11urn:lsid:arphahub.com:pub:C004A564-9D6A-5F9F-B058-6A3815DFE9C3MycoKeysMC1314-40571314-4049Pensoft Publishers10.3897/mycokeys.32.2283822838Research ArticleBasidiomycotaTaxonomyAmericasCantharellusviolaceovinosus, a new species from tropical Quercus forests in eastern MexicoHerreraMariana1BandalaVictor M.victor.bandala@inecol.mxhttps://orcid.org/0000-0001-7093-06231MontoyaLeticia1Red Biodiversidad y Sistemática, Instituto de Ecología A.C., P.O. Box 63, Xalapa, Veracruz, 91000, MéxicoInstituto de Ecología A.C.XalapaMexico
Corresponding author: Victor M. Bandala (victor.bandala@inecol.mx)
Academic editor: M.P. Martín
2018200320183291109FFEAC450-AF63-C664-9524-FFCA855CF66712225530712201701032018Mariana Herrera, Victor M. Bandala, Leticia MontoyaThis 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.
During explorations of tropical oak forests in central Veracruz (eastern Mexico), the authors discovered a Cantharellus species that produces basidiomes with strikingly violet pileus and a hymenium with yellow, raised gill-like folds. It is harvested locally and valued as a prized edible wild mushroom. Systematic multiyear sampling of basidiomes allowed the recording of the morphological variation exhibited by fresh fruit bodies in different growth stages, which supports the recognition of this Cantharellus species from others in the genus. Two molecular phylogenetic analyses based on a set of sequences of species of all major clades in Cantharellus, one including sequences of the transcription elongation factor 1-alpha (tef-1α) and a combined tef-1α and nLSU region (the large subunit of the ribosome), confirm the isolated position of the new species in a clade close to C.lewisii from USA, in the subgenus Cantharellus. Detailed macroscopic and microscopic descriptions, accompanied by illustrations and a taxonomic discussion are presented.
Herrera M, Bandala VM, Montoya L (2018) Cantharellusviolaceovinosus, a new species from tropical Quercus forests in eastern Mexico. MycoKeys 32: 91–109. https://doi.org/10.3897/mycokeys.32.22838
Introduction
The diversity of species in Cantharellus Adans.: Fr., in combination with their ectomycorrhizal nature (mycobionts of several plant lineages), as well as their highly prized value as edible wild mushrooms, have attracted the attention of specialists from different fields worldwide (Smith and Morse 1947, Trappe 1962, Chandra 1989, Molina et al. 1992, Homola 1993, Thoen 1993, Watling 1997, Danell 1999, Pilz et al. 2002, Lee et al. 2003,Yun and Hall 2004, Boa 2005, Agerer 2006, Arora and Dunham 2008, Kumari et al. 2011, Wilson et al. 2012).
Cantharellus encompasses fungi with long-lived, gymnocarpic, fleshy, variedly coloured, trumpet-shaped basidiomes with nearly smooth, veined, gill-like folded to distinctly lamellate hymenophore, pileipellis poorly differentiated, cystidia lacking, smooth and thin-walled spores, with or without clamps (Wilson et al. 2012, Buyck et al. 2014). In many cases, basidiomes of members of closely related species or inclusive, unrelated look-alike species are difficult to identify in a strict sense, especially if there is not an accurate record of the variation of morpho-anatomical characters and colours in fresh condition. Few microscopic features in the genus had been considered discriminative, especially clamps (presence or absence), wall thickness of the terminal elements of the pileipellis hyphae and the basidiospore features (size and form).
The two former features are considered amongst the most taxonomically informative at subgeneric level and the latter used to distinguish species (Eyssartier and Buyck 2001, Buyck et al. 2014). Additionally, it has been hypothesized that there are cryptic species still undefined taxonomically, even amongst the best known Cantharellus species, especially from tropical regions but also from temperate regions (Smith and Morse 1947, Corner 1966, Heinemann 1958, 1966, Smith 1968, Petersen and Ryvarden 1971, Petersen 1976, Bigelow 1978, Petersen and Mueller 1992, Buyck and Hofstetter 2011, Buyck et al. 2012, Eyssartier et al. 2003, De Kesel et al. 2011, Tian et al. 2012, Wartchow et al. 2012, Buyck and Randrianjohany 2013, Foltz et al. 2013, Buyck et al. 2014, 2016a, c, Leacock et al. 2016).
Taxonomic research on Cantharellus has increased substantially in the last decade, especially by combining DNA and morphological information to support the definition of early recognised species and others recently discovered (Buyck and Hofstetter 2011, Buyck et al. 2011, Tibuhwa et al. 2012, Wilson et al. 2012, Buyck et al. 2012, 2013, 2014, 2015, 2016a, 2016b, Foltz et al. 2013, Shao et al. 2014, Shao et al. 2016, Leacock et al. 2016).
The earliest description of Cantharellus in Mexico dates from Fries (1855), who proposed C.mexicanus based on a specimen with “… pileo carnoso turbinato-infundibuliformi glabro griseofusco… lamellis augustissimis longe decurrentibus”. It was collected by F.M. Liebmann at El Mirador, Veracruz and, years later, considered by Corner (1966) as “… incert. sed. (? Gomphus)…” no longer recorded in the literature. From the same region (Orizaba, relatively near to the current study site), Craterellusconfluens Berk. & M.A. Curtis was described by Berkeley (1867). This species is characterised by its yellow basidiomes, it is closely related to Cantharelluslateritius (Berk.) Singer, with which it has been confused or even with other yellow chanterelles, such as C.cibarius Fr. and Craterellusodoratus (Schwein.) Fr. (Heim 1954, Corner 1966, Petersen 1979, Guzmán and Sampieri 1984, Buyck and Hofstetter 2011). After such descriptions of new Cantharellus species from Mexico, only some additional records of about ten species described from other latitudes have been mentioned to occur in different forest ecosystems in the country, including Quercus forests (Guzmán and Sampieri 1984, Guzmán 1985, Guevara et al. 2004, Perez-Moreno et al. 2008, Garibay-Orijel et al. 2009). The identity of these records, however, has not been confirmed with molecular evidence. Recently, C.coccolobae Buyck, Moreau & Courtecuisse was described from the Caribbean (Guadeloupe), including two collections from Yucatán, Mexico (Buyck et al. 2016c).
During the authors’ long term explorations in tropical oak forests in central Veracruz, a Cantharellus species was found with a striking habit, distinctive when compared to the previous records from Mexico. In fact, this fungus is unique because the fresh basidiomes in different growth stages possess a strikingly violet pileus and yellow, raised gill-like folded hymenophore, in combination with ellipsoid basidiopores and terminal elements of pileipellis slightly thick-walled. The macro- and micromorphological features depicted in this fungus, as well as its distinct position in two phylogenetic analyses, one of tef-1α and other of a combined tef-1α+nLSU sequences datasets, allowed its recognition as a new species. This Cantharellus species is locally considered a prized edible mushroom.
Materials and methodsSampling and morphological study
Cantharellus basidiomes were collected during June-October, through six consecutive years (2012–2017) including some collections in 2009 and 2011, in tropical oak forests from Zentla (837–850 m a.s.l.) and Alto Lucero (400–500 m a.s.l.) counties in central Veracruz (eastern Mexico). In these oak forests, Quercusoleoides is dominant and even forms pure stands. In the Zentla locality, however, Q.glaucescens and Q.sapotifolia are also present and, at times, also form monodominant small patches. Descriptions of morpho-anatomical features were achieved based on fresh samples and following Largent (1973). The colour notations indicated in the descriptions follow Kornerup and Wanscher (1978) and Munsell colour chart (1994). Basidiomes were dried in a hot air dehydrator (45 °C). Microscopic features were observed and measured after tissues were rehydrated in 3 % potassium hydroxide (KOH) and stained with 1 % Congo red or analysed in Melzer´s solution. At least thirty-five basidiospores per collection were measured in length and width. Mean ranges denoted as X–m and the length/width ratio (Q–) of basidiospores, in side view, are given as an interval of mean values per collection (n=15 collections). The form of the basidiospores was interpreted after calculating the Q values, following Bas (1969). Line drawings were made with the aid of a drawing tube. Collections are part of XAL Herbarium (Thiers B. [continuously updated] Index Herbariorum: a global directory of public herbaria and associate staff. New York Botanical Garden`s Virtual Herbarium. http://sweetgum.nybg.org/science/ih/).
DNA extraction, PCR and sequencing
DNA was isolated from fresh material using DNAeasy Plant Mini Kit (QIAGEN, Hilden, Germany) following the manufacturer´s recommendations. The transcription elongation factor 1-alpha (tef-1α) was amplified using the primers tef1F and tef1R (Morehouse et al. 2003) and the large subunit of ribosome (nLSU) using the primers LR0R and LR7 (Vilgalys and Hesler 1990). PCR conditions were performed with an initial denaturation at 94 °C for 3 min; 35 cycles of 1 min 94 °C, 1 min at 55 °C and 2 min at 72 °C; and final elongation at 72 °C for 7 min. Amplified PCR products were purified with the DNA Clean & Concentrator Kit (Zymo Research, USA) following the manufacturer’s instructions. Cycle sequencing reactions were made using BigDye Terminator 3.1 Cycle Sequencing kit (Applied Biosystems, USA); reactions were purified with ZR DNA Sequencing Clean-up Kit (Zymo Research, USA) and run in a sequencer, ABIPrism 310 Genetic Analyzer (Applied Biosystems). Sequences obtained were assembled and edited in BioEdit (Hall 1999) and deposited at GenBank database (Benson et al. 2017) (Table 1).
Cantharellus species included in this study: samples, location and accession number for tef-1α and nLSU sequences.
Taxon
Voucher Specimen
Location
GenBank
tef-1α
nLSU
C.addaiensis
BB 98.033
Tanzania
JX192992
KF294667
C.afrocibarius
BB 96.236
Zambia
JX192993
KF294668
C.albidolutescens
BB 08.057
Madagascar
KF294752
KF294645
C.albidolutescens
BB 08.080
Madagascar
JX192982
–
C.ambohitantelyensis
BB 08.336
Madagascar
JX192989
–
C.amethysteus
BB 07.284
Slovakia
GQ914953
KF294639
C.amethysteus
BB 07.309
Slovakia
GQ914954
KF294642
C.appalachiensis
BB 07.123
USA
GQ914979
KF294565
C.cascadensis
BB 13.251
USA
KX857044
–
C.chicagoensis
JJ/MO-CANT1
USA
KX857025
–
C.cibarius
BB 07.300
Slovakia
GQ914950
KF294641
C.cibarius
GE 07.025
France
GQ914949
–
C.cinnabarinus
BB 07.053
USA
GQ914984
KF294630
C.cinnabarinus
BB 07.001
USA
GQ914985
KF294624
C.congolensis
BB 98.039
Tanzania
JX193015
KF294609
C.congolensis
BB 98.058
Tanzania
JX192996
KF294673
C.corallinus
JJ/MO-CANT2
USA
KX857031
–
C.corallinus
JJ/MO-CANT5
USA
KX857034
–
C.deceptivus
JJ/NC-CANT5
USA
KX857029
–
C.decolorans
BB 08.278
Madagascar
GQ914968
–
C.decolorans
BB 08.243
Madagascar
JX192987
–
C.densifolius
BB 98.013
Tanzania
JX193014
KF294616
C.ferruginascens
BB 07.283
Slovakia
GQ914952
KF294638
C.fistulosus
DT 43
Tanzania
JX192997
KF294674
C.flavolateritius
VH 1076
USA
KX857027
–
C.flavolteritius
VH1078
USA
KX857029
–
C.gracilis
BB 98.234
Tanzania
JX192970
–
C.humidicolus
BB 98.036
Tanzania
JX193005
KF294666
C.ibityensis
BB 08.203
Madagascar
JX192985
KF294651
C. isabellinus var. parvisporus
BB 98.020
Tanzania
JX192972
KF294614
C.iuventateviridis
SH13/7/2012
USA
KX857063
–
C.iuventateviridis
SH14/7/2012
USA
KX857064
–
C.lateritius
BB 07.025
USA
GQ914957
KF294628
C.lateritius
BB 07.058
USA
GQ914959
KF294633
C.lewisii
BB 02.197
USA
GQ914961
KF294623
C.lewisii
BB 07.003
USA
GQ914962
–
C.lilacinopruinatus
BB 07.221
Slovakia
GQ914951
KF294637
C.minor
BB 07.002
USA
JX192978
KF294625
C.minor
BB 07.057
USA
JX192979
KF294632
C.pallens
BB 09.441
Italy
KX857013
–
C.pallens
BB 12.082
Italy
KX857035
–
C.paucifurcatus
BB 08.320
Madagascar
KF294655
JK192988
C.persicinus
MH 15.001
USA
KX857080
–
C.phasmasis
CO57
USA
JX030417
–
C.phasmasis
CO74
USA
JX030418
–
C.platyphyllus
BB 98.012
Tanzania
GQ914969
KF294617
C.platyphyllussubsp.bojeriensis
BB 08.160
Madagascar
JX192984
KF294648
C.pseudominimus
JV 00.663
Portugal
JX192991
KF294657
C.quercophilus
BB 07.097
USA
JX192981
KF294644
C.sebosus
BB 08.234
Madagascar
JX192986
KF294652
C.spectaculus
C081
USA
JX030414
–
C.cfsubamethysteus
AV 12.003
Thailand
KX857062
–
C.subcyanoxanthus
BB 00.1137
Madagascar
JX192990
–
C.subincarnatussubsp.rubrosalmoneus
BB 06.080
Madagascar
JX192962
KF294601
C.subincarnatussubsp.rubrosalmoneus
BB 06.096
Madagascar
JX192963
KF294602
C.symoensii
BB 98.011
Tanzania
GQ914970
KF294618
C.symoensii
BB 98.113
Tanzania
JX192974
KF294619
C.tabernensis
BB 07.119
USA
GQ914976
KF294634
C.tabernensis
BB 07.020
USA
GQ914971
–
C.tanzanicus
BB 98.040
Tanzania
JX192977
KF294622
C.tenuithrix
BB 14.008
USA
KX857045
–
C.tenuithrix
BB 14.009
USA
KX857045
–
C.tomentosus
BB 98.038
Tanzania
GQ914965
KF294610
C.vellutinus
VH 1583
USA
KX857070
–
C.vellutinus
WR WV 07.074
USA
KX857068
–
C.versicolor
Tian 160
China
KM893857
–
C.versicolor
Yu 24
China
KM893856
–
C.violaceovinosus*
Bandala 4513
Mexico
MF616520
MF616524
C.violaceovinosus*
Corona 648
Mexico
MF616521
MF616525
C.violaceovinosus*
Herrera125
Mexico
MF616522
MF616526
Craterellustubaeformis
BB 07.293
Slovakia
GQ914989
KF294640
Hydnumrepandum
BB 07.341
Slovakia
JX192980
KF294643
*samples and sequences obtained here
Phylogenetic analysis
Six tef-1α and nLSU sequences obtained in this study, together with 113 sequences of Cantharellus species from all major clades across the genus (after Buyck et al. 2014) and with the highest similarity scores from the results of BLAST (Altschul et al. 1997) were downloaded from GenBank (http://www.ncbi.nlm.nih.gov/) and used to construct two datasets. One dataset consisted of tef-1α and other combined tef-1α+nLSU sequences. Craterellustubaeformis and Hydnumrepandum were included as outgroup taxa (Table 1 and alignment in TreeBASE S21920). Both datasets were assembled in the data editor PhyDE v.0.995 programme (Müller et al. 2010). They were aligned using Muscle (Edgar 2004) with inconsistencies corrected manually. A phylogeny of each dataset was constructed under maximum likelihood (ML) and Bayesian Inference (BI) methods. The best evolutionary model for both datasets was calculated with Mega 6.06 (Tamura et al. 2013). ML analyses were also performed using Mega 6.06 with 500 replicates of bootstrap. BI analyses were implemented with MrBayes on XSEDE (3.2.6) on CIPRES portal (Miller et al. 2010) with settings as described in Montoya et al. (2014). The phylogenies from ML and BI analyses were displayed using Mega 6.06 and FigTree v 1.3.1 (Rambaut 2009), respectively.
Results
Sixty fresh collections were obtained of the violet Cantharellus species, including basidiomes in different growth stages, most of them detected between August-October, in both localities explored. Six new tef-1α and nLSU sequences from three collections were generated in this study (Table 1). In the inferred molecular phylogenies (from tef-1α and tef-1α+nLSU sequences datasets) (Figs 1–2), the generated sequences from the Mexican specimens, clustered in a terminal clade, strongly supported only bootstrap values ≥70 and posterior probabilities ≥0.90 were considered and indicated (BS/BPP) on the branches of each tree. Both trees were congruent and the sequences of the Mexican Cantharellus cluster in a sister clade to C.lewisii from USA, in the subgenus Cantharellus (Buyck et al. 2014). Based on the distinctive morphological features and colour variation of the studied Cantharellus specimens, as well as the isolated position of the samples in the phylogenies obtained, it was concluded that this should be proposed as a new Cantharellus species, which inhabits the tropical Quercus forests in eastern Mexico.
Molecular phylogenetic analysis by maximum likelihood of tef-1α sequences dataset of Cantharellus species. Posterior probabilities and Bootstrap values (BPP/BS) are indicated on the tree branches.
Molecular phylogenetic analysis by maximum likelihood of tef-1α+nLSU sequences dataset of Cantharellus species. Posterior probabilities and Bootstrap values (BPP/BS) are indicated on the tree branches.
https://binary.pensoft.net/fig/192389Description of the new speciesFungiCantharellalesCantharellaceaeEEE4806B-DD02-53A9-817B-91136658B750CantharellusviolaceovinosusMycoBank: MB823600M. Herrera, Bandala & Montoyasp. nov.Figs 3, 4, 5Holotype.
MEXICO. Veracruz: Municipality of Zentla, around town of Zentla, 850 m a.s.l., gregarious in soil, under Quercusoleoides Schltdl. & Cham., 5 July 2012, Corona 648 (XAL).
Diagnosis.
Differing from other Cantharellus species by: uniformly dark violet, violet-grey to violet-wine or violet-reddish pileus; yellow, gill-like folded hymenophore and ellipsoid basidiospores 7–10 (–11) × (4.5–) 5–6.5 (–7) µm. X–m = 7.8–9× 5.1–6.3 µm, Q–= 1.31–1.66, basidia (40–) 45–114 (–125) × (6–) 7–11 (–12) µm, with (1–) 2–5 sterigmata, and terminal elements of the pileipellis 4–6 µm diam, slightly thick-walled.
Gene sequences ex-holotype.
MF616521 (tef-1α), MF616525 (nLSU).
Etymology.
Referring to the dark violaceous, becoming wine to reddish pileus.
Pileus (15–) 25–113 mm diam, convex to broadly-convex with margin incurved when young, expanding to plane or subplane, often shallowly depressed or finally broadly infundibuliform, surface dry, not hygrophanous, dull, smooth, glabrescent, surface at times breaking in faintly tesselate-rimose-like pattern, then appearing appressed fibrillose with age and not forming scales; surface uniformly dark violet (15D4, 15F2–7, 16D3–4, 16D6, 16F4–5) to pale violet with age (15DE5–7) or violet-grey (16D3–4, 16D6), lilac or greyish-lilac (15A3, 15C3–4, 16C2–3), becoming violet-wine or violet-reddish (14E5–8, 14EF4–5), wine (12D4), fading with age and sun exposure, developing pinkish, lilac and reddish tints, especially towards the margin (13A3–4, 13D3–4, 15A4–5), naked parts showing the yellow context (4A2–3); margin incurved or straight, entire or slightly crenate, undulate or irregular, often incised, rarely lobed, not striated. Hymenophore with well-defined gill-like folds, up to 3 mm deep, decurrent, subdistant, in some specimens almost straight and inclusive thin, in other materials with faintly sinuous or irregular thicker folds, frequently forking at different levels or only towards the pileus margin, with lower irregular anastomosis amongst the folds, in some specimens the anastomosis occur practically in the whole hymenophore, while in others only at some areas, especially at pileus margin, some specimens (specially towards the stipe) with irregular low veins amongst the folds or the folds become as low and sinuous vein-like; butter-yellow or yellow (2.5Y 8/4,10YR 8/6; 4A3–4). Stipe (20–) 25–75 × 5–18 mm, equal and only slightly swollen at base or widening above and tapering gradually downwards, solid, surface glabrous, concolorous with hymenophore, often staining ochraceous or rusty orange colour when handled, occasionally with whitish, small rhizomorphs at base. Context whitish to yellow (4A2–3), at times wax-like, odour mild, agreeable, at time fruity somewhat to apricot; taste mild, agreeable.
Basidiospores 7–10 (–11) × (4.5–) 5– 6.5 (–7) µm, [X–m = 7.8–9 × 5.1–6.3 µm, Q–= 1.31–1.66, (n=13)], ellipsoid, smooth, thin-walled, hyaline, inamyloid, devoid of granular contents or refractive droplets. Basidia (40–) 45–114 (–125) × (6–) 7–11(–12) µm, narrowly clavate to subcylindrical, with (1–) 2–5 sterigmata 8–10 µm long, thin-walled, hyaline; subhymenium composed of cylindrical hyphae 4–5 µm diam. Cystidia absent. Pileipellis a cutis composed of hyphae 4–6 µm diam, intermingled in a compact arrangement, cylindrical, hyaline to yellowish, inamyloid, often some of them with pale brownish contents, these decidedly brown coloured in group; distinctive terminal elements 4–6 µm broad, slightly thick-walled (<1 µm thick), smooth, hyaline, some pale brownish, scattered on the surface. Pileus trama composed of cylindrical to inflated hyphae, 3–12 µm diam, slightly thick-walled (<1 µm thick), hyaline, yellowish in mass, some of the hyphal segments completely filled with darker contents. Hymenophoral trama composed of hyphae 3–5 µm diam, thin-walled, some with weakly refringent contents. Clamp connections present on hyphae in all tissues.
Habitat.
Solitary to gregarious, in soil, at tropical oak forest, under Quercusoleoides, less frequently also under both Q.glaucescens Bonpl. and Q.sapotifolia Liebm. June-October, known in the coastal plain of central Veracruz State, east coast of Mexico.
Specimen examined.
MEXICO. Veracruz, Zentla Co., Road Puentecilla-La Piña, 837 m a.s.l., 2 Jul 2009, Del Moral 427, Ramos 216; 27 Oct 2009, García 20, García 22; 16 Jun 2011, Bandala 4490; 21 Jul 2012, Herrera 25; 31 Jul 2012, Bandala 4513; 20 Sep 2012, Bandala 4550, Corona 743; 4 Oct 2012, Bandala 4569, 4573; 4 Jul 2013, Gutiérrez 23; 12 Jul 2013, Bandala 4671; 20 Sep 2013, Herrera 67; 15 Sep 2015, Herrera 135. Around town of Zentla, 850 m a.s.l., 5 Jul 2012, Corona 648; 25 Jun 2013, Herrera 60, 61; 15 Sep 2015, Herrera 137, Santillan 16; 1 Oct 2015, Herrera 151; 30 Jun 2016, Herrera 172; 6 Jul 2016, Herrera 184; 12 Jul 2016, Herrera 187; 22 Sep 2016, Herrera 200, 201, 202, 203; 5 Oct 2016, De la Cruz 14,15; 13 Oct 2016, De la Cruz 42; 27 Oct 2016, Herrera 210, 211; 6 Jul 2017, Garay 350; 3 Aug 2017, Garay 364; 31 Aug 17, Garrido 79; 7 Sep 2017, Herrera 214, 215, 216; 15 Sep 17, Montoya 5403; 21 Sep 17, Corona 1420; 5 Oct 17, Mateo 5. Alto Lucero Co., NE Mesa de Venticuatro, 450–500 m a.s.l., 2 Jul 15, Herrera 125, Herrera 126; 17 Sep 2015, Herrera 138; 2 Aug 2016, Herrera 191; 10 Aug 2016, Herrera 192; 20 Sep 2016, Herrera 195, 196, 197, 198; 27 Sep 2016, Herrera 205, 206, 207; 4 Oct 2016, Herrera 208, 209; 22 Aug 17, Herrera 214; 12 Sep 2017, Garay 375; 19 Sep 2017, Garay 392; 2 Oct 17, Mateo 1 (all at XAL).
Discussion
Distinctive features of this species include the medium to large size basidiomes, with pileus practically homogeneously violet pigmented (only fading with age), smooth, with surface free of scales, at times with disrupted pileus surfaces due to age, hymenophore bearing yellow gill-like folds, ellipsoid, medium-sized basidiospores [7–10 (–11) × (4.5–) 5– 6.5 (–7) µm], medium to large basidia [(40–) 45–114 (–125) × (6–) 7–11(–12) µm] and terminal elements of pileipellis 4–6 µm diam, slightly thick-walled (<1 µm thick). Molecular phylogenetic analyses support that the species is genetically distinct from other Cantharellus taxa, in both analyses, C.violaceovinosus was nested in an isolated and well-supported clade (95–99/1) (Figs 1–2).
Cantharellus species with basidiomata having violet pileus are rare but occur in various regions worldwide (Eyssartier et al. 2009; Buyck et al. 2012). Amongst about 45 species of the genus known from USA, Mexico, Central and South America (Guzmán and Sampieri 1984, Guzmán 1985, Eyssartier et al. 2003, Guevara et al. 2004, Henkel et al. 2006, Wartchow et al. 2012, Wilson et al. 2012, Pinheiro and Wartchow 2013, Wartchow et al. 2013, Nascimento et al. 2014, Buyck et al. 2016c), C.lewisii Buyck & V. Hofst., C.atrolilacinus Eyssart., Buyck & Halling and the new C.violaceovinosus are, up to now, the species known to produce basidiomes with violet tints in the Americas.
Cantharelluslewisii grows in floodplain hardwoods, in Water Oak plots next to a Taxodium swamp, in beech-magnolia-loblolly pine forests and also under beech-white oak-loblolly pine-magnolia forests in the south of USA (Buyck and Hofstetter 2011). In the inferred phylogeny, it appears as sister of C.violaceovinosus, but differs because its pileus is pale yellow, dull to greyish-yellow or ochre to pale brownish-orange, sometimes reddish-brown near the margin, with a surface covered with dark purplish-lilac appressed fibrils (in young stages, C.lewisii is often entirely dark lilac-purple) and with terminal elements of pileipellis conspicuously thick-walled (mostly 1–1.5 μm thick) (Buyck and Hofstetter 2011). According to the original description, C.lewisii also differs by its ellipsoid or often somewhat reniform and narrower basidiospores [(7.08–) 7.16–7.62–8.07 (–8.96) × (4.17–) 4.24–4.58–4.93 (–5.21) μm; Q= (1.42–) 1.45–1.57–1.70 (–1.80)] and by 5–6-spored and shorter basidia (60–75 × 7–8 μm) (Buyck and Hofstetter 2011). Two Texan collections of C.lewisii (holotype BB 07.003 and BB 02.197, both at PC) were studied. Based on observations, it was confirmed that this later species differs from the Mexican C.violaceovinosus, because of its markedly reniform, narrower basidiospores, then tending to be “more ellipsoid” [BB 07.003, holotype: 7.5–9.5 × (4–) 4.5–5.5 µm, X–m = 8.4 × 5 µm, Q–= 1.68; BB 02.197: 7.5–10 (–11) × 4–5.5 µm, X–m = 9 × 4.8 µm, Q–= 1.87).
Cantharellusatrolilacinus was described from Costa Rica, growing under Quercuscorrugata Hook.) and Q. sp. (Eyssartier et al. 2003). According to the data on this species (R. Halling, www.nybg.org/bsci/res/hall/canlilac.html; Eyssartier et al. 2003), it differs from C.violaceovinosus because its pileus colours tend to be darker, even blackish, dark lilac-grey or brown-lilac, with tomentose surface at the disc, with strong radial, adnate fibrils at the margin, and the stipe whitish with lilac tints. Microscopically, C.atrolilacinus has basidiospores (7–) 7.5–8–8.5 (–9) × 4.5–5–5.5 (–6) μm, tending to be more ellipsoid (Eyssartier et al. 2003, fig. 1:2) and having wider pileipellis hyphae [(4–) 5–10 (–15) μm] with a very thick wall (“..très nettement épaissies..”).
Although Cantharellusamethysteus (Quél.) Sacc. (subg.Cantharellus) from Europe, may appear superficially similar to some forms of C.violaceovinosus, the former however, especially has a pileus surface covered with vinous or lilac, small scales. The authors studied two specimens of C.amethysteus from France (BB 07.284 and BB 07.309 at PC) displaying elongate basidiospores, 9.5–12 (–12.5) × 5–7 µm (X–m = 11–11.2 × 5.9–6.4 µm; Q–= 1.76–1.86), as Eyssartier and Buyck (2000) reported [(9–) 9.5–10.37–11.5 (–12.5) × 6–6.5–7 µm], resulting in being larger and more elongate than in the Mexican species. Also, it is interesting that one sequence of tef-1α of a specimen from Thailand (GB coded KX857062, Table 1) identified as “C.cf.subamethysteus”, appeared close to C.violaceovinosus (Fig. 1). Cantharellussubamethysteus indeed is phylogenetically related to C.lewisii (Buyck et al. 2014) and differs from C.violaceovinosus in the shorter basidiomes (pileus 20–65 mm; stipe 42–57 × 5–11 mm), deep and bright yellow pileus surface, covered with squamules even with rather brown to dark brown tinges, shorter basidiospores [7–8 (8.75) × (4.75) 5–6 μm] and wider pileipellis elements (8–15 μm width) (Eyssartier et al. 2009). Additionally, the hymenophore of this species is rugose to faintly veined (as depicted in the picture accompanying the description).
Cantharellusgoossensiae (Beeli) Heinem., C.cyanoxanthus R. Heim ex Heinem., C.subcyanoxanthus Buyck, Randrianjohany & Eyssart. and C.longisporus Heinem. represent African species with basidiomes displaying violaceous tinges (Buyck et al. 2012) therefore, at some stages their basidiomes could resemble those of C.violaceovinosus. However, the three former species have the pileipellis with thin-walled hyphal extremities, thus differing from members of subgenus Cantharellus, including the new species here described. Moreover, the four African taxa have basidiospores distinctly narrowly ellipsoid to elongate (Q>1.70) and often slightly reniform, curved or even somewhat peanut-shaped (Beeli 1928, Heinemann 1958, 1959, Buyck et al. 2012).
Cantarellusviolaceovinosus was recorded as a common fleshy mushroom, during the multiyear sampling developed in the tropical Quercus forests studied. It was found in ectomycorrhizal association with native trees of Quercus species. This mushroom was very often recorded in pure stands of Q.oleoides and less frequently in Q.glaucescens and Q.sapotifolia patches. This violet pigmented chanterelle shares the same habit preferences as C.lateritius, also found in the study sites. A similar co-ocurrence has been reported between C.lewisii (the sister relative of C.violaceovinosus) and C.lateritius in the State of Texas in the USA (Buyck et al. 2011). Basidiomes of C.violaceovinosus and C.lateritius are abundant in the local oak forests studied and both are considered choice wild mushrooms although the latter is more highly prized. They are even more appreciated than species of Amanita or Lactarius, representing an income source for wild mushroom collectors. Benefits from mushrooms harvesting, as well as other ecosystemic services, are motivating some owners to conserve relicts of the tropical Quercus forest of the region.
Cantharellusviolaceovinosus (Corona 648, holotype): a basidiospores b terminal elements of the pileipellis c basidia d pileipellis. Scale bars: 5 µm (a); 10 µm (b, c); 25 µm (d).
We recognise the support given by CONACYT (CB 252431) to study the EcM fungi associated with tropical species of Quercus in Veracruz, Mexico. M. Herrera is grateful for the scholarship grant from CONACYT (261413). Thanks are given to Dr. B. Buyck (PC) for the loan of herbarium specimens. We appreciate the assistance in the field and in the laboratory to Biol. D. Ramos (Instituto de Ecología, A.C.). Biol. J.C. Corona collaborated in some explorations. We acknowledge the support given by CONACYT (225382) to the Laboratorio de Presecuenciación, Red Biodiversidad y Sistemática, INECOL. Dra. Edith Garay and IBT Bertha Pérez assisted us in some molecular procedures.
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