the margin, smooth at the center, volval remnants present as one (large) to a few (small) warts, warts membranous, white at first, turning greyish (2.5Y 7/4), context creamy white, fragile, thicker at the center ( 2‒3 mm ). Lamellae free, close, pruinose, especially in young specimens, broader ( 5‒7 mm ) at the center, lamellar edge even, creamy (7.5Y 9/4) at first, with a pinkish tone (2.5Y 9/4) at maturity, fragile, lamellulae rare, short, mostly in a single tier, rarely in three tiers, of varying lengths. Stipe 80‒100 × 8‒12 mm , slightly thickening downward, basal bulb absent, central, creamy white (10Y 9/4) to pale yellowish tan (2.5Y 9/4), fibrillose especially at the base, fibrils whitish to greyish white in young specimens, browning (10YR 5/6) with age, Annulus absent. Volva saccate, non-coherent in the upper part, white at first, turning beige (10Y 9/4) upon maturity, reduced in mature specimens. Odour not observed. Lamellar trama bilateral, mediostratum 30–50 μm wide composed of ellipsoid to sub-fusiform inflated cells (25–50 × 10–15 μm) with abundant interwoven filamentous hyphae, 5‒7 μm wide, vascular hyphae rare, 3‒7 μm in diam. Lateral stratum composed of ellipsoid to sub-fusiform hyphae (25‒40 × 10‒15 μm) with abundant interwoven filamentous hyphae which are 3‒7 μm wide and often branching, vascular hyphae rare, 3‒7 μm. Sub-hymenium 2‒3 layers, composed of subglobose to ovoid or rarely short ellipsoidal cells, 20‒30 × 10–20 μm, inflated cells rare, 3–7 μm. Basidia 40‒45 × 10‒15 μm, club-shaped, 2‒4 spored, basal clamp connections absent. Basidiospores [n/b/p] [60/3/3] (9.6‒) 10‒12 (‒12.5) × (8‒) 8.5‒10 (‒10.5) μm, L × W = 11 × 9 μm, Q = (1.1‒) 1.2‒1.3 (‒1.5), Qe = 1.22, broadly ellipsoid to ellipsoid, inamyloid. Lamellar edge sterile, 200–250 μm wide, composed of subglobose to ovoid cells (20–50 × 15‒30 μm), filamentous hyphae abundant, 3‒7 μm in diam., irregularly arranged, hyaline or rarely with a brownish pigment. Pileipellis 90‒130 μm thick, upper layer 30‒50 μm thick, gelatinized, composed of moderately compact hyaline cells, 2‒5 μm in diam., lower layer 60‒80 μm thick, composed of radial, compact, filamentous hyphae 3‒5 μm wide, hyaline or with a brownish pigment in KOH, vascular hyphae rare 2‒5μm wide. Stipe trama composed of longitudinally arranged filamentous hyphae, 5‒10 μm wide, hyaline in KOH, terminal cells clavate to broadly clavate 100‒300 × 15‒30 μm. Volval remnants on the pileus composed of filamentous hyphae 3‒5 μm wide, frequently branching, interwoven, thin-walled, hyaline, scattered inflated cells, variable, subglobse to globose, rarely ellipsoid to ovoid, 25‒30 × 18‒25 μm wide, mostly single, terminal, thin-walled, hyaline in KOH. Volva at stipe base two-layered, outer volval layer: filamentous hyphae (4–7 μm diameter) abundant to very abundant, hyaline, thin walled, compact, scattered, infrequent, globose to ellipsoidal inflated cells; inner volval layer: inflated cells abundant to fairly abundant, frequent, globose to subglobose to ovoid (45‒50 × 55‒60 μm), usually single and terminal. Clamp connections absent in all tissues. FIGURE 4. Amanita griseofusca ( SWAT000137! ). A. basidiospores; B. volval remnants from pileus surface; C. basidia and subhymenium; D. stipitipellis. E. outer layer of volval elements at stipe base F. inner layer of volval elements at stipe base. Scale bars: A = 8.7μm; B = 15 μm; C = 8.5 μm; D = 20 μm; E= 20 μm. Drawings by Munazza Kiran. Habit and Habitat:—solitary on soil under Quercus dilatata Royle. Distribution:—currently only collected from Kishawra village, Malam Jabba Valley, Swat district, Khyber Pakhtunkhwa province , Pakistan . Additional materials examined:— PAKISTAN . Khyber Pakhtunkhwa , Swat District, Malam Jabba valley, 1950 m a.s.l., under Quercus dilatata in mixed coniferous forest, 10 August 2016 , Junaid Khan MJ1630 ( LAH35366! , Genbank no. MH241055 for ITS & MH241056 for LSU) 31August 2016 , Junaid Khan MJ1664 ( SWAT000138! , Genbank no. MH241059 for ITS).