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Cacaoporus, a new Boletaceae genus, with two new species from Thailand
expand article infoSanthiti Vadthanarat, Saisamorn Lumyong§, Olivier Raspé|
‡ Chiang Mai University, Chiang Mai, Thailand
§ Academy of Science, The Royal Society of Thailand, Bangkok, Thailand
| Botanic Garden Meise, Meise, Belgium
¶ Fédération Wallonie-Bruxelles, Service général de l’Enseignement universitaire et de la Recherche scientifique, Bruxelles, Belgium
Open Access

Abstract

We introduce a new genus, Cacaoporus, characterised by chocolate brown to dark brown basidiomata and hymenophore, tubes not separable from the pileus context, white to off-white basal mycelium, reddening when bruised, amygdaliform to ovoid spores and dark brown spore deposit. Phylogenetic analyses of a four-gene dataset (atp6, tef1, rpb2 and cox3) with a wide selection of Boletaceae showed that the new genus is monophyletic and sister to the genera Cupreoboletus and Cyanoboletus in the Pulveroboletus group. Two new species in the genus, C. pallidicarneus and C. tenebrosus are described from northern Thailand. Full descriptions and illustrations of the new genus and species are presented. The phylogeny also confirmed the reciprocal monophyly of Neoboletus and Sutorius, which further support the separation of these two genera.

Keywords

3 new taxa, atp6, Boletales, cox3, Fungal Diversity, multigene phylogeny, Neoboletus, Pulveroboletus group, Taxonomy

Introduction

In the last decade or so, since molecular techniques and phylogenetic analyses have been used in taxonomy and systematics of the Boletaceae, many new species and genera have been described worldwide (e.g. Halling et al. 2012, 2016; Zeng et al. 2012; Arora and Frank 2014; Gelardi et al. 2014, 2015; Li et al. 2014, Zhao et al. 2014b, Zeng et al. 2014; Wu et al. 2015, 2016; Zhu et al. 2015). In Thailand, although the Boletaceae have been studied for a long time, only a few new Boletaceae species and a new genus have recently been described (Desjardin et al. 2009; Neves et al. 2012; Halling et al. 2014; Raspé et al. 2016; Vadthanarat et al. 2018). At the same time, many new species and genera have been described from southern and south-western China, an area with a climate and forests similar to Thailand (e.g. Li et al. 2011; Wu et al. 2015, 2016; Zhu et al. 2015). Similarly, a high number of new species and possibly new genera are expected to occur in Thailand (Hyde et al. 2018)

During our survey on the diversity of boletes in Thailand, several collections of brown to chocolate to dark brown boletes were obtained. Some collections bearing resemblance to Sutorius Halling, Nuhn & N.A. Fechner species, which typically have brown or reddish to purplish-brown basidiomata with reddish to purplish-brown hymenophore, reddish-brown spore deposit and narrowly ellipsoid to ellipsoid basidiospores (Halling et al. 2012). However, our chocolate brown bolete collections also showed differences, in particular in having a darker hymenophore, as well as in some microscopic characters like spore shape. We therefore performed a family-wide phylogeny, which showed that those brown to chocolate to dark brown boletes belong in a generic lineage, different from Sutorius. Consequently, we introduce the new Boletaceae genus Cacaoporus and describe two new species, C. pallidicarneus and C. tenebrosus, with full descriptions and illustrations.

Materials and method

Specimens collecting

Fresh basidiomata were collected in Chiang Mai Province, northern Thailand during the rainy season in 2013 to 2018. The specimens were photographed in situ, wrapped in aluminium foil and taken to the laboratory. After description of macroscopic characters, the specimens were dried in an electric drier at 45–50 °C. Examined specimens were deposited in the herbaria CMUB, MFLU, BKF and BR (listed in Index Herbariorum; Thiers, continuously updated).

Morphological studies

Macroscopic descriptions were made, based on detailed field notes and photos of fresh basidiomata. Colour codes were taken from Kornerup and Wanscher (1978). Macrochemical reactions (colour reactions) of pileus, pileus context, stipe, stipe context and hymenophore were determined using 10% aqueous potassium hydroxide (KOH) and 28–30% ammonium hydroxide (NH4OH). Microscopic structures were observed from dried specimens, using 5% KOH, NH4OH, Melzer’s reagent or stained with 1% ammoniacal Congo red. A minimum of 50 basidiospores, 20 basidia and 20 cystidia were randomly measured at 1000× with a calibrated ocular micrometer using an Olympus CX51 compound microscope. The notation ‘[m/n/p]’ represents the number of basidiospores “m” measured from “n” basidiomata of “p” collections. Dimensions of microscopic structures are presented in the following format: (a–)b–c–d(–e), in which “c” represents the average, “b” the 5th percentile, “d” the 95th percentile, “a” the minimum and “e” the maximum. Q, the length/width ratio, is presented in the same format. A section of the pileus surface was radially and perpendicularly cut to the surface at a point halfway between the centre and margin of the pileus. Sections of stipitipellis were taken from halfway up the stipe and longitudinally cut, perpendicularly to the surface (Hosen et al. 2013; Li et al. 2011). All microscopic features were drawn by free hand using an Olympus Camera Lucida model U−DA fitted to the microscope cited above. For scanning electron microscopy (SEM), a spore print was mounted on to an SEM stub with double-sided tape. The samples were coated with gold, then examined and photographed with a JEOL JSM–5910 LV SEM.

DNA isolation, PCR amplification and DNA sequencing

Genomic DNA was extracted from fresh tissue preserved in CTAB or about 10–15 mg of dried tissue using a CTAB isolation procedure adapted from Doyle and Doyle (1990). Portions of the genes atp6, tef1, rpb2 and cox3 were amplified by polymerase chain reaction (PCR) and sequenced by Sanger sequencing. The primer pairs ATP6-1M40F/ATP6-2M (Raspé et al. 2016), EF1-983F/EF1-2218R (Rehner and Buckley 2005) and bRPB2-6F/bRPB2-7.1R (Matheny 2005) were used to amplify atp6, tef1 and rpb2, respectively. Part of the mitochondrial gene cox3 was amplified with the newly designed primers COX3M1-F (5’-ATYGGAGCWGTAATGTWYATGC-3’) and COX3M1-R (5’-CCWACTAWTACRTGRATWCCATG-3’), using the following PCR programme: 2 min 30 s at 95 °C; 35 cycles of 25 s at 95 °C, 30 s at 48 °C, 30 s at 72 °C; 3 min at 72 °C. PCR products were purified by adding 1 U of Exonuclease I and 0.5 U FastAP Alkaline Phosphatase (Thermo Scientific, St. Leon-Rot, Germany) and incubated at 37 °C for 1 h, followed by inactivation at 80 °C for 15 min. Standard Sanger sequencing was performed in both directions by Macrogen Europe (The Netherlands) with PCR primers, except for atp6, for which universal primers M13F-pUC(-40) and M13F(-20) were used; for tef1, additional sequencing was performed with two internal primers, EF1-1577F and EF1-1567R (Rehner and Buckley 2005).

Alignment and phylogeny inference

The sequences were assembled in GENEIOUS Pro v. 6.0.6 (Biomatters) and introns were removed prior to alignment based on the amino acid sequence of previously published sequences. All sequences, including sequences from GenBank, were aligned using MAFFT (Katoh and Standley 2013) on the server accessed at http://mafft.cbrc.jp/alignment/server/.

Maximum Likelihood (ML) phylogenetic inference was performed using RAxML (Stamatakis 2006) on the CIPRES web portal (RAxML-HPC2 on XSEDE; Miller et al. 2009). The phylogenetic tree was inferred by a single analysis with three partitions (one for each gene), using the GTRCAT model with 25 categories, two Buchwaldoboletus and nine Chalciporus species from sub-family Chalciporoideae were used as outgroup since Chalciporoideae always appeared as sister to the remainder of the Boletaceae in recent phylogenetic analyses (e.g. Nuhn et al. 2013; Wu et al. 2014, 2016). Statistical support of clades was obtained with 1,000 rapid bootstrap replicates.

For Bayesian Inference (BI), the best-fit model of substitution amongst those implementable in MrBayes was estimated separately for each gene using jModeltest (Darriba et al. 2012) on the CIPRES portal, based on the Bayesian Information Criterion (BIC). The selected models were HKY+I+G for atp6 and rpb2 and GTR+I+G for cox3 and tef1. Partitioned Bayesian analysis was performed with MrBayes 3.2 (Ronquist et al. 2012) on the CIPRES portal. Two runs of five chains were run for 15,000,000 generations and sampled every 500 generations. The chain temperature was decreased to 0.02 to improve convergence. At the end of the run, the average deviation of split frequencies was 0.008147.

Results

Phylogenetic analysis

A total of 325 sequences were newly generated and deposited in GenBank (Table 1). The alignment contained 1,013 sequences from four genes (186 for atp6, 358 for tef1, 326 for rpb2, 143 for cox3) from 362 voucher specimens and was 2946 characters long (TreeBase number 23886).

The four-gene analyses retrieved the six subfamilies (Austroboletoideae, Boletoideae, Chalciporoideae, Leccinoideae, Xerocomoideae, Zangioideae) as monophyletic (Fig. 1). The genera belonging to the Pulveroboletus group of Wu et al. (2014, 2016) did not form a monophyletic group. The new genus, Cacaoporus was monophyletic (BS=100% and PP=1) within a clade containing the genera Cupreoboletus Simonini, Gelardi & Vizzini and Cyanoboletus Gelardi, Vizzini & Simonini and one undescribed taxon, Boletus p.p. sp., clade 2 (specimen voucher JD0693) with high support (BS=94% and PP=0.99). The macromorphologically most similar genus, Sutorius, formed another clade (BS=100% and PP=1) sister to Neoboletus Gelardi, Simonini & Vizzini, with 67% BS and 0.97 PP support, in another clade of the Pulveroboletus group.

Figure 1. 

Phylogenetic tree inferred from the four-gene dataset (atp6, cox3, rpb2 and tef1), including Cacaoporus species and selected Boletaceae using Maximum Likelihood and Bayesian Inference methods (ML tree is presented). The two Buchwaldoboletus and nine Chalciporus species in subfamily Chalciporoideae were used as outgroup. Most of the taxa not belonging to the Pulveroboletus group were collapsed into subfamilies. All genera clades in Pulveroboletus group that were highly supported were also collapsed. Bootstrap support values (BS ≥ 70%) and posterior probabilities (PP ≥ 0.90) are shown above the supported branches.

Our phylogeny also showed that thirteen Sutorius species including S. brunneissimus (W.F. Chiu) G. Wu & Zhu L. Yang, S. ferrugineus G. Wu, Fang Li & Zhu L. Yang, S. flavidus G. Wu & Zhu L. Yang, S. hainanensis (T.H. Li & M. Zang) G. Wu & Zhu L. Yang, S. junquilleus (Quél.) G. Wu & Zhu L. Yang, S. magnificus (W.F. Chiu) G. Wu & Zhu L. Yang, S. obscureumbrinus (Hongo) G. Wu & Zhu L. Yang, S. rubriporus G. Wu & Zhu L. Yang, S. sanguineoides G. Wu & Zhu L. Yang, S. sanguineus G. Wu & Zhu L. Yang, S. tomentulosus (M. Zang, W.P. Liu & M.R. Hu) G. Wu & Zhu L. Yang and S. venenatus (Nagas.) G. Wu & Zhu L. Yang clustered in the Neoboletus clade with high support (85% BS and 0.95 PP), while the true Sutorius, including the typus generis S. eximius (Peck) Halling, Nuhn & Osmundson, formed a different well-supported clade (BS=100% and PP=1).

List of collections used for DNA analyses, with origin, GenBank accession numbers and reference(s).

Species Voucher Origin atp6 cox3 tef1 rpb2 Reference(s)
Afroboletus aff. multijugus JD671 Burundi MH614651 MH614794 MH614700 MH614747 This study
Afroboletus costatisporus ADK4644 Togo KT823958 MH614795* KT824024 KT823991 Raspé et al. 2016; *This study
Afroboletus luteolus ADK4844 Togo MH614652 MH614796 MH614701 MH614748 This study
Aureoboletus catenarius HKAS54467 China KT990711 KT990349 Wu et al. 2016
Aureoboletus duplicatoporus HKAS50498 China KF112230 KF112754 Wu et al. 2014
Aureoboletus gentilis ADK4865 Belgium KT823961 MH614797* KT824027 KT823994 Raspé et al. 2016; *This study
Aureoboletus mirabilis HKAS57776 China KF112229 KF112743 Wu et al. 2014
Aureoboletus moravicus VDKO1120 Belgium MG212528 MH614798* MG212573 MG212615 Vadthanarat et al. 2018; *This study
Aureoboletus nephrosporus HKAS67931 China KT990720 KT990357 Wu et al. 2016
Aureoboletus projectellus AFTOL-ID-713 USA DQ534604* AY879116 AY787218 *Binder and Hibbett 2006; Binder et al., Unpublished
Aureoboletus shichianus HKAS76852 China KF112237 KF112756 Wu et al. 2014
Aureoboletus sp. HKAS56317 China KF112239 KF112753 Wu et al. 2014
Aureoboletus sp. OR0245 China MH614653 MH614799 MH614702 MH614749 This study
Aureoboletus sp. OR0369 Thailand MH614654 MH614800 MH614703 MH614750 This study
Aureoboletus thibetanus HKAS76655 China KF112236 KF112752 Wu et al. 2014
Aureoboletus thibetanus AFTOL-ID-450 China DQ534600* DQ029199 DQ366279 *Binder and Hibbett 2006; Unpublished
Aureoboletus tomentosus HKAS80485 China KT990715 KT990353 Wu et al. 2016
Aureoboletus viscosus OR0361 Thailand MH614655 MH614801 MH614704 MH614751 This study
Aureoboletus zangii HKAS74766 China KT990726 KT990363 Wu et al. 2016
Austroboletus cf. dictyotus OR0045 Thailand KT823966 MH614802* KT824032 KT823999 Raspé et al. 2016; *This study
Austroboletus cf. subvirens OR0573 Thailand MH614656 MH614803 MH614705 MH614752 This study
Austroboletus eburneus REH9487 Australia JX889708 Halling et al. 2012b
Austroboletus olivaceoglutinosus HKAS57756 China KF112212 KF112764 Wu et al. 2014
Austroboletus sp. HKAS59624 China KF112217 KF112765 Wu et al. 2014
Austroboletus sp. OR0891 Thailand MH614657 MH614804 MH614706 MH614753 This study
Baorangia major OR0209 Thailand MG897421 MK372295* MG897431 MG897441 Phookamsak et al. 2019; *This study
Baorangia pseudocalopus HKAS63607 China KF112167 KF112677 Wu et al. 2014
Baorangia pseudocalopus HKAS75739 China KJ184570 KM605179 Wu et al. 2015
Baorangia pseudocalopus HKAS75081 China KF112168 KF112678 Wu et al. 2014
Baorangia rufomaculata BOTH4144 USA MG897415 MH614805* MG897425 MG897435 Phookamsak et al. 2019; *This study
Boletellus ananas NY815459 Costa Rica KF112308 KF112760 Wu et al. 2014
Boletellus ananas K(M)123769 Belize MH614658 MH614807 MH614707 MH614754 This study
Boletellus aff. emodensis OR0061 Thailand KT823970 MH614806* KT824036 KT824003 Raspé et al. 2016; *This study
Boletellus sp. HKAS59536 China KF112306 KF112758 Wu et al. 2014
Boletellus sp. OR0621 Thailand MG212529 MH614808* MG212574 MG212616 Vadthanarat et al. 2018; *This study
Boletus aereus VDKO1055 Belgium MG212530 MH614809* MG212575 MG212617 Vadthanarat et al. 2018; *This study
Boletus albobrunnescens OR0131 Thailand KT823973 MH614810* KT824039 KT824006 Raspé et al. 2016; *This study
Boletus botryoides HKAS53403 China KT990738 KT990375 Wu et al. 2016
Boletus edulis HMJAU4637 Russia KF112202 KF112704 Wu et al. 2014
Boletus edulis VDKO0869 Belgium MG212531 MH614811* MG212576 MG212618 Vadthanarat et al. 2018; *This study
Boletus p.p. sp JD0693 Burundi MH645583 MH645591 MH645599 This study
Boletus p.p. sp. OR0832 Thailand MH645584 MH645605 MH645592 MH645600 This study
Boletus p.p. sp. OR1002 Thailand MH645585 MH645606 MH645593 MH645601 This study
Boletus pallidus BOTH4356 USA MH614659 MH614812 MH614708 This study
Boletus pallidus TDB-1231-Bruns AF002142 AF002154 Kretzer and Bruns 1999
Boletus reticuloceps HKAS57671 China KF112201 KF112703 Wu et al. 2014
Boletus s.s. sp. OR0446 China MG212532 MH614813* MG212577 KF112703 Vadthanarat et al. 2018; *This study
Boletus sp. HKAS59660 China KF112153 KF112664 Wu et al. 2014
Boletus sp. HKAS63598 China KF112152 KF112663 Wu et al. 2014
Boletus violaceofuscus HKAS62900 China KF112219 KF112762 Wu et al. 2014
Borofutus dhakanus HKAS73789 Bangladesh JQ928576 JQ928597 Hosen et al. 2013
Borofutus dhakanus OR0345 Thailand MH614660 MH614814 MH614709 MH614755 This study
Buchwaldoboletus lignicola HKAS76674 China KF112277 KF112819 Wu et al. 2014
Buchwaldoboletus lignicola VDKO1140 Belgium MH614661 MH614815 MH614710 MH614756 This study
Butyriboletus appendiculatus VDKO0193b Belgium MG212537 MH614816* MG212582 MG212624 Vadthanarat et al. 2018; *This study
Butyriboletus cf. roseoflavus OR0230 China KT823974 MH614819* KT824040 KT824007 Raspé et al. 2016; *This study
Butyriboletus frostii NY815462 USA KF112164 KF112675 Wu et al. 2014
Butyriboletus pseudoregius VDKO0925 Belgium MG212538 MH614817* MG212583 MG212625 Vadthanarat et al. 2018; *This study
Butyriboletus pseudospeciosus HKAS63513 China KT990743 KT990380 Wu et al. 2016
Butyriboletus roseoflavus HKAS54099 China KF739779 KF739703 Wu et al. 2014
Butyriboletus roseopurpureus BOTH4497 USA MG897418 MH614818* MG897428 MG897438 Phookamsak et al., 2019; *This study
Butyriboletus sp. HKAS52661 China KF112169 KF112676 Wu et al. 2014
Butyriboletus sp. HKAS52525 China KF112163 KF112671 Wu et al. 2014
Butyriboletus sp. HKAS57774 China KF112155 KF112670 Wu et al. 2014
Butyriboletus sp. HKAS59814 China KF112199 KF112699 Wu et al. 2014
Butyriboletus sp. HKAS63528 China KF112156 KF112673 Wu et al. 2014
Butyriboletus sp. MHHNU7456 China KT990741 KT990378 Wu et al. 2016
Butyriboletus subsplendidus HKAS50444 China KT990742 KT990379 Wu et al. 2016
Butyriboletus yicibus HKAS55413 China KF112157 KF112674 Wu et al. 2014
Cacaoporus pallidicarneus OR0681 Thailand MK372259 MK372296 MK372283 This study
Cacaoporus pallidicarneus OR0683 Thailand MK372260 MK372297 MK372284 This study
Cacaoporus pallidicarneus OR1306 Thailand MK372261 MK372298 MK372272 MK372285 This study
Cacaoporus pallidicarneus SV0221 Thailand MK372262 MK372299 MK372273 MK372286 This study
Cacaoporus pallidicarneus SV0451 Thailand MK372263 MK372300 MK372274 MK372287 This study
Cacaoporus sp. SV0402 Thailand MK372270 MK372281 MK372293 This study
Cacaoporus tenebrosus OR0654 Thailand MK372264 MK372301 MK372275 MK372288 This study
Cacaoporus tenebrosus OR1435 Thailand MK372265 MK372302 MK372276 MK372289 This study
Cacaoporus tenebrosus SV0223 Thailand MK372266 MK372303 MK372277 MK372290 This study
Cacaoporus tenebrosus SV0224 Thailand MK372267 MK372304 MK372278 MK372291 This study
Cacaoporus tenebrosus SV0422 Thailand MK372268 MK372305 MK372279 This study
Cacaoporus tenebrosus SV0452 Thailand MK372269 MK372306 MK372280 MK372292 This study
Caloboletus aff. calopus HKAS74739 China KF112166 KF112667 Wu et al. 2014
Caloboletus calopus ADK4087 Belgium MG212539 MH614820 KJ184566 KP055030 Vadthanarat et al. 2018; Zhao et al. 2014a, b; This study
Caloboletus inedulis BOTH3963 USA MG897414 MH614821* MG897424 MG897434 Phookamsak et al. 2019; *This study
Caloboletus panniformis HKAS55444 China KF112165 KF112666 Wu et al. 2014
Caloboletus radicans VDKO1187 Belgium MG212540 MH614822* MG212584 MG212626 Vadthanarat et al. 2018; *This study
Caloboletus sp. HKAS53353 China KF112188 KF112668 Wu et al. 2014
Caloboletus sp. OR0068 Thailand MH614662 MH614823 MH614711 MH614757 This study
Caloboletus yunnanensis HKAS69214 China KJ184568 KT990396 Zhao et al. 2014a; Wu et al. 2016
Chalciporus aff. piperatus OR0586 Thailand KT823976 MH614824* KT824042 KT824009 Raspé et al. 2016; *This study
Chalciporus aff. rubinus OR0139 China MH614663 MH614712 MH614758 This study
Chalciporus africanus JD517 Cameroon KT823963 MH614825* KT824029 KT823996 Raspé et al. 2016; *This study
Chalciporus piperatus VDKO1063 Belgium MH614664 MH614826 MH614713 MH614759 This study
Chalciporus rubinus AF2835 Belgium KT823962 KT824028 KT823995 Raspé et al. 2016
Chalciporus sp. HKAS53400 China KF112279 KF112821 Wu et al. 2014
Chalciporus sp. HKAS74779 China KF112278 KF112820 Wu et al. 2014
Chalciporus sp. OR0363 Thailand MH645586 MH645607 MH645594 MH645602 This study
Chalciporus sp. OR0373 Thailand MH645587 MH645608 MH645595 MH645603 This study
Chiua sp. OR0141 China MH614665 MH614827 MH614714 MH614760 This study
Chiua virens OR0266 China MG212541 MH614828* MG212585 MG212627 Vadthanarat et al. 2018; *This study
Chiua viridula HKAS74928 China KF112273 KF112794 Wu et al. 2014
Crocinoboletus cf. laetissimus OR0576 Thailand KT823975 MH614833* KT824041 KT824008 Raspé et al. 2016; *This study
Crocinoboletus rufoaureus HKAS53424 China KF112206 KF112710 Wu et al. 2014
Cupreoboletus poikilochromus GS10070 Italy KT157072 KT157068 Gelardi et al. 2015
Cupreoboletus poikilochromus GS11008 Italy KT157071 KT157067 Gelardi et al. 2015
Cyanoboletus brunneoruber HKAS80579_1 China KT990763 KT990401 Wu et al. 2016
Cyanoboletus brunneoruber OR0233 China MG212542 MH614834* MG212586 MG212628 Vadthanarat et al. 2018; *This study
Cyanoboletus instabilis HKAS59554 China KF112186 KF112698 Wu et al. 2014
Cyanoboletus pulverulentus RW109 Belgium KT823980 MH614835* KT824046 KT824013 Raspé et al. 2016; *This study
Cyanoboletus sinopulverulentus HKAS59609 China KF112193 KF112700 Wu et al. 2014
Cyanoboletus sp. HKAS52639 China KF112195 KF112701 Wu et al. 2014
Cyanoboletus sp. HKAS76850 China KF112187 KF112697 Wu et al. 2014
Cyanoboletus sp. OR0257 China MG212543 MH614836* MG212587 MG212629 Vadthanarat et al. 2018; *This study
Cyanoboletus sp. HKAS90208_1 China KT990766 KT990404 Wu et al. 2016
Cyanoboletus sp. OR0322 Thailand MH614673 MH614837 MH614722 MH614768 This study
Cyanoboletus sp. OR0491 China MH614674 MH614838 MH614723 MH614769 This study
Cyanoboletus sp. OR0961 Thailand MH614675 MH614839 MH614724 MH614770 This study
Fistulinella prunicolor REH9880 Australia MH614676 MH614840 MH614725 MH614771 This study
Gymnogaster boletoides NY01194009 Australia KT990768 KT990406 Wu et al. 2016
Harrya atriceps REH7403 Costa Rica JX889702 Halling et al. 2012b
Harrya chromapes HKAS50527 China KF112270 KF112792 Wu et al. 2014
Harrya moniliformis HKAS49627 China KT990881 KT990500 Wu et al. 2016
Heimioporus cf. mandarinus OR0661 Thailand MG212545 MH614841* MG212589 MG212631 Vadthanarat et al. 2018; *This study
Heimioporus japonicus OR0114 Thailand KT823971 MH614842* KT824037 KT824004 Raspé et al. 2016; *This study
Heimioporus retisporus HKAS52237 China KF112228 KF112806 Wu et al. 2014
Heimioporus sp. OR0218 Thailand MG212546 MG212590 MG212632 Vadthanarat et al. 2018
Hemileccinum depilatum AF2845 Belgium MG212547 MH614843* MG212591 MG212633 Vadthanarat et al. 2018; *This study
Hemileccinum impolitum ADK4078 Belgium MG212548 MH614844* MG212592 MG212634 Vadthanarat et al. 2018; *This study
Hemileccinum indecorum OR0863 Thailand MH614677 MH614845 MH614726 MH614772 This study
Hemileccinum rugosum HKAS84970 China KT990773 KT990412 Wu et al. 2016
Hortiboletus amygdalinus HKAS54166 China KT990777 KT990416 Wu et al. 2016
Hortiboletus rubellus VDKO0403 Belgium MH614679 MH614847 MH614774 This study
Hortiboletus sp. HKAS50466 China KF112183 KF112694 Wu et al. 2014
Hortiboletus sp. HKAS51239 China KF112184 KF112695 Wu et al. 2014
Hortiboletus sp. HKAS51292 China KF112181 KF112692 Wu et al. 2014
Hortiboletus sp. HKAS76673 China KF112182 KF112693 Wu et al. 2014
Hortiboletus subpaludosus HKAS59608 China KF112185 KF112696 Wu et al. 2014
Hourangia cf. pumila OR0762 Thailand MH614680 MH614848 MH614728 MH614775 This study
Hourangia cheoi HKAS74744 China KF112285 KF112772 Wu et al. 2014
Hourangia cheoi Zhu108 China KP136979 KP136928 Zhu et al. 2015
Hourangia nigropunctata HKAS 57427 China KP136927 KP136978 Zhu et al. 2015
Hymenoboletus luteopurpureus HKAS46334 China KF112271 KF112795 Wu et al. 2014
Imleria badia VDKO0709 Belgium KT823983 MH614849* KT824049 KT824016 Raspé et al. 2016; *This study
Imleria obscurebrunnea OR0263 China MH614681 MH614850 MH614729 MH614776 This study
Imleria subalpina HKAS74712 China KF112189 KF112706 Wu et al. 2014
Lanmaoa angustispora HKAS74759 China KM605155 KM605178 Wu et al. 2015
Lanmaoa angustispora HKAS74765 China KF112159 KF112680 Wu et al. 2014
Lanmaoa angustispora HKAS74752 China KM605154 KM605177 Wu et al. 2015
Lanmaoa asiatica HKAS54094 China KF112161 KF112682 Wu et al. 2014
Lanmaoa asiatica HKAS63516 China KT990780 KT990419 Wu et al. 2016
Lanmaoa asiatica OR0228 China MH614682 MH614851 MH614730 MH614777 This study
Lanmaoa carminipes BOTH4591 USA MG897419 MH614852* MG897429 MG897439 Phookamsak et al. 2019, *This study
Lanmaoa flavorubra NY775777 Costa Rica KF112160 KF112681 Wu et al. 2014
Lanmaoa pallidorosea BOTH4432 USA MG897417 MH614853* MG897427 MG897437 Phookamsak et al. 2019, *This study
Lanmaoa sp. HKAS52518 China KF112162 KF112683 Wu et al. 2014
Lanmaoa sp. OR0130 Thailand MH614683 MH614854 MH614731 MH614778 This study
Lanmaoa sp. OR0370 Thailand MH614684 MH614855 MH614732 MH614779 This study
Leccinellum aff. crocipodium HKAS76658 China KF112252 KF112728 Wu et al. 2014
Leccinellum aff. griseum KPM-NC-0017832 Japan KC552164 JN378450* unpublished, *Orihara et al. 2012
Leccinellum corsicum Buf4507 USA KF030435 Nuhn et al. 2013
Leccinellum cremeum HKAS90639 China KT990781 KT990420 Wu et al. 2016
Leccinellum crocipodium VDKO1006 Belgium KT823988 MH614856* KT824054 KT824021 Raspé et al. 2016; *This study
Leccinellum sp. KPM-NC-0018041 Japan KC552165 KC552094 Orihara et al. 2016
Leccinellum sp. OR0711 Thailand MH614685 MH614733 MH614780 This study
Leccinum monticola HKAS76669 China KF112249 KF112723 Wu et al. 2014
Leccinum quercinum HKAS63502 China KF112250 KF112724 Wu et al. 2014
Leccinum scabrum RW105a Belgium KT823979 MH614857* KT824045 KT824012 Raspé et al. 2016; *This study
Leccinum scabrum VDKO0938 Belgium MG212549 MH614858* MG212593 MG212635 Vadthanarat et al. 2018; *This study
Leccinum scabrum KPM-NC-0017840 Scotland KC552170 JN378455 Orihara et al. 2016, 2012
Leccinum schistophilum VDKO1128 Belgium KT823989 MH614859* KT824055 KT824022 Raspé et al. 2016; *This study
Leccinum variicolor VDKO0844 Belgium MG212550 MH614860* MG212594 MG212636 Vadthanarat et al. 2018; *This study
Mucilopilus castaneiceps HKAS75045 China KF112211 KF112735 Wu et al. 2014
Neoboletus brunneissimus HKAS50538 China KM605150 KM605173 Wu et al. 2015
Neoboletus brunneissimus HKAS52660 China KF112143 KF112650 Wu et al. 2014
Neoboletus brunneissimus HKAS57451 China KM605149 KM605172 Wu et al. 2015
Neoboletus brunneissimus OR0249 China MG212551 MH614861* MG212595 MG212637 Vadthanarat et al. 2018; *This study
Neoboletus erythropus VDKO0690 Belgium KT823982 MH614864* KT824048 KT824015 Raspé et al. 2016; *This study
Neoboletus ferrugineus HKAS77718 China KT990789 KT990431 Wu et al. 2016
Neoboletus ferrugineus HKAS77617 China KT990788 KT990430 Wu et al. 2016
Neoboletus flavidus HKAS59443 China KU974136 KU974144 Wu et al. 2016
Neoboletus flavidus HKAS58724 China KU974137 KU974145 Wu et al. 2016
Neoboletus hainanensis HKAS63515 China KT990808 KT990449 Wu et al. 2016
Neoboletus hainanensis HKAS74880 China KT990790 KT990432 Wu et al. 2016
Neoboletus hainanensis HKAS90209 China KT990809 KT990450 Wu et al. 2016
Neoboletus hainanensis HKAS59469 China KF112175 KF112669 Wu et al. 2014
Neoboletus junquilleus AF2922 France MG212552 MH614862* MG212596 MG212638 Vadthanarat et al. 2018; *This study
Neoboletus magnificus HKAS54096 China KF112149 KF112654 Wu et al. 2014
Neoboletus magnificus HKAS74939 China KF112148 KF112653 Wu et al. 2014
Neoboletus multipunctatus HKAS76851 China KF112144 KF112651 Wu et al. 2014
Neoboletus multipunctatus OR0128 Thailand MH614686 MH614863 MH614734 MH614781 This study
Neoboletus obscureumbrinus OR0553 Thailand MK372271 MK372282 MK372294 This study
Neoboletus obscureumbrinus HKAS63498 China KT990791 KT990433 Wu et al. 2016
Neoboletus obscureumbrinus HKAS77774 China KT990792 KT990434 Wu et al. 2016
Neoboletus obscureumbrinus HKAS89014 China KT990793 KT990435 Wu et al. 2016
Neoboletus obscureumbrinus HKAS89027 China KT990794 KT990436 Wu et al. 2016
Neoboletus rubriporus HKAS57512 China KF112151 KF112656 Wu et al. 2014
Neoboletus rubriporus HKAS83026 China KT990795 KT990437 Wu et al. 2016
Neoboletus sanguineoides HKAS57766 China KT990799 KT990440 Wu et al. 2016
Neoboletus sanguineoides HKAS74733 China KT990800 KT990441 Wu et al. 2016
Neoboletus sanguineoides HKAS55440 China KF112145 KF112652 Wu et al. 2014
Neoboletus sanguineus HKAS80823 China KT990802 KT990442 Wu et al. 2016
Neoboletus tomentulosus HKAS77656 China KT990806 KT990446 Wu et al. 2016
Neoboletus tomentulosus HKAS53369 China KF112154 KF112659 Wu et al. 2014
Neoboletus venenatus HKAS57489 China KF112158 KF112665 Wu et al. 2014
Neoboletus venenatus HKAS63535 China KT990807 KT990448 Wu et al. 2016
Neoboletus sp. HKAS76660 China KF112180 KF112731 Wu et al. 2014
Octaviania asahimontana KPM-NC-17824 Japan KC552154 JN378430 Orihara et al. 2016, 2012
Octaviania asterosperma AQUI3899 Italy KC552159 KC552093 Orihara et al. 2016
Octaviania celatifilia KPM-NC-17776 Japan KC552147 JN378416 Orihara et al. 2016, 2012
Octaviania cyanescens PNW-FUNGI-5603 USA KC552160 JN378438 Orihara et al. 2016, 2012
Octaviania decimae KPM-NC17763 Japan KC552145 JN378409 Orihara et al. 2016, 2012
Octaviania tasmanica MEL2128484 Australia KC552157 JN378437 Orihara et al. 2016, 2012
Octaviania tasmanica MEL2341996 Australia KC552156 JN378436 Orihara et al. 2016, 2012
Octaviania zelleri MES270 USA KC552161 JN378440 Orihara et al. 2016, 2012
Parvixerocomus pseudoaokii OR0155 China MG212553 MH614865 MG212597 MG212639 This study
Phylloporus bellus OR0473 China MH580778 MH614866* MH580798 MH580818 Chuankid et al. 2019; *This study
Phylloporus brunneiceps OR0050 Thailand KT823968 MH614867* KT824034 KT824001 Raspé et al. 2016; *This study
Phylloporus castanopsidis OR0052 Thailand KT823969 MH614868* KT824035 KT824002 Raspé et al. 2016; *This study
Phylloporus imbricatus HKAS68642 China KF112299 KF112786 Wu et al. 2014
Phylloporus luxiensis HKAS75077 China KF112298 KF112785 Wu et al. 2014
Phylloporus maculatus OR0285 China MH580780 MH580800 MH580820 Chuankid et al. 2019
Phylloporus pelletieri WU18746 Austria MH580781 MH614869* MH580801 MH580821 Chuankid et al. 2019; *This study
Phylloporus pusillus OR1158 Thailand MH580783 MH614870* MH580803 MH580823 Chuankid et al. 2019; *This study
Phylloporus rhodoxanthus WU17978 USA MH580785 MH614871* MH580805 MH580824 Chuankid et al. 2019; *This study
Phylloporus rubeolus OR0251 China MH580786 MH614872* MH580806 MH580825 Chuankid et al. 2019; *This study
Phylloporus rubiginosus OR0169 China MH580788 MH614873* MH580808 MH580827 Chuankid et al. 2019; *This study
Phylloporus sp. OR0896 Thailand MH580790 MH614874* MH580810 MH580829 Chuankid et al. 2019; *This study
Phylloporus subbacillisporus OR0436 China MH580792 MH614875* MH580812 MH580831 Chuankid et al. 2019; *This study
Phylloporus subrubeolus BC022 Thailand MH580793 MH614876* MH580813 MH580832 Chuankid et al. 2019; *This study
Phylloporus yunnanensis OR0448 China MG212554 MH614877* MG212598 MG212640 Vadthanarat et al. 2018; *This study
Porphyrellus castaneus OR0241 China MG212555 MH614878* MG212599 MG212641 Vadthanarat et al. 2018; *This study
Porphyrellus cf. nigropurpureus ADK3733 Benin MH614687 MH614879 MH614735 MH614782 This study
Porphyrellus nigropurpureus HKAS74938 China KF112246 KF112763 Wu et al. 2014
Porphyrellus porphyrosporus MB97 023 Germany DQ534609 GU187734 GU187800 Binder and Hibbett 2006; Binder et al. 2010
Porphyrellus sp. HKAS53366 China KF112241 KF112716 Wu et al. 2014
Porphyrellus sp. JD659 Burundi MH614688 MH614880 MH614736 MH614783 This study
Porphyrellus sp. OR0222 Thailand MH614689 MH614881 MH614737 MH614784 This study
Pulveroboletus aff. ravenelii HKAS50203 China KT990810 KT990451 Wu et al. 2016
Pulveroboletus aff. ravenelii ADK4360 Togo KT823957 MH614882* KT824023 KT823990 Raspé et al. 2016; *This study
Pulveroboletus aff. ravenelii ADK4650 Togo KT823959 MH614883* KT824025 KT823992 Raspé et al. 2016; *This study
Pulveroboletus aff. ravenelii HKAS53351 China KF112261 KF112712 Wu et al. 2014
Pulveroboletus brunneopunctatus HKAS52615 China KT990813 KT990454 Wu et al. 2016
Pulveroboletus brunneopunctatus HKAS55369 China KT990814 KT990455 Wu et al. 2016
Pulveroboletus brunneopunctatus HKAS74926 China KT990815 KT990456 Wu et al. 2016
Pulveroboletus fragrans OR0673 Thailand KT823977 MH614884* KT824043 KT824010 Raspé et al. 2016; *This study
Pulveroboletus macrosporus HKAS57628 China KT990812 KT990453 Wu et al. 2016
Pulveroboletus ravenelii REH2565 USA KU665635 MH614885* KU665636 KU665637 Raspé et al. 2016; *This study
Pulveroboletus sp. HKAS74933 China KF112262 KF112713 Wu et al. 2014
Pulveroboletus sp. HKAS57665 China KF112264 KF112715 Wu et al. 2014
Retiboletus aff. nigerrimus OR0049 Thailand KT823967 MH614886* KT824033 KT824000 Raspé et al. 2016; *This study
Retiboletus brunneolus HKAS52680 China KF112179 KF112690 Wu et al. 2014
Retiboletus fuscus HKAS59460 China JQ928580 JQ928601 Hosen et al. 2013
Retiboletus fuscus OR0231 China MG212556 MH614887* MG212600 MG212642 Vadthanarat et al. 2018; *This study
Retiboletus fuscus HKAS63624 China KT990829 KT990466 Wu et al. 2016
Retiboletus fuscus HKAS74756 China KT990830 KT990467 Wu et al. 2016
Retiboletus griseus MB03 079 USA KT823964 MH614888* KT824030 KT823997 Raspé et al. 2016; *This study
Retiboletus griseus HKAS63590 China KF112178 KF112691 Wu et al. 2014
Retiboletus kauffmanii OR0278 China MG212557 MH614889* MG212601 MG212643 Vadthanarat et al. 2018; *This study
Retiboletus nigerrimus HKAS53418 China KT990824 KT990462 Wu et al. 2016
Retiboletus sinensis HKAS59832 China KT990827 KT990464 Wu et al. 2016
Retiboletus zhangfeii HKAS59699 China JQ928582 JQ928603 Hosen et al. 2013
Rhodactina himalayensis CMU25117 Thailand MG212558 MG212602, MG212603 Vadthanarat et al. 2018
Rhodactina rostratispora SV170 Thailand MG212560 MG212605 MG212645 Vadthanarat et al. 2018
Rossbeevera cryptocyanea KPM-NC17843 Japan KT581441 KC552072 Orihara et al. 2016
Rossbeevera eucyanea TNS-F-36986 Japan KC552115 KC552068 Orihara et al. 2016
Rossbeevera griseovelutina TNS-F-36989 Japan KC552124 KC552076 Orihara et al. 2016
Rossbeevera pachydermis KPM-NC23336 New Zealand KJ001064 KP222912 Orihara et al. 2016
Rossbeevera vittatispora OSC61484 Australia KC552109 JN378446 Orihara et al. 2016, 2012
Royoungia reticulata HKAS52253 China KT990786 KT990427 Wu et al. 2016
Royoungia rubina HKAS53379 China KF112274 KF112796 Wu et al. 2014
Rubroboletus latisporus HKAS80358 China KP055020 KP055029 Zhao et al. 2014b
Rubroboletus legaliae VDKO0936 Belgium KT823985 MH614890* KT824051 KT824018 Raspé et al. 2016; *This study
Rubroboletus rhodosanguineus BOTH4263 USA MG897416 MH614891* MG897426 MG897436 Phookamsak et al. 2019, *This study
Rubroboletus rhodoxanthus HKAS84879 Germany KT990831 KT990468 Wu et al. 2016
Rubroboletus satanas VDKO0968 Belgium KT823986 MH614892* KT824052 KT824019 Raspé et al. 2016; *This study
Rubroboletus sinicus HKAS68620 China KF112146 KF112661 Wu et al. 2014
Rubroboletus sinicus HKAS56304 China KJ619483 KP055031 Zhao et al. 2014a; Zhao et al. 2014b
Rubroboletus sp. HKAS68679 China KF112147 KF112662 Wu et al. 2014
Rugiboletus brunneiporus HKAS68586 China KF112197 KF112719 Wu et al. 2014
Rugiboletus brunneiporus HKAS83009 China KM605146 KM605169 Wu et al. 2015
Rugiboletus brunneiporus HKAS83209 China KM605144 KM605168 Wu et al. 2015
Rugiboletus extremiorientalis HKAS76663 China KM605147 KM605170 Wu et al. 2015
Rugiboletus extremiorientalis OR0406 Thailand MG212562 MH614893* MG212607 MG212647 Vadthanarat et al. 2018; *This study
Rugiboletus sp. HKAS55373 China KF112303 KF112804 Wu et al. 2014
Singerocomus inundabilis TWH9199 Guyana MH645588 MH645609 MH645596 LC043089* *Henkel et al. 2016; This study
Singerocomus rubriflavus TWH9585 Guyana MH645589 MH645610 MH645597 This study
Spongiforma thailandica DED7873 Thailand MG212563 MH614894** KF030436* MG212648 *Nuhn et al. 2013; Vadthanarat et al. 2018; **This study
Strobilomyces atrosquamosus HKAS55368 China KT990839 KT990476 Wu et al. 2016
Strobilomyces echinocephalus OR0243 China MG212564 MG212608 MG212649 Vadthanarat et al. 2018
Strobilomyces mirandus OR0115 Thailand KT823972 MH614896* KT824038 KT824005 Raspé et al. 2016; *This study
Strobilomyces strobilaceus MB03 102 USA DQ534607* AY883428 AY786065 *Binder and Hibbett 2006, Unpublished
Strobilomyces strobilaceus RW103 Belgium KT823978 MH614895* KT824044 KT824011 Raspé et al. 2016; *This study
Strobilomyces verruculosus HKAS55389 China KF112259 KF112813 Wu et al. 2014
Strobilomyces sp. OR0259 China MG212565 MH614897* MG212609 MG212650 Vadthanarat et al. 2018; *This study
Strobilomyces sp. OR0319 Thailand MH614690 MH614898 MH614738 MH614785 This study
Strobilomyces sp. OR0778 Thailand MG212566 MH614899* MG212610 MG212651 Vadthanarat et al. 2018; *This study
Strobilomyces sp. OR1092 Thailand MH614691 MH614900 MH614739 MH614786 This study
Suillellus amygdalinus 112605ba USA JQ327024 Halling et al. 2012a
Suillellus luridus VDKO0241b Belgium KT823981 MH614901* KT824047 KT824014 Raspé et al. 2016; *This study
Suillellus queletii VDKO1185 Belgium MH645590 MH645611 MH645598 MH645604 This study
Suillellus subamygdalinus HKAS57262 China KF112174 KF112660 Wu et al. 2014
Suillellus subamygdalinus HKAS53641 China KT990841 KT990478 Wu et al. 2016
Suillellus subamygdalinus HKAS74745 China KT990843 KT990479 Wu et al. 2016
Sutorius aff. eximius HKAS52672 China KF112207 KF112802 Wu et al. 2014
Sutorius aff. eximius HKAS56291 China KF112208 KF112803 Wu et al. 2014
Sutorius australiensis REH9441 Australia MG212567 MK386576** JQ327032* MG212652 *Halling et al. 2012a; Vadthanarat et al. 2018; **This study
Sutorius eximius HKAS59657 China KT990887 KT990505 Wu et al. 2016
Sutorius eximius REH9400 USA MG212568 MH614902** JQ327029* MG212653 *Halling et al. 2012a; Vadthanarat et al. 2018; **This study
Sutorius eximius HKAS50420 China KT990750 KT990387 Wu et al. 2016
Sutorius sp. OR0378B Thailand MH614692 MH614903 MH614740 MH614787 This study
Sutorius sp. OR0379 Thailand MH614693 MH614904 MH614741 MH614788 This study
Tengioboletus glutinosus HKAS53425 China KF112204 KF112800 Wu et al. 2014
Tengioboletus reticulatus HKAS53426 China KF112313 KF112828 Wu et al. 2014
Tengioboletus sp. HKAS76661 China KF112205 KF112801 Wu et al. 2014
Turmalinea persicina KPM-NC18001 Japan KC552130 KC552082 Orihara et al. 2016
Turmalinea yuwanensis KPM-NC18011 Japan KC552138 KC552089 Orihara et al. 2016
Tylocinum griseolum HKAS50281 China KF112284 KF112730 Wu et al. 2014
Tylopilus alpinus HKAS55438 China KF112191 KF112687 Wu et al. 2014
Tylopilus atripurpureus HKAS50208 China KF112283 KF112799 Wu et al. 2014
Tylopilus balloui s.l. OR0039 Thailand KT823965 MH614905* KT824031 KT823998 Raspé et al. 2016; *This study
Tylopilus brunneirubens HKAS53388 China KF112192 KF112688 Wu et al. 2014
Tylopilus felleus VDKO0992 Belgium KT823987 MH614906* KT824053 KT824020 Raspé et al. 2016; *This study
Tylopilus ferrugineus BOTH3639 USA MH614694 MH614907 MH614742 MH614789 This study
Tylopilus otsuensis HKAS53401 China KF112224 KF112797 Wu et al. 2014
Tylopilus sp. HKAS74925 China KF112222 KF112739 Wu et al. 2014
Tylopilus sp. HKAS50229 China KF112216 KF112769 Wu et al. 2014
Tylopilus sp. JD598 Gabon MH614695 MH614908 MH614743 MH614790 This study
Tylopilus sp. OR0252 China MG212569 MH614909* MG212611 MG212654 Vadthanarat et al. 2018; *This study
Tylopilus sp. OR0542 Thailand MG212570 MH614910* MG212612 MG212655 Vadthanarat et al. 2018; *This study
Tylopilus sp. OR0583 Thailand MH614696 MH614744 This study
Tylopilus sp. OR1009 Thailand MH614697 MH614911 MH614791 This study
Tylopilus vinaceipallidus HKAS50210 China KF112221 KF112738 Wu et al. 2014
Tylopilus vinaceipallidus OR0137 China MG212571 MH614912* MG212613 MG212656 Vadthanarat et al. 2018; *This study
Tylopilus violaceobrunneus HKAS89443 China KT990886 KT990504 Wu et al. 2016
Tylopilus virens KPM-NC-0018054 Japan KC552174 KC552103 Unpublished
Veloporphyrellus alpinus HKAS68301 China JX984515 JX984550 Li et al. 2014b
Veloporphyrellus conicus REH8510 Belize MH614698 MH614913 MH614745 MH614792 This study
Veloporphyrellus gracilioides HKAS53590 China KF112210 KF112734 Wu et al. 2014
Veloporphyrellus pseudovelatus HKAS59444 China JX984519 JX984553 Li et al. 2014b
Veloporphyrellus velatus HKAS63668 China JX984523 JX984554 Li et al. 2014b
Xanthoconium affine NY00815399 USA KT990850 KT990486 Wu et al. 2016
Xanthoconium porophyllum HKAS90217 China KT990851 KT990487 Wu et al. 2016
Xanthoconium sinense HKAS77651 China KT990853 KT990488 Wu et al. 2016
Xerocomellus chrysenteron VDKO0821 Belgium KT823984 MH614914* KT824050 KT824017 Raspé et al. 2016; *This study
Xerocomellus cisalpinus ADK4864 Belgium KT823960 MH614915* KT824026 KT823993 Raspé et al. 2016; *This study
Xerocomellus communis HKAS50467 China KT990858 KT990494 Wu et al. 2016
Xerocomellus corneri HKAS90206 Philippines KT990857 KT990493 Wu et al. 2016
Xerocomellus porosporus VDKO0311 Belgium MH614678 MH614846 MH614727 MH614773 This study
Xerocomellus ripariellus VDKO0404 Belgium MH614699 MH614916 MH614746 MH614793 This study
Xerocomellus sp. HKAS56311 China KF112170 KF112684 Wu et al. 2014
Xerocomus aff. macrobbii HKAS56280 China KF112265 KF112708 Wu et al. 2014
Xerocomus fulvipes HKAS76666 China KF112292 KF112789 Wu et al. 2014
Xerocomus magniporus HKAS58000 China KF112293 KF112781 Wu et al. 2014
Xerocomus s.s. sp. OR0237 China MH580796 MH580816 MH580835 Chuankid et al. 2019
Xerocomus s.s. sp. OR0443 China MH580797 MH614917* MH580817 MH580836 Chuankid et al. 2019; *This study
Xerocomus sp. OR0053 Thailand MH580795 MH614918* MH580815 MH580834 Chuankid et al. 2019; *This study
Xerocomus subtomentosus VDKO0987 Belgium MG212572 MH614919* MG212614 MG212657 Vadthanarat et al. 2018; *This study
Zangia citrina HKAS52684 China HQ326850 HQ326872 Li et al. 2011
Zangia olivacea HKAS45445 China HQ326854 HQ326873 Li et al. 2011
Zangia olivaceobrunnea HKAS52272 China HQ326857 HQ326876 Li et al. 2011
Zangia roseola HKAS51137 China HQ326858 HQ326877 Li et al. 2011
Zangia roseola HKAS75046 China KF112269 KF112791 Wu et al. 2014

Taxonomy

Cacaoporus Raspé & Vadthanarat, gen. nov.

MycoBank No: MB829655

Etymology

Refers to the dark, chocolate brown hymenophore and overall colour of basidiomata.

Diagnosis

Similar to the genus Sutorius in having brown basidiomata with brown encrustations in the flesh but differs from Sutorius in having the following combination of characters: brown to chocolate brown or greyish-brown to dark brown or blackish-brown basidiomata, without violet tinge, chocolate brown to dark brown hymenophore, tubes not separable from the pileus context, white to off-white basal mycelium which turns reddish-white to pale red when bruised, amygdaliform to ovoid with subacute apex in side view to ovoid basidiospores and dark brown spore deposit.

Description

Basidiomata stipitate-pileate with poroid hymenophore, small to medium-sized, dull, brown to greyish-brown to dark brown or blackish-brown. Pileus convex when young becoming plano-convex to slightly depressed with age, with deflexed to inflexed margin; surface even to subrugulose, minutely tomentose or slightly cracked at the centre; context soft, yellowish to greyish off-white then slightly greyish-orange to dull orange to greyish-brown when exposed to the air, patchy or marmorated with greyish-brown to dark brown, sometimes with scattered small dark brown to brownish-black encrustations, not or inconsistently reddening when cut. Hymenophore tubulate, adnate, subventricose to ventricose, slightly depressed around the stipe; tubes brown to greyish-brown to dark brown, not separable from the pileus context; pores regularly arranged, mostly roundish at first becoming slightly angular with age, sometimes irregular, elongated around the stipe, dark brown to greyish-brown at first, becoming brown to chocolate brown with age. Stipe central, terete to sometimes slightly compressed, cylindrical to sometimes slightly wider at the base; surface even, minutely tomentose, dull, dark brown to greyish-brown, basal mycelium white to off-white becoming reddish-white to pale red when touched; context solid, yellowish to orange white to yellowish-grey to pale orange to dull orange to reddish-grey, marmorated or virgated with brownish-grey to greyish-brown to dark brown, sometimes scattered with small reddish-brown to brownish-black fine encrustations, unchanged or inconsistently reddening when cut. Spore print dark brown.

Basidiospores amygdaliform to ovoid or ovoid with subacute apex in side view, thin-walled, smooth, slightly reddish to brownish hyaline in water, slightly yellowish to greenish hyaline in KOH or NH4OH, inamyloid. Basidia 4-spored, clavate to narrowly clavate without basal clamp connection. Cheilocystidia fusiform or cylindrical with obtuse apex, sometimes bent or sinuate, thin-walled, often scattered with small brownish-yellow to yellowish-brown crystals on the walls in KOH or NH4OH. Pleurocystidia narrowly fusiform with obtuse apex or cylindrical to narrowly subclavate, sometimes bent or sinuate, thin-walled, densely covered with small reddish-brown to brownish dark encrustations on the walls when observed in H2O, which are discoloured then dissolved in KOH or NH4OH. Pileipellis a trichoderm becoming tangled trichoderm to tomentum, composed of thin-walled hyphae; terminal cells mostly slightly sinuate cylindrical to irregular with rounded apex or clavate to elongated clavate. Stipitipellis a trichoderm to tangled trichoderm or disrupted hymeniderm, composed of loosely to moderately interwoven cylindrical hyphae anastomosing at places. Clamp connections not seen in any tissue.

Typus generis

Cacaoporus tenebrosus

Distribution

Currently known from Thailand.

Notes

Sutorius most closely resembles the new genus. In the field, Cacaoporus is easily distinguished from the Sutorius by the following combination of characters: chocolate brown to dark brown to blackish-brown basidiomata, which are darker than in Sutorius and never purplish-brown like in Sutorius species; chocolate brown to dark brown hymenophore, which is much darker than in Sutorius and never reddish- to purplish-brown like in Sutorius; tubes that are not separable from the pileus context but can be separated in Sutorius; off-white basal mycelium that more or less turns red when bruised, which is never the case in Sutorius.

Cacaoporus pallidicarneus Vadthanarat, Raspé & Lumyong, sp. nov.

MycoBank No: MB829657
Figs 2a, 3a, 4a and 5

Etymology

Refers to the context, which is paler than in the other species, especially at the stipe base and in the pileus.

Type

THAILAND, Chiang Mai Province, Mae On District, 18°52'37"N, 99°18'23"E, elev. 860 m, 15 August 2015, Santhiti Vadthanarat, SV0221 (CMUB!, isotype BR!).

Diagnosis

Cacaoporus pallidicarneus is characterised by having a paler context than the other species and basidiospores that are amygdaliform or elongated amygdaliform to ovoid in side view, sometimes with subacute apex, shorter basidia and fusiform to narrowly bent fusiform to narrowly fusiform hymenophoral cystidia.

Description

Basidiomata small to medium-sized. Pileus (1.6)2.4–5.5 cm in diameter, convex when young becoming plano-convex with age; margin deflexed to inflexed, slightly exceeding (1–2 mm), surface even to subrugulose, minutely tomentose, dull, at first brown to greyish-brown to blackish-brown (8F3–4) sometimes paler (8C2) at places, becoming paler to greyish-brown (8E3–5) with age; context 4–9 mm thick half-way to the margin, soft, yellowish to greyish off-white then slightly pale orange to greyish-orange (6A3 to 6B3) when exposed to the air, with patchy or marmorated with greyish-brown (8E3) especially when young, scattered with reddish-brown to brownish-black of fine encrustations at places, slightly reddening when cut. Stipe central, terete or sometimes slightly compressed, cylindrical with slightly wider base, (2.0)2.8–3.7 × 0.4–0.7 cm, surface even, minutely tomentose, dull, greyish-brown to dark brown (8 E/F 3–4 to 8F2), basal mycelium white to off-white becoming pale red (7A3) when bruised; context solid, yellowish to greyish off-white then orange white to pale orange (5A2–3) when exposed to the air, virgate to marmorate with brownish-grey (8F2), less so at the stipe base, at places scattered with brownish-black fine encrustations, unchanged to slowly slightly reddening when cut. Hymenophore tubulate, adnate, subventricose, slightly depressed around the stipe. Tubes (2)4–6 mm long half-way to the margin, brown to greyish-brown (8F3), not separable from the pileus context. Pores 0.4–1.5 mm wide at mid-radius, regularly arranged, mostly roundish to elliptical at first, becoming slightly angular with age, slightly elongated around the stipe, colour distribution even, dark brown to chocolate brown (9F4 to 10F3) at first, becoming chocolate brown to brown (10F4 to 7–8F4–5) with age. Odour rubbery. Taste slightly bitter at first, then mild. Spore print dark brown (8F4/5) in mass.

Macrochemical reactions. KOH, orange brown on cap, yellowish-black on stipe, yellowish-black on the pileus context and stipe context, brownish-black on hymenium; NH4OH, yellowish-brown on cap, yellowish-orange on stipe, orangey yellow to yellowish-orange on the pileus context, stipe context and hymenium.

Basidiospores [437/7/5] (6.5–)6.7–7.7–8.6(–11.5) × (3.8–)4–4.6–5.1(–5.5) µm Q = (1.4–)1.48–1.68–1.9(–2.44). From the type (3 basidiomata, N = 177) (6.8–)7–7.8–8.5(–9.1) × (4–)4.2–4.6–5(–5) µm, Q = (1.49–)1.5–1.69–1.9(–2.21), amygdaliform or elongated amygdaliform sometimes to ovoid with subacute apex in side view, ovoid in front view, thin-walled, smooth, slightly reddish to brownish hyaline in water, slightly yellowish to greenish hyaline in KOH or NH4OH, inamyloid. Basidia 4-spored, (25.3–)25.4–29.7–33.8(–33.8) × (7.3–)7.3–8.4–9.8(–10) µm, clavate without basal clamp connection, slightly yellowish to brownish hyaline in KOH or NH4OH; sterigmata up to 5 µm long. Cheilocystidia (16–)16.3–23.4–32.8(–34) × (5.5–)5.8–7.3–9(–9) µm, frequent, fusiform, thin-walled, yellowish to brownish hyaline to brown in KOH or NH4OH. Pleurocystidia (44–)44.2–54.7–67.6(–68) × (5–)5–6–7(–7) µm, frequent, usually narrowly bent fusiform to narrowly fusiform with obtuse apex, thin-walled, yellowish to brownish hyaline in KOH or NH4OH. Hymenophoral trama subdivergent to divergent, 62–175 µm wide, with 25–100 µm wide, regular to subregular mediostratum, composed of cylindrical, 4–7(11) µm wide hyphae, yellowish to brownish hyaline in KOH or NH4OH. Pileipellis a trichoderm to tangled trichoderm at first, becoming a tomentum to tangled trichoderm with age, 65–110 µm thick, composed of firmly to moderately interwoven thin-walled hyphae; terminal cells 12–55 × 4–6 µm, slightly bent cylindrical with rounded apex, at places clavate to sub-clavate to elongated clavate, 16–34 × 8–10 µm, slightly dark to reddish to brownish dark in water, yellowish to brownish hyaline to yellowish-brown to slightly dark at places in KOH or NH4OH. Pileus context made of moderately interwoven, thin-walled, hyaline hyphae, 6–12 µm wide. Stipitipellis a disrupted hymeniderm, 55–95 µm thick, composed clavate cells, 11–37 × 5–8 µm, yellowish-brown to slightly dark in KOH or NH4OH mixed with caulocystidia. Caulocystidia (17–)17–23.6–31(–31) × (5–)5–6.3–7(–7) µm, frequent, thin-walled, mostly yellowish-brown to slightly dark at places in KOH or NH4OH. Stipe context composed of parallel, 3–7 µm wide hyphae, brownish hyaline to yellowish pale brown in KOH or NH4OH. Clamp connections not seen in any tissue.

Habitat and Distribution

solitary to gregarious up to 4 basidiomata, on soil in hill evergreen forest dominated by Fagaceae trees, with a few Dipterocarpus spp. and Shorea spp. or in Dipterocarp forest dominated by Dipterocarpus spp. and Shorea spp. with a few Lithocarpus sp., Castanopsis sp. and Quercus sp. Currently known only from Chiang Mai Province, Northern Thailand.

Additional specimens examined

THAILAND, Chiang Mai Province, Mae Taeng District, 23 km marker (Ban Tapa), 19°08'50"N, 98°46'50"E, elev. 930 m, 2 August 2013, Olivier Raspé & Anan Thawthong, OR0681; Ban Mae Sae, 19°14'70"N, 98°38'70"E, elev. 960 m, 3 August 2013, Olivier Raspé & Anan Thawthong, OR0683; Muang District, Doi Suthep-Pui National Park, 18°48'37"N, 98°53'33"E, elev. 1460 m, 14 July 2016, Olivier Raspé, OR1306; Mae On District, 18°52'35"N, 99°18'16"E, elev. 860 m, 6 June 2018, Santhiti Vadthanarat, SV0451.

Remarks

We observed some small yellowish to reddish to brownish dark particles or crystals covering the cell walls in pileipellis, stipitipellis and on the hymenium, especially the cystidia and basidia when observed in water. The small particles or crystals were mostly dissolved in KOH.

Cacaoporus pallidicarneus differs from C. tenebrosus by its basidiomata context colour which is paler, especially at the stipe base. A combination of the following characters are also distinctive: spore shape which is amygdaliform or elongated amygdaliform or sometimes ovoid with subacute apex in side view and ovoid in front view, while C. tenebrosus has ovoid spores, shorter basidia and differently shaped hymenophoral cystidia (see note under C. tenebrosus). Cacaoporus pallidicarneus has a stipitipellis which is a disrupted hymeniderm composed of caulocystidia and clavate cells, while the other species has a loose trichoderm or tangled trichoderm. Interestingly, one collection (SV0402) had a slightly paler context than C. tenebrosus but not as pale as C. pallidicarneus. The phylogenetic analyses indicated that this collection might be a species different from C. pallidicarneus and C. tenebrosus. However, the specimen was immature and, therefore, more collections are needed before the species can be formally recognised.

Figure 2. 

Habit of Cacaoporus species. a C. pallidicarneus (SV0221) b–d C. tenebrosus (b - SV0223, c - SV0224, d - SV0422). Scale bars: 1 cm (a–d).

Cacaoporus tenebrosus Vadthanarat, Raspé & Lumyong, sp. nov.

MycoBank No: MB829656
Figs 2b–d, 3b–c, 4b and 6

Etymology

Refers to the overall darkness of basidiomata, including the context.

Type

THAILAND, Chiang Mai Province, Mae On District, 18°52'37"N, 99°18'32"E, elev. 940 m, 15 August 2015, Santhiti Vadthanarat, SV0223 (holotype CMUB!, isotype BR!).

Diagnosis

Cacaoporus tenebrosus is characterised by having a darker context than the other species, longer basidia and cylindrical to narrowly subclavate hymenophoral cystidia.

Description

Basidiomata medium-sized. Pileus (2.3)3.1–5(9) cm in diameter, convex when young becoming plano-convex to slightly depressed with age; margin inflexed to deflexed, slightly exceeding (1–2 mm); surface even to subrugulose, minutely tomentose, slightly cracked at the centre, dull, greyish-brown (10F3) to dark brown to blackish-brown (8F4–5) to the margin; context 5–10 mm thick half-way to the margin, soft, marmorated, greyish-brown to dark brown (10F3–5) with greyish-brown (9B/D3), scattered with reddish-brown to brownish-black, fine encrustations at places, slightly reddening in paler spots when cut. Stipe central, terete, cylindrical to sometimes with slightly wider base, 4.3–7.0 × 0.7–1.4 cm, surface even, minutely tomentose, dull, dark brown to greyish-brown (9F4 to 10F3), basal mycelium white to off-white becoming reddish-white to pale red (7A3–4) when bruised; context solid, greyish-brown to dark brown (9–10F3–5) marmorated with reddish-grey (7/10B2), usually scattered with small reddish-brown to brownish-black fine encrustations, slightly reddening when cut. Hymenophore tubulate, adnate, subventricose to ventricose, slightly depressed around the stipe. Tubes (4)7–13 mm long half-way to the margin, brown to dark brown (8F3 to 9F4), not separable from the pileus context. Pores 0.8–2 mm wide at mid-radius, regularly arranged, mostly roundish at first, becoming slightly angular with age, sometime irregular, elongated around the stipe; colour distribution even, greyish-brown to dark brown (9F4) at first, becoming chocolate brown to brown (10F3 to 7–8F4–5) with age. Odour mild fungoid. Taste slightly bitter at first, then mild. Spore print dark brown (8/9F4) in mass.

Macrochemical reactions. KOH, yellowish then brown to black on cap, stipe, pileus context, stipe context and hymenium; NH4OH, yellowish then orange to brown on cap, stipe, pileus context, stipe context and hymenium.

Basidiospores [290/8/6] (7.4–)7.7–8.4–9.2(–10) × (4.5–)5–5.3–5.7(–6.1) µm Q = (1.25–)1.44–1.57–1.77(–2). From the type (2 basidiomata, N = 134) (7.5–)7.7–8.2–9(–9.9) × (4.9–)5–5.4–5.7(–5.9) µm, Q = (1.41–)1.43–1.54–1.71(–1.9), ovoid, thin-walled, smooth, slightly reddish to brownish hyaline in water, slightly yellowish to greenish hyaline in KOH or NH4OH, inamyloid. Basidia 4-spored, (33.6–)34.3–38.8–45.8(–47) × (7.7–)7.8–9.5–10.8(–10.9) µm, clavate to narrowly clavate without basal clamp connection, yellowish to brownish hyaline to slightly dark in KOH or NH4OH; sterigmata up to 5 µm long. Cheilocystidia (22–)22.1–28.7–37(–37) × (3–)3.1–4.4–5(–5) µm, frequent, cylindrical with obtuse apex, sometimes bent or sinuate, thin-walled, yellowish-brown to dark brown in KOH or NH4OH, often scattered with small brownish-yellow to yellowish-brown crystals on the walls in KOH or NH4OH. Pleurocystidia (62–)62.5–81.5–99(–99) × (7–)7–8–9(–9) µm, frequent, cylindrical to narrowly subclavate, sometimes bent or sinuate, thin-walled, with yellowish-brown to slightly dark content in KOH or NH4OH, densely covered with small reddish-brown to brownish dark encrustations on the walls when observed in H2O, with some scattered small brownish-yellow to yellowish-brown crystals on the walls in KOH or NH4OH. Hymenophoral trama subdivergent to divergent, 80–170 µm wide, with 60–80 µm wide of subregular mediostratum, composed of cylindrical, 4–8(11) µm wide hyphae, slightly yellowish to brownish hyaline in KOH or NH4OH. Pileipellis a tangled trichoderm to tomentum at places, 70–110 µm thick, composed of moderately interwoven thin-walled hyphae; terminal cells 12–48 × 4–7 µm mostly slightly sinuate, cylindrical to irregular with rounded apex, at places clavate to elongated clavate terminal cells 18–33 × 7–9 µm, slightly dark to reddish to brownish dark in water, yellowish-brown to slightly dark in KOH or NH4OH, scattered with small brownish-yellow to yellowish-brown crystals on the walls in KOH or NH4OH. Pileus context made of moderately interwoven, thin-walled, hyaline hyphae, 7–12 µm wide. Stipitipellis a trichoderm to tangled trichoderm, 70–120 µm thick, composed of loosely to moderately interwoven cylindrical hyphae anastomosing at places, brownish dark to dark in KOH or NH4OH. Caulocystidia (17–)17.6–29.4–46.3(–47) × (4–)4.1–5.5–6.9(–7) µm, clavate to cylindrical with obtuse apex, thin-walled, yellowish to brownish dark in KOH or NH4OH. Stipe context composed of parallel, 4–6(12) µm wide hyphae, brownish hyaline to yellowish pale brown in KOH or NH4OH. Clamp connections not seen in any tissue.

Habitat and distribution

Gregarious (up to 9 basidiomata) to fasciculate or solitary, on soil in hill evergreen forest dominated by Fagaceae trees, with a few Dipterocarpus spp. and Shorea spp. or in Dipterocarp forest dominated by Dipterocarpus spp., Shorea spp. with a few Lithocarpus sp., Castanopsis sp. and Quercus sp. Currently known only from Chiang Mai Province, Northern Thailand.

Additional specimens examined

THAILAND, Chiang Mai Province, Mae Taeng District, 19°07'15"N, 98°43'55"E, elev. 910 m, 29 July 2013, Olivier Raspé & Benjarong Thongbai, OR0654; ibid. 19°7'29"N, 98°40'59"E, elev. 1010 m, 24 May 2018, Santhiti Vadthanarat, SV0422; Mae On District, 18°52'37"N, 99°18'19"E, elev. 850 m, 15 August 2015, Santhiti Vadthanarat, SV0224; ibid., 18°52'35"N, 99°18'16"E, elev. 860 m, 15 July 2017, Olivier Raspé , OR1435; ibid., 6 June 2018, Santhiti Vadthanarat, SV0452.

Remarks

There were many small yellowish to reddish to dark brownish particles or crystals on the walls of pileipellis, stipitipellis and hymenium cells, especially on the cystidia and basidia when observed in water. The small particles or crystals are somewhat dissolved and discoloured in KOH.

Microscopically, Cacaoporus tenebrosus differs from C. pallidicarneus by having a darker context, longer basidia (33.6–47 µm vs. 25.3–33.8 µm, respectively), longer and larger hymenophoral cystidia, which also differ in shape (cylindrical to narrowly subclavate in C. tenebrosus but fusiform to narrowly fusiform in C. pallidicarneus). Phylogenetically, all Cacaoporus collections with a dark context formed a clade sister to C. pallidicarneus (BS = 85% and PP = 0.88), but some (SV0224 and SV0422) were genetically somewhat distant from the other collections. However, we could not find any difference in morphology. Consequently, we consider them as the same species (C. tenebrosus). Study of more collections is needed to confirm or infirm that they belong to the same species.

Figure 3. 

Close-ups of hymenium/pileus context transition zone in Cacaoporus species, illustrating the non-separability of both tissues a C. pallidicarneus (OR0681) b C. tenebrosus (OR0654) c C. tenebrosus (SV0452). The transition between both tissues is particularly unmarked in C. pallidicarneus (a) Scale bars: 3 mm (a); 5 mm (b–c).

Figure 4. 

Scanning electron micrographs of Cacaoporus basidiospores a C. pallidicarneus (SV0221) b C. tenebrosus (SV0223). Scale bars: 1 µm (a–b).

Figure 5. 

Microscopic features of Cacaoporus pallidicarneus a basidiospores b basidia c cheilocystidia d pleurocystidia e caulocystidia f pileipellis g stipitipellis. Scale bars: 10 µm (a–b); 25 µm (c–e); 50 µm (f–g). All drawings were made from the type (SV0221).

Figure 6. 

Microscopic features of Cacaoporus tenebrosus a basidiospores b basidia c cheilocystidia d pleurocystidia e caulocystidia f pileipellis g stipitipellis. Scale bars: 10 µm (a–b); 25 µm (c–e); 50 µm (f–g). All drawings were made from the type (SV0223).

Discussion

Morphologically, Cacaoporus is most similar to Sutorius, with which it shares the overall brown colour of basidiomata and encrustations in the flesh. However, the genus Cacaoporus has darker basidiomata, especially the hymenophore and pore surface and is more chocolate brown, not reddish-brown or purplish-brown like Sutorius, tubes that are not separable from the pileus context whereas they are easily separable in Sutorius, white to off-white basal mycelium which becomes reddish when bruised, whereas in Sutorius, the basal mycelium is more or less white and unchanging. Cacaoporus also produces dark brown spore deposits whereas in Sutorius, spore deposits are reddish-brown (Halling et al. 2012). Microscopically, the two genera differ in the shape of basidiospores, which is amygdaliform to ovoid or ovoid with subacute apex in side view in Cacaoporus, whereas Sutorius produces narrowly ellipsoid to ellipsoid or subfusoid to fusoid basidiospores. Phylogenetically, Cacaoporus and Sutorius are not closely related - the two genera belong in two different clades of the Pulveroboletus group.

Some species in Porphyrellus E.-J. Gilbert also have brown to dark brown to umber basidiomata similar to Cacaoporus. However, Porphyrellus differs from the new genus in having white to greyish-white hymenophore when young, becoming greyish-pink to blackish-pink with age, white to pallid context in pileus and stipe variably staining blue and/or reddish when cut and white basal mycelium that does not turn red when bruised (Wolfe 1979; Wu et al. 2016). Some species in Strobilomyces Berk also share some characters with Cacaoporus, including dark brown basidiomata, white to off-white basal mycelium that turns red when bruised and the context turning red when cut. However, Strobilomyces species clearly differ from Cacaoporus, especially in the pileus surface, which is coarsely fibrillose or shows conical to patch-like scales, in the hymenophore, which is whitish-cream or greyish-brown or vinaceous drab and stains reddish then blackish when bruised and also basidiospores, which are subglobose to obtusely ellipsoid with reticulation or longitudinally striate (Gelardi et al. 2012; Antonín et al. 2015; Wu et al. 2016). Moreover, Porphyrellus and Strobilomyces were phylogenetically inferred to belong in subfamily Boletoideae (Wu et al. 2014, 2016; Vadthanarat et al. 2018) distinct from Cacaoporus.

Phylogenetically, Cacaoporus was monophyletic and clustered in a well-supported clade with the genera Cyanoboletus and Cupreoboletus and one undescribed taxon, Boletus p.p. sp. (specimen voucher JD0693), belonging to the Pulveroboletus group of Wu et al. (2014, 2016). Cyanoboletus and Cupreoboletus, however, differ from Cacaoporus in important morphological characters. The former two genera have a yellow hymenophore and yellowish context and tissues instantly discolouring dark blue when injured, and olive-brown spore deposits (Gelardi et al. 2014, 2015; Wu et al. 2016). The undescribed taxon represented by the voucher specimen JD0693, which clustered within the same clade as Cacaoporus, Cyanoboletus and Cupreoboletus, is also morphologically very different from Cacaoporus, in having yellow tubes, reddish pores, pale yellow to off-white context and reddish-brown pileus and stipe.

Our survey on the diversity of Boletes in the north of Thailand has been conducted since 2012 and no Cacaoporus has been found in the forests at elevations lower than 850 m. Cacaoporus was found only between 850 m and 1460 m elevation. However, more collections are needed to confirm that the distribution of the genus is restricted to mid- to high-elevation forests and does not occur in the lower elevation, drier forests. Most collections were made from Fagaceae-dominated, evergreen hill forests. The dominant trees in these forests belong to the Fagaceae, including Lithocarpus, Castanopsis and Quercus, but some Dipterocarpaceae may also occur. At the lower end of its elevation range, however, Cacaoporus was also found in Dipterocarpaceae-dominated forests (in which Fagaceae, especially Quercus spp., also occurs). The Dipterocarpaceae trees include Dipterocarpus, namely D. tuberculatus, D. obtusifolius and Shorea, namely S. obtusa and S. siamensis. The listed trees have previously been reported as ectomycorrhizal hosts of Boletaceae (Moser et al. 2009; Desjardin et al. 2009, 2011; Hosen et al. 2013; Arora and Frank 2014; Halling et al. 2014; Wu et al. 2018) and presumably are also the hosts for Cacaoporus.

Interestingly, our phylogeny indicated that the genera Neoboletus and Sutorius formed two different clades, both with high support (BS = 85% and PP = 0.95 for Neoboletus; BS = 100% and PP = 1 for Sutorius). Recently, Wu et al. (2016) synonymised Neoboletus with Sutorius because, in their phylogeny based on a four-gene dataset (28S+tef1+rpb1+rpb2), Boletus obscureumbrinus, a species morphologically more similar to Neoboletus than to Sutorius, seemed to cluster with Sutorius rather than with the Neoboletus species, although with neither ML nor BI support. Moreover, the Neoboletus clade was not supported either. Later, Chai et al. (2019) treated the two genera as different genetic lineages based on morphology and phylogeny (28S+ITS+tef1+rpb2), in which B. obscureumbrinus clustered with the other Neoboletus species in a well-supported clade. Our phylogenetic analyses, based on a different set of genes (atp6+ tef1+rpb2+cox3), confirm the separation of the two genera Neoboletus and Sutorius. The differences in gene trees obtained could be explained by a long-branch attraction artefact in datasets with different taxon and gene samplings and/or problems in the dataset (e.g. suboptimal alignment). Neoboletus obscureumbrinus is quite atypical amongst Neoboletus species and its phylogenetic affinities within this genus remain unclear (Fig. 7).

Cacaoporus is the second novel bolete genus described from Thailand, the first one being Spongiforma Desjardin, Manfr. Binder, Roekring & Flegel, described in 2009 (Desjardin et al.). However, fungal diversity in Thailand is high and still poorly known (Hyde et al. 2018), with a large number of species and possibly genera that remain to be described.

Figure 7. 

Sub-tree of the phylogram in Fig. 1, showing the well-supported Sutorius and Neoboletus clades and the unsupported sister relationship of Neoboletus obscureumbrinus.

Acknowledgements

Financial support from the Graduate School, Chiang Mai University, is appreciated. The work was partly supported by a TRF Research Team Association Grant (RTA5880006) to SL and OR. OR is grateful to the Fonds National de la Recherche Scientifique (Belgium) for travel grants. The authors are grateful for the permit number 0907.4/4769 granted by the Department of National Parks, Wildlife and Plant Conservation, Ministry of Natural Resources and Environment for collecting in Doi Suthep-Pui National Park. Beatriz Ortiz-Santana (CFMR), Roy E. Halling (NY), and Terry W. Henkel are gratefully acknowledged for the loan of specimens.

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