Biodiversity Data Journal 11: e95945 OO) doi: 10.3897/BDJ.11.e95945 open access Taxonomy & Inventories Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from China Xiao-Hong Ji*, Longfei Fan§ + College of Pharmacy and Life Sciences, Jiujiang University, Jiujiang, China § College of Plant Protection, Gansu Agricultural University, Lanzhou, China Corresponding author: Xiao-Hong Ji (xhji2010@163.com) Academic editor: Alfredo Vizzini Received: 03 Oct 2022 | Accepted: 02 Feb 2023 | Published: 09 Feb 2023 Citation: Ji X-H, Fan L (2023) Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from China. Biodiversity Data Journal 11: e€95945. https://doi.org/10.3897/BDJ.11.e95945 Abstract Background Fomitiporella is an important genus of wood-decaying fungi. Many new species were revealed in the last five years, based on morphological characters and molecular data. During a study on the taxonomy of Fomitiporella, two specimens from China were investigated, which have morphological characteristics close to Fomitiporella. After morphological examinations and phylogenetic analyses, a new species was confirmed to be a member of the Fomitiporella clade. New information Fomitiporella crystallina sp. nov. is described and illustrated as a new species, based on morphological characters and molecular evidence. It has perennial, irregular, pileate basidiocarps, an indistinct subiculum (ultrathin to almost lacking), lack of any kind of setae, has brownish, thick-walled basidiospores and causes a white rot. A molecular study, based on the combined ITS (internal transcribed spacer region) and nrLSU (the large nuclear ribosomal RNA _ subunit) dataset, supports the new species in Fomitiporella. The differences between the new species and phylogenetically related and morphologically © Ji X, Fan L. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 2 Ji X, Fan L similar species are discussed. A key to species with pileate to effused-reflexed basidiocarps of Fomitiporelila is given. Keywords Taxonomy, phylogeny, wood-decaying fungi Introduction Fomitiporella Murrill (1907), typified by F umbrinella Murrill, was proposed to accommodate polypores with perennial, resupinate and adnate basidiocarps, a_ thin subiculum, stratified tubes and subglobose and brown basidiospores. However, it was not treated as an independent genus, but as a synonym of Phellinus Quel. by most mycologists (Ryvarden and Johansen 1980, Larsen and Cobb-Poulle 1990, Ryvarden 1991, Ryvarden and Gilbertson 1994, Dai 1999, Nuhez and Ryvarden 2000) until Wagner and Fischer (2002) confirmed Fomitiporella as a distinct genus in Hymenochaetaceae, based on nrLSU DNA sequence data. Fomitiporella was accepted as a monophyletic genus by molecular phylogeny (Zhou 2014). Ji et al. (2017) broadened its concept to accommodate species with resupinate to effused reflexed and annual basidiocarps. Recently, Arambarria Rajchenb. & Pildain, Phellinotus Drechsler-Santos, Robledo & Rajchenb. and Rajchenbergia Salvador-Montoya were established and formed three clades in phylogeny of Fomitiporella (Rajchenberg et al. 2015, Drechsler-Santos et al. 2016, Salvador-Montoya et al. 2020). However, based on molecular data and morphological re-interpretation, these genera were treated as synonyms of Fomitiporelia (Wu et al. 2022). Furthermore, several species with pileate basidiocarps were introduced into Fomitiporella (Wu et al. 2022). During a study on the taxonomy of Fomitiporella, two collections from China were investigated. After morphological examinations and phylogenetic analyses, a new species was confirmed to be member of the Fomitiporella clade. In this paper, we describe and illustrate the new species. In addition, an identification key to the worldwide species with pileate to effused-reflexed basidiocarps of Fomitiporelia is provided. Materials and methods Morphology Studied specimens are deposited in the herbaria of Southwest Forestry University (SWFC) and the Mycological Herbarium of Jiujiang University (MHJU). Microscopic examination follows Dai (2010) and colour terms follow Petersen (1996). Microscopic structures were photographed using a Nikon Digital Sight DS-Fi1 camera. Microscopic measurements were made from slide preparations stained with Cotton Blue and Melzer’s reagent. In the text, the following abbreviations were used: KOH = 5% potassium hydroxide, IKI = Melzer’s reagent, IKI— = neither amyloid nor dextrinoid, CB = Cotton Blue, CB(+) = cyanophilic after Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from ... 3 12 hours stained with Cotton Blue, CB— = acyanophilic, L = mean spore length, W = mean spore width, Q = variation in the ratios of L/W between specimens studied, n = number of spores measured from given number of specimens. Molecular phylogeny DNA extraction, amplification and sequencing A CTAB rapid plant genome extraction kit (Aidlab Biotechnologies Co., Ltd., Beijing, China) was used to extract total genomic DNA from dried specimens following the manufacturer’s instructions with some modifications (Chen et al. 2016). The DNA was amplified with the primers: ITS4 and ITS5 for ITS (White et al. 1990) and LROR and LR7 for nrLSU. The PCR protocol for ITS was as follows: initial denaturation at 95°C for 3 min, followed by 35 cycles at 94°C for 40 s, 54°C for 45 s and 72°C for 1 min and a final extension of 72°C for 10 min. The PCR procedure for nrLSU was as follows: initial denaturation at 94°C for 1 min, followed by 35 cycles at 94°C for 30 s, 50°C for 1 min and 72°C for 1.5 min and a final extension of 72°C for 10 min. The PCR products were purified and sequenced at the Changsha Genomics Institute, China, with the same primers. The resulted ITS and nrLSU sequences were submitted to GenBank (http:/Awww.ncbi.nim.nih.gov/genbank). Phylogenetic analysis Besides the newly-generated sequences, additional sequences from a_ previous phylogenetic study on Fomitiporella (Wu et al. 2022) were included in the current dataset (Table 1). The dataset, outgrouped by /nocutis dryophila (Berk.) Fiasson & Niemela (VVu et al. 2022), was aligned using BioEdit (Hall 1999) and ClustalX 1.83 (Chenna et al. 2003). Alignment was manually adjusted to allow maximum alignment and to minimise gaps. Maximum Likelihood (ML) and Bayesian Inference (BI) were employed to perform phylogenetic analysis of the aligned dataset. Table 1. Information on the sequences used in this study. Type specimens are shown in bold. Species Location Sample no. GenBank accession no. ITS nLSU Fomitiporella austroasiana China Dai 16244 MG657328 MG657320 F. austroasiana China Dai 16168 MG657329 MG657321 F. austroasiana Singapore Dai 17879 MG657330 MG657324 F. badia Mexico JV 1707/22-J MW928042 — F. badia USA JV 0205/1J MW928043 — F. caryophylli India CBS 448.76 AY558611 AY059021 F. cavicola UK N 153 — AY059052 F. caviphila China LWZ 2013081 2-1 — KE729937 Species F. chinensis F. chinensis F. chinensis F. coruscans F. coruscans F. coruscans F. coruscans F. crassa F. crassa F. crystallina F. crystallina F. destruens F. destruens F. destruens F. destruens F. inermis F. inermis F. inermis F. mangrovei F. mangrovei F. neoarida F. pertenuis F. piptadeniae F. piptadeniae F. queenslandica F. queenslandica F. resupinata F. sinica F. sinica Fomitiporella sp. Fomitiporella sp. Fomitiporella sp. Fomitiporella sp. Fomitiporella sp. Location China China China USA USA USA USA Australia Australia China China Uruguay Uruguay Argentina Chile USA USA USA USA France Brazil Brazil Brazil Brazil Australia Australia Cameroon China China South Africa South Africa China China China Ji X, Fan L Sample no. Cui 11097 LWZ 20130713-7 LWZ 20130916-3 JV 1312/E2J JV 1407/46 JV 0409/6J JV 1207/6.1J Dai 18787 Dai 18716 CL Zhao 9453 CL Zhao 9567 CGP 473 CGP 474 CIEFAPcc 192 Fv.Ch-7 JV 0509/57K JV 1109/19A JV 1009/56 JV 1008/60 JV 1612/25-J URM 80362 PPT 111 URM 80322 URM 80345 Dai 18849 Dai 18844 Douanla-Meli 476 LWZ 20140625-2 LWZ 20140624-5 STE-U7109 STE-U7136 Cui 6557 Cui 11352 LWZ 20140721-2 GenBank accession no. ITS KX181310 KJ787817 KJ787818 KX181294 KX181295 KX181296 KX181297 MW924064 MW924063 ON493552 ON493553 KY907683 KY907682 AY072033 KX181305 KX181304 KX181306 KX181313 MG657331 KM211294 MG806101 KM211290 KM211291 KJ787822 KX181301 KX181302 KP279300 JQ038896 KX181303 KX181315 KX181316 nLSU KX181342 KJ787808 KJ787809 KX181333 KX181332 KX181331 KX181330 MW892735 MW892736 ON493576 ON493577 KY907687 KY907692 KP347520 DQ459301 KX181346 KX181347 KX181334 MG657325 KM211286 MG806100 KM211282 KM211283 MW892737 MW892738 JF712935 KX181320 KX181321 KX181338 KX181337 Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from ... Species Location Sample no. GenBank accession no. ITS nLSU Fomitiporella sp. Thailand LWZ 20140729-22 KX181317 KX181339 Fomitiporella sp. Ethiopia AM 12 JF895466 JQ910908 Fomitiporella sp. Ethiopia AM 15 JF895467 JQ910909 Fomitiporella sp. Ethiopia AM 18 JF895468 JQ910910 Fomitiporella sp. Ethiopia AM 04 KX181318 KX181335 Fomitiporella sp. South Africa STEU 7147 JQ038897 a Fomitiporella sp. South Africa STEU 7042 JQ038893 = Fomitiporelila sp. South Africa STEU 7155 KP279297 — Fomitiporella sp. South Africa STEU 7154 JQ038894 a Fomitiporella sp. South Africa STEU 7148 KP279296 —_ Fomitiporella sp. South Africa STEU 7141 KP279295 = Fomitiporella sp. South Africa STEU 7798 KP279299 — Fomitiporella sp. South Africa STEU 7039 JQ038892 — Fomitiporella sp. South Africa STEU 7038 JQ038891 — Fomitiporella sp. South Africa STEU 7799 KP279298 = Fomitiporella sp. USA 0509/114 KX181314 KX181336 Fomitiporella sp. USA CBS 303.66 — AY059036 F, subinermis China Dai 15114 KX181308 KX181344 F. subinermis China Dai 15131 KX181307 KX181345 F. tenuissima China Dai 12365 KC456244 KC999901 F. tenuissima China Dai 12245 KC456242 KC999902 F. tenuissima China Dai 12255 KC456243 KC999903 F. umbrinella USA JV 0312/26.6J KX181291 — F. umbrinella USA JV 0904/149J KX181293 KX181329 F. vietnamensis Vietnam Dai 18377 MG657332 MG657326 F. vietnamensis Vietnam Dai 18382 MG657333 MG657327 Inocutis dryophila USA L(61)5-20-A AM269783 AM269846 |. dryophila USA SP 25 AM269782 AM269845 The two phylogenetic analysis algorithms generated nearly congruent topologies for the dataset and, thus, only the topology from the ML analysis is presented along with statistical values from the ML and BI algorithms (BS not less than 50% and BPP not less than 0.9) at the nodes. The tree was visualised in TreeView 1.6.6 (Page 1996). 6 Ji X, Fan L Taxon treatment Fomitiporella crystallina X.H. Ji, sp. nov. ° MycoBank 844077 Materials Holotype: a. scientificName: Fomitiporella crystallina; acceptedNameUsage: Fomitiporella crystallina X.H. Ji, 2022, sp. nov.; parentNameUsage: Fomitiporella, Murrill, 1907; kingdom: Fungi; phylum: Basidiomycota; class: Agaricomycetes; order: Hymenochaetales; family: Hymenochaetaceae; taxonRank: species; verbatinlaxonRank: species; genus: Fomitiporella; specificEpithet: Crystallina; scientificNameAuthorship: X.H. Ji; continent: Asia; country: China; stateProvince: Yunnan Province; municipality: Puer; locality: Huilianghe,Puer, Yunnan Province,China; verbatimElevation: 1400 m; locationRemarks: label transliteration: "Yunnan, Jingdong Yi Autonomous County, Huilianghe, 2019 1.5, Zhao Changlin et al."; [GHA Rat 2019.01.05, &X+1 Fomitiporella mangrovei JV 1612/25-) USA 9711.00 Fomitiporella mangrovei JV 1008/60 USA Fomitiporella pertenuis PPT 111 Brazil Holotype Fomitiporella tenuissima Dai 12245 China Famitiporella tenuissima Dai 12365 China Fomitiporella tenuissima Dai 12255 China tow't.oe Fomitiporella crassa Dat 18787 Australia Fomitiporella crassa Dai 18716 Australia 62026 1001.00) Fomitiporella piptadeniae URM 80322 Brazil 940.99) Fomitiporella piptadeniae URM 80345 Brazil Fomitiporella neoarida URM 80362 Brazil iou/t.oeg Fomitiporiella badia JV 1707/22-J Mexico Fomitiporiella badia IV 0205/1) USA 891.00. 960.98 8) LOO 90% pes 101 Oo 100/100) sn0.o07 Fomitiporella sp, AM 15 Ethiopia Fomitiporella sp. AM 18 Ethiopia Fomitiporella caviphila LWZ 20130812-1 China Fomitiporella sp. AM 12 Ethiopia : Fomitiporella cavicola N 153 UK 961.00! + Fomitiporella crystallina CL. Zhao 9567 China Fomitiporella crystallina CL. Zhao 9453 China 96.00 Fomitiporella sp. CBS 303.66 USA Fomitiporella sp. 0509/114 USA Fomitiporella sp. STEU 7042 South Africa 72. Fomitiporella queenstandica Dai 18849 Australia $9-1- Fomitiporella queenslandica Dai 18844 Australia 1001 1.0¢ Fomitiporella sp. LWZ_ 20140721-2 China Fomitiporella sp. Cui 11352 China 0S 0. Fomitiporella sp. LWZ 20140729-22 Thailand Fomitiporella austroasiana Dai 16244 China L004 Fomitiporella austroasiana Dai 16168 China Fomitiporella austroasiana Dai 16244 China Fomitiporella sp. AM 04 Ethiopia 404 Inocutis dryophila SP25 USA Inocutis dryophila L(61)S-20-A USA 0.1 Figure 1. El Phylogeny of Fomitiporella inferred from the ITS and nrLSU dataset. Topology is from ML tree and statistical values (ML/BI) are indicated for each node that simultaneously received BS from ML not below 50%, and BPP from BI not below 0.9. Ji X, Fan L Hyphal structure: Hyphal system dimitic; generative hyphae simple septate; skeletal hyphae dominant; tissue darkening but otherwise unchanged in KOH. Figure 2. EESI Basidiocarp of Fomitiporella crystallina (CL Zhao 9453). Scalebar: 1 cm. Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from ... SS) Tubes: Generative hyphae frequent, hyaline to pale yellow, thin- to slightly thick-walled, occasionally branched, frequently simple septate 1.5—2.5 um in diam; skeletal hyphae golden yellow to pale brown, thick-walled with a wide to narrow lumen, unbranched, aseptate, interwoven, 2—3.5 um in diam; setae and cystidioles absent; hymenium collapsed in the studied material, basidia and basidioles not seen; small or large rhomboid crystals abundant. Spores: Basidiospores broadly ellipsoid, golden yellow, thick-walled, smooth, IKI-, CB(+), (4.1-)4.3-4.7(-4.9) * (3-)3.2-3.9(—4) um, L = 4.50 um, W = 3.42 um, Q = 1.20- 1.35 (n = 60/2) (Fig. 3). Etymology Crystallina, referring to the species having abundant crystals. Figure 3. EES] Microscopic structures of Fomitiporella crystallina (CL Zhao 9453). A Basidiospores; B Rhomboid crystals; C Hyphae from trama. Scale bars: 5 um. 10 Ji X, Fan L Identification keys An identification key to the accepted species with pileate to effused-reflexed basidiocarps of Fomitiporella. 10 Basidiocarps pileate Basidiocarps effused-reflexed Basidiospores 4—5 ym long Basidiospores 5—7.5 ym long Pores 8-9 per mm Pores 5-8 per mm Context with a granular core, black line absent Context without a granular core, black line present Baisiospores 5—5.5 x 3.5—4.5 um Baisiospores 7—7.5 x 5.5-6 um Basidiocarps annual to biennial Basidiocarps perennial Context homogeneous; hyphal system monomitic; Neotropical species Context duplex; hyphal system dimitic; Asian species Pores 1—4 per mm Pores 4—9 per mm Basidiospores 5.2—6.5 ym long Basidiospores 7.5—9.5 ym long Pores 7—9 per mm; basidiospores 3—4 ym long 4 F. queenslandica F. crystallina F. neoarida (Drechsler-Santos & Robledo) Y.C. Dai & F. Wu F. piptadeniae (Teixeira) Y.C. Dai & F. Wu F. badius (Cooke) Y.C. Dai & F. Wu 7 F. destruens (Rajchenb. & Pildain) Y.C. Dai F. chinensis (Pilat) Y.C. Dai, X.H. Ji & Vlasak 10 F. cyclobalanopsidis (T.T. Chang & W.N. Chou) Y.C. Dai & F. Wu F. shoushana (T.T. Chang & W.N. Chou) Y.C. Dai & F. Wu F. caryophylli (Racib.) T. Wagner & M. Fisch. Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from ... 11 P 4-7 : idi 4-4. a aay per mm; basidiospores éyum F. vietnamensis Y.C. Dai, X.H. Ji & J. Viasak Analysis A total of 68 fungal collections, representing 22 species and 22 taxa of Fomitiporella, were included in the phylogenetic analyses and two samples of genus /nocutis were used as outgroups. The final alignment comprised a total of 1667 base pairs (bp) including 804 of ITS and 863 of nrLSU. Two sampled specimens of the new species, Fomitiporella crystallina, formed a well-supported lineage (BS/PP values 98/1.0), indicating that they are phylogenetically distinct from other species in Fig. 1. Discussion Fomitiporella crystallina has unique morphological characters in Fomitiporella and forms a distinct lineage within the Fomitiporella clade. Morphologically, F. crystallina is similar to F. queenslandica Y.C. Dai & F. Wu in sharing perennial, pileate basidiocarps, a dimitic hyphal structure and the same size of basidiospores (4.1-5 x 3-4 um), whereas the latter has dimidiate pilei and smaller pores (5-8 per mm; VVu et al. (2022)). Moreover, the presence of the cystidioles and a distinct subiculum in F queens/andica makes it different from F. crystallina. Phylogenetically, Fomitiporella crystallina is closely related to F. cavicola (Kottl. & Pouzar) T. Wagner & M. Fisch. by sharing the perennial basidiocarps, a dimitic hyphal system and the same size range of pores (5-8 per mm), but F cavicola has resupinate basidiocarps and larger basidiospores (4.7-5.5 x 4—4.5 um). In addition, F cavicola is known from Europe only (Kotlaba and Pouzar 1995). In conclusion, both morphology and phylogeny support that the specimens, collected from China, are a new species within the genus Fomitiporella. Acknowledgements We are grateful to Dr. Chang-Lin Zhao (Kunming, China) who allowed us to study his specimens. The research is supported by the National Natural Science Foundation of China (Project Nos. 32000014) and National Natural Science Foundation of Jiangxi Province (Project No. 20202BABL213043). References ° Chen JJ, Cui BK, Dai YC (2016) Global diversity and molecular systematics of Wrightoporia s. | (Russulales, Basidiomycota). Persoonia 37: 21-36. https://doi.org/ 10.3767/003158516X689666 12 Ji X, Fan L Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD (2003) Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Research 31: 3497-3500. https://doi.org/10.1093/nar/gkg500 Dai YC (1999) Phellinus sensu lato (Aphyllophorales, Hymenochaetaceae) in East Asia. Acta Botanica Fennica 166: 1-115. Dai YC (2010) Hymenochaetaceae (Basidiomycota) in China. Fungal Diversity 45: 131-343. https://doi.org/10.1007/s13225-010-0066-9 Drechsler-Santos ER, Robledo GL, Lima-Junior NC, Malosso E, Reck MA, Gibertoni TB, Queiroz Cavakcanti MA, Rajchenberg M (2016) Phellinotus, a new neotropical genus in the Hymenochaetaceae (Basidiomycota, Hymenochaetales). Phytotaxa 261: 218-239. https://doi.org/10.11646/phytotaxa.261.3.2 Hall TA (1999) Bioedit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98. Ji XH, Vlasak J, Zhou LW, Wu F, Dai YC (2017) Phylogeny and diversity of Fomitiporella (Hymenochaetales, Basidiomycota). Mycologia 109: 308-322. htips://doi.org/10.1080/ 00275514.2017.1305943 Kotlaba F, Pouzar Z (1995) Phellinus cavicola, a new xanthochroic setae-less polypore with coloured spores. Czech Mycology 48: 155-159. https://doi.org/10.33585/cmy.48205 Larsen MJ, Cobb-Poulle LA (1990) Phellinus (Hymenochaetaceae). A survey of the world taxa. Synopsis Fungorum 3: 1-206. Murrill WA (1907) Polyporaceae 1. North American Flora 9: 1-72. Nufez M, Ryvarden L (2000) East Asian polypores 1. Ganodermataceae and Hymenochaetaceae. Synopsis Fungorum 13: 1-168. Page RMD (1996) Treeview: An application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences 12: 357-358. Petersen JH (1996) Farvekort. The Danish Mycological Society’s color-chart. Foreningen til Svampekundskabens Fremme, Greve, 6 pp. Rajchenberg M, Pildain MB, Bianchinotti MV, Barroetavena C (2015) The phylogenetic position of poroid Hymenochaetaceae (Hymenochaetales, Basidiomycota) from Patagonia, Argentina. Mycologia 107: 754-767. https://doi.org/10.3852/14-170 Ryvarden L, Johansen | (1980) A preliminary polypore flora of East Africa. Fungiflora, Oslo, 636 pp. Ryvarden L (1991) Genera of polypores. Nomenclature and taxonomy. Synopsis Fungorum 5: 1-363. Ryvarden L, Gilbertson RL (1994) European polypores. 2. Meripilus-Tyromyces. Synopsis Fungorum 7: 394-743. Salvador-Montoya CA, Popof OF, Goés-Neto A, Drechsler-Santos ER (2020) Global phylogenetic and morphological reassessment of Fomitiporella s.|. (Hymenochaetales, Basidiomycota): Taxonomic delimitation of Fomitiporella s.s. and segregation of Rajchenbergia, gen. nov. Plant Systematics Evolution 306: 1-27. httos://doi.org/10. 1007/sO0606-020-01648-w Wagner T, Fischer M (2002) Proceedings towards a natural classification of the worldwide taxa Phellinus s.|. and Inonotus s.|., and phylogenetic relationships of allied genera. Mycologia 94: 998-1016. https://doi.org/10.1080/15572536.2003.11833156 White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols315-322. https://doi.org/10. 1016/b978-0-12-372180-8.50042-1 Fomitiporella crystallina sp. nov. (Basidiomycota, Hymenochaetales) from ... 13 Wu F, Zhou LW, Vilasak J, Dai YC (2022) Global diversity and systematics of Hymenochaetaceae with poroid hymenophore. Fungal Diversity 113: 1-192. https:// doi.org/10.1007/s13225-021-00496-4 Zhou LW (2014) Fomitiporella caviphila sp. nova (Hymenochaetaceae Basidiomycota) from Eastern Chain, with a preliminary discussion on the taxonomy of Fomitiporella. Annales Botanici Fennici 51: 279-284. https://doi.org/10.5735/085.051.0503