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Gerhardtia Sinensis (Agaricales, Lyophyllaceae), a New Species and a Newly Recorded Genus for China

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Gerhardtia Sinensis (Agaricales, Lyophyllaceae), a New Species and a Newly Recorded Genus for China Phytotaxa 332 (2): 172–180 ISSN 1179-3155 (print edition) http://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2017 Magnolia Press Article ISSN 1179-3163 (online edition) https://doi.org/10.11646/phytotaxa.332.2.4 Gerhardtia sinensis (Agaricales, Lyophyllaceae), a new species and a newly recorded genus for China TING LI, TAIHUI LI*, CHAOQUN WANG, WANGQIU DENG & BIN SONG State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open laboratory of Applied Microbiology, Guangdong Institute of Microbiology, Guangzhou, 510070, China * Corresponding author: [email protected] Abstract Gerhardtia sinensis, a new species discovered from southern China, is described, illustrated and compared with morphologi- cally similar and phylogenetically related species. Morphologically, it is characterized by its white to yellowish or partially pale yellow pileus with faint striae, subdistant or nearly subdistant intervenose lamellae, and slightly verruculose basidio- spores that are (4.7–)5.2–6.2(–6.6) × (2.5–)3.0–3.4(–3.8) μm in size. Molecular phylogenetic analyses of the new species and related taxa were performed based on the sequences of nuclear ribosomal large subunit (nrLSU) and nuclear ribosomal internal transcribed spacer (ITS). Key words: Lyophyllaceae, phylogenetics, subtropical fungus, taxonomy Introduction Gerhardtia Bon (1994: 66) is a fungal genus in the family Lyophyllaceae, with G. borealis (Fr.) Contu & A. Ortega (2001: 176) [= G. incarnatobrunnea (Ew. Gerhardt) Bon (1994: 66)] as the type species (Bon 1994). It was once regarded as a subgenus Lyophyllopsis Sathe & J.T. Daniel (1981: 87) under Lyophyllum P. Karst. (1881: 29)(Gerhardt 1982), and then was raised to an independent genus with the new name Gerhardtia by Bon (1994) who emphasized the presence of minutely verruculose basidiospores, siderophilous granulation in the basidia, a pileipellis organized as a cutis, trichoderm or hymeniderm and an absence of clamp-connections as the generic characterizing features which distinguish Gerhardtia from other genera in Lyophyllaceae. Vizzini et al. (2017) redefined the characters of this genus, with a particular emphasis on basidiospores that were irregular and undulate to nodulose but not verrucose in outline. To date, only nine species in Gerhardtia have been validated excluding synonyms and varieties (Baroni 1981; Bon 1994; Contu & Ortega 2001; Contu & Consiglio 2004; Kalamees 2008; Cooper 2014; Vizzini et al. 2015, 2017; Matheny et al. 2017). Recently, a new species of Gerhardtia was found in southern China, which was the first discovery of the genus in China and even in Asia. Therefore, full description and illustration of the new species are formally presented as follows. Materials and methods Morphological methods Fresh basidiomata were collected from several field sites in southern China. All samples were dried with a food dehydrator and deposited in the Fungal Herbarium of Guangdong Institute of Microbiology (GDGM), Guangzhou, China, except for some small pieces of basidiomata which were dried with silica gel for molecular study. The photographs of the basidiomata were taken in the field with digital cameras in natural light. Microscopic characters were observed with a light microscope (Zeiss Axio Lab. A1). After rehydrating and CO2 critical point drying processes, several tiny pieces of hymenophore were coated with gold/palladium, and basidiospore images were taken with a 172 Accepted by Genevieve Gates: 30 Nov. 2017; published: 19 Dec. 2017 scanning electron microscope (SEM) (HITACHIS-3000N) at 15kV with a working distance of 10 mm. The observed macro-morphological features were based on fresh materials and field photographs; and the microscopic structures were measured and drawn from revived sections of dried specimens, which were hand-sectioned and mounted in 5% KOH solution, 1% Congo Red or Melzer’s reagent, and 0.05% Cotton Blue in lactic acid solution following Kühner (1938), Grund & Marr (1965) and Matheny et al. (2017) in order to examine siderophilous granulation in the basidia. Colour descriptions were made according to Kornerup & Wanscher (1978). The notations of basidiospores (n/m/p) indicate that the measurements were made on ‘n’ randomly selected basidiospores from ‘m’ basidiomata of ‘p’ collections. Basidiospore dimensions are given as: (minimum–) average minus standard deviation–average plus standard deviation (–maximum) of length × (minimum–) average minus standard deviation-average plus standard deviation (–maximum) of width. Q refers to the length/width ratio of an individual basidiospore; Qm refers to the average Q value of all basidiospores ± sample standard deviation. Molecular methods Total DNA was isolated from 15 mg of voucher specimen dried with silica gel using the Sangon Fungus Genomic DNA Extraction kit (Sangon Biotech Co., Ltd., Shanghai, China). Primers LROR & LR5 (Vilgalys & Hester 1990, Vilgalys lab. http://www.botany.duke.edu/fungi/mycolab) were used for the LSU rDNA amplification and primers ITS1 & ITS4 for the ITS region amplification (White et al. 1990; Gardes & Bruns 1993). Amplification reactions were performed in 25 μL reaction mixtures with the following total amounts: 5 ng DNA, 2×PCR EasyTaq SuperMix (TransGen Biotech, Beijing) 10 μL, 1 mM of each primer and added to 25 μL with ddH2O. The cycling parameters were as follows: 5 min at 94℃ for 1 cycle; 30 s at 94℃, 30 s at 55℃, 60 s at 72℃ for 35 cycles, 8 min at 72℃ for 1 cycle, pause at 22℃. PCR products were electrophoresed on a 1% agarose gel and visualized by staining with ethidium bromide. The PCR products were sequenced by the Beijing Genomics Institute (BGI, Shenzhen). The obtained sequences were submitted to GenBank; and the other referred sequences of related taxa were downloaded from GenBank. The nrLSU data matrix includes 19 fungal sequences and consists of 8 taxa with 904 nucleotide sites (gaps included); Tricholoma sinoacerbum T.H. Li, Iqbal Hosen & Ting Li (2016: 234) served as the outgroup. The ITS dataset includes 21 fungal sequences vesting in 8 taxa with 655 nucleotide sites (gaps included), and the sequence of T. sinoacerbum was rooted as the outgroup. Sequence alignment was carried out with ClustalX (Thompson et al. 1997). Alignments were edited with Bioedit software (Hall 1999). Maximum Likelihood (ML) analyses were performed with MEGA version 5.2 (Hall 2013). Bootstrap analysis was implemented with 1000 replicates. Branches corresponding to partitions reproduced in less than 50% bootstrap replicates were collapsed. All positions containing gaps and missing data were implemented from the dataset. Results Taxonomy Gerhardtia sinensis T.H. Li, T. Li, C.Q. Wang & W.Q. Deng, sp. nov. Figs. 1–3 MycoBank: MB 819692 Diagnosis:—Differs from G. highlandensis (Hesler & A.H. Sm.) Consiglio & Contu (2004: 158) in having white to yellowish pileus with faint striae, more distant, intervenose lamellae, more lamellulae between two complete lamellae, and slightly larger basidiospores measuring (4.7–)5.2–6.2(–6.6) × (2.5–)3.0–3.4(–3.8) μm. Etymology:—The epithet “sinensis” refers to the type locality in China. Typification:—CHINA. Guangdong Province, Zhaoqing, Dinghushan Biosphere Reserve, at 112°34′ E, 23°10′ N; 8 August 2010, coll. Chunying Deng & Taihui Li (Holotype, GDGM 29981). GERHARDTIA SINENSIS (AGARICALES, LYOPHYLLACEAE) Phytotaxa 332 (2) © 2017 Magnolia Press • 173 FIGURE 1. Gerhardtia sinensis. a. Basidiomata of GDGM 29981 (holotype) b. Basidiomata of GDGM 46394 c. Basidiomata of GDGM 42158 d. Basidiomata of GDGM 45221 Scale bars: a–d = 2 cm. Photos by: Ting Li & Chaoqun Wang. FIGURE 2. Gerhardtia sinensis. a. Cyanophilous bodies in basidia of GDGM 29981 (holotype). b. Cyanophilic basidiospores of GDGM 29981. c–d. Basidiospores of GDGM 29981 under SEM. e–f. Basidiospores of GDGM 46393 under SEM. Scale bars: a–b = 10 µm; c = 2 µm; d–f = 1 µm. Photos by: Ting Li. 174 • Phytotaxa 332 (2) © 2017 Magnolia Press LI ET AL. FIGURE 3. Microscopic features of Gerhardtia sinensis (GDGM 29981, holotype). a. Basidiospores. b. Basidia. c. Pileipellis. Scale bars: a–c 10 µm. Drawings by: Ting Li. Description:—Basidiomata small to medium. Pileus 32–56 mm broad, convex at first, expanding to plano-convex or applanate, depressed over disc when matured, sometimes slightly broadly umbonate or umbilicate at centre; margin involute when young, expanding to inflexed or at times uplifted in places when mature, sometimes flexuous or even somewhat lobed; surface nearly white, yellowish white to pale yellow (1A2–3, 3A3) at disc, snow white, satin white to yellowish white (1A1, 2A1–2, 3A2) near margin, dry, dull, somewhat hygrophanous with moisture loss, glabrous, usually with faint to moderately obvious striation; context 2.5–4.5 mm thick at stipe, less than 1 mm thick at near pileus margin, white to yellowish white (2A1–2), unchanged when cut or injured. Lamellae adnate to slightly sinuate, GERHARDTIA SINENSIS (AGARICALES, LYOPHYLLACEAE) Phytotaxa 332 (2) © 2017 Magnolia Press • 175 subdistant or close to subdistant, with 24–32 complete lamellae and 5–8 lamellulae of different lengths between two complete lamellae, 4–7 mm broad, concolorous with pileus to somewhat yellowish white (1A2–3, 2A2–3), slightly intervenose, not or occasionally forked; edge entire or eroded, concolorous. Stipe 20–60 × 5–12 mm, central or slightly eccentric, cylindrical to subcylindrical, often curved, white to milk white (1A1–2) or yellowish white (2A2), glabrous to finely
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