Phallus Fuscoechinovolvatus (Phallaceae, Basidiomycota), a New Species with a Dark Spinose Volva from Southern China
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Phytotaxa 334 (1): 019–027 ISSN 1179-3155 (print edition) http://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2018 Magnolia Press Article ISSN 1179-3163 (online edition) https://doi.org/10.11646/phytotaxa.334.1.3 Phallus fuscoechinovolvatus (Phallaceae, Basidiomycota), a new species with a dark spinose volva from southern China BIN SONG1, TING LI1, TAIHUI LI*, QIUJU HUANG & WANGQIU DENG State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collec- tion and Application, Guangdong Open laboratory of Applied Microbiology, Guangdong Institute of Microbiology, Guangzhou 510070, China * Corresponding author: [email protected] 1 BIN SONG and TING LI contributed equally to this work. Abstract Phallus fuscoechinovolvatus is described as a new species from southern China, mainly characterized by its dark brown vol- va with white spines. Molecular phylogenetic analyses based on the sequences of nuclear ribosomal large subunit (nrLSU) and nuclear ribosomal internal transcribed spacer (ITS) confirm its species status within Phallaceae. Detailed morphological description, colour photographs, line drawings of the new species and comparisons with similar taxa are presented. Key words: Phallales, phylogenetics, subtropical fungus, taxonomy Introduction Phallus Junius ex L. (1753: 1178) is a fungal genus under the family Phallaceae (Phallales, Phallomycetidae, Agaricomycotina, Basidiomycota) with P. impudicus L. (1753: 1178) as the type species, and it includes taxa typically characterized by their noticeably foetid, a bell-shaped or campanulate to conical receptacle, an erect to curved spongy pseudostipe, presence or absence of an indusium (a pseudoparenchymatous reticulum) hanging from the pseudostipe apex beneath the receptacle, and a volva with rhizomorphs most of the time (Linnæi & Salvius 1753; Cunningham 1944; Zeller 1949; Baseia et al. 2003; Liu et al. 2005; Li et al. 2016). The genus Dictyophora Desv. (1809: 92) was later split from Phallus, containing the taxa with an indusium (Desvaux 1809). Although it was accepted by many mycologists, the taxonomic independency of Dictyophora was still controversial and questioned by some taxonomists, especially in recent decades when some researchers have advocated with new scientific evidences that the genus Dictyophora should be merged into Phallus again (Andersson 1989; Kreisel 1996; Calonge 2005; Baseia et al. 2006; Dentinger et al. 2010; Zhao et al. 2011; Li et al. 2014; Adamčík et al. 2015). Excluding formae, varieties and synonyms, there are around 90 Phallus species documented worldwide, according to Index Fungorum. Twenty-eight species have been reported as Dictyophora or Phallus in China (Zang et al. 1976; Zang & Ji 1985; Zang et al. 1988; Li et al. 2002; Li et al. 2003; Li et al. 2005; Liu et al. 2005; Li et al. 2014; Li et al. 2016), including some popular edible and medicinal species, e.g. P. duplicatus Bosc (1811: 86), P. echinovolvatus (M. Zang, D.R. Zheng & Z.X. Hu) Kreisel (1996: 277), P. indusiatus Vent. (1798: 520), P. multicolor (Berk. & Broome) Cooke (1882: 58), P. luteus (Liou & L. Hwang) T. Kasuya (2008: 8) and P. rubrovolvatus (M. Zang, D.G. Ji & X.X. Liu) Kreisel (1996: 280) (Dai & Yang 2008; Dai et al. 2010). Recently, a distinct species of Phallus with a long white indusium and a spinose dark volva was found from southern China. Detailed morphological observations and molecular phylogenetic analyses proved that it is a species new to science. Morphological and phylogenetic descriptions of the new species are therefore formally presented as follows. Accepted by Genevieve Gates: 14 Dec. 2017; published: 12 Jan. 2018 19 Materials and methods Morphological study Fresh specimens were collected from Chebaling National Nature Reserve in Shixing county and Jigongshan Mountain in Lianshan county of Guangdong province, southern China. Fungal samples were dried with an electronic dryer and deposited in the Fungal Herbarium of Guangdong Institute of Microbiology (GDGM), Guangzhou, China. Some small pieces of basidiomata were dried, using silica gel, for molecular genetics analysis. Basidiomata photographs were taken in situ with a digital camera (PowerShot G16, Canon) in natural light. The macro-morphological descriptions were made based on fresh materials and field photographs. Microscopic features were observed under a light microscope (Zeiss Axio Lab. A1), and micro-morphological structures were measured with revived sections of dried specimens, which were hand-sectioned and mounted in 5% KOH solution, 1% Congo Red or Melzer’s reagent. Colour descriptions were 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 were 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 referred to length/width ratio of an individual basidiospore; Qm referred to the average Q value of all basidiospores ± sample standard deviation. TABLE 1. Species used in the molecular phylogenetic analyses, taxa information, vouchers, and GenBank accession numbers. Taxon Voucher Locality nrLSU ITS Source Phallus atrovolvatus MEL:2382871 Australia KP012745 KP012745 GenBank P. atrovolvatus MEL:2382962 Australia KP012823 KP012823 GenBank P. calongei AH31862 Pakistan FJ785522 - Moreno et al. (2009) P. cinnabarinus INPA:255835 - - KJ764821 Cabral et al. (2015) P. costatus MB02040 - DQ218513 - Hosaka et al. (2006) P. duplicatus OSC38819 - DQ218481 - Hosaka et al. (2006) P. echinovolvatus ASI 32008 - - AF324166 GenBank P. echinovolvatus ASI 32010 - - AF324167 GenBank P. fuscoechinovolvatus GDGM 43465 China MF039584 MF039580 This study P. fuscoechinovolvatus GDGM 48589 China MF039585 MF039581 This study P. fuscoechinovolvatus GDGM 48663 China MF039586 MF039582 This study P. fuscoechinovolvatus GDGM 48676 China MF039587 MF039583 This study P. hadriani AFTOL-ID 683 - AY885165 DQ404385 GenBank P. hadriani AH39161 Pakistan - KF481956 Moreno et al. 2013 P. hadriani TNS Kasuya B2045 Japan KP222544 KP222542 GenBank P. hadriani TNS-F-70036 Japan KU516107 KU516100 GenBank P. haitangensis HKAS 88197 China - KU705383 Li et al. (2016) P. haitangensis HKAS 88199 China - KU705384 Li et al. (2016) P. impudicus ASI 25008 - - AF324171 GenBank P. impudicus KA13-1262 - - KR673719 Kim et al. (2015) P. impudicus KH-TGB11-1034 (TNS) - KF783249 - Trierveiler-Pereira et al. (2014) P. impudicus TNS-F-70035 - KU516106 KU516099 GenBank P. impudicus TNS-F-70037 Japan KU516108 KU516101 GenBank P. indusiatus Mushroom Observer # 181359 Mexico - MF428417 GenBank P. luteus TNS Kasuya B218 Japan KP222545 KP222543 GenBank P. mengsongensis HKAS 78344 China - KF052626 GenBank P. mengsongensis HKAS 78342 China - KF052627 GenBank P. multicolor MEL:2382891 Australia KP012762 KP012762 GenBank P. ravenelii UMO(USA-MO):0001 USA KP779906 - GenBank P. rubrovolvatus YZS046 - - KF939506 Lu et al. 2014 P. rubrovolvatus YZS047 - - KF939507 Lu et al. 2014 P. rugulosus ASI 32004 - - AF324169 GenBank P. rugulosus ASI 25007 - - AF324170 GenBank P. serratus HKAS 78341 China - KF052623 GenBank P. ultraduplicatus HMAS:253050 China KJ591586 KJ591584 Adamcik et al. (2015) P. ultraduplicatus HMAS:253051 China KJ591587 KJ591585 Adamcik et al. (2015) Mutinus zenkeri MA-2013 JD781 - KC128654 KC128650 Degreef et al. (2013) 20 • Phytotaxa 334 (1) © 2018 Magnolia Press SONG ET AL. Molecular phylogenetic study Total DNA were isolated from about 10mg of four dried voucher specimens (GDGM 43465, GDGM 48589, GDGM 48663, GDGM 48676) using the Sangon Fungus Genomic DNA Extraction kit (Sangon Biotech Co., Ltd., Shanghai, China). For the amplification of nrLSU gene, the LROR/LR5 primers were chosen (Vilgalys & Hester 1990) and ITS1/ ITS4 primers were selected for the ITS gene amplification (White et al. 1990; Gardes & Bruns 1993). PCR reaction reagents and programs were set according to Hosen et al. (2016). PCR products were electrophoresed on a 1% agarose gel and visualized by staining with Gold View (SBS Genetech, Beijing, China). The PCR products were sequenced by Majorbio Co., Ltd. (Shanghai, China). Sequences of the four specimens were submitted to GenBank, and the other referred sequences were obtained from GenBank (Table 1). The nrLSU data matrix included 21 fungal sequences from 12 taxa meanwhile ITS dataset included 32 fungal sequences from 16 taxa, Mutinus zenkeri (Henn.) E. Fisch. (1900: 24) (nrLSU: KC128654 and ITS: KC128650) was rooted as outgroup (Table 1). All sequences were aligned using ClustalX software (Thompson et al. 1997), and edited with Bioedit software (Hall 1999). Each individual dataset, either nrLSU and ITS, was analyzed using Maximum Likelihood (ML), Maximum Parsimony (MP) and Bayesian Inference (BI) methods. ML and MP were performed using MEGA version 5.2 (Hall 2013). Bootstrap analysis was implemented with 1000 replicates. BI was executed in MrBayes v.3.1.2 (GTR+I+Gmodel) to evaluate Posterior probabilities (PP) (Ronquist & Huelsenbeck 2003). Results Taxonomy Phallus fuscoechinovolvatus T.H. Li, B. Song & T. Li, sp. nov. Figs. 1–2 FIGURE 1. Plate of Phallus fuscoechinovolvatus. A: Mature fruiting body of GDGM 48589 (holotype), B: Receptacle of GDGM 48589, C: Indusium and pseudostipe of GDGM 48589. D and E: Immature basidiomata of GDGM 48663 and GDGM 48629, respectively. F: