Abstract
Two new species of Polyporales, Cerrenacaulinicystidiata and Polyporusminutissimus, are illustrated and described on the basis of morphological studies and phylogenetic analyses from southern China and Vietnam. C.caulinicystidiata is characterized by annual, resupinate, sometimes effused-reflexed basidiocarps, greyish orange to brownish orange pore surface, irregular pores (3–8 per mm), a trimitic hyphal system, pyriform to ventricose cystidia, and subglobose basidiospores 3.2–4.5 × 2.8–3.5 µm in size. P.minutissimus is characterized by annual, solitary, fan-shaped with a depressed center or infundibuliform basidiocarps, obvious black stipe, cream to buff yellow pileal surface with glabrous, occasionally zonate and radially aligned stripes, angular pores (6–9 per mm), a dimitic hyphal system, and cylindrical basidiospores, 5–9.2 × 2.2–4 μm. Detailed descriptions and illustrations of the two new species are provided. The differences between the two new species and their morphologically similar and phylogenetically related species are discussed.
Key words: Cerrenaceae, phylogeny, Polyporaceae, taxonomy, wood-decaying fungi
Introduction
The order Polyporales presents a great diversity of basidiocarp types and hymenophore configurations (Binder et al. 2013). It is an important group of fungi, as Polyporales species can cause wood-decay and thus play an essential role in the carbon cycle. In addition, some species of Polyporales may have medicinal properties (Dai 1996, 1999, 2012; Dai et al. 2009). The order has long been the subject of research on taxonomic diversity, distribution patterns, and ecological functions (Hibbett et al. 2014). As of early 2024, more than 1,800 species are recognized in the order (Martinez et al. 2004; Martinez et al. 2009; Kirk et al. 2008; Grigoriev et al. 2013; Zhao et al. 2015; Justo et al. 2017). Due to its great diversity, the order is intensively studied worldwide (Justo et al. 2017).
Cerrena Gray is the type genus of Cerrenaceae within the Polyporales (Ryvarden and Gilbertson 1993; Justo et al. 2017), and it is typified by C.unicolor (Bull.) Murrill. It is widely distributed throughout the world. The genus is characterized by resupinate, effused-reflexed or pileate basidiocarps, irregular hymenophore, dimitic or trimitic hyphal systems, cylindric to ellipsoid basidiospores, and white rot (Ryvarden and Gilbertson 1993; Ryvarden et al. 2022). The genus Cerrena published in 1821 has priority above Trametes Fr., published in 1838, and the two genera were considered as a single taxon in several studies. Thus, a huge number of new combinations in Cerrena. (Cunningham and Cunningham1965; Gilbertson and Ryvarden 1987; Corner 1989; Ryvarden and Melo 2022). However, Ryvarden (1991) recommended to keep Cerrena as a separate genus based on their anatomical characters. According to Index Fungorum (http://www.indexfungorum.org) and Yuan (2014), Cerrena currently comprises around 10 species.
Polyporus P. Micheli ex Adans., the type genus of the Polyporaceae, is a well-known polypore genus (Gilbertson and Ryvarden 1987). Given that Micheli (1729) did not originally select a type species for Polyporus, there is no consensus on the selection of the type. Since Donk (1933) selected P.tuberaster (Jacq. ex Pers.) Fr. as the type species, this lectotype was accepted by most subsequent mycologists (Cunningham and Cunningham 1965; Singer 1986; Niemelä and Kotiranta 1991; Ryvarden 1991; Sotome et al. 2008; Ji et al. 2022). Morphologically, Polyporus is a heterogeneous genus including many species belonging to six morphological groups described by Núñez and Ryvarden (1995), viz, the Polyporus group, the Favolus group (= Favolus Fr.), the Melanopus group (= Melanopus Pat.), the Polyporellus group (= Polyporellus Karst.), the Admirabilis group, and the Dendropolyporus group (= Dendropolyporus (Pouz.) Jülich). Phylogenetically, Polyporus s. str. is known as a polyphyletic genus (Krüger et al. 2006; Sotome et al. 2008, 2011; Ji et al. 2022). Phylogenetic analyses of Polyporus did not conform to the six morphological groups, for which further in-depth study of the group is needed (Sotome et al. 2008; Zhou et al. 2016).
During investigations on wood-decaying polypores from South China and Vietnam, specimens that morphologically fit the definitions of Cerrena and Polyporus were collected. Phylogenetically, these samples formed two distinct lineages within Cerrena and Polyporus, respectively, and they are different from their morphologically similar and phylogenetically related species. Therefore, we describe and illustrate two new species, Cerrenacaulinicystidiata sp. nov. and Polyporusminutissimus sp. nov. within the Polyporales on the basis of morphological studies and phylogenetic analyses.
Materials and methods
Morphological studies
The studied specimens are deposited in the Fungarium of Beijing Forestry University (BJFC) and the Institute of Applied Ecology of the Chinese Academy of Sciences (IFP). Macro-morphological descriptions were based on field notes and voucher herbarium specimens. Microscopic measurements and drawings were made from slides prepared from voucher tissues and stained with Cotton Blue and Melzer’s reagent. The following abbreviations were used: KOH = 5% potassium hydroxide; CB = Cotton Blue; CB+ = cyanophilous in Cotton Blue; CB– = acyanophilous in Cotton Blue; IKI = Melzer’s; IKI– = neither amyloid nor dextrinoid in Melzer’s reagent; L = mean basidiospore length (arithmetic average of basidiospores); W = mean basidiospore width (arithmetic average of basidiospores); Q = variation in the L/W ratios between specimens studied; n (a/b) = number of basidiospores (a) measured from the given number of specimens (b). When we present basidiospore size variation, 5% of measurements were excluded from each end of the range. These excluded values are given in parentheses. Special color terms follow Anonymous (1969) and Petersen (1996).
DNA extraction and sequencing
A CTAB rapid plant genome extraction kit (Aidlab Biotechnologies, Co., Ltd., Beijing, China) was used to obtain DNA products from voucher specimens following the manufacturer’s instructions with some modifications (Wu et al. 2020, 2022). The following primer pairs were used to amplify the DNA: ITS5 and ITS4 for the internal transcribed spacer (ITS) region (White et al. 1990) and LR0R and LR7 for the nuclear large subunit (nLSU) rDNA gene (Vilgalys and Hester 1990).
The procedures for DNA extraction and polymerase chain reaction (PCR) used in this study were the same as described by Wu et al. (2022). The PCR products were purified and sequenced by Beijing Genomics Institute (BGI), China. All newly generated sequences in this study were deposited in GenBank (Sayers et al.2024; http://www.ncbi.nlm.nih.gov/genbank/) and listed in Table 1.
Table 1.
Taxa information and GenBank accession numbers of the sequences used in this study.
Species | Specimen No. | Country | ITS | LSU |
---|---|---|---|---|
Cerrenaalbocinnamomea | Miettinen 10511 | China | OR262168 | OR262168 |
Cerrenaalbocinnamomea | NIBRFG0000102423 | South Korea | FJ821532 | – |
Cerrenaalbocinnamomea | Dai 12892 | China | KC485522 | KC485539 |
Cerrenaalbocinnamomea | KUC20121102-06 | South Korea | KJ668561 | – |
Cerrenacaulinicystidiata | Yuan 12664 | Vietnam | MT269762 | MT259328 |
Cerrenacaulinicystidiata | Yuan 12666 | Vietnam | MT269763 | MT259329 |
Cerrenacaulinicystidiata (Cerrena sp. 1) | BJ2-11 | China | KX527879 | – |
Cerrenacaulinicystidiata (Cerrena sp. 1) | G1669 | China | MK247953 | – |
Cerrenacaulinicystidiata (Cerrena sp. 1) | Otu0185 | China | MT908560 | – |
Cerrenacaulinicystidiata* | Wu 661 | China | PP035831 | PP035828 |
Cerrenacystidiata | 548/17 | Brazil | MZ649034 | MZ649034 |
Cerrenagilbertsonii | JV 1609/29 | Guadeloupe | OR262202 | – |
Cerrenagilbertsonii | Vandevender 94-144 | Mexico | OR262171 | OR262171 |
Cerrenamultipileata | JV 1407/63 | Costa Rica | OR262201 | OR262201 |
Cerrenamultipileata | Ryvarden 43881 | Costa Rica | OR262155 | OR262155 |
Cerrenamultipileata | Kout A36 | Guatemala | OR262203 | – |
Cerrena sp. 2 | F12 | China | OP022000 | – |
Cerrena sp. 2 | 7-SU-3-B-77(M)-B | Indonesia | KJ654531 | – |
Cerrena sp. 2 | NTOU5117 | Taiwan | MN592928 | – |
Cerrenaunicolor | B2 | Antarctica | HM589361 | – |
Cerrenaunicolor | D.T6.5_2 | Argentina | MH019790 | – |
Cerrenaunicolor | CBS 154.29 | Canada | MH855029 | – |
Cerrenaunicolor | He6082 | China | OM100740 | OM083972 |
Cerrenaunicolor | GSM-10 | China | JQ798288 | – |
Cerrenaunicolor | Han 849 | China | MW467890 | – |
Cerrenaunicolor | CU2 | Czech | FJ821536 | – |
Cerrenaunicolor | H:Otto Miettinen 9443 | Finland | FN907915 | FN907915 |
Cerrenaunicolor | MUT<ITA_:5063 | Italy | MK581063 | – |
Cerrenaunicolor | FCG-1937 | Japan | LC415531 | – |
Cerrenaunicolor | Pertti Uotila 47558 (H) | Kyrgyzstan | OR262167 | – |
Cerrenaunicolor | Feketic | Serbia | MW485440 | – |
Cerrenaunicolor | KA17-0024 | South Korea | MN294859 | – |
Cerrenaunicolor | 3115 | Sweden | JN710525 | JN710525 |
Cerrenaunicolor | CUZFVG179 | Turkey | MK120293 | – |
Cerrenaunicolor | K(M):249944 | UK | MZ159683 | – |
Cerrenaunicolor | FD-299 | USA | KP135304 | KP135209 |
Cerrenaunicolor | TASM: YG/PS79 | Uzbekistan | MT526291 | – |
Cerrenazonata | Gates 2008-4-17 (H) | Australia | OR262160 | OR262160 |
Cerrenazonata | Otto Miettinen 9773 (H) | China | OR262157 | OR262157 |
Cerrenazonata | Otto Miettinen 9889 (H) | China | OR262158 | OR262158 |
Cerrenazonata | Otto Miettinen 13798 (H) | Indonesia | OR262166 | OR262166 |
Cerrenazonata | WS36_1_2_B_As | Japan | LC631683 | – |
Cerrenazonata | PDD:95790 | New Zealand | HQ533016 | – |
Cerrenazonata | KA17-0224 | South Korea | MN294861 | – |
Cerrenazonata | LE-BIN 4492 | Vietnam | OP985107 | – |
Datroniellascutellata | RLG9584T | USA | JN165004 | JN164792 |
Datroniellatropica | Dai 13147 | China | KC415181 | KC415189 |
Echinochaetebrachypora | TFM:F 24996 | Japan | AB462321 | AB462309 |
Echinochaeterussiceps | TFM:F 15716 | Japan | AB462310 | AB368065 |
Echinochaeterussiceps | TFM:F 24250 | Japan | AB462313 | AB462301 |
Favolusacervatus | Cui 11053 | China | KU189774 | KU189805 |
Favolusacervatus | Dai 10749b | China | KX548953 | KX548979 |
Favolusgracilisporus | Cui 4292 | China | KX548970 | KX548992 |
Favolusgracilisporus | Li 1938 | China | KX548971 | KX548993 |
Hexagoniaglabra | Dai 10691 | China | JX569733 | JX569750 |
Hexagoniatenuis | Cui 8468 | China | JX559277 | JX559302 |
Irpexlatemarginatus | Dai 8289 | China | KY131835 | – |
Lentinuslongiporus | DAOM:229479 | Canada | AB478880 | LC052217 |
Lentinuslongiporus | WD2579 | Japan | AB478879 | LC052218 |
Lentinussubstrictus | Wei 1582 | China | KU189767 | KU189798 |
Lentinussubstrictus | Wei 1600 | China | KC572022 | KC572059 |
Microporusaffinis | Cui 7714 | China | JX569739 | JX569746 |
Microporusflabelliformis | Dai 11574 | China | JX569740 | JX569747 |
Mycoboniaflava | CulTENN10256 | Costa Rica | AY513570 | AJ487934 |
Mycoboniaflava | TENN59088 | Argentina | AY513571 | AJ487933 |
Neodatroniagaoligongensis* | Cui 8055 | China | JX559269 | JX559286 |
Neodatroniasinensis* | Dai 11921 | China | JX559272 | JX559283 |
Neofavoluscremeoalbidus | Cui 12412 | China | KX899982 | KX900109 |
Neofavoluscremeoalbidus* | TUMH:50009 | Japan | AB735980 | AB735957 |
Neofavolusmikawai | Cui 11152 | China | KU189773 | KU189804 |
Neofavolusmikawai | Dai 12361 | China | KX548975 | KX548997 |
Physisporinuslineatus | JV_1008_18 | Costa Rica | OM669902 | – |
Physisporinuslineatus | JV_1407_37 | Costa Rica | OM669903 | – |
Physisporinusvinctus | JV0610_A31B-1 | Mexico | JQ409460 | – |
Physisporinusvinctus | JV0610_A31B-2 | Mexico | JQ409461 | – |
Picipesailaoshanensis | Cui 12585 | China | KX900068 | KX900183 |
Picipesailaoshanensis* | Cui 12578 | China | KX900067 | KX900182 |
Picipesamericanus | JV 0809-104 | USA | KC572003 | KC572042 |
Picipesamericanus* | JV 0509-149 | USA | KC572002 | KC572041 |
Picipesannularius* | Cui 10123 | China | KX900060 | KX900176 |
Picipesatratus | Dai 13375 | China | KX900042 | KX900158 |
Picipesatratus* | Cui 11289 | China | KX900043 | KX900159 |
Picipesauriculatus | Yuan 4221 | China | KX900064 | KX900180 |
Picipesauriculatus* | Cui 13616 | China | KX900063 | KX900179 |
Picipesbadius | Cui 10853 | China | KU189780 | KU189811 |
Picipesbadius | Cui 11136 | China | KU189781 | KU189812 |
Picipesbaishanzuensis | Cui 11395 | China | KU189763 | KU189794 |
Picipesbaishanzuensis* | Dai 13418 | China | KU189762 | KU189793 |
Picipesbrevistipitatus | Cui 11345 | China | KX900074 | KX900188 |
Picipesbrevistipitatus* | Cui 13652 | China | KX900075 | KX900189 |
Picipescf.dictyopus | Cui 11109 | China | KX900025 | KX900145 |
Picipescf.dictyopus | Cui 11092 | China | KX900026 | KX900146 |
Picipesconifericola | Cui 9950 | China | KU189783 | KU189814 |
Picipesconifericola* | Dai 11114 | China | JX473244 | KC572061 |
Picipesdictyopus | TENN 59385 | Belize | AF516561 | AJ487945 |
Picipesfraxinicola | Dai 2494 | China | KC572023 | KC572062 |
Picipesfraxinicola | Wei 6025 | China | KC572024 | KC572063 |
Picipesmelanopus | H 6003449 | Finland | JQ964422 | KC572064 |
Picipesmelanopus | MJ 372-93 | Czech | KC572026 | KC572065 |
Picipesnigromarginatus* | Cui 8113 | China | KX900062 | KX900178 |
Picipespumilus | Cui 5464 | China | KX851628 | KX851682 |
Picipespumilus | Dai 6705 | China | KX851630 | KX851684 |
Picipesrhizophilus | Dai 11599 | China | KC572028 | KC572067 |
Picipesrhizophilus | Dai 16082 | China | KX851634 | KX851687 |
Picipessubdictyopus | Cui 11220 | China | KX900057 | KX900173 |
Picipessubdictyopus | Cui 12539 | China | KX900058 | KX900174 |
Picipessubmelanopus | Dai 13294 | China | KU189770 | KU189801 |
Picipessubmelanopus | Dai 13296 | China | KU189771 | KU189802 |
Picipessubtropicus | Li 1928 | China | KU189758 | KU189790 |
Picipessubtropicus* | Cui 2662 | China | KU189759 | KU189791 |
Picipessubtubaeformis | Cui 10793 | China | KU189753 | KU189785 |
Picipessubtubaeformis* | Dai 11870 | China | KU189752 | KU189784 |
Picipestaibaiensis | Dai 5741 | China | JX489169 | KC572071 |
Picipestaibaiensis* | Dai 5746 | China | KX196783 | KX196784 |
Picipestibeticus | Cui 12225 | China | KU189756 | KU189788 |
Picipestibeticus* | Cui 12215 | China | KU189755 | KU189787 |
Picipestubaeformis | Niemela 6855 | Finland | KC572036 | KC572073 |
Picipestubaeformis | JV 0309-1 | USA | KC572034 | KC572072 |
Picipesulleungus | Cui 12410 | China | KX900022 | KX900142 |
Picipesvirgatus | CulTENN11219 | Argentina | AF516581 | AJ488122 |
Picipesvirgatus | CulTENN11406 | Argentina | AF516582 | AJ488122 |
Picipeswuyishanensis* | Dai 7409 | China | KX900061 | KX900177 |
Podofomesmollis | RLG6304sp | USA | JN165002 | JN164791 |
Podofomesstereoides | Holonen | Finland | KC415179 | KC415196 |
Polyporusauratus* | Dai 13665 | China | KX900056 | KX900172 |
Polyporusaustrosinensis | Cui 11140 | China | KX900046 | KX900162 |
Polyporusaustrosinensis* | Cui 11126 | China | KX900045 | KX900161 |
Polyporuscuticulatus | Cui 8637 | China | KX851614 | KX851668 |
Polyporuscuticulatus | Dai 13141 | China | KX851613 | KX851667 |
Polyporusguianensis | TENN 58404 | Venezuela | AF516566 | AJ487948 |
Polyporusguianensis | TENN 59093 | Argentina | AF516564 | AJ487947 |
Polyporushapalopus* | Yuan 5809 | China | KC297219 | KC297220 |
Polyporushemicapnodes | Cui 11259 | China | KX851625 | KX851679 |
Polyporushemicapnodes | Dai 13403 | China | KX851627 | KX851681 |
Polyporuslamelliporus | Dai 12327 | China | KX851622 | KX851676 |
Polyporuslamelliporus* | Dai 15106 | China | KX851623 | KX851677 |
Polyporusleprieurii | TENN 58579 | Costa Rica | AF516567 | AJ487949 |
Polyporusmangshanensis* | Dai 15151 | China | KX851796 | KX851797 |
Polyporusminutissimus | Wu 970 | China | PP035829 | PP035826 |
Polyporusminutissimus* | Wu 971 | China | PP035830 | PP035827 |
Polyporusparvovarius | Yuan 6639 | China | KX900049 | KX900165 |
Polyporusparvovarius | Dai 13948 | China | KX900050 | KX900166 |
Polyporusradicatus | DAOM198916 | Canada | AF516584 | AJ487955 |
Polyporusradicatus | TENN 58831 | USA | AF516585 | AJ487956 |
Polyporus sp.1 | Cui 11071 | China | KX851642 | KX851695 |
Polyporus sp.1 | Cui 11045 | China | KX851643 | KX851696 |
Polyporus sp.2 | Dai 13585A | China | KX900055 | KX900171 |
Polyporussquamosus | Cui 10394 | China | KX851635 | KX851688 |
Polyporussquamosus | Cui 10595 | China | KU189778 | KU189809 |
Polyporussubvarius | WD2368 | Japan | AB587643 | AB587638 |
Polyporussubvarius* | Yu 2 | China | AB587632 | AB587621 |
Polyporustuberaster | Dai 11271 | China | KU189769 | KU189800 |
Polyporustuberaster | Dai 12462 | China | KU507580 | KU507582 |
Polyporusumbellatus | Pen 13513 | China | KU189772 | KU189803 |
Polyporusvarius | Cui 12249 | China | KU507581 | KU507583 |
Polyporusvarius | Dai 13874 | China | KU189777 | KU189808 |
Pseudofavoluscucullatus | Dai 13584A | China | KX900071 | KX900185 |
Pseudofavoluscucullatus | WD2157 | Japan | AB587637 | AB368114 |
Trametesconchifer | FP106793sp | USA | JN164924 | JN164797 |
Trameteselegans | FP105679sp | USA | JN164944 | JN164799 |
Trametespolyzona | Cui 11040 | China | KR605824 | KR605767 |
Notes: New sequences are in bold; “–” represents missing data; * represents type specimens
Phylogenetic analysis
Phylogenetic trees of Cerrena and Polyporus were constructed using the two concatenated ITS1-5.8S-ITS2-nLSU sequences dataset, respectively, and phylogenetic analyses were performed with Maximum Likelihood (ML) and Bayesian Inference (BI) methods. New sequences generated in this study and reference sequences retrieved from GenBank (Table 1) were partitioned to ITS1, 5.8S, ITS2, nLSU and then aligned separately using MAFFT v.74 (Katoh et al. 2019; http://mafft.cbrc.jp/alignment/server/) with the G-INS-I iterative refinement algorithm and optimised manually in BioEdit v.7.0.5.3 (Hall 1999). The separate alignments were then concatenated using PhyloSuite v.1.2.2 (Zhang et al. 2020). Unreliably aligned sections were removed before the analyses, and efforts were made to manually inspect and improve the alignment. The data matrix was edited in Mesquite v3.70. Irpexlatemarginatus (Durieu & Mont.) C.C. Chen & Sheng H. Wu was used as an outgroup in the phylogenetic analysis of Cerrena (Parmasto and Hallenberg 2000). Trametesconchifer (Schwein.) Pilát, T.elegans (Spreng.) Fr. and T.polyzona (Pers.) Justo were selected as outgroups in the phylogenetic analysis of Polyporus (Ji et al. 2022). The final alignments and the retrieved topologies were deposited in TreeBASE (http://www.treebase.org) under accessions 31102, 31103.
RAxML 7.2.8 was used to infer ML trees for both datasets with the GTR+I+G model of site substitution, including estimation of Gamma-distributed rate heterogeneity and a proportion of invariant sites (Stamatakis 2006). The branch support was evaluated with a bootstrapping method of 1,000 replicates (Hillis and Bull 1993).
For BI, the best-fit partitioning scheme and substitution model were determined by using ModelFinder (Kalyaanamoorthy et al. 2017) via the “greedy” algorithm, branch lengths estimated as “linked” and AICc. The BI was conducted with MrBayes 3.2.6 in two independent runs, each of which had four chains for 20 million generations and started from random trees (Ronquist et al. 2012). Trees were sampled every 1,000 generations. The first 25% of the sampled trees were discarded as burn-in and the remaining ones were used to reconstruct a majority rule consensus and calculate Bayesian Posterior Probabilities (BPP) of the clades.
Phylogenetic trees were visualized using FigTree version 1.4.4 (Rambaut 2018). Branches that received bootstrap support (BS) for ML and BPPs greater than or equal to 75% (ML) and 0.95 (BPP) were considered significantly supported, respectively.
Results
Phylogenetic analyses
In the phylogenetic analysis of Cerrena (Fig. 1), the combined ITS1-5.8S-ITS2-nLSU dataset included sequences from 50 fungal collections representing 11 taxa, and one sample of Irpexlatemarginatus was used as an outgroup. ModelFinder proposed models were HKY+F+G4 for ITS1, GTR+F+I+G4 for 5.8s, HKY+F+G4 for ITS2 and GTR+F+I for nLSU, for Bayesian analysis. The BI analysis resulted in an average standard deviation of split frequencies = 0.008865. As both ML and BI trees resulted in similar topologies, only the topology of the ML analysis is presented together with the statistical values of the ML (≥75%) and BPP (≥0.90) algorithms (Fig. 1). The phylogeny inferred from ITS1-5.8S-ITS2-nLSU sequences (Fig. 1) showed that our three newly sequenced samples together with three samples defined as Cerrena sp. 1 by Miettinen et al. (2023) formed an independent lineage with strong support (97/0.98, Fig. 1). The lineage is defined as the new species Cerrenacaulinicystidiata.
Figure 1.
Maximum Likelihood (ML) phylogenetic tree illustrating the phylogeny of Cerrena and related genera in five families based on the combined ITS1-5.8S-ITS2-nLSU dataset. Branches are labeled with maximum likelihood bootstrap values (ML) higher than 75% and Bayesian posterior probabilities above 0.90. The new species is given in bold.
In the phylogenetic analysis of Polyporus (Fig. 2), the combined ITS1-5.8S-ITS2-nLSU dataset included sequences from 113 fungal collections representing 71 species, and three samples of Trametes were used as outgroups. ModelFinder suggested models were GTR+F+I+G4 for ITS1, K2P+I+G4 for 5.8s, K2P+I+G4 for ITS2 and GTR+F+I+G4 for nLSU, for Bayesian analysis. The BI analysis resulted in an average standard deviation of split frequencies = 0.009675. The ML and BI trees were similar in topology, and only the topology of the ML analysis is presented along with the statistical values of the ML (≥75%) and BPP (≥0.90) algorithms (Fig. 2). The phylogeny inferred from the combined ITS1-5.8S-ITS2-nLSU sequences (Fig. 2) revealed that a new lineage with high support (100/1.00,) nests in the squamosus clade in Polyporus, namely Polyporusminutissimus. The new species is closely related to P.hemicapnodes Berk. & Broome and P.parvovarius H. Lee, N.K. Kim & Y.W. Lim.
Figure 2.
Maximum Likelihood (ML) phylogenetic tree illustrating the phylogeny of Polyporus and related genera based on the combined ITS1-5.8S-ITS2-nLSU dataset. Branches are labeled with maximum likelihood bootstrap values (ML) higher than 75% and Bayesian posterior probabilities (BPPs) more than 0.90. The new species is given in bold.
Taxonomy
. Cerrena caulinicystidiata
T. Cao, F. Wu & H.S. Yuan sp. nov.
161AF221-198F-5B22-A0A7-DA9653A2DC30
853719
Figure 3.
Basidiocarps of Cerrenacaulinicystidiata (Holotype, Wu 661).
Figure 4.
Microscopic structures of Cerrenacaulinicystidiata (Wu 661) a basidiospores b basidia and basidioles c cystidia d encrusted cystidia e hyphae from subiculum f hyphae from trama.
Holotype.
China • Zhejiang Province, Hangzhou, Xiaoshan District, Yangjingwu Forest Park; 30°4'1"N, 120°19'35"E; 134 m a.s.l.; 27 Mar. 2023; on fallen angiosperm branch; F. Wu leg., Wu 661 (BJFC040654).
Etymology.
Caulinicystidiata (Lat.): Refers to the cystidia with a tapering base.
Description.
Basidiocarps. Annual, resupinate, sometimes effused-reflexed, continuous, easily separable, without special odor or taste when fresh, corky when dry, up to 10 cm long, 3 cm wide and 0.5 mm thick. Pore surface greyish orange to brownish orange; pores irregular, 3–8 per mm, partly split up to 2 mm long; dissepiments thin. Sterile margin finely fimbriated. Subiculum very thin, yellowish white, ca. 0.5 mm thick, a very thin brownish red crust present in the bottom next to wood. Tubes concolorous with pore surface, corky, 0.5–1 mm long.
Hyphal structure. Hyphal system trimitic, generative hyphae with clamp connections; skeletal and binding hyphae CB+, IKI–; tissues unchanged in KOH.
Subiculum. Generative hyphae thin- to slightly thick-walled, hyaline, clamped, frequently branched, 2–5 µm in diam; skeletal hyphae dominant, thick-walled to subsolid, unbranched, interwoven, 2.5–6 µm in diam; binding hyphae hyaline, thick-walled to subsolid, tortuose, moderately branched, 1.5–2.5 μm diam. The thin crust made up of subsolid, brownish and strongly agglutinated hyphae.
Tubes. Generative hyphae infrequent, hyaline, thin- to slightly thick-walled, clamped, rarely branched, 2–3 µm diam; skeletal hyphae dominant, hyaline, thick-walled to subsolid, rarely branched, sometimes with septate, interwoven, 2–4 µm in diam; binding hyphae rare. Cystidia clavate to pyriform to ventricose, mostly thin-walled, occasionally thick-walled, smooth, 13–20 × 6–12 µm; encrusted cystidia numerous, clavate, originated from and tightly embedded in trama, 10–25 × 7–15 µm (with encrustation). Basidia short clavate, with four sterigmata and a basal clamp, 8–11 × 4–5 µm, basidioles in shape similar to basidia, but slightly smaller.
Basidiospores. Basidiospores broadly-ellipsoid to ovoid, hyaline, thin-walled, smooth, CB–, IKI–, (3–)3.2–4.5(–4.8) × (2.5–)2.8–3.5(–3.9) µm, L = 3.94 µm, W = 2.84 µm, Q = 1.38–1.44 (n = 60/2).
Additional specimens examined
(paratypes). Vietnam • Lam dong Province (Lat.), Lac Duong District, Bidoup Nui Ba National Park; 12°11'8"N, 108°40'41"E; 1495 m a.s.l.; 15 Oct. 2017; on fallen angiosperm branch; H.S. Yuan leg., Yuan 12666 (IFP 019379), Yuan 12664 (IFP 019378).
. Polyporus minutissimus
Q.Y. Zhang, Z.W. Zheng & F. Wu sp. nov.
3C5B563B-D60C-5F05-A739-E3B1EC5F46A6
853720
Figure 5.
Basidiocarps of Polyporusminutissimus (Holotype, Wu 971).
Figure 6.
Microscopic structures of Polyporusminutissimus (Wu970/Wu 971) a basidiospores b basidia and basidioles c hyphae from trama.
Holotype.
China • Zhejiang Province, Hangzhou, Yuhang District, Luniao Town; 30°25'50"N, 119°42'38"E; 158.47 m a.s.l.; 9 Jun. 2023; on ground of Bamboo forest; F. Wu leg., Wu 971 (BJFC040963, holotype).
Etymology.
Minutissimus (Lat.): Referring to the species having tiny basidiocarps.
Description.
Basidiocarps. Annual, centrally stipitate, solitary, fleshy to soft leathery when fresh, becoming fragile when dry. Pilei flat with a depressed center or infundibuliform, up to 1.5 cm in diam and 0.5–1 mm thick. Pileal surface cream to buff yellow when dry, glabrous, occasionally zonate and with radially aligned stripes; margin sharp, incurved upon drying. Pore surface cream when dry; pores angular, 6–9 per mm; dissepiments thin, entire. Context buff cream to pale neutral when dry, fragile upon drying, up to 0.5 mm thick. Tubes white to cream when dry, decurrent, up to 0.5 mm thick. Stipe dark violet, glabrous, 0.3–0.5 cm long and 1–2 mm in diam.
Hyphal structure. Hyphal system dimitic; generative hyphae bearing clamp connections, thin-walled, hyaline; skeleton-binding hyphae thick-walled with a wide lumen, with arboriform branches, IKI–, CB+; tissue unchanged in KOH.
Context. Generative hyphae frequent, colorless, thin-walled, 2.5–4 μm in diam; skeleto-binding hyphae dominant, colorless, thick-walled with a wide lumen, moderately branched, strongly interwoven, 2–4.5 μm diam.
Tubes. Generative hyphae frequent, colorless, thin-walled, 2–3 μm in diam; skeleto-binding hyphae dominant, colorless, thick-walled with a wide lumen, moderately branched, interwoven, 1–3 μm in diam. Cystidia and cystidioles absent. Basidia clavate, with four sterigmata and a basal clamp connection, 22–28 × 7–9 μm; basidioles in shape similar to basidia, but slightly smaller.
Stipe. Generative hyphae frequent, colorless, thin-walled, rarely branched, 3–4 μm in diam; skeleto-binding hyphae dominant, colorless, thick-walled with a wide lumen, moderately branched, interwoven, 1.5–4 μm in diam.
Basidiospores. Basidiospores cylindrical to oblong, colorless, thin-walled, smooth, IKI–, CB–, 5–9.2(–10) × (2–)2.2–4(–4.2) μm, L = 7.30 μm, W = 3.23 μm, Q = 2.25–2.27 (n = 60/2).
Additional specimen examined
(paratype). China • Zhejiang Province, Hangzhou, Yuhang District, Luniao Town; 30°25'50"N, 119°42'38"E; 155.11 m a.s.l; on ground of bamboo forest, 9 Jun. 2023; F. Wu leg., Wu 970 (BJFC040962).
Discussion
In this study, two new species of the Polyporales - Cerrenacaulinicystidiata and Polyporusminutissimus - are proposed based on morphological and phylogenetic evidence. Our three newly sequenced Cerrena samples together with three samples which were defined as Cerrena sp. 1 by Miettinen et al. (2023) formed an independent well-supported lineage in our phylogeny (Fig. 1). The lineage is proposed as the new species C.caulinicystidiata. Another lineage which is defined as Cerrena sp. 2 in our phylogeny is closely related to C.caulinicystidiata, but we didn’t collect one specimen within the lineage, so the lineage is considered to be Cerrena sp.
Cerrenacaulinicystidiata is characterized by its resupinate, sometimes effused-reflexed basidiocarps, greyish orange to brownish orange pore surface, 3–8 per mm pores, and subglobose basidiospores, 3.2–4.5 × 2.8–3.5 µm in size. C.albocinnamomea (Y.C. Dai & Niemelä) H.S. Yuan originally described from Northeast China resembles C.caulinicystidiata by sharing resupinate and easily separable basidiocarps. However, C.albocinnamomea differs from C.caulinicystidiata by its clavate to pyriform cystidia, slightly smaller ellipsoid basidiospores (2.8–3.5 × 2–3 µm vs. 3.2–4.5 × 2.8–3.5 µm), and a dimitic hyphal system (Yuan 2014).
In addition, Rigidoporusvinctus (Berk.) Ryvarden [≡ Physisporinusvinctus (Berk.) Murrill, Wu et al. 2017] resembles C.caulinicystidiata by having resupinate basidiocarps, ochraceous pore surface, ventricose cystidia with a subcylindric appendage, encrusted cystidia, and subglobose basidiospores, but it can be distinguished from the latter species by its smaller pores (6–12 per mm vs. 3–8 per mm) and generative hyphae with simple septa (Ryvarden and Johansen 1980).
The genus Cerrena is widely distributed and has diverse morphological characteristics. Currently, there are 13 records according to Index Fungorum (http://www.indexfungorum.org). However, C. ‘gilbertsonii’ Ryvarden cannot be distinguished from C.cystidiata Rajchenb. & De Meijer by morphological characteristics, and C. ‘multipileata’ (C.L. Leite & J.E. Wright) Miettinen cannot be distinguished from C.zonata (Berk.) H.S. Yuan (Miettinen et al. 2023). Cerrenaaurantiopora J.S. Lee & Y.W. Lim is a synonym of C.albocinnamomea (Lee and Lim 2010; Miettinen et al. 2023). Therefore, we provide a Key to 11 undisputed Cerrena species including the new species.
Key to species of the Cerrena
1 | Paleotropical or temperate-boreal species | 2 |
– | Neotropical (South American) species | 9 |
2 | Basidiocarp poroid, occasionally lacerate | 3 |
– | Basidiocarp irpicoid | 8 |
3 | Pore surface umbrinous to bay or blackish | C.subglabrescens |
– | Pore surface white, light orange to brown | 4 |
4 | Pores umber, round, 1–2 per mm | C.drummondii |
– | Pores round to angular, > 3 per mm | 5 |
5 | Basidiospores narrowly ellipsoid, 7.5–10 × 2.5–3.5 µm | C.caperata |
– | Basidiospores ellipsoid to broadly-ellipsoid, < 6 µm in length | 6 |
6 | Pores angular, dissepiments even or lacerate | C.albocinnamomea |
– | Pores rounded to irregular | 7 |
7 | Basidiocarps coriaceous, imbricate | C.fulvocinerea |
– | Basidiocarps resupinate, sometimes effused-reflexed | C.caulinicystidiata |
8 | Pore surface white to cream | C.unicolor |
– | Pore surface first white to pale ochraceous | C.zonata |
9 | Cystidia present | C.cystidiata |
– | Cystidia absent | 10 |
10 | Pore surface pale cinnamon to brown | C.sclerodepsis |
– | Pore surface dark brown to almost black | C.hydnoides |
In the phylogenetic analysis of Polyporus, P.minutissimus was assigned to the squamosus clade with strong support (100/1.00, Fig. 2). The squamosus clade has always been supported by phylogenetic analysis based on the ITS+nLSU or eight-gene datasets, but the species within this clade cannot be combined into a monophyletic genus because they manifest greatly diverse morphology (Ji et al. 2022). Phylogenetically, P.minutissimus is closely related to P.hemicapnodes and P.parvovarius (Fig. 2). P.hemicapnodes was described from Dolosbagey (Sri Lanka). For some time, it was treated as a synonymy of P.leprieurii (Núñez and Ryvarden 1995), which differs from P.minutissimus by its larger basidiocarps (up to 10 cm vs. up to 1.5 cm), cream to tan pore surface, and longer stipe (up to 5 cm vs. up to 0.5 cm, Berkeley and Broome 1873). Polyporusparvovarius has microscopic features similar to P.minutissimus. However, P.parvovarius differs by its smaller basidiocarps (up to 0.35 cm vs. up to 1.5 cm) and light buff to brown pileal surface (Tibpromma et al. 2017).
Macro-morphologically, Polyporusminutissimus has a depressed center or infundibuliform basidiocarps and black stipe, cream to buff yellow pileal surface, and 6–9 per mm pores. Microscopically, it has a dimitic hyphal system, strongly branched skeleton-binding hyphae in both trama and context, and cylindrical basidiospores. Morphologically, P.lamelliporus B.K. Cui, Xing Ji & J.L. Zhou is similar to P.minutissimus by sharing depressed center or infundibuliform basidiocarps, cream to buff yellow pileal surface, and similar-sized basidiospores, but the former differs through its larger basidiocarps (up to 5.2 cm vs. up to 1.5 cm), longer stipe (1–3.5 cm vs. 0.3–0.5 cm), and larger pores (0.5–1 per mm vs. 6–9 per mm, Ji et al. 2022). In addition, Picipesbaishanzuensis J.L. Zhou & B.K. Cui, which is similar to P.minutissimus and shares infundibuliform basidiocarps and a black stipe, has also been reported from Baishanzu nature reserve, which is the type producing area of our new species. However, P.baishanzuensis differs from P.minutissimus by its larger basidiocarps (up to 5.5 cm vs. up to 1.5 cm) and smaller basidiospores (6.6–7.9 × 2.5–3.1 µm vs. 5–9.2 × 2.2–4 µm; Zhou et al. 2016).
Polyporus is a very complicated genus with more than 3000 records according to the Index Fungorum. However, studies on Polyporus species in China are gradually being carried out, with some Chinese species having been described in Cui et al. (2019) and Ji et al. (2022). Therefore, we provide a Key to Chinese Polyporus species including the new species.
Key to species of Polyporus in China
1 | Stipe absent | P.megasporoporus |
– | Stipe present | 2 |
2 | Stipe bearing black cuticle | 3 |
– | Stipe white to ochraceous | 7 |
3 | Pileal surface covered with dark-brown to reddish-brown squamules | P.squamosus |
– | Pileal surface glabrous | 4 |
4 | Pores more than 5 per mm | 5 |
– | Pores less than 5 per mm | 6 |
5 | Pileal surface concentrically zonate; basidiospores 5.4–7.6 × 2.9–3.8 μm | P.hemicapnodes |
– | Pileal surface azonate; basidiospores 7.5–9 × 2.5–3.3 μm | P.varius |
6 | Pores 3–5 per mm | P.mangshanensis |
– | Pores 1–2 per mm | P.subvarius |
7 | Stipes numerous and branched | P.umbellatus |
– | Stipes usually single and not branched | 8 |
8 | Basidiospores < 8 μm in length | 9 |
– | Basidiospores > 8 μm in length | 11 |
9 | Basidiocarps imbricate | P.hapalopus |
– | Basidiocarps solitary | 10 |
10 | Pores angular, 2–3 per mm | P.brumalis |
– | Pores round, 4–5 per mm | P.ciliatus |
11 | Pileal surface with radial stripes | 12 |
– | Pileal surface without radial stripes | 13 |
12 | Pores 2–5 per mm | P.cuticulatus |
– | Pores 6–9 per mm | P.minutissimus |
13 | Basidiospores usually < 10 μm in length | 14 |
– | Basidiospores usually > 10 μm in length | P.tuberaster |
14 | Cystidioles absent | 15 |
– | Cystidioles infrequent | P.austrosinensis |
15 | Basidia < 27 μm in length | 16 |
– | Basidia > 27 μm in length | P.lamelliporus |
16 | Basidiospores smaller, 6–8.3 × 2.2–3 μm | P.arcularius |
– | Basidiospores larger, 7.7–10 × 3–3.9 μm | P.auratus |
Polyporales is a large group of Basidiomycota with diverse morphology and phylogeny. There have been over 577 taxonomic proposals in the Polyporales and 2,183 publications with the keyword ‘Polyporales’ over the past decade (Binder et al. 2013; Justo et al. 2017). However, the species in the order are still not sufficiently investigated in Asia, especially in the subtropics and tropics (Li et al. 2016; Hyde 2022). New DNA sequencing techniques have revolutionized fungal taxonomy and diversity, with multi-marker datasets. In the present study, two new polypore species, C.caulinicystidiata and P.minutissimus were found in subtropical regions, which enriches our understanding of the fungal diversity of the Polyporales in Asia.
Supplementary Material
Citation
Zheng Z-W, Zhang Q-Y, Zhang L-R, Yuan H-S, Wu F (2024) Morphological and molecular data reveal Cerrena caulinicystidiata sp. nov. and Polyporus minutissimus sp. nov. in Polyporales from Asia. MycoKeys 106: 1–21. https://doi.org/10.3897/mycokeys.106.121840
Funding Statement
This study was financed by the National Natural Science Foundation of China (Project Nos. 32070006, 32270011, 31701978), the Tibet Autonomous Region Science and Technology Project (XZ202201ZY0006N), and the Fundamental Research Funds for the Central Universities (No. QNTD202307).
Footnotes
Zi-Wei Zheng, Qiu-Yue Zhang contributed equally to this work.
Additional information
Conflict of interest
The authors have declared that no competing interests exist.
Ethical statement
No ethical statement was reported.
Funding
This study was financed by the National Natural Science Foundation of China (Project Nos. 32070006, 32270011, 31701978), the Tibet Autonomous Region Science and Technology Project (XZ202201ZY0006N), and the Fundamental Research Funds for the Central Universities (No. QNTD202307).
Author contributions
Data curation: LRZ. Investigation: HSY, QYZ, FW, ZWZ. Methodology: QYZ, ZWZ. Resources: HSY. Supervision: LRZ, FW. Validation: QYZ, ZWZ. Writing - original draft: ZWZ, QYZ. Writing - review and editing: FW.
Author ORCIDs
Zi-Wei Zheng https://orcid.org/0009-0006-8442-408X
Qiu-Yue Zhang https://orcid.org/0000-0001-9458-3566
Hai-Sheng Yuan https://orcid.org/0000-0001-7056-140X
Data availability
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Supplementary materials
31102 Treebase
This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Zi-Wei Zheng, Qiu-Yue Zhang, Li-Rong Zhang, Hai-Sheng Yuan, Fang Wu
Data type
nxs
31103 Treebase
This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Zi-Wei Zheng and Qiu-Yue Zhang, Li-Rong Zhang, Hai-Sheng Yuan, Fang Wu
Data type
nxs
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Associated Data
This section collects any data citations, data availability statements, or supplementary materials included in this article.
Supplementary Materials
31102 Treebase
This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Zi-Wei Zheng, Qiu-Yue Zhang, Li-Rong Zhang, Hai-Sheng Yuan, Fang Wu
Data type
nxs
31103 Treebase
This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
Zi-Wei Zheng and Qiu-Yue Zhang, Li-Rong Zhang, Hai-Sheng Yuan, Fang Wu
Data type
nxs
Data Availability Statement
All of the data that support the findings of this study are available in the main text or Supplementary Information.