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Research Article
Species diversity and taxonomy of Vararia (Russulales, Basidiomycota) with descriptions of six species from Southwestern China
expand article infoYinglian Deng‡, Sana Jabeen§, Changlin Zhao‡
‡ Southwest Forestry University, Kunming, China
§ University of Education, Lahore, Pakistan

Corresponding author: Changlin Zhao ( fungichanglinz@163.com )

Academic editor: María P. Martín
© 2024 Yinglian Deng, Sana Jabeen, Changlin Zhao.
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.
Citation: Deng Y, Jabeen S, Zhao C (2024) Species diversity and taxonomy of Vararia (Russulales, Basidiomycota) with descriptions of six species from Southwestern China. MycoKeys 103: 97-128. https://doi.org/10.3897/mycokeys.103.118980
Open Access

Abstract

Vararia is a species-rich genus in the family Peniophoraceae and has been shown to be polyphyletic. In this study, sequences of ITS and LSU rRNA markers of the studied samples were generated and phylogenetic analyses were performed with the maximum likelihood, maximum parsimony, and Bayesian inference methods. Seventeen lineages including six new species from China, i.e., V. fissurata, V. lincangensis, V. punctata, V. isabellina, V. sinensis, and V. yaoshanensis were recognized, in which V. fissurata is characterized by the brittle basidiomata with pruinose and cracking hymenophore having white to olivaceous buff hymenial surface, the clamped generative hyphae, presence of the two types gloeocystidia; V. lincangensis is characterized by the simple-septa generative hyphae, and thick-walled skeletal hyphae, and ellipsoid basidiospores; V. punctata is delimited by its thin to slightly thick-walled generative hyphae, and thick-walled skeletal hyphae, present thick-walled, clavate to cylindrical gloeocystidia; V. isabellina is characterized by having the cream to isabelline to slightly brown hymenial surface, thin to slightly thick-walled generative hyphae, and sub-fusiform to navicular basidiospores; V. sinensis is distinguishable by its white to slightly pink hymenial surface, thick-walled skeletal hyphae, and sub-fusiform to navicular basidiospores; V. yaoshanensis is characterized by cream to pinkish buff to cinnamon-buff hymenial surface, slightly thick-walled generative hyphae, the presence of two types gloeocystidia, and slightly thick-walled, ellipsoid basidiospores. Phylogram based on the ITS+nLSU rDNA gene regions included nine genera within the family Peniophoraceae as Amylostereum, Asterostroma, Baltazaria, Dichostereum, Michenera, Peniophora, Scytinostroma and Vararia, in which the six new wood-inhabiting fungi species were grouped into genus Vararia. The phylogenetic tree inferred from the combined ITS and LSU tree sequences highlighted that V. fissurata was found to be the sister to V. ellipsospora with strong supports. Additionally, V. lincangensis was clustered with V. fragilis. Furthermore, V. punctata was retrieved as a sister to V. ambigua. Moreover, V. sinensis was grouped with five taxa as V. breviphysa, V. pirispora, V. fusispora, V. abortiphysa and V. insolita. The new species V. isabellina formed a monophyletic lineage, in which it was then grouped closely with V. daweishanensis, and V. gracilispora. In addition, V. yaoshanensis was found to be the sister to V. gallica with strong supports. The present results increased the knowledge of Vararia species diversity and taxonomy of corticioid fungi in China. An identification key to 17 species of Vararia in China is provided.

Key words

Biodiversity, China, phylogenetic analyses, taxonomy, wood-inhabiting fungi, Yunnan Province

Introduction

Fungi represent one of the most diverse groups of organisms on the earth, with an indispensable role in the processes and functioning of forest ecosystems (Hyde 2022). The genus Vararia P. Karst. belongs to the family Peniophoraceae of the order Russulales (Larsson and Larsson 2003; Miller et al. 2006; Larsson 2007). The Russulales is a well-known order that contains morphologically diverse mushrooms (Miller et al. 2006). Species from this order comprise many representative wood-inhabiting fungal taxa, including hydnoid, corticioid, and polyporoid basidiomes with diverse hymenophoral and cystidial morphology (Yurchenko and Wu 2016; Riebesehl and Langer 2017; Yurchenko et al. 2017; Cui et al. 2019; Riebesehl et al. 2019; Jiang et al. 2021; Wu et al. 2022).

The genus Vararia is a corticioid wood-inhabiting fungal genus with a wide distribution, typified by V. investiens (Schwein.) P. Karst. It was first described by Karsten as a subgenus of Xerocarpus P. Karst. for Xerocarpus alutarius (Berk. & M. A. Curtis) P. Karst., which was later found to be a synonym of Radulum investiens Schwein. Karsten raised Xerocarpus subgen. Vararia to the generic rank (Karasinski 2010). The genus is characterized by the resupinate basidiomata, a dimitic hyphal structure with simple-septate or clamped generative hyphae and often dextrinoid dichohyphae in Melzer’s reagent, the presence of gloeocystidia, and variously shaped smooth basidiospores with or without an amyloid reaction (Karnste 1898; Boidin and Lanquetin 1975; Boidin 1980; Bernicchia and Gorjón 2010). The species of Vararia are found on fallen angiosperm branches, dead woody or herbaceous stems or occasionally on gymnosperm wood (Yurchenko et al. 2017). Based on the MycoBank database (http://www.mycobank.org, accessed on 17 January 2024) and the Index Fungorum (http://www.indexfungorum.org, accessed on 17 January 2024), Vararia has registered 99 specific and infraspecific names, and the actual number of the species has reached up to 76, currently known, and they occur mainly in the tropical and subtropical areas of the world (Cunningham 1955; Gilbertson 1965; Boidin 1967; Pouzar 1982; Boidin and Lanquetin 1987; Stalpers 1996; Boidin and Gilles 1999; Larsson and Larsson 2003; Bernicchia and Gorjón 2010; Duhem and Buyck 2012; Sanyal et al. 2012; Nakasone 2015; Liu and He 2016; Dai et al. 2021; Zou et al. 2022; Deng and Zhao 2023).

Classification of the kingdom of fungi has been updated continuously, based on the frequent inclusion of data from DNA sequences in many phylogenetic studies (Yurchenko et al. 2020). These pioneering research studies into the family Peniophoraceae were just the prelude to the molecular systematics period (Zou et al. 2022). The phylogenetic diversity displayed by corticioid fungal species, based on ITS1-5.8S-ITS2-nrLSU nuclear rDNA, revealed that the taxa of Peniophoraceae were nested in the russuloid clade, which holds a considerable share of the phylogenetic framework, and included the genera of Asterostroma Massee, Baltazaria Leal-Dutra, Dentinger & G.W. Griff., Dichostereum Pilát, Gloiothele Bres., Lachnocladium Lév., Michenera Berk. & M.A. Curtis, Peniophora Cooke, Scytinostroma Donk, Vesiculomyces E. Hagstr. and Vararia (Larsson and Larsson 2003; Larsson and Larsson 2004; Larsson 2007; Leal-Dutra et al. 2018; Zou et al. 2022; Li et al. 2023). Morphologically, Scytinostroma was similar to Vararia, which usually differed in having the typical dichohyphae (Bernicchia and Gorjón 2010). The taxonomic distinction between Scytinostroma and Vararia has been questioned (Hallenberg 1985; Boidin and Lanquetin 1987; Stalpers 1996; Boidin et al 1998). However, there has been general agreement that the two genera were closely related and that they together made up a natural group. Larsson and Larsson (2003) strongly suggested that neither skeletal hyphae nor their branching patterns have any predictive power in a phylogenetic context.

During investigations on the wood-inhabiting fungi in the Yunnan province of China, the samples representing six additional species belonging to genera Vararia were collected. To clarify the placement and relationships of the six species, we carried out a phylogenetic and taxonomic study on Vararia, based on the ITS and LSU sequences.

Materials and methods

Morphology

Fresh fruiting bodies of the fungi were collected from Dali, Dehong, Lincang, Puer, Yuxi and Zhaotong of Yunnan Province, P.R. China. Specimens were dried in an electric food dehydrator at 40 °C, then sealed and stored in an envelope bag and deposited in the herbarium of the Southwest Forestry University (SWFC), Kunming, Yunnan Province, P.R. China. Macromorphological descriptions are based on field notes and photos captured in the field and lab. Color terminology follows Petersen (Petersen 1996). Micromorphological data were obtained from the dried specimens when observed under a light microscope following the previous study (Zhao et al. 2023; Guan et al. 2023). The following abbreviations are used: KOH = 5% potassium hydroxide water solution, CB = Cotton Blue, CB– = acyanophilous, IKI = Melzer’s Reagent, IKI– = both inamyloid and indextrinoid, L = mean spore length (arithmetic average for all spores), W = mean spore width (arithmetic average for all spores), Q = variation in the L/W ratios between the specimens studied and n = a/b (number of spores (a) measured from given number (b) of specimens).

Molecular phylogeny

The EZNA HP Fungal DNA Kit (Omega Biotechnologies Co., Ltd., Kunming, China) was used to extract DNA with some modifications from the dried specimens. The nuclear ribosomal ITS region was amplified with primers ITS5 and ITS4 (White et al. 1990). The PCR procedure for ITS was as follows: initial denaturation at 95 °C for 3 min, followed by 35 cycles at 94 °C for 40 s, 58 °C for 45 s and 72 °C for 1 min, and a final extension of 72 °C for 10 min. The nuclear LSU region was amplified with primer pair LR0R and LR7 (Vilgalys and Hester 1990; Rehner and Samuels 1994). The PCR procedure for LSU was as follows: initial denaturation at 94 °C for 1 min, followed by 35 cycles at 94 °C for 30 s, 48 °C for 1 min and 72 °C for 1.5 min, and a final extension of 72 °C for 10 min. The PCR procedure for ITS and LSU followed a previous study (Zhao and Wu 2017). All of the newly generated sequences were deposited in NCBI GenBank (https://www.ncbi.nlm.nih.gov/genbank/) (Table 1).

Table 1.
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List of species, specimens and GenBank accession numbers of sequences used in this study. [* Indicates type materials].

Species name Specimen No. GenBank accession No. Country References
ITS nLSU
Amylostereum chailletii NH8031 AF506406 AF506406 Sweden Larsson and Larsson 2003
A. laevigatum NH12863 AF506407 AF506407 Sweden Larsson and Larsson 2003
Asterostroma bambusicola He4132 KY263865 KY263871 Thailand Liu et al. 2017
A. cervicolor He2314 KY263859 KY263869 China Unpublished
A. cervicolor He4020 KY263860 KY263868 Thailand Unpublished
A. muscicola He4397 MK625630 MK625563 China Unpublished
Baltazaria galactina He4999 MK625618 MK625547 China Unpublished
B. octopodites FLOR63715 MH260042 MH260060 United Kingdom Leal-Dutra et al. 2018
Confertobasidium olivaceoalbum FP90196 AF511648 AF511648 Sweden Larsson and Larsson 2003
Dichostereum boidinii He4410 MH538315 MH538331 China Vu et al. 2019
D. boidinii He5026 MH538324 MH538330 China Liu et al. 2019
D. pallescens CBS:718.81 MH861456 MH873198 USA Vu et al. 2019
Metulodontia nivea NH13108 AF506423 AF506423 Sweden Larsson and Larsson 2003
Michenera artocreas GHL-2016-Oct MH204688 MH204691 USA Liu et al. 2019
M. incrustata He5368 MH204689 MH204690 China Liu et al. 2019
Peniophora cinerea CBS:261.37 MH855905 MH867412 Belgium Vu et al. 2019
P. cinerea He3725 MK588769 MK588809 China Unpublished
P. incarnata CBS 430.72 MH860518 MH872230 Netherlands Vu et al. 2019
P. incarnata NH10271 AF506425 AF506425 Sweden Larsson and Larsson 2003
P. nuda LZ15-07 MT859929 — China Unpublished
P. quercina CBS 407.50 MH856687 MH868204 France Vu et al. 2019
P. quercina CBS:410.50 MH856690 MH868207 France Vu et al. 2019
Scytinostroma acystidiatum He5646 MK625568 MK625494 China Unpublished
S. alutum CBS:762.81 MH861482 MH873221 France Vu et al. 2019
S. beijingensis He7768 OQ731943 OQ729731 China Li et al. 2023
S. boidinii He6911 OQ731934 OQ729724 China Li et al. 2023
S. duriusculum He3590 MK625571 MK625499 China Unpublished
S. hemidichophyticum CBS:702.84 MH861818 MH873509 Belgium Vu et al. 2019
S. renisporum CBS:771.86 MH862051 MH873738 Bali Vu et al. 2019
S. subrenisporum He4792 MK625566 MK625493 China Unpublished
Vararia abortiphysa CBS:632.81 MH861387 — Gabon Vu et al. 2019
V. ambigua CBS 634.81 MH861388 MH873137 France Vu et al. 2019
V. amphithallica CBS:635.81 MH861389 MH873138 Gabon Vu et al. 2019
V. amphithallica CBS:687.81 MH861431 — France Vu et al. 2019
V. aurantiaca CBS:641.81 MH861393 — France Vu et al. 2019
V. aurantiaca CBS:642.81 MH861394 — Gabon Vu et al. 2019
V. breviphysa CBS:643.81 MH873144 MH873144 Gabon Vu et al. 2019
V. breviphysa CBS:644.81 MH861396 — Gabon Vu et al. 2019
V. calami CBS:646.81 MH861398 — France Vu et al. 2019
V. calami CBS:648.81 MH861399 — France Vu et al. 2019
V. callichroa CBS:744.91 MH874000 MH874000 France Vu et al. 2019
V. cinnamomea CBS:641.84 MH861794 — Madagascar Vu et al. 2019
V. cinnamomea CBS:642.84 MH873488 MH873488 Madagascar Vu et al. 2019
V. cremea CBS:651.81 MH873147 MH873147 France Vu et al. 2019
V. daweishanensis CLZhao 17911 OP380613 — China Zou et al. 2022
V. daweishanensis CLZhao 17936 OP380614 — China Zou et al. 2022
V. dussii CBS:652.81 MH873148 MH873148 France Vu et al. 2019
V. dussii CBS:655.81 MH861405 — France Vu et al. 2019
V. ellipsospora HHB-19503 MW740328 — New Zealand Zou et al. 2022
V. fissurata CLZhao 10118 PP083288 — China Present study
V. fissurata CLZhao 10181 PP083289 — China Present study
V. fissurata CLZhao 22538 PP083290 — China Present study
V. fissurata CLZhao 4614 PP083283 — China Present study
V. fissurata CLZhao 5218 OQ025218 OR539502 China Present study
V. fissurata CLZhao 6070 PP083284 — China Present study
V. fissurata CLZhao 8171* OQ025219 OR539503 China Present study
V. fissurata CLZhao 9618 PP083285 — China Present study
V. fissurata CLZhao 9668 PP083286 — China Present study
V. fissurata CLZhao 9697 PP083287 — China Present study
V. fragilis CLZhao 16475 OP380612 — China Zou et al. 2022
V. fragilis CLZhao 2628 OP380611 — China Zou et al. 2022
V. fusispora PDD:119539 OL709443 — New Zealand Zou et al. 2022
V. gallica CBS 234.91 MH862250 MH873932 Canada Vu et al. 2019
V. gallica CBS 656.81 MH861406 MH873152 France Vu et al. 2019
V. gillesii CBS:660.81 MH873153 MH873153 Cote d’Ivoire Vu et al. 2019
V. gomezii CBS:661.81 MH873154 MH873154 France Vu et al. 2019
V. gracilispora CBS:663.81 MH861411 — Gabon Vu et al. 2019
V. gracilispora CBS:664.81 MH861412 — Gabon Vu et al. 2019
V. insolita CBS:668.81 MH861413 — France Vu et al. 2019
V. intricata CBS:673.81 MH861418 — France Vu et al. 2019
V. investiens FP-151122ITS MH971976 — USA Liu et al. 2019
V. investiens UC2023140 KP814286 — USA Rosenthal et al. 2017
V. isabellina CLZhao 22852 OR048789 OR506350 China Present study
V. isabellina CLZhao 22887 OR048788 OR506351 China Present study
V. lincangensis CLZhao 22791* OR048819 OR506348 China Present study
V. lincangensis CLZhao 22799 OR048818 OR506349 China Present study
V. malaysiana CBS:644.84 MH873490 MH873490 Singapore Vu et al. 2019
V. minispora CBS:682.81 MH861426 — France Vu et al. 2019
V. ochroleuca CBS:465.61 MH858109 — France Vu et al. 2019
V. ochroleuca JS24400 AF506485 AF506485 Norway Larsson and Larsson 2003
V. parmastoi CBS:879.84 MH861852 MH861852 Uzbekistan Vu et al. 2019
V. pectinata CBS:685.81 MH861429 — Cote d’Ivoire Vu et al. 2019
V. perplexa CBS:695.81 MH861438 — France Vu et al. 2019
V. pirispora CBS:720.86 MH862016 — France Vu et al. 2019
V. punctata CLZhao 22423 OR048813 OR539685 China Present study
V. punctata CLZhao 22439* OR048812 OR510675 China Present study
V. rhombospora CBS:743.81 MH861470 — France Vu et al. 2019
V. rosulenta CBS:743.86 MH862028 — France Vu et al. 2019
V. rugosispora CBS:697.81 MH861440 — Gabon Vu et al. 2019
V. sigmatospora CBS:748.91 MH874001 MH874001 Netherlands Vu et al. 2019
V. sinensis CLZhao 25160* OR102494 OR510678 China Present study
V. sinensis CLZhao 25161 OR102495 OR510679 China Present study
V. sphaericospora CBS:700.81 MH873185 MH873185 Gabon Vu et al. 2019
V. sphaericospora CBS:703.81 MH861446 — Gabon Vu et al. 2019
V. sphaericospora He4847 MK625592 MK625521 China Unpublished
V. trinidadensis CBS:650.84 MH873495 MH873495 Madagascar Vu et al. 2019
V. trinidadensis CBS:651.84 MH861803 — Madagascar Vu et al. 2019
V. tropica CBS 704.81 MH861447 MH873189 France Vu et al. 2019
V. vassilievae UC2022892 KP814203 — USA Unpublished
V. verrucosa CBS:706.81 MH861449 MH861449 France Vu et al. 2019
V. yaoshanensis CLZhao 20528 PP091673 — China Present study
V. yaoshanensis CLZhao 20531 PP091674 — China Present study
V. yaoshanensis CLZhao 20565 PP091675 PP091683 China Present study
V. yaoshanensis CLZhao 20605 PP091676 — China Present study
V. yaoshanensis CLZhao 20608 PP091677 — China Present study
V. yaoshanensis CLZhao 20617 PP091678 — China Present study
V. yaoshanensis CLZhao 20619 PP091679 — China Present study
V. yaoshanensis CLZhao 20624 PP091680 — China Present study
V. yaoshanensis CLZhao 20646 PP091681 — China Present study
V. yaoshanensis CLZhao 20656 PP091682 — China Present study
V. yaoshanensis CLZhao 20669 PP091666 — China Present study
V. yaoshanensis CLZhao 20677 PP091667 — China Present study
V. yaoshanensis CLZhao 20693* PP091665 PP091684 China Present study
V. yaoshanensis CLZhao 20697 PP091668 — China Present study
V. yaoshanensis CLZhao 20709 PP091669 — China Present study
V. yaoshanensis CLZhao 20713 PP091670 — China Present study
V. yaoshanensis CLZhao 20717 PP091671 — China Present study
V. yaoshanensis CLZhao 20724 PP091672 — China Present study

The sequences were aligned in MAFFT version 7 (Katoh et al. 2019) using the G-INS-i strategy. The alignment was adjusted manually using AliView version 1.27 (Larsson 2014). Sequences of Confertobasidium olivaceoalbum (Bourdot & Galzin) (AF511648) Jülich and Metulodontia nivea (P. Karst.) Parmasto () retrieved from GenBank were used as the outgroups in the ITS+LSU analysis (Fig. 1); Sequences of Peniophora incarnata (Pers.) P. Karst. (AF506425) and Peniophora nuda (Fr.) Bres. (MT859929) retrieved from GenBank were used as the outgroups in the ITS analysis (Fig. 2) (Leal-Dutra et al. 2018; Zhao et al. 2021).

Figure 1. 

Maximum parsimony strict consensus tree illustrating the phylogeny of Vararia and related genera in the family Peniophoraceae based on ITS+LSU sequences. Branches are labelled with maximum likelihood bootstrap values > 70%, parsimony bootstrap values > 50% and Bayesian posterior probabilities > 0.95, respectively.

Figure 2. 

Maximum parsimony strict consensus tree illustrating the phylogeny of the two new species and related species in Vararia, based on ITS sequences. Branches are labelled with maximum likelihood bootstrap values > 70%, parsimony bootstrap values > 50% and Bayesian posterior probabilities > 0.95, respectively.

Maximum parsimony (MP), Maximum Likelihood (ML), and Bayesian Inference (BI) analyses were applied to the combined three datasets following a previous study (Zhao and Wu 2017). All characters were equally weighted and gaps were treated as missing data. Trees were inferred using the heuristic search option with TBR branch swapping and 1,000 random sequence additions. Max-trees were set to 5,000, branches of zero length were collapsed and all parsimonious trees were saved. Clade robustness was assessed using bootstrap (BT) analysis with 1,000 pseudo replicates (Felsenstein 1985). Descriptive tree statistics – tree length (TL), composite consistency index (CI), composite retention index (RI), composite rescaled consistency index (RC) and composite homoplasy index (HI) – were calculated for each maximum parsimonious tree generated. The combined dataset was also analysed using Maximum Likelihood (ML) in RAxML-HPC2 through the CIPRES Science Gateway (Miller et al. 2012). Branch support (BS) for the ML analysis was determined by 1000 bootstrap pseudo replicates.

MrModeltest 2.3 (Nylander 2004) was used to determine the best-fit evolution model for each dataset for the purposes of Bayesian inference (BI) which was performed using MrBayes 3.2.7a with a GTR+I+G model of DNA substitution and a gamma distribution rate variation across sites (Ronquist et al. 2012). A total of four Markov chains were run for two runs from random starting trees for 1.2 million generations for ITS+LSU (Fig. 1); and 4 million generations for ITS (Fig. 2) with trees and parameters sampled every 1,000 generations. The first quarter of all the generations were discarded as burn-ins. A majority rule consensus tree was computed from the remaining trees. Branches were considered as significantly supported if they received a maximum likelihood bootstrap support value (BS) of > 70%, a maximum parsimony bootstrap support value (BT) of > 70% or a Bayesian posterior probability (BPP) of > 0.95.

Results

Molecular phylogeny

The ITS+LSU dataset (Fig. 1) comprised sequences from 45 fungal specimens representing 38 taxa. The dataset had an aligned length of 2,304 characters, of which 1,181 characters were constant, 346 were variable and parsimony-uninformative and 777 (50%) were parsimony-informative. Maximum parsimony analysis yielded 3 equally parsimonious trees (TL = 5,051, CI = 0.3985, HI = 0.6015, RI = 0.5522 and RC = 0.2201). The best model of nucleotide evolution for the ITS+LSU dataset estimated and applied in the Bayesian analysis was found to be GTR+I+G. Bayesian analysis and ML analysis resulted in a similar topology as in the MP analysis. The Bayesian analysis had an average standard deviation of split frequencies = 0.004451 (BI) and the effective sample size (ESS) across the two runs is double the average ESS (avg. ESS) = 324. The phylogram based on the ITS+LSU rDNA gene regions (Fig. 1) included eight genera within Peniophoraceae (Russulales), which were Asterostroma, Amylostereum, Baltazaria, Dichostereum, Michenera, Peniophora, Scytinostroma and Vararia, in which six new species were grouped into the genera Vararia.

The ITS dataset (Fig. 2) comprised sequences from 79 fungal specimens representing 38 taxa. The dataset had an aligned length of 849 characters, of which 199 characters were constant, 65 were variable and parsimony-uninformative and 585 (50%) were parsimony-informative. Maximum parsimony analysis yielded 1 equally parsimonious tree (TL = 4,058, CI = 0.3233, HI = 0.6767, RI = 0.7299 and RC = 0.2360). The best model of nucleotide evolution for the ITS dataset estimated and applied in the Bayesian analysis was found to be GTR+I+G. Bayesian analysis and ML analysis resulted in a similar topology as in the MP analysis. The Bayesian analysis had an average standard deviation of split frequencies = 0.001947 (BI) and the effective sample size (ESS) across the two runs is double the average ESS (avg. ESS) = 888. The phylogenetic tree (Fig. 2), inferred from the ITS sequences, highlighted that V. fissurata was the sister to V. ellipsospora G. Cunn. with strong supports. The new species V. lincangensis was clustered with V. fragilis L. Zou & C.L. Zhao. Furthermore, V. punctata was retrieved as a sister to V. ambigua Boidin, Lanq. & Gilles. Moreover, V. isabellina formed a monophyletic lineage, and it was then grouped closely with V. daweishanensis L. Zou & C.L. Zhao, and V. gracilispora Boidin & Lanq. The species V. sinensis was grouped with five taxa as Vararia breviphysa Boidin & Lanq., V. pirispora Boidin, Gilles & Lanq., V. fusispora G. Cunn., V. abortiphysa Boidin & Lanq., and V. insolita Boidin & Lanq. In addition, V. yaoshanensis was sister to V. gallica (Bourdot & Galzin) Boidin with strong supports.

Taxonomy

Vararia fissurata Y.L. Deng & C.L. Zhao, sp. nov.

MycoBank No: MB851793
Figs 3, 4

Holotype

China. Yunnan Province, Yuxi, Xinping County, the Ancient Tea Horse Road, 23°57'10"N, 101°30'41"E, altitude 2600 m a.s.l., on the trunk of angiosperm, leg. C.L. Zhao, 21 August 2018, CLZhao 8171 (SWFC).

Figure 3. 

Basidiomata of Vararia fissurata (holotype). Scale bars: 1 cm (A); 1 mm (B).

Etymology

Fissurata (Lat.): referring to the cracking hymenial surface.

Figure 4. 

Microscopic structures of Vararia fissurata (holotype) A a section of hymenium B basidiospores C basidia D basidioles E gloeocystidia subulate F gloeocystidia subglobose G dichohyphae. Scale bars: 10 µm (A–G).

Description

Basidiomata annual, resupinate, adnate, pruinose, brittle, without odor or taste when fresh, up to 12 cm long, 2.5 cm wide, and 100 µm thick. Hymenial surface smooth, white to olivaceous buff when fresh, and olivaceous buff upon drying, sparsely and deeply cracked with age. Sterile margin distinct, white, and up to 2 mm wide.

Hyphal system dimitic, generative hyphae with clamp connections, colorless, thin-walled, moderately branched, interwoven, 2–3 µm in diameter; IKI–, CB–, tissues unchanged in KOH. Dichohyphae predominate, yellowish, capillary, frequently branched, 1.5 µm in diameter, thick-walled, dichotomously to irregularly branched with main branches and acute tips, weakly to moderately dextrinoid in Melzer’s reagent, CB–, tissues unchanged in KOH; subhymenial hyphae densely covered by a lot of bulk crystals.

Gloeocystidia empty or filled with refractive flocculent matter, two types: (1) Gloeocystidia subglobose, colorless, thin-walled, smooth, 11–23 × 6–12 µm; (2) Gloeocystidia subulate, usually containing refractive materials; slightly constricted at the neck, colorless, thin-walled, smooth, 25.5–43 × 7–11 µm. Basidia cylindrical, with four sterigmata and a basal clamp connection, 20–27 × 4–8 µm; basidioles dominant, in shape similar to basidia but slightly smaller.

Basidiospores ellipsoid to broadly ellipsoid, colorless, thin-walled, smooth, IKI–, CB–, 5–10 × 3–7 µm, L = 7.37 µm, W = 5.22 µm, Q = 1.38–1.44 (n = 150/5).

Additional specimens examined

(paratypes). China. Yunnan Province, Yuxi, Xinping County, the Ancient Tea Horse Road, 23°57'10"N, 101°30'41"E, altitude 2600 m a.s.l., on fallen angiosperm branch, leg. C.L. Zhao, 13 January 2018, CLZhao 5218 (SWFC); Puer, Zhenyuan County, Heping Town, Damoshan, 23°56'21"N, 101°25'32"E, altitude 2240 m a.s.l., on fallen angiosperm branch, leg. C.L. Zhao, 16 January 2018, CLZhao 6070 (SWFC); Dali, Weishan Country, Qinghua Town, Green Peacock Nature Reserve, 25°23'35"N, 100°31'39"E, altitude 1500 m a.s.l., on the fallen branch of angiosperm, leg. C.L. Zhao, 18 July 2022, CLZhao 22538 (SWFC); Puer, Jingdong County, Wuliangshan National Nature Reserve, 24°34'45"N, 100°830'03"E, altitude 2000 m a.s.l., on fallen angiosperm branch, leg. C.L. Zhao, 6 October 2017, CLZhao 4614 (SWFC); 6 January 2019, CLZhao 9618, CLZhao 9668 and CLZhao 9697 (SWFC); Dali, Nanjian County, Lingbaoshan National Forest Park, 24°78'26"N, 100°51'30"E, altitude 2500 m a.s.l., on fallen angiosperm branch, leg. C.L. Zhao, 9 January 2019, CLZhao 10118, and CLZhao 10181 (SWFC).

Vararia isabellina Y.L. Deng & C.L. Zhao, sp. nov.

MycoBank No: MB851798
Figs 5, 6

Holotype

China. Yunnan Province, Lincang, Fengqing County, 24°67'18"N, 100°19'67"E, altitude 1660 m a.s.l., on the fallen angiosperm branch, leg. C.L. Zhao, 20 July 2022, CLZhao 22852 (SWFC).

Figure 5. 

Basidiomata of Vararia isabellina (holotype). Scale bars: 1 cm (A); 1 mm (B).

Etymology

Isabellina (Lat.): referring to the isabelline to yellowish-brown basidiomata.

Figure 6. 

Microscopic structures of Vararia isabellina (holotype) A basidiospores B basidioles C basidia D dichohyphae E gloeocystidia F a section of hymenium. Scale bars: 10 µm (A–F).

Description

Basidiomata annual, membranous, soft, and adnate, without odor or taste when fresh, up to 90 mm long, 10 mm wide, and 50–90 µm thick. Hymenial surface smooth, cream to isabelline when fresh, isabelline to slightly brown when dry. Sterile margin thinning out, cream to isabelline, and up to 1 mm wide.

Hyphal system dimitic, generative hyphae bearing simple-septa, colorless, thin to slightly thick-walled, frequently branched, 2.5–4 µm in diameter, IKI–, CB–, tissues unchanged in KOH. Dichohyphae predominant, yellowish, distinctly thick-walled, dichotomously to irregularly branched with main branches up to 4 μm in diameter and with acute tips, moderately dextrinoid in Melzer’s reagent, CB–, tissues unchanged in KOH; dichohyphae in hymenium similar to those in subiculum but more branched, with more narrow and shorter branches, with slightly curved tips and stronger.

Gloeocystidia spindle to subcylindrical, smooth, colorless, thin-walled, usually containing refractive materials, 38–47 × 8–13 μm. Basidia subcylindrical, slightly constricted at the neck, with four sterigmata and a basal simple septum connection, 33–39 × 7–9 μm; basidioles dominant, in shape similar to basidia, but slightly smaller.

Basidiospores sub-fusiform to navicular, colorless, smooth, with numerous oil-drops, thin-walled, IKI–, CB–, 9–13 × 5–8 µm, L = 11.66 µm, W = 6.69 µm, Q = 1.68–1.78 (n = 60/2).

Additional specimen examined

(paratype). China. Yunnan Province, Lincang, Fengqing County, 24°67'18"N, 100°19'67"E, altitude 1660 m a.s.l., on the fallen angiosperm branch, leg. C.L. Zhao, 20 July 2022, CLZhao 22887 (SWFC).

Vararia lincangensis Y.L. Deng & C.L. Zhao, sp. nov.

MycoBank No: MB851794
Figs 7, 8

Holotype

China. Yunnan Province, Lincang, Fengqing County, Yaojie Township, Xingyuan Village, 24°61'44"N, 100°17'21"E, altitude 1660 m a.s.l., on the fallen angiosperm branch, leg. C.L. Zhao, 20 July 2022, CLZhao 22791 (SWFC).

Figure 7. 

Basidiomata of Vararia lincangensis (holotype). Scale bars: 1 cm (A); 1 mm (B).

Etymology

Lincangensis (Lat.): referring to the locality (Lincang) of the type specimen.

Figure 8. 

Microscopic structures of Vararia lincangensis (holotype) A basidiospores B basidioles C basidia D gloeocystidia E dichohyphae F a section of hymenium. Scale bars: 10 µm (A–F).

Description

Basidiomata annual, resupinate, membranous, soft and adnate, without odor or taste when fresh, up to 90 mm long, 20 mm wide, and 70–150 µm thick. Hymenial surface smooth, white to cream when fresh, cream upon drying, cracking with age. Sterile margin distinct, narrow, whitish, attached, and up to 1 mm wide.

Hyphal system dimitic, generative hyphae bearing simple-septa, rarely branched, colorless, thin-walled, 2–3 μm in diameter, IKI–, CB–, tissues unchanged in KOH; subhymenial hyphae densely covered by some crystals. Dichohyphae predominate, white to cream, capillary, thick-walled, frequently branched, dichotomously to irregularly branched with main branches and acute tips, 1–1.5 µm diameter, weakly to moderately dextrinoid in Melzer’s reagent, CB–, tissues unchanged in KOH, subiculum composed of colorless. Skeletal hyphae colorless, thick-walled, 2–3 µm in diameter, IKI–, CB–, tissues unchanged in KOH.

Gloeocystidia subglobose, and clavate to fusiform, usually containing refractive materials, colorless, smooth, thin-walled, 6.5–16 × 3–5 µm. Basidia clavate, with four sterigmata and a basal simple septum, thin-walled, smooth, 11–17.5 × 2–4 μm; basidioles in shape similar to basidia, but slightly smaller.

Basidiospores ellipsoid, colorless, thin-walled, smooth, occasionally acyanophilous, CB–, (3–)3.5–5.5(–6) × (2–)2.5–4 µm, L = 4.18 µm, W = 3.11 µm, Q = 1.33–1.36 (n = 60/2).

Additional specimen examined

(paratype). China. Yunnan Province, Lincang, Fengqing County, Yaojie Township, Xingyuan Village, 24°61'44"N, 100°17'21"E, altitude 1660 m a.s.l., on the fallen angiosperm branch, leg. C.L. Zhao, 20 July 2022, CLZhao 22799 (SWFC).

Vararia punctata Y.L. Deng & C.L. Zhao, sp. nov.

MycoBank No: MB851795
Figs 9, 10

Holotype

China. Yunnan Province, Dali, Weishan Country, Qinghua Town, Green Peacock Nature Reserve, 25°23'35"N, 100°31'39"E, altitude 1500 m a.s.l., on the fallen branch of angiosperm, leg. C.L. Zhao, 18 July 2022, CLZhao 22439 (SWFC).

Figure 9. 

Basidiomata of Vararia punctata (holotype). Scale bars: 1 cm (A); 1 mm (B).

Etymology

Punctata (Lat.): referring to the species having cushion-shaped basidioma.

Figure 10. 

Microscopic structures of Vararia punctata (holotype) A a section of hymenium B basidia C basidioles D gloeocystidia E basidiospores F dichohyphae. Scale bars: 10 µm (A–F).

Description

Basidiomata annual, membranous, soft, adnate, without odor or taste when fresh, up to 50 mm long, 15 mm wide, and 90–150 µm thick. Hymenial surface smooth, and white to cream when fresh, cream when dry. Sterile margin thin, distinct, narrow, whitish, attached, and up to 1 mm.

Hyphal system dimitic, generative hyphae bearing simple-septa, colorless, thin to slightly thick-walled, rarely branched, interwoven, 2–3 µm in diameter, IKI–, CB–, tissues unchanged in KOH. Dichohyphae predominate, white to cream, capillary, frequently branched, thick-walled, 1 µm in diameter, dichotomously to irregularly branched with main branches and acute tips, weakly to moderately dextrinoid in Melzer’s reagent, CB–, tissues unchanged in KOH. Skeletal hyphae colorless, thick-walled, 2–3 µm in diameter, IKI–, CB–, tissues unchanged in KOH; subhymenial hyphae densely covered by bulk crystals.

Gloeocystidia clavate to cylindrical, usually containing oil droplets, colorless, smooth, thick-walled, and 12–21 × 5–9 µm. Basidia subcylindrical, with four sterigmata and a basal simple septum, 11–25 × 4–7 µm; basidioles in shape similar to basidia, but slightly smaller.

Basidiospores ellipsoid, colorless, thin-walled, smooth, with oil drops, amyloid, CB–, 6–10 × 4–6(–6.5) µm, L = 7.81 µm, W = 5.1 µm, Q = 1.51–1.56 (n = 120/4).

Additional specimen examined

(paratype). China. Yunnan Province, Dali, Weishan Country, Qinghua Town, Green Peacock Nature Reserve, 25°23'35"N, 100°31'39"E, altitude 1500 m a.s.l., on the fallen branch of angiosperm, leg. C.L. Zhao, 18 July 2022, CLZhao 22423 (SWFC).

Vararia sinensis Y.L. Deng & C.L. Zhao, sp. nov.

MycoBank No: MB851796
Figs 11, 12

Holotype

China. Yunnan Province, Lincang, Yun County, Dumu Village, 24°39'79"N, 100°18'17"E, altitude 1960 m a.s.l., on the fallen angiosperm branch, leg. C.L. Zhao, 20 October 2022, CLZhao 25160 (SWFC).

Figure 11. 

Basidiomata of Vararia sinensis (holotype). Scale bars: 1 cm (A); 1 mm (B).

Etymology

Sinensis (Lat.): referring to the locality (China) of the type specimen.

Figure 12. 

Microscopic structures of Vararia sinensis (holotype) A basidiospores B basidioles C basidia D dichohyphae E gloeocystidia subulate F a section of hymenium. Scale bars: 10 µm (A–F).

Description

Basidiomata annual, membranous, and adnate, up to 70 mm long, 35 mm wide, and 80–160 µm thick. Hymenial surface smooth, white to slightly pink when fresh, pink upon drying. Sterile margin thinning out, narrow, whitish, attached, and up to 1 mm.

Hyphal system dimitic, generative hyphae bearing simple-septa, colorless, thin-walled, branched, 2.5–3 µm diameter, IKI–, CB–, tissues unchanged in KOH. Dichohyphae predominant, yellowish, thick-walled, dichotomously to irregularly branched with main branches up to 1.4 μm in diameter and with acute tips, moderately dextrinoid in Melzer’s reagent, CB–, tissues unchanged in KOH, dichohyphae in hymenium similar to those in subiculum but more branched, with more narrow and shorter branches, with slightly curved tips and stronger, subhymenial hyphae densely covered by crystals. Skeletal hyphae rarely branched, interwoven, colorless, thick-walled, 2–3 µm in diameter, IKI–, CB–, tissues unchanged in KOH.

Gloeocystidia subulate, smooth, colorless, thin-walled, filled with refractive oil-like matter, 17–35 × 6–7 μm. Basidia clavate, with four sterigmata and a basal simple septum connection, 25–35 × 6–7 μm; basidioles dominant, in shape similar to basidia, but slightly smaller.

Basidiospores sub-fusiform to navicular, with a beaklike extension, colorless, smooth, with numerous oil-drops, thin-walled, IKI–, CB–, 6–11 × 4–6 µm, L = 8.21 µm, W = 4.88 µm, Q = 1.66–1.71 (n = 60/2).

Additional specimen examined

(paratype). China. Yunnan Province, Lincang, Yun County, Dumu Village. GPS coordinates: 24°39'79"N, 100°18'17"E, altitude 1960 m a.s.l., on the fallen angiosperm branch, leg. C.L. Zhao, 20 October 2022, CLZhao 25161 (SWFC).

Vararia yaoshanensis Y.L. Deng & C.L. Zhao, sp. nov.

MycoBank No: MB851797
Figs 13, 14

Holotype

China. Yunnan Province, Zhaotong, Qiaojia County, Yao Shan National Nature Reserve, 26°89'62"N, 102°95'04"E, altitude 2500 m a.s.l., on fallen branch of angiosperm, 21 August 2020, CLZhao 20693 (SWFC).

Figure 13. 

Basidiomata of Vararia yaoshanensis (holotype). Scale bars: 1 cm (A); 1 mm (B).

Etymology

Yaoshanensis (Lat.): referring to the provenance (Yaoshan) of the type specimen.

Figure 14. 

Microscopic structures of Vararia yaoshanensis (holotype) A basidiospores B basidia C basidioles D dichohyphae E gloeocystidia subglobose F gloeocystidia clavate to fusiform G a section of hymenium. Scale bars: 10 µm (A–G).

Description

Basidiomata annual, membranous, adnate, without odor or taste when fresh, up to 8 cm long, 4 cm wide, 80–120 µm thick. Hymenial surface smooth, cream to cinnamon-buff when fresh, pinkish buff to cinnamon-buff upon drying, cracking with age. Sterile margin thin, pinkish buff, up to 1 mm.

Hyphal system dimitic, generative hyphae bearing simple-septa, rarely branched, slightly thick-walled, 2–4 μm in diameter, IKI–, CB–, tissues unchanged in KOH. Dichohyphae, predominant, capillary, frequently branched, distinctly thick-walled, 1.6 µm diameter, dichotomously to irregularly branched with main branches and acute tips, weakly to moderately dextrinoid in Melzer’s reagent, CB–, tissues unchanged in KOH.

Gloeocystidia with two types, (1) Gloeocystidia fusiform, colorless, thick-walled, smooth, tapered or gradually elongated apex, 28.5–50 × 6–12.5 µm; (2) Gloeocystidia subglobose, usually containing refractive materials, colorless, thick-walled, smooth, 11–27 × 7–11 µm. Basidia are subclavate to subcylindrical, thin-walled, with four sterigmatas and a basal simple septum, 23–46 × 5–8 µm; basidioles dominant, in shape similar to basidia, but slightly smaller.

Basidiospores ellipsoid, colorless, slightly thick-walled, smooth, amyloid, CB–, (7.5–)7.6–10.8(–10.9) × (5.3–)5.7–7.8(–7.9) µm, L = 9.52 µm, W = 6.61 µm, Q = 1.4–1.5 (n = 210/7).

Additional specimens examined

(paratypes). China. Yunnan Province, Zhaotong, Qiaojia County, Yao Shan National Nature Reserve, 26°89'62"N, 102°95'04"E, altitude 2500 m a.s.l., on fallen branch of angiosperm, 21 August 2020, CLZhao 20669, CLZhao 20677, CLZhao 20697, CLZhao 20709, CLZhao 20713, CLZhao 20717 and CLZhao 20724 (SWFC), 22 August 2020, CLZhao 20528, CLZhao 20531, CLZhao 20565, CLZhao 20605, CLZhao 20608, CLZhao 20617, CLZhao 20619, CLZhao 20624, CLZhao 20646 and CLZhao 20656 (SWFC).

Discussion

Many recently described wood-inhabiting fungal taxa have been reported worldwide, including in the genera Vararia (Larsson 2007; Bernicchia and Gorjón 2010; Duhem and Buyck 2012; Sanyal et al. 2012; Nakasone 2015; Liu and He 2016; Leal-Dutra et al. 2018; Liu 2019; Dai et al. 2021; Zou et al. 2022; Deng and Zhao 2023; Li et al. 2023). Prior to this study, the following eleven Vararia species were reported from China, V. amphithallica Boidin, Lanq. & Gilles, V. bispora S.L. Liu & S.H. He, V. breviphysa, V. cinnamomea Boidin, Lanq. & Gilles, V. daweishanensis, V. fragilis, V. investiens, V. montana S.L. Liu & S.H. He, V. racemosa (Burt.) D.P. Rogers & H.S. Jacks., V. sphaericospora Gilb. and V. yunnanensis Y.L. Deng & C.L. Zhao (Dai 2011; Liu 2019; Dai et al. 2021; Zou et al. 2022; Deng and Zhao 2023). The present study (Figs 1, 2) reports six new species in Vararia, based on a combination of morphological features and molecular evidences.

Phylogenetically, based on the multiple loci in Scytinostroma s.s., nine genera, Asterostroma, Baltazaria, Dichostereum, Gloiothele, Lachnocladium, Michenera, Peniophora, Vesiculomyces and Vararia were divided in the family Peniophoraceae (Larsson and Larsson 2003, 2004; Larsson 2007; Leal-Dutra et al. 2018; Liu and He 2018; Zou et al. 2022; Li et al. 2023). In the present study, based on the ITS+LSU data (Fig. 1), Vararia was grouped with Asterostroma, Baltazaria, Dichostereum, and Peniophora, in which six new species were grouped into the genus Vararia. From the phylogram inferred from the ITS+LSU data (Fig. 1), the four new species V. fissurata, V. punctata, V. isabellina and V. sinensis were retrieved as a sister to V. ellipsospora, V. ambigua, V. investiens and V. breviphysa, respectively. Furthermore, the two new species Vararia lincangensis and V. yaoshanensis formed a monophyletic lineage respectively, and then V. yaoshanensis was clustered with V. ellipsospora and V. tropica. The species V. lincangensis was grouped closely with V. ambigua, V. gallica and V. punctata. However, morphologically, V. investiens can be delimited from V. isabellina by having the resupinate basidiomata with the yellowish cream to ochraceous hymenial surface, thin-walled, clamped generative hyphae, longer fusiform gloeocystidia (40–80 × 4–8 µm), longer basidia (30–50 × 4–5 µm), and smaller fusoid basidiospores measuring as 8–12 × 3–3.5 µm (Boidin and Lanquetin 1975). The taxon V. ellipsospora is different from V. yaoshanensis by having the smaller cylindrical basidia (24–30 × 5–6 µm), longer gloeocystidia (40–56 × 8–10 µm), and narrower basidiospores (8–12 × 5.5–6.5 µm; Cunningham 1955), and V. tropica is distinguished from V. yaoshanensis by its smaller subcylindrical gloeocystides (20–42 × 6.5–10 µm), and larger basidia (35–50 × 7–8.5 µm; Welden 1965). In addition, V. ambigua differs from V. lincangensis by having both larger gloeocystidia (15–32 × 3.5–7 µm), and basidiospores measuring as 6–7.3 × 3.4–5 µm (Boidin et al. 1980); V. gallica is different from V. lincangensis by its larger fusiform gloeocystidia (15–36 × 3.5–6.5 µm) and basidiospores (9–12 × 3.5–5 µm; Boidin and Lanquetin 1975; Grosse-Brauckmann and Kummer 2004).

Based on ITS topology (Fig. 2), the present study highlighted that V. fissurata was found to be the sister to V. ellipsospora with strong supports, and morphologically V. ellipsospora is different from V. fissurata by the fimbriate basidiomata, thick-walled generative hyphae, larger flexuous-cylindrical gloeocystidia (40–56 × 8–10 µm), longer basidia (24–30 × 5–6 µm), and longer oblong ellipsoid basidiospores (8–12 × 5.5–6.5 µm; Cunningham 1955). In addition, V. lincangensis was clustered with V. fragilis, but morphologically V. fragilis is distinguished from V. lincangensis by the brittle basidiomata, with a buff to ochraceous hymenial surface and elliptical to ovoid gloeocystidia, both larger subulate gloeocystidia (16.5–27 × 4–7 µm) and subcylindrical basidia (13–23.5 × 3–4.5 µm; Zou et al. 2022). Furthermore, V. punctata was retrieved as a sister to V. ambigua, but morphologically V. ambigua differs from V. punctata by its cream to buff hymenophore, and larger fusiform gloeocystidia measuring as 15–32 × 3.5–7 µm (Boidin et al. 1980). Further, V. isabellina formed a monophyletic lineage and then was grouped closely with V. daweishanensis and V. gracilispora Boidin & Lanq. However, morphologically V. daweishanensis is distinguishable from V. isabellina by its pale yellowish hymenial surface, clamped generative hyphae, and smaller gloeocystidia (9–23 × 7–10.5 µm), longer basidia measuring as 26–46 × 5–8 µm, narrower allantoid basidiospores (9–13 × 3.5–5 µm; Zou et al. 2022). Moreover, V. sinensis was grouped with five taxa: V. breviphysa, V. pirispora, V. fusispora, V. abortiphysa, and V. insolita, however, morphologically, V. breviphysa is distinguishable from V. sinensis by having light yellow to light brown basidiomata, larger subcylindrical gloeocystides (50–65 × 6–8.5 µm), larger basidia (30–38 × 5.5–7 µm), and longer fusiform basidiospores (16–20 × 4–5 µm, Boidin and Lanquetin 1975; Liu et al. 2019); the species V. pirispora is distinct from V. sinensis by its larger subcylindrical gloeocystides (40–65 × 6–8 µm), longer basidia measuring as 36–52 × 6–7 µm, larger pyriform basidiospores (10–16.5 × 5–7 µm; Boidin et al. 1987); V. fusispora can be delimited from V. sinensis by having larger cylindrical gloeocystidia (40–60 × 5–6 µm) and oval to fusiform gloeocystidia (24–60 × 6–12 µm), subclavate basidia (35–56 × 6–9 µm), and larger fusiform basidiospores measuring as 14–17 × 4–6 µm (Cunningham 1955); V. abortiphysa is distinct from V. sinensis by its plagio and subcylindrical gloeocystides measuring as 25–45 × 4.5–9 µm, and longer cylindrical basidiospores (14–17 × 2.2–2.8 µm; Boidin and Lanquetin 1975); V. insolita is distinguishable from V. sinensis by having larger gloeocystidia measuring as 60–80 × 5–8 µm, longer subcylindrical basidia (30–78 × 5.5–6.5 µm), and longer subfusiform basidiospores (12–16 × 4.2–5.75 µm; Boidin and Lanquetin 1975). Then V. yaoshanensis was found to be the sister to V. gallica (Bourdot & Galzin) Boidin with strong supports. However, morphologically, V. gallica can be delimited from V. yaoshanensis by its thin-walled generative hyphae, smaller thin-walled fusiform gloeocystidia (15–36 × 3.5–6.5 µm), and thin-walled, narrower basidiospores measuring as 9–12 × 3.5–5 µm (Boidin and Lanquetin 1975; Grosse-Brauckmann and Kummer 2004).

Based on our phylogenetic and morphological research results, 17 species have been reported from China, including newly described in the present study and other recently published papers in this country (Dai 2011; Liu and He 2016; Liu 2019; Dai et al. 2021; Zou et al. 2022; Deng and Zhao 2023). It seems that the species diversity of Vararia is rich in China. Although Vararia taxa are well studied in the present paper, the species diversity, taxonomy and phylogeny of Vararia and related genera are still unresolved. A comprehensive study on this issue is urgently needed.

A key to 17 species of Vararia s.l. in China

1 Generative hyphae with clamp connections 2
– Generative hyphae bearing simple-septa 3
2 Basidia with 2 sterigmatas 4
– Basidia with 4 sterigmatas 5
3 Present thick-walled skeletal hyphae 6
– Absent thick-walled skeletal hyphae 7
4 Subcylindrical to fusiform basidiospores measuring as (10.5–)12–17(–20) × 4.5–5.5(–6.5) µm, slightly thick-walled, subglobose gloeocystidia (15–30(–35) × 6–8(–10) µm), and subcylindrical or gradually narrower gloeocystidia (25–40(–65) × 4.5–6(–18) µm) V. amphithallica
– Fusiform to cylindrical basidiospores measuring as (16–)18–22(–14) × 6–7.2(–8) µm, thick-walled, ventricose, gloeocystidia with an apical papilla (20–40 × 9–12 µm) V. bispora
5 Thin to thick-walled generative hyphae, subcylindrical basidia (26–46 × 5–8 µm), allantoid basidiospores measuring as (8.5–) 9–13 (–14) × 3.5–5 µm, and ellipsoid to ovoid to subcylindrical gloeocystidia (9–23 × 7–10.5 µm) V. daweishanensis
– Thin-walled generative hyphae 8
6 Thin to slightly thick-walled generative hyphae, thick-walled, clavate to cylindrical gloeocystidia (12–21 × 5–9 µm), subcylindrical basidia (11–25 × 4–7 µm), and ellipsoid basidiospores (6–10 × 4–6(–6.5) µm) V. punctata
– Thin-walled generative hyphae, clavate basidia 9
7 Slightly thick-walled generative hyphae 10
– Thin-walled generative hyphae 11
8 Gloeocystidia two kinds V. fissurata
– Gloeocystidia one kinds 12
9 Ellipsoid basidiospores measuring as (3–)3.5–5.5(–6) × (2–)2.5–4 µm, subglobose, clavate to fusiform gloeocystidia (6.5–16 × 3–5 µm) V. lincangensis
– Subfusiform to navicular basidiospores (6–11 × 4–6 µm), subulate gloeocystidia (17–35 × 6–7 µm) V. sinensis
10 Slightly thick-walled, ellipsoid basidiospores measuring as (7.5–)7.6–10.8(–10.9) × (5.3–)5.7–7.8(–7.9) µm, thick-walled, fusiform gloeocystidia (28.5–50 × 6–12.5 µm), globose gloeocystidia (11–27 × 7–11 µm), subclavate to subcylindrical basidia (23–46 × 5–8 µm) V. yaoshanensis
– Thin-walled basidiospores, subcylindrical basidia 13
11 Slightly thick-walled, ellipsoid basidiospores measuring as (5.1–)5.9–11.5(–11.8) × (4.3–)4.7–8.6(–9) µm, cylindrical basidia (17.5–32 × 5–9.5 µm), thin- to slightly thick-walled, subcylindrical gloeocystidia (16.5–58.5 × 4–10 µm), fusiform gloeocystidia (18.5–43.5 × 7–9 µm), tapering gloeocystidia (27.5–42 × 5.5–9 µm) V. yunnanensis
– Thin-walled basidiospores 14
12 Basidiospores < 5 µm in diameter V. investiens
– Basidiospores > 5 µm in diameter 15
13 Broad ellipsoid to ellipsoid basidiospores measuring as 3.5–5.5(–6) × 2.5–3.5 µm, elliptical to ovoid gloeocystidia (5.8–16 × 3.5–7 µm), subulate gloeocystidia (16.5–27 × 4–7 µm) V. fragilis
– Sub-fusiform to navicular basidiospores with numerous oil-drops measuring as 9–13 × 5–8 µm, spindle to subcylindrical gloeocystidia (38–47 × 8–13 µm) V. isabellina
14 Rose to orange subfusiform basidiospores measuring as (14–)16–19(–21.5) × 4.2–6 µm, cylindrical basidia (30–53 × 6.5–7.5 µm), thick-walled, subcylindrical Gloeocystides (50–65 × 6–7(–8.5) µm) V. breviphysa
– Colorless basidiospores 16
15 Broadly ellipsoid basidiospores measuring as (11–)12–16(–17) × (7.5–)9.5–13(–14) µm, clavate basidia (70–110 × 10–16 µm), clavate gloeocystidia (50–100 × 4–9 µm) V. montana
– Spherical basidiospores measuring as 8–10 × 7.5–8.5 µm, cylindrical to clavate basidia (40–45 × 6–7.5 µm), subcylindrical to fusiform gloeocystides (48–80(–105) × 7–11(–14) µm) V. sphaericospora
16 Subcylindrical to fusiform gloeocystides (26–40 × 4.5–9 µm), cylindrical basidiospores (6–8 × 2–3 µm), cylindrical basidia (30–40 × 4–5 µm) V. racemosa
– Absent gloeocystides, oblong to subellipsoid basidiospores measuring as 9–13 × 5–7.2 µm, and subcylindrical basidia (45–65 × 8–10 µm) V. cinnamomea

Additional information

Conflict of interest

The authors have declared that no competing interests exist.

Ethical statement

No ethical statement was reported.

Funding

The research was supported by the National Natural Science Foundation of China (Project Nos. 32170004, U2102220), and the High-level Talents Program of Yunnan Province (YNQR-QNRC-2018-111), and Forestry Innovation Programs of Southwest Forestry University (Grant No: LXXK-2023Z07).

Author contributions

Conceptualization: CZ. Data curation: YD, CZ. Formal analysis: CZ, YD, SJ. Funding acquisition: CZ. Investigation: YD, CZ, SJ. Methodology: SJ, YD, CZ. Project administration: CZ. Resources: CZ, YD. Software: YD, CZ. Supervision: YD, SJ, CZ. Validation: CZ. Visualization: CZ. Writing – original draft: YD, CZ, SJ. Writing – review and editing: YD, CZ.

Author ORCIDs

Yinglian Deng https://orcid.org/0000-0002-8220-508X

Sana Jabeen https://orcid.org/0000-0001-8839-7716

Changlin Zhao https://orcid.org/0000-0002-8668-1075

Data availability

All of the data that support the findings of this study are available in the main text.

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