A New Species of Fuscoporia (Hymenochaetaceae, Basidiomycota) from the Brazilian Atlantic Rainforest
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A new species of Fuscoporia (Hymenochaetaceae, Basidiomycota) from the Brazilian Atlantic Rainforest Viviana Motato-Vásquez(1), Ricardo Matheus Pires, & Adriana de Mello Gugliotta (1) Instituto de Botânica, Núcleo de Pesquisa em Micologia, Av. Miguel Stéfano 3687, 04301-902, São Paulo, SP, Brazil. Corresponding author: [email protected] Abstract: Fuscoporia atlantica sp. nov. is Brazil, an unknown species of Fuscoporia was found described and illustrated based on specimens and it is described as new. The phylogenetic collected in Atlantic Rainforest areas of São Paulo inference of its relationship based on ribosomal ITS state, Brazil, and the inferred phylogenetic and LSU sequence data demonstrated that it forms a relationships are based on sequence data from monophyletic terminal clade, distinct from all the the ribosomal ITS and LSU regions. The new other Fuscoporia species for which DNA sequence species is characterized by an effused-reflexed to data are available. The species is described here as F. pileate basidioma, golden-yellow-ferruginous atlantica. pore surface, tiny pores, dimitic hyphal system with thin-walled hyphae at dissepiments edges MATERIALS AND METHODS encrusted, mostly uncinate or hooked hymenial Morphological study setae and broadly ellipsoid to ellipsoid Specimens were collected in Atlantic Rainforest basidiospores. areas in the São Paulo state, Brazil, between 2012 Key words: basidiomycetes, neotropics, and 2013. For microscopic analysis, free hand phylogeny, taxonomy sections of the basidiomata were mounted on microscope slides with a drop of 3% KOH solution INTRODUCTION and 1% aqueous phloxine solution. Amyloid and Traditionally the species of the genus dextrinoid reactions were observed in Melzer’s Fuscoporia Murrill have been included in reagent. The specimens were deposited in Herbário, Phellinus s.l., a morphologically heterogeneous Instituto de Botânica, São Paulo, Brazil (SP). and phylogenetically polyphyletic group (Fiasson Sequencing & Niemelä 1984, Dai 1999, Wagner & Fischer Total DNA was extracted from dried basidiomata. 2001). However, molecular phylogenetic analyses Extractions were carried out using the Sigma-Aldrich have shown that Fuscoporia species form a clade Gen EluteTM Plant Genomic DNA Miniprep Kit and independent from Phellinus s.s. (Wagner & later amplified with Sigma-Aldrich ReadMixTM Taq Fischer 2002, Larsson et al. 2006). PCR P4600 (Sigma-Aldrich Corporation, St. Louis, MO, The genus Fuscoporia was described to USA), following the manufacturer’s accommodate species occurring principally on recommendations. PCR reactions were performed in coniferous and deciduous trees, with resupinate a thermal cycler (C1000 Touch™ Thermal Cycler Bio- to pileate basidiomes; monomitic to dimitic Rad). Primers pairs used were ITS1 and ITS4 for the hyphal system; encrusted generative hyphae at ITS region (including ITS1, 5.8S and ITS2) and LR0R the dissepiments (which is described as a and LR7 for nLSU (White et al. 1990). PCR products distinctive feature within Phellinus s.l.), hymenial were sequenced in both directions using the same setae and cylindric to ellipsoid, hyaline, primers as the amplification. Nucleotide sequences inamyloid, indextrinoid, thin-walled and smooth were determined using an Applied Byosystems basidiospores (Murrill 1907, Fiasson & Niemelä 3730xl DNA Analyser (Macrogen, Korea). Edited 1984, Dai 1999, 2010; Wagner & Fischer 2001). sequences have been deposited in GenBank. During the study of wood-inhabiting fungi in Phylogenetic analyses Atlantic Rainforest areas of the São Paulo state, All sequences were matched against the entire 21ª Reunião Anual do Instituto de Botânica - Resumo Expandido - GenBank database using BLAST Etymology: atlantica – refers to the Brazilian (http://blast.ncbi.nlm.nih.gov/), and their Atlantic Rainforest biome, the only known habitat of pairwise identity was recorded. Sequences this species. retrieved from GenBank (NCBI) included in the Basidioma annual, pileate, sessile to effuse- study were selected based on the quality, reflexed, flexible when fresh to leathery when dry, identity, length and alignment test to build the in- solitary to imbricate, without odor or taste. Pileus group. The selected out-group for ITS analyses broadly attached, semicircular, applanate, up to was Phellinidium fragrans (M.J. Larsen & (5.0–)8.0–9.0 × 4.0–6.0 × 0.1–0.4 cm; upper surface Lombard) M. Fisch., P. weirii (Murrill) Y.C. Dai, concentrically zonate, radially wrinkled, glabrous to and P. sulphurascens (Pilát) Y.C. Dai, based in the tomentose, brown to yellowish-brown, with a matte results of the LSU region, that places Phellinidium dark brown crust at the base. MARGIN thin, obtuse, as a sister group of Fuscoporia; and for LSU entire, not involute when dry, sterile, up to 0.2 cm, analyses was Bondarzewia montana (Quél.) yellowish-brown to black brown. Context Singer, Russula violacea Quél., and Stereum homogeneous, dense, azonate, golden-yellow- hirsutum (Willd.) Pers., based in previous work ferruginous. Pore surface golden-yellow-ferruginous (Wagner & Fischer 2002). Nucleotide sequences (N30A99M30), pores rounded, invisible to the naked were aligned automatically with MAFFT v.7 eye, 7–9 per mm, tubes concolor with the pore (http://mafft.cbrc.jp/aligment/server/), and then surface, up to 0.2 cm deep, dissepiments entire to optimized manually with BioEdit 7.2.0 (Hall slightly lacerated, thin to slightly thick. Hyphal 1999). Best models of evolution were estimated system dimitic, generative hypha simple-septate, by using JModeltest 2.c1.4 (Darriba et al. 2012). hyaline, golden yellow to rusty brown, thin to thick- The substitution model used in the LSU dataset walled, straight, sparsely branched, 3.0–6.5 µm was TrN+I+G and for ITS dataset was TVM+G. diam., rarely branched, thin-walled hyphae at Bayesian analysis was performed based on dissepiments edges encrusted; skeletal hyphae thick- independent Markov chain Monte Carlo (MCMC). walled, straight, unbranched, dark-brown, 3.0–6.0 The run consisted of five million generations, two µm. Hymenial setae abundant, subulate to independent runs, and four independent chains. ventricose, mostly uncinate or hooked, dark brown The sampling frequency was every 500 and thick-walled, 20–45 × 7–10 µm. Cystidioles generations. The first ten percent of the analysis abundant. Basidia clavate, hyaline, 4-sterigmated, (burn-in phase) was discarded. Posterior thin-walled, 9–11 × 5–7 µm. Basidiospores abundant, probabilities (PP) were determined in MrBayes broadly ellipsoid to ellipsoid, hyaline to pale yellow, 3.2.2 (Ronquist et al. 2012) to test the support of smooth, thin-walled, inamyloid, 4–4.5 × (2–)3–3.5 the branches’ nodes. PP values ≥0.95 were µm (Xm = 4.2 × 2.8 µm), n = 60/2, Q = (1.1–)1.3–2, considered statistically significant, hence, all Qm = 1.5. branches supported below this value were Additional specimens examined — Fuscoporia disregarded. Maximum likelihood (ML) analysis atlantica: BRAZIL. SÃO PAULO STATE: Parque Estadual was performed using RaxML 7.5.4-standard da Serra do Mar, Núcleo Santa Virginia, 11 May 2013, (Stamatakis 2006) with 1000 rapid bootstrap R.M. Pires 8 (SP465829; GenBank KP058514, replicates. Trees from ML analysis were KPKP058516). Fuscoporia gilva: BRAZIL. SÃO PAULO compared with those resulting from the Bayesian STATE: Parque Estadual da Cantareira, 21 October analysis and bootstrap values >50% were used to 2011, V. Motato-Vásquez & A.M. Gugliotta 22 support the respective nodes compatible (SP417971); 16 February 2012, M. Capelari & J.J.S. between two trees. Oliveira 4683 (SP445350). Fuscoporia rhabarbarina: BRAZIL. SANTA CATARINA STATE: 26 December 1988, F. RESULTS AND DISCUSSION Furlani & C.L. Leite 186 (FLOR 10929). CANADA. on wood, Dr. Richardson 4 (K187843, lectotype). Taxonomy Fuscoporia atlântica Motato-Vásquez, R.M. Pires Comments — Fuscoporia atlantica is a distinct & Gugliotta, sp. nov. species that produces effused-reflexed to pileate Types: Brazil. São Paulo state: Parque Estadual basidiomata, radially wrinkled and tomentose pileus, da Cantareira, 23º3236S 46º3759W, 27 June with a matte dark brown crust at the base, a 2012, V. Motato-Vásquez, M.C. Westphalen & yellowish-brown when fresh to black brown after dry A.C. Bolaños 230 (Holotype, SP445618; GenBank margin, tiny pores, hooked hymenial setae and KP058515, KP058517). broadly ellipsoid to ellipsoid basidiospores. 2 21ª Reunião Anual do Instituto de Botânica - Resumo Expandido - Fuscoporia atlantica is very similar to F. gilva VMV also gratefully acknowledges the financial (Schwein.) T. Wagner & M. Fisch., which differs support received from the Programa Estudantes- microscopically by its straight setae and smaller Convênio de Pós-Graduação – PEC-PG, from basidiospores, and macroscopically by its CAPES/CNPq – Brazil. RMP also gratefully crustless pileus base and its dark purplish brown acknowledges the financial support received from pore surface (Loguercio-Leite & Wright 1995, Dai Fundação de Amparo à Pesquisa do Estado de São 1999). Fuscoporia atlantica is also Paulo – FAPESP (12/25493-7). morphologically similar to F. rhabarbarina (Berk.) Groposo et al., which can be easily distinguished LITERATURE CITED macroscopically by a glabrous pileus, with sulcate Dai, Y.C. 1999: Phellinus sensu lato (Aphyllophorales, Hymenochaetaceae) in East Asia. Acta Bot. Fenn. 166: zones and a distinctive black crust covering the 1–115. entire pileus surface, yellowish brown context; Dai, Y.C. 2010: Hymenochaetaceae (Basidiomycota) in and microscopically