Broadening the Knowledge of Mexican Boletes: Addition of a New Genus, Seven New Species, and Three New Combinations

Boletes are one of the most common groups of fungi in temperate, subtropical, and tropical ecosystems. In Mexico, the northern region has mainly been explored in terms of bolete diversity. This study describes a new genus and seven new species based on macromorphological, micromorphological, molecular, phylogenetic, and ecological data. Garcileccinum gen. nov. is typified with G. salmonicolor based on multigene phylogenetic analysis of nrLSU, RPB2, and TEF1, and it is closely related to Leccinum and Leccinellum. Garcileccinum viscosum and G. violaceotinctum are new combinations. Boletellus minimatenebris (ITS, nrLSU, and RPB2), Cacaoporus mexicanus (RPB2 and ATP6), Leccinum oaxacanum, Leccinum juarenzense (nrLSU, RPB2, and TEF1), Tylopilus pseudoleucomycelinus (nrLSU and RPB2), and Xerocomus hygrophanus (ITS, nrLSU, and RPB2) are described as new species. Boletus neoregius is reclassified as Pulchroboletus neoregius comb. nov. based on morphological and multigene phylogenetic analysis (ITS and nrLSU), and its geographic distribution is extended to Central Mexico, since the species was only known from Costa Rica. Furthermore, T. leucomycelinus is a new record from Mexico. This study contributes to increasing our knowledge of boletes and expands the diversity found in Mexican forests.


Introduction
Many ecosystems thrive in Mexico ranging between tropical, subtropical, temperate, subalpine, and alpine, which develop in different soil types, weather conditions, and contrasting altitudes due to the complex orography and geological history of the country [1,2].These conditions have produced communities where fungi can easily diversify [3,4].Consequently, more than 2000 fungal species have been reported in the country [3].Despite the relatively high diversity recorded in Mexico, studies are far from complete.Currently, most of the records have come from temperate forests.However, tropical and subtropical forests harbor abundant species of the families Casuarinaceae, Dipterocarpaceae, Fagaceae, Fabaceae, Mimosaceae, Nictaginaceae, Pinaceae, and Polygonaceae, which establish ectomycorrhizal symbiosis and, therefore, may potentially contain high fungal diversity.

Alignments, Model Selection, and Phylogenetic Analyses
The consensus sequences were compared with those in the GenBank database of the National Center for Biotechnology Information (NCBI) using BLASTN v. 2.2.19 [65].In order to estimate the phylogenetic position of the specimens studied in this work, Bayesian inference and maximum likelihood analyses were performed on the following molecular matrices: (1) ITS, nrLSU, and RPB2 of Boletellus Murrill species with Hemileccinum rugosum G. Wu and Zhu L. Yang as outgroup (Table 1); (2) RPB2 and ATP6 of Cacaoporus Raspé and Vadthanarat, species, with specimens of Cyanoboletus Gelardi, Vizzini, and Simonini, Chalciporus Bataille, Lanmaoa G. Wu and Zhu L. Yang, and Rubinoboletus rubinus (W.G.Sm.) Pilát and Dermek as outgroup, according to Vadthanarat et al. [66] (Table 2); (3) nrLSU, RPB2, and TEF1 of several members of the subfamily Leccinoideae, including Leccinum, Leccinellum, the new genus Garcileccinum, Rugiboletus, Ionosporus, Spongiforma, Pseudoaustroboletus, and Retiboletus, with Tylocinum as outgroup [14,15] (Table 3) (additionally, ITS, nrLSU, and RPB2 sequences were obtained and deposited in the Genbank); (4) ITS and nrLSU of Pulchroboletus Gelardi, Vizzini, and Simonini species with Alessioporus Gelardi, Vizzini, and Simonini, as outgroup (Table 4); (5) ITS and nrLSU of Tylopilus P. Karst.species, with Xanthoconium sinense as outgroup [26] (Table 5); and (6) ITS, nrLSU, and RPB2 of Xerocomus Quél.species, with Phylloporus imbricatus as outgroup [67] (Table 6).All sequences, except for those produced for this study, were retrieved from the GenBank in an attempt to represent most taxa from those groups.All matrices were aligned using the MUSCLE algorithm [68] in MEGA v. 11 with default parameters [69] and visually inspected in BioEdit v. 7.2.5 [70] for corrections and trimming.For Bayesian inference (BI) analysis, we selected DNA substitution models according to the Akaike information criterion (AIC) using jModelTest v. 2.1.7[71].A majority-rule BI consensus tree was produced in MrBayes v. 3.2.6 [72,73] with the following parameters: substitution model retrieved from jModelTest v. 2.1.7,with two independent runs each running for 10 million generations sampling every 1000 generations with one cold chain and three hot chains with a temperature of 0.2 and a final burn-in fraction of 0.25.For the concatenated matrices, each region was treated as an independent partition, and the following parameters were unlinked: transition/transversion rate, state frequency, and shape.The rate model was set to "variable", and the remaining parameters were used as default.Additionally, a maximum likelihood (ML) analysis was carried out in RAxML v. 8.2.12 [74] using the GTR+GAMMA substitution model treating each partition as independent.In addition, ML bootstrap (BS) was performed based on 1000 replicates.Both BI and ML analyses were implemented through CIPRES Science Gateway v. 3.3 [75].
Table 1.Specimens and sequences used for the molecular phylogenetic analyses in Figure 1.Sequences newly generated for this study are highlighted in bold.

Fungal Taxon
Specimen Voucher ITS nrLSU RPB2   Table 6.Specimens and sequences used for the molecular phylogenetic analyses in Figure 6.Sequences newly generated for this study are highlighted in bold.

Boletellus Phylogeny
The concatenated matrix of ITS, nrLSU, and RPB2 consisted of 31 accessions and 2478 positions, of which 809 (32.6%) were variable and 612 (24.7%) were parsimony-informative.The phylogram obtained from the BI analysis shows that two specimens of the new species, B. minimatenebris, are included in clade A (1BI/98ML), together with B. ananiceps, B. aurocontextus, B. ananas, and two unidentified specimens of Boletellus.One of them (HKAS122526) forms a well-supported clade with the two specimens of B. minimatenebris (1BI/98ML).Clade A is a sister to B. brunoflavus (0.97BI) (Figure 1).

Cacaoporus Phylogeny
The concatenated matrix of RPB2-ATP6 genes consisted of 44 accessions and 1483 positions, of which 457 (30.8%) were variable and 371 (25.0%) were parsimony-informative.In the phylograms of both BI and ML analyses, Cacaoporus was recovered as monophyletic (1BI/99ML) and sister to a clade, which includes a paraphyletic Cyanoboletus with Cupreoboletus nested within it.This sister relationship is moderately well supported (1BI/79ML).A monophyletic Lanmaoa (0.99BI/98ML) is sister to both clades.The four sequences of Cacaoporus mexicanus (Figure 2) form a monophyletic species (1BI/99ML) that

Boletellus Phylogeny
The concatenated matrix of ITS, nrLSU, and RPB2 consisted of 31 accessions and 2478 positions, of which 809 (32.6%) were variable and 612 (24.7%) were parsimonyinformative.The phylogram obtained from the BI analysis shows that two specimens of the new species, B. minimatenebris, are included in clade A (1BI/98ML), together with B. ananiceps, B. aurocontextus, B. ananas, and two unidentified specimens of Boletellus.One of them (HKAS122526) forms a well-supported clade with the two specimens of B. minimatenebris (1BI/98ML).Clade A is a sister to B. brunoflavus (0.97BI) (Figure 1).

Cacaoporus Phylogeny
The concatenated matrix of RPB2-ATP6 genes consisted of 44 accessions and 1483 positions, of which 457 (30.8%) were variable and 371 (25.0%) were parsimonyinformative.In the phylograms of both BI and ML analyses, Cacaoporus was recovered as monophyletic (1BI/99ML) and sister to a clade, which includes a paraphyletic Cyanoboletus with Cupreoboletus nested within it.This sister relationship is moderately well supported (1BI/79ML).A monophyletic Lanmaoa (0.99BI/98ML) is sister to both clades.The four sequences of Cacaoporus mexicanus (Figure 2) form a monophyletic species (1BI/99ML) that is sister to an unidentified specimen of Cacaoporus (SV0402) (1BI/95ML).The most closely related species to this clade is a monophyletic Cacaoporus pallidicarneus, whose sister relationship is moderately supported (71ML).

Pulchroboletus
The phylogenetic analysis of the genus Pulchroboletus (Figure 4) was performed with a concatenated matrix of the nrLSU and ITS markers of 17 accessions.The matrix had 1450 positions, of which 278 (19.2%) were variable and 154 (10.6%) were parsimonyinformative. Pulchroboletus was found to be monophyletic with high support (1BI/100ML).Inside the genus, a dichotomy is observed: one clade is composed of one sequence of P. rubricitrinus (0.98BI/91ML) and a monophyletic group of five sequences of P. roseoalbidus (1BI/94ML).The other clade (0.95BI/82ML) is composed of three specimens of unidentified Pulchroboletus, a clade of P. sclerotiorum, and the new combination here proposed: P. neoregius.Both species are monophyletic, highly supported (1BI/100ML), and are sister taxa.

Tylopilus Phylogeny
Two independent phylogenetic analyses were performed for Tylopilus, and another was performed with ITS and nrLSU, due to the missing sequences, to produce the four markers' concatenated matrix.The two-marker matrix consisted of 105 accessions and 2168 positions, of which 1193 (55.0%) were variable and 828 (35.2%) were parsimonyinformative.The phylogram (Figure 5) demonstrated low resolution and/or support at the deepest nodes.Tylopilus balloui is highly polyphyletic, appearing in several lineages.The specimen described here as a new species, T. pseudoleucomycelinus, is sister to another specimen identified as T. balloui (FMNH1073250) with high support (0.99BI/99ML).This clade is sister to a monophyletic T. leucomycelinus with good support (0.98BI/85ML).The lineage related to the type locality of T. balloui (USA) has good support outside this clade.
Diagnosis: basidiomata: pink salmon, grayish, pearl gray, grayish orange, then cinnamon brown to mustard brown to brown-orange color.Hymenophore: cream, yellowish white, grayish orange; context whitish, bruising pale grayish, vinaceous to dark violet, gray-violet, pale blue-green to greenish blue to deep blue at the context stipe base, and often slowly developing scattered orange-pink or coral pink stains; interior: white, changing to pinkish at apex, bluish green in the base.Stipe: finely floccose upper half, pruinose to scabrous, dry, white, apricot yellow, at first, becoming a pale caramel color, grayish orange.Basidiospores: smooth, fusoid to subfusoid.Pileipellis: composed of an ixotrichodermium embedded in a gelatinous matrix.
Diagnosis: this species is characterized by its medium-sized basidiomata; brownish red, reddish brown to cinnamon pileus with tomentose surface; and whitish, changing to greenish blue, or grayish violet context.After it is cut, the context of the stipe turns bluish Notes: Garcileccinum salmonicolor differs from other species in the genus due to its pileate-stipitate, pink salmon, salmon, apricot yellow to gray-reddish basidiomata, with pink salmon to salmon context edge; due to the surface of its pileus, that is subtomentose or subrugulose to light viscid when humid or whitish, pale yellow to salmon, and immutable when the pileus context is cut; due to the whitish and pale yellow to salmon stipe context; due to the green (28B5) to blue-gray (23C5, 22D7) base; and due to the length (10-) 11.2-16.5 × 3.5 -4.5 (-5.5) µm) of its basidiospores.Meanwhile, the pileus surface of Garcileccinum viscosum is always viscid, subrugulose to rugulose at first, and becomes reticulate-pitted, grayish orange, apricot yellow, cinnamon brown to mustard brown, and white changing to pinkish at its apex.The base stipe context is bluish green, and the basidiospores are 12.6-17.5× 4.9-6.3µm.G. viscosum occurs under Quercus peduncularis Née and Pinus caribaea Morelet, while G. salmonicolor is distributed in mixed forest putatively associated with Quercus rugosa at altitudes between 1530 and 1540 m.a.s.l.Garcileccinum salmonicolor differs from Garcileccinum violaceotinctum by its pearl gray to cinnamon brown color.When cut, it turns turquoise-tinted.Its basidiomata are pale grayish and vinaceous to dark violet.Its basidiospores are 12-13.6(-16) × 4-4.8 (-5.6) µm.G. viscosum and G. violaceotinctum were originally described from Belize, found under Pinus caribaea and Quercus sp.[17].In Mexico, G. viscosum is distributed in a Pinus-Quercus mixed forest occurring under Pinus teocote, P. oocarpa, and Quercus rugosa.
Holotype species: Mexico, Oaxaca State, Santiago Zacatepec District, Mixistlán de la Reforma Municipality, Santa María Mixistlán Town, 1 km, 29 June 2022, N 17 Etymology: the name refers to the state where the species was identified.
Habitat and distribution: solitary, distributed in disturbed cloud forest, associated putatively to Arbutus Xalapensis Kunth.Currently, it has been identified only in Mexico, Oaxaca state, Mixistlan town.
Notes: Leccinum oaxacanum is phylogenetically found in the Leccinum s.s.clade with 100 ML/1IB support.It shares macromorphological characteristics, mainly the pileus color, with the rest of the species in the genus.However, Leccinum oaxacanum differs from other species due to its medium-sized basidiomata, tomentose pileus surface, whitish context that becomes greenish blue to grayish violet when cut, stipe context that turns bluish gray to greenish blue, and basidiospores that are (10−) 11-14 (-15) × 4-5 µm, 2-5 µm.Morphologically, this species is related to L. manzanitae (NY14041), described by Thiers [77] and associated with Arbutus menziesii Pursh and Arctostaphyllus spp. in the coastal areas of California, USA.L. manzanitae is characterized by its large basidiomata, dark red, viscid, reticulate, and occasionally tomentose pileus surface, white context that changes to fuscous (especially when young), and the fact that it is never reddish.In addition, it is characterized by subellipsoid, fusoid, subcylindrical to inequilateral basidiospores of 13-17 × 4-5.5 µm, a trichoderm pileipellis that is composed of free, tangled hyphal tips, and elongated terminal cells that are often tapered and possess ochraceous contents.Another related species is L. monticola, which differs due to its sterile flap margin pileus; white with brown to black scabrous stipe; context that is white, changing to fuscous and blue-green; basidiospores that are 15-18.9× (3-) 4.9-5.6 µm; and its association with Comarostaphylis arbutoides Lindley in Costa Rica [31].
Description: pileus: 16-38 mm in diameter, plane-convex, citrus yellow (3A8), wet surface, hygrophanous when young, pale brown, ferrugineus to brown at maturity with straight to incurved margins.Hymenophore: adhered, pores of 1-2 mm in diameter, angular, yellow (3A8) when young, pale brown at maturity; tubes: 3-6 mm long, concolorous to the pores, unchanging when young, pale green at maturity.Context: 2-6 mm thick, whitish to pale yellow when cut, changing to pale green (28A3) at maturity; context of stipe: white to brown at base.Stipe: 25−30 × 6−7 mm, cylindrical to subclaviform, smooth to fibrillose at maturity, whitish to pale yellow when young, yellow and pale brown to brown at maturity.Basal mycelium: whitish.Odor: fungoid.Taste: pleasant.Chemical reaction: pileus, hymenium, and stipe turn pale orange (6A3), dark brown (6F8), and orange-brown (5B4), respectively, in KOH.The pileus and hymenophore turn brown in NH Notes: the phylogram (ITS-nrLSU-RPB2) shows that Xerocomus hygropanus is a new species whose sequences nested with a sequence of an unidentified Xerocomus sp.(MAN061) from Costa Rica [35].Xerocomus illudens is a sister species of X. hygrophanus with support of 0.96 BI/70ML.Additionally, the sequence named Xerocomus illudens (DD9854) described from North Carolina and Virginia, USA, is separated from the clade of X. illudens s.s., which indicates that X. illudens is probably a species complex [82], which requires further study.Morphologically, X. illudens and X. hygrophanus have similarities in their pale yellow coloration, but X. hygrophanus differs in the size of its basidiomata; its pale yellow to olive yellow hymenophore (which does not change color when cut); its white to pale yellow color (which changes to pale green when it is cut); its pileus context of (8−) 9−13 (15) × 3−5 µm; its elliptical to subfusiform basidiospores; and its ornamentation, which is barely visible on scanning electron microscopy.On the other hand, X. illudens has a coarsely reticulate stipe, extending from two-thirds to its entire length, of 10-12 (16) × 4-5 (6) µm, with ellipsoid to subfusiform basidiospores [83].

Discussion
In the last five years, a number of new species of Boletaceae from Mexico have been described, demonstrating the great and unexplored diversity of this fungal group in the country [2,39,40,48,52,84].Additionally, recently new genera have been described world-wide [13,27,34], for example, Cacaoporus [66], and others remain obscure, including genera belonging to the subfamily Leccinoideae [14,15].
The subfamily Leccinoideae currently includes hypogeous genera, such as Octaviania, Chamonixia, Rossbeevera, and Turmalinea; epigeous genera, such as Kaziboletus, Leccinum s.s.Leccinellum s.s., and Spongispora; and undefined Leccinum clades [12,85].On the basis of morphological, molecular, and phylogenetic analyses, Garcileccinum is proposed as a new genus in the subfamily Leccinoideae.Garcileccinum salmonicolor gen.nov.sp.nov., its type species, is distributed in Oaxaca, Mexico, occurring under Quercus rugosa.Meanwhile, G. viscosum and G. violaceotinctum have been recorded in Belize [17,18].Therefore, this genus is currently known only from Mesoamerica.Leccinum oaxacanum is found in the clade of Leccinum s.s.while Leccinum juarenzense is found in an undefined clade with L. talamancae Halling, L.D. Gómez and Lannoy.Our multi-locus analysis shows that Leccinum and Leccinellum are polyphyletic, as mentioned in previous works [15,16], and our phylogram shows undefined clades for some American sequences.
Pulchroboletus is a small genus, which currently includes only four species: P. begoniinus N.K. Zeng, Chang Xu and Zhi Q. Liang, described from China; P. roseoalbidus (Alessio and Littini) Gelardi, Vizzini, and Simonini, recorded in Italy; and P. rubricitrinus (Murrill) Farid and A.R. Franck and P. sclerotiorum M.E.Sm., Bessette and A.R. Bessette, described from the USA [86,87].In this work, we added P. neoregius, distributed in Costa Rica and Mexico, as a fifth species in the genus.
Tylopilus balloui (Peck) Singer is a complex of species with the holotype being described from New York, USA.It has also been recorded in Australia, Mexico, Thailand [24], India [86], and China [9].However, its phylogenetic analysis shows the complexity of the species since it has no basal support [9].Halling [24] mentioned that T. balloui is actually a polymorphic species and that a more detailed study is needed to determine if it is a cryptic group of related species.Several authors have mentioned that obligate ectomycorrhizal fungi such as the genus Tylopilus migrated along with their hosts [24,88]; therefore, the origins of Pinaceae [89] and Fagaceae [90] in Mexico share similarities to those of the Asian species.Halling [24] carried out a distribution analysis of the T. complex balloui and considered that Mexico shares more similarity with Asian, Australian, and Central American specimens than with those from the USA.Gelardi et al. [26] mentioned that several authors consider that T. balloui s.s. is distributed in the eastern USA and Mexico and that the sequences reported from Central America belong to T. leucomycelinus.In our work, we consider that T. leucomycelinus is distributed in the Dominican Republic, Belize, and Mexico [1,26,81].In Mexico, it is distributed in tropical Quercus forests, including Q. oleoides and Q. sapotifolia, while in Central America, it occurs in Pinus forests (including P. oocarpa, P. caribaea, and P. occidentalis) and, possibly, in Quercus forests.Mexico has a great diversity of Quercus, which includes more than 160 species, indicating, therefore, a great diversity of taxa within the Tylopilus balloui complex, which requires further study.Tylopilus pseudoleucomycelinus is distributed in Pinus-Quercus forests, forming a putatively ectomycorrhizal association with Pinus teocote and P. oaxacanus at 1737 to 1926 m.a.s.l.The hypothesis is that it is distributed from the Sierra Madre Oriental to the Coastal Plain of the Gulf of Mexico, according to the biogeographical classification proposed by Morrone [1,91].
Xerocomus s.l. has few sequences compared to the listed species in Index Fungorum [85].The genus requires extensive study to resolve the identity of many species [35].Our concatenated phylogram of nrLSU, ITS, and RPB2 (Figure 6) shows that X. hygrophorus belongs to the section Xerocomus s.s.[82].Xerocomus illudens is a sister species described from the USA [83] and Mexico.In Mexico, it possesses biocultural importance for the Otomi-Hñähñu native culture from Queretaro state, where it is referred to as the Ixka hyethe (yellow or sulfureous) mushroom [92].Xerocomus hygrophanus has been recorded from Mexico and Costa Rica (MAN2011-b-MAN061) [35].In Mexico, it is putatively associated with Quercus rugosa in mixed forests.Previously, only Xerocomus illudens and X. tenax [2] were described as from Mexico, and in this work, we added X. hygrophorus.

Figure 1 .
Figure 1.Phylogram of genus Boletellus with gene ITS, nrLSU, and RPB2.Maximum likelihood (ML)/Bayesian inference (BI) analyses were performed; the phylogram presented is the result of Bayesian inference.The bootstrap values (≥50%) and posterior probabilities (BI ≥ 0.90) are shown at the supported branches.Hemileccinum rugosum (HKAS84355) was used as the outgroup.The cluster where the new species is included is marked with the letter A. Newly generated sequences are indicated in bold.

Figure 1 .
Figure 1.Phylogram of genus Boletellus with gene ITS, nrLSU, and RPB2.Maximum likelihood (ML)/Bayesian inference (BI) analyses were performed; the phylogram presented is the result of Bayesian

Figure 2 .
Figure 2. Phylogram of genus Cacaoporus with RPB2 and ATP6.Maximum likelihood (ML)/Bayesian Inference (BI) analyses were performed; the phylogram presented is the result of Bayesian inference.The bootstrap values (≥50%) and posterior probabilities (BI ≥ 0.90) are shown at the supported branches.Cyanoboletus, Chalciporus africanus, Lanmaoa, and Rubinoboletus rubinus were used as the outgroup.Newly generated sequences are indicated in bold.

Figure 2 .
Figure 2. Phylogram of genus Cacaoporus with RPB2 and ATP6.Maximum likelihood (ML)/Bayesian Inference (BI) analyses were performed; the phylogram presented is the result of Bayesian inference.The bootstrap values (≥50%) and posterior probabilities (BI ≥ 0.90) are shown at the supported branches.Cyanoboletus, Chalciporus africanus, Lanmaoa, and Rubinoboletus rubinus were used as the outgroup.Newly generated sequences are indicated in bold.

Figure 4 .
Figure 4. Phylogram of Pulchroboletus genus with gene ITS and nrLSU.Maximum likelihood (ML)/Bayesian inference (BI) analyses were performed; the phylogram presented is the result of Bayesian inference.The bootstrap values (≥50%) and posterior probabilities (BI ≥ 0.90) are shown at the supported branches.Alessioporus ichnusanus (MG549a) and A. rubriflavus (JLF2561) were used as the outgroup.Newly generated sequences are indicated in bold.

Figure 4 .
Figure 4. Phylogram of Pulchroboletus genus with gene ITS and nrLSU.Maximum likelihood (ML)/Bayesian inference (BI) analyses were performed; the phylogram presented is the result of Bayesian inference.The bootstrap values (≥50%) and posterior probabilities (BI ≥ 0.90) are shown at the supported branches.Alessioporus ichnusanus (MG549a) and A. rubriflavus (JLF2561) were used as the outgroup.Newly generated sequences are indicated in bold.

Figure 5 .
Figure 5. Phylogram of Tylopilus with ITS and nrLSU.Maximum likelihood (ML)/Bayesian inference (BI) analyses were performed.The phylogram presents Bayesian inference, bootstrap values (≥50%), and posterior probabilities (BI ≥ 0.90) at the supported branches.Xanthoconium was used as the outgroup.Newly generated sequences are indicated in bold.The symbol * shows sequences of T. leucomycelinus generated in this work as a new record for Mexico.

Figure 14 .
Figure 14.Leccinum juarenzense (MEXU HO, holotype): (A) basidiospores; (B) basidia; (C) cheilocystidia; (D) pleurocystidia; (E) terminal elements of the pileipellis; and (F) caulocystidia.Scale bars: 10 µm.Habitat and distribution: solitary, occurring in Quercus sp. and Quercus-Pinus forests, in October, at 3000 m.a.s.l.Currently only identified in Oaxaca, Mexico.Notes: Leccinum juarenzense is characterized by its very rugulose pileus (which is grayish magenta to grayish brown and purplish gray at the center), its whitish hymenophore (when touched, it turns vinaceous), its whitish context (when cut, it changes to pale red to brown vinaceous), and its turquoise base stipe.In addition, its basidiospores are (11-) 13-18 (20) × 5-6 (-7) µm and elliptical to fusiform, and its pileipellis is composed of an epithelium containing hyphae that are globose, subglobose, cylindrical, clavate, and, in some cases, isodiametric.Phylogenetically, L. juarenzense is the sister to L. talamancae described from a Quercus forest, from Costa Rica by[28].L. juarenzense differs due to its brownish gray and cocoa brown to dark reddish brown pileus; its tomentose, pitted, and rugose pileus surface when young; its areolate pileus surface when mature; its white context (which changes slowly to pink to reddish orange); its dark blue base stipe; its basidiospores of 17.5-22.4× 4.9-6.3µm (these are subfusoid to ellipsoid); and its pileipellis, which is composed of an epithelium containing subisodiametric to spherical cells (in rare cases, these are subcylindric) and hyaline to brown pigment.L. juarenzense can be confused

Table 3 .
Specimens and sequences of Leccinoideae subfamily used for the molecular phylogenetic analyses in Figure3.Sequences newly generated for this study are highlighted in bold.

Table 4 .
Specimens and sequences used for the molecular phylogenetic analyses in Figure4.Sequences newly generated for this study are highlighted in bold.

Table 7 .
Comparative morphology of species in Tylopilus complex balloui.