Multigene Phylogeny and Morphology Reveal Unexpectedly High Number of New Species of Cantharellus Subgenus Parvocantharellus (Hydnaceae, Cantharellales) in China

The genus Cantharellus, commonly known as chanterelles, has recently been divided into six subgenera; however, wider sampling approaches are needed to clarify the relationships within and between these groups. A phylogenetic overview of Cantharellus subgenus Parvocantharellus in China was inferred based on the large subunit nuclear ribosomal RNA gene (nrLSU), the DNA-directed RNA polymerase II subunit 2 (rpb2), and the transcription elongation factor 1-alpha (tef1). A total of nine species from China were assigned to the subgenus, including seven novel species, namely Cantharellus aurantinus, C. austrosinensis, C. galbanus, C. luteolus, C. luteovirens, C. minioalbus, and C. sinominior, and two known species, namely C. albus and C. zangii. The detailed descriptions and illustrations were provided based on the newly obtained data, with the comparisons to closely related species. C. zangii was restudied based on the paratype specimens and multiple new collections from the type locality. Futhermore, the Indian species C. sikkimensis was identified as a synonym of C. zangii based on the morphological and molecular analyses. A key to the Chinese species belonging to the subg. Parvocantharellus is also provided.


Introduction
Cantharellus Adans. ex Fr., typified by C. cibarius Fr., is an important genus of wild edible mushrooms and is renowned for its high culinary, economic, and ecological value. Chantarelles have a global distribution and are especially rich in subtropical to tropical zones, demonstrating ectomycorrhizal associations with a variety of plants [1]. Approximately 300 species have been estimated to exist worldwide, and nearly 180 species have been described thus far [1,2]. Cantharellus species possess a colourful pileus, nearly smooth to evidently lamellate-folded hymenophore with variously forked or anastomosing veins, and smooth basidiospores [1, 3,4]. It is easy to recognise Cantharellus species at the genus level in the field solely based on their morphological features. However, the determination of their taxonomic positions at the species level is extremely complicated owing to overlaps in phenotypic variation among species. With the development of molecular biology, molecular-based studies have provided a basis for species identification and taxonomic development, especially for the molecular review of some type specimens and for re-classification of some old species based on new collections, so as to make species recognition more effective and accurate [5][6][7][8][9][10]. Molecular phylogenetic studies have delimited several species and revealed an unexpected species diversity. The tef 1 gene has been identified as a suitable DNA barcoding marker to determine terminal relationships among closely related Cantharellus species [1, 6,[11][12][13][14]. Phylogenetic analyses were performed following the methods that were described by Zhang et al. [42]. A maximum likelihood (ML) analysis was performed using RAxML v.7.2.6 [43] and Bayesian inference (BI) was performed using MrBayes 3.1.2 [44]. For both the ML and BI analyses, the most suitable substitution model for each gene partition was determined based on the Akaike Information Criterion (AIC) using MrModeltest v2.3 [45]. The default parameters were included for the ML analysis, except for selecting GTRGAMMAI as the model, and the statistical data were obtained by performing rapid non-parametric bootstrapping with 1000 replicates. A BI analysis using four chains was conducted using 30 million generations and the stoprul command with stopval set to 0.01. Bayesian trees were sampled every 100 generations, the first 25% of the generations were discarded as a burn-in, and the Bayesian posterior probabilities (BPP) were calculated from the posterior distribution of the retained Bayesian trees. The bootstrap support (BS) of ≥50% in the ML tree and BPP of ≥0.90 indicated statistical significance. The phylogenetic trees were visualised using FigTree v1.4.23.

Molecular Phylogeny
In the concatenated dataset (LSU + tef 1 + rpb2), 304 sequences (108 for LSU, 101 for tef 1, and 95 for rpb2) from 114 fungal collections were included. The alignment length was 3135 characters including gaps (1536 characters for LSU, 723 characters for tef 1, and 876 characters for rpb2), of which 1847 characters were conserved, 226 were variable and parsimony-uninformative, and 1062 were parsimony-informative. Craterellus cornucopioides (L.) Pers. and Cr. tubaeformis were selected as outgroups based on recent studies [1, 2,46]. The best models for the BI analysis of the concatenated dataset were GTR + I + G for LSU, K2P + I for tef 1, and GTR + I for rpb2, respectively. The ML analysis resulted in a similar topology to the Bayesian analysis, and only the ML topology has been depicted in Figure 1. The phylogenetic tree of the representative species of Cantharellus that was inferred from a multigene (LSU + tef1 + rpb2) dataset by means of both ML and BI methods. Craterellus cornucopioides BB 07_293 and Cr. tubaeformis AFTOL ID_286 were used as outgroups. The maximum likelihood tree is depicted. The bootstrap supports (BS ≥ 50%) and Bayesian posterior probabilities (BPP ≥ 0.90) are shown on the supported branches. The species generated in this study are in black bold. Phylogenetic analyses that were based on the multi-locus dataset (LSU + tef 1 + rpb2) showed that C. subg. Parvocantharellus formed a distinct clade in the genus Cantharellus, and seven new well-supported lineages were nested in this subgenus. Lineage I formed a wellsupported terminal clade (100% BS and 1.00 BPP) and was closely related to C. appalachiensis R.H. Petersen and C. tabernensis Feib. & Cibula. Lineages II, III, and IV formed three isolated terminal branches with robust evidence (100% BS and 1.00 BPP). Lineage V formed sister relationships with C. parvoflvus M. Herrera, Bandala, & Montoya and C. minor Peck, and was also closely related to C. romagnesianus Eyssart. & Buyck. Lineage VI formed a sister relationship with C. albus S.P. Jian & B. Feng. Finally, Lineage VII was closely related to C. himalayensis D. Kumari, Ram. Upadhyay & Mod.S. Reddy and C. curvatus Buyck, R. Ryoo & Antonín. In addition, two known species, C. albus and C. zangii, that were originally reported from China, were strongly supported (100% BS and 1.00 BPP) in the phylogenetic trees, but a sequence named C. sikkimensis K. Das, Buyck, D. Chakr., Baghela, S.K. Singh & V. Hofst. was clustered with C. zangii in the multi-locus phylogenetic tree.
In the tef 1 dataset, 74 sequences from the 18 species were selected for the phylogenetic analyses. The length of the dataset was 706 characters including gaps, of which 448 characters were conserved, 22 were variable and parsimony-uninformative, and 236 were parsimony informative. Cantharellus cinnabarinus (Schwein.) Schwein. was selected as the outgroup based on the above multi-locus analyses. K2P + G4 was selected as the best model for BI. The ML and Bayesian analyses produced similar estimates of tree topologies, and only the tree that was inferred from the ML analysis is displayed ( Figure 2). Species in the C. subg. Parvocantharellus formed three main subclades but without significant support. The seven new lineages were also strongly revealed in the phylogenetic tree and generated similar results with the multi-locus phylogenetic analysis.   Basidiomata small-sized. Pileus 18-42 mm broad, convex when young, then gradually to nearly applanate with a central shallow depression or broadly infundibuliform at maturity; surface dry, with appressed fibrillose squamules, white to yellowish white (4A1-4A2); margin wavy, incurved when young, decurved to slightly upturned at maturity, slightly changing to yellowish when handled, yellowish white to pale orange when dried (4A2-5A2, 4A3-5A3). Context white, 1-3 mm thick in the center of the pileus, sharply attenuate towards margin, unchanging or slightly changing to yellowish when cut. Hymenophore decurrent, close, poorly developed, composed of bifurcate and strongly interconnected low veins, usually less than 1 mm high, white to yellowish white, unchanging or slightly changing to yellowish when bruised. Stipe 20-60 × 2-8 mm, central, cylindrical or slightly tapering towards base, solid, smooth or with faintly scaly, concolourous with pileus, but in the lower part usually yellowish, slightly changing to yellowish when handled. All parts of basidioma becoming yellowish with 5% KOH solution. Odour milk fragrance, pleasant. Taste a little spicy. Basidiomata small-sized. Pileus 18-42 mm broad, convex when young, then gradually to nearly applanate with a central shallow depression or broadly infundibuliform at maturity; surface dry, with appressed fibrillose squamules, white to yellowish white (4A1-4A2); margin wavy, incurved when young, decurved to slightly upturned at maturity, slightly changing to yellowish when handled, yellowish white to pale orange when dried (4A2-5A2, 4A3-5A3). Context white, 1-3 mm thick in the center of the pileus, sharply attenuate towards margin, unchanging or slightly changing to yellowish when cut. Hymenophore decurrent, close, poorly developed, composed of bifurcate and strongly interconnected low veins, usually less than 1 mm high, white to yellowish white, unchanging or slightly changing to yellowish when bruised. Stipe 20-60 × 2-8 mm, central, cylindrical or slightly tapering towards base, solid, smooth or with faintly scaly, concolourous with pileus, but in the lower part usually yellowish, slightly changing to yellowish when handled. All parts of basidioma becoming yellowish with 5% KOH solution. Odour milk fragrance, pleasant. Taste a little spicy.  Notes-Cantharellus albus was recently described from southwest China [2] and exhibits small white basidiomata and slightly changes to a yellowish colour when it is bruised or treated with 5% KOH solution. They have poorly-developed gill-like folds with strongly bifurcate and interconnected low veins, a distinct creamy aroma and a slightly spicy taste. They have a white-coloured basidiomata that changes to yellowish-white to pale orange colour when it is dried, and have broadly ellipsoid to subglobose basidiospores. The distinct morphological characteristics and the well-supported monophyletic lineage render it easily distinguished from other the Cantharellus species. In the present study, C. albus was redescribed based on the specimens that were from Guangdong province, which were compared to the description of C. albus in Jian et al. [2], and the macro-and micro-characteristics were almost identical. However, the size of the basidiospores in Jian's specimens [6-8 × 5-7 µm, L m × W m = 6.9 (±0.48) × 5.92 (±0.62) µm] were larger than those in our specimens [5.5-7.5 The minor difference in the size of the basidiospores in Jian et al. [2] and the present study could be explained by small quantitative differences between the geographically distant populations or the number of measured basidiospores; this has often been noted in other Etymology-refers to the greyish-orange pileus colour. Diagnosis-This species is characterized by its small basidiomata, light orange pileus, relatively well-developed hymenophore, and broadly ellipsoid basidiospores (6.5)7-9 × (4.5)5-6 µm in size.
Basidiomata small-sized. Pileus 15-40 mm broad, convex when young, then gradually to nearly applanate with a central shallow depression at maturity; surface dry, smooth, light yellow, light orange, greyish yellow to greyish orange (2A5-6A5, 2B5-6B5), margin even, incurved when young, decurved to slightly upturned at maturity, unchanging when handled. Context white to yellowish white, 1.5-2.5 mm thick in the center of the pileus, sharply attenuate towards margin, unchanging when exposed. Hymenophore decurrent, relatively well developed, composed of bifurcate and interconnected low veins, in particular toward the cap margin, usually less than 1 mm high, pale yellow (2A3-4A3), unchanging when bruised. Stipe 20-40 × 8-12 mm, central, cylindrical or slightly tapering towards base, solid, smooth or with faintly scaly, pale yellow to pale orange (2A3-5A3), unchanging when handled. Odour not distinct.  Notes-The distinctive morphological features of C. aurantinus are the light orange to greyish-orange pileus, the pale yellow, gill-like folds with bifurcate and interconnected low veins, the broadly ellipsoid basidiospore, and the thin-walled hyphae of the pileipellis. The phylogenetic analyses supported C. aurantinus as an isolated lineage (Lineage VII) that is closely related to C. curvatus and C. himalayensis. However, C. curvatus, recently reported from South Korea, differs by its small and slender basidiomata, dull yellow to orangishyellow pileus, and shorter basidia (42-55 × 9.5-12 µm) [33]. Cantharellus himalayensis, that is reported from India, differs by its large basidiomata, yellowish pileus with pecan-brown scales at the center, relatively small basidiospores (6-8 × 4.5-6 µm), and partially gelatinous pileipellis [38].
In the field, C. aurantinus is easily misidentified as C. cibarius, as both species share a yellow-orange pileus. However, C. cibarius belongs to the subg. Cantharellus, and differs by its relatively large basidiomata, well-developed hymenophore of up to 3 mm in depth, longer basidia (80-105 × 7-9 µm) and thick-walled pileipellis hyphae [9,18]. Diagnosis-This species is characterized by its small basidiomata, pastel yellow to greyish-yellow pileus with a greyish-orange to brownish-orange center, pale yellow to light yellow hymenophore that is composed of bifurcate and interconnected low veins, elliptical to broadly elliptical basidiospores 6-8 × 4.8-6 µm, and the thin-walled hyphae of the pileipellis.
Basidiomata small-sized. Pileus 12-30 mm broad, applanate with center depressed, not perforate, margin incurved when young, applanate or slightly reflexed with age, obscure striated on surface; subfleshy to slightly membranous; surface dry, glabrous or tomentosus at central, pastel yellow, light yellow to greyish yellow at mass (3A5-5A5, 3B5-4B5), with a greyish orange to brownish orange center (5B5-6B5, 5C5-6C5), often with reddish brown tinge in some specimens (8D7-9D7). Context thin, 0.5-1.5 mm thick in the center of pileus, fibrous, pale yellow to light yellow (3A3-3A5), unchanging when bruised. Hymenophore decurrent, but with a clearly delimitation from the stipe surface, gill-like, well or poorly developed, ridges 1-2 mm high, composed of bifurcate and interconnected low veins, pale yellow to light yellow (3A3-4A3, 3A5-4A5), unchanging when bruised. Stipe 10-40 mm long, 2-5 mm thick, subcylindrical, enlarged downward, smooth or with faintly scaly, hollow, concolourous with pileus, darker and more somber than lamellae. Odour not special. Taste mild. 45(1.6), Q m = 1.36 ± 0.097, elliptical to broadly elliptical. Basidia 50-55 × 7-9 µm, clavate, with 5-6(-7) sterigmata. Pileipellis a cutis, composed of interwoven hyphae 5-12 µm in diam., colourless, thin-walled. Hymenophoral trama composed of cylindrical hyphae 7-10 µm in diam. Stipitipellis a cutis of cylindrical, parallel hyphae, 4-12 µm wide, branched, septate. Clamp connections abundant in all tissues. Notes-The presence of small basidiomata, a pastel yellow to greyish-yellow pileus with a greyish-orange to brownish-orange centre, thin-walled hyphae, and abundant clamp connections enable the classification and placement of C. austrosinensis in the subg. Notes-The presence of small basidiomata, a pastel yellow to greyish-yellow pileus with a greyish-orange to brownish-orange centre, thin-walled hyphae, and abundant clamp connections enable the classification and placement of C. austrosinensis in the subg. Parvocantharellus. In the phylogenetic trees, the new species was closely related to C. appalachiensis, C. koreanus Buyck, Antonín & Ryoo, and C. tabernensis. However, C. appalachiensis and C. tabernensis, that are both described from North America, can be distinguished by their relatively large and more robust basidiomata, with a pileus that is usually up to 50 mm in width. Additionally, C. appalachiensis differs in the existence of its drab yellow to dull brown pileus, its relatively large and narrow basidiospores (6.6-8.9 × 4.4-5.9 µm or 6-10.5 × 4-6 µm), and its association with oaks and other hardwoods [47,48]. Cantharellus tabernensis also differs in the presence of its dull orange-yellow to yellowish-brown pileus, vivid orange-yellow hymenophore, its well-developed gills that are up to 3 mm in depth, vivid orange-yellow stipe that is up to 8 mm in diameter, narrow basidiospores with a large Q value (1.49-1.52), and narrow hymenophoral trama hyphae (3-6 µm in diameter) [49]. Cantharellus koreanus, originally described from the temperate region of the Republic of Korea, differs in the presence of its dirty yellow-brown to pale brown pileus with a brown to dark brown centre, yellow to greyish stipe, relatively narrow basidiospores (4.2-5.5 µm in breadth), and its association with various deciduous trees (Carpinus laxiflora, Castanea crenata, and Quercus mongolica) mixed with coniferous trees (Pinus densiflora) [16]. In contrast, C. austrosinensis is distributed in subtropical regions of China and is currently only known to be associated with Pinus massoniana.
Basidiospores ( Notes-Cantharellus luteovirens is characterised by the presence of its small pastel yellow to yellowish-orange basidiomata, poorly-developed hymenophore with bifurcate and strongly interconnected low veins, broadly ellipsoid to subglobose basidiospores, and thin-walled pileipellis hyphae. These traits enable the classification and placement of C. luteovirens into the subg. Parvocantharellus. The molecular phylogenetic analyses showed that all of the C. luteovirens specimens formed a distinct lineage (Lineage III) close to C. minioalbus (Lineage II). However, C. luteovirens is morphologically different from C. minioalbus by its pastel yellow to yellowish-orange basidiomata, poorly-developed hymenophore, and relatively large basidiospores.
Cantharellus galbanus is extremely morphologically similar to C. luteovirens. However, C. galbanus differs by its small basidiomata, greenish pileus colour, relatively distant and well-developed hymenophore, and tropical distribution. In the phylogenetic trees, the two species formed two distinct monophyletic taxa and could easily be separated from each other owing to their branch lengths.
Additionally, C. koreanus, C. minor, and C. tabernensis are also morphologically similar to C. luteovirens, owing to the presence of small basidiomata and a yellowish to orange tinct pileus. However, the former three species can be distinguished from C. luteovirens in the field by their relatively distant and well-developed hymenophores. Additionally, C. koreanus differs by the presence of its dirty yellowish-brownish to pale brown pileus, with a brown centre, and relatively large but narrow basidiospores [6-8(-9) × 4.2-5.5(-6.5) μm] [16]. Cantharellus minor differs by the presence of its egg-yellow to orange pileus, long stipe, and large basidiospores 8-11 × 5-7 μm [51][52][53]. Cantharellus tabernensis differs by the presence of its dull orange-yellow to yellowish-brown pileus, vivid orange-yellow hymenophore and stipe, relatively large basidiospores (6-9 × 4-6 μm), and small basidia (35-55 × 5-8 μm) [49]. MycoBank: MB840654 Etymology-refers to the small white basidiomata. Diagnosis-This species can be easily distinguished from others in Cantharellus by its small and white-coloured basidiomata, gill-like hymenophore that is well-developed with   Basidiomata small-sized. Pileus 3-10 mm broad, convex when young, then grad to nearly applanate with a central shallow depression or finally broadly infundibuli at maturity; surface dry, with appressed fibrillose scales, white to yellowish white, m wavy, incurved when young, decurved to slightly upturned with maturity, unchan when handled. Context yellowish white, unchanging when exposed. Hymenophor current, but with a clearly delimitation from the stipe surface, distant, with well-de gill-like folds, relatively well developed, frequently forking towards pileus margin, lower irregular anastomosis amongst the folds, white to pale yellow, unchanging w bruised. Stipe 15-30 × 1.5-2.5 mm, central, cylindrical or slightly inflated towards solid, smooth or faintly scaly, concolourous with pileus, unchanging when han Odour not distinct.
Basidiospores ( Notes-Cantharellus minioalbus is characterised by the presence of its small white basidiomata, broadly infundibuliform pileus that is covered with fibrillose scales, distant and well-defined gill-like hymenophore that are frequently forking towards the pileus margin, with the existence of few abnormal anastomosis among the folds, broadly ellipsoid to subglobose basidiospores, and thin-walled hyphae of pileipellis. These traits enable the classification and placement of C. minioalbus into the subg. Parvocantharellus. The molecular phylogenetic analyses showed that the new species formed an isolated lineage in the subg. Parvocantharellus and was genetically distinct from all of the other Cantharellus taxa with sequence data.
Cantharellus albus is similar to C. minioalbus, as both species share white basidiomata. However, C. albus, redescribed above, differs by the alteration of the basidiomata colour from white to yellow when it is bruised, and by the presence of a poorly-developed hymenophore with variously forked or strongly anastomosing veins, and relatively large basidiospores [5.5-7.5  Etymology-refers to the species described from China and is similar to C. minor.
Diagnosis-This species is characterized by its small and light yellow basidiomata with a relatively longer stipe, the stipe is usually longer than the diameter of the pileus. It has well-developed gill-like ridges that are mostly forked at the margin, as well as elliptical to elongate elliptical basidiospores, and thin-walled hyphae of the pileipellis.
Habitat and distribution-Growing in groups or gregariously under mixed forests that are dominated by Keteleeria sp. and Picea sp. Currently known from southwest China.
Notes-Cantharellus sinominor is one of the most commonly documented Cantharellus species in subtropical coniferous forests in southwest China, and it can be found in local wild edible mushroom markets. In the field, C. sinominor is easily confused with C. minor, a species with small yellow basidiomata. The molecular phylogenetic analyses showed that they are closely related, but independent species. Cantharellus minor differs by the presence of its glabrous, bright yellow-orange to orange pileus, that is usually fading to pale orange-buff or pale orange, and its relatively large basidiospores (8-11 × 5-7 µm) [51][52][53]. Additionally, C. minor is reportedly associated with oaks and other hardwoods, whereas C. sinominor is associated with coniferous trees.

Discussion
In our multi-locus phylogenetic analyses, the ingroup sequences resulted in the formation of six main subgenera that is largely consistent with the most recent phylogenetic studies [1, 2,8,9,26,34,46]. Thus, we adopted the treatment of Buyck et al.
[1] and treated C. subg. Parvocantharellus as a monophyletic group, sister to the subg. Cinnabarinus. Apart from C. aff. subcyanoxanthus, the species of the subg. Parvocantharellus formed a wellsupported (93% BS and 0.99 BPP) clade in the multi-locus phylogenetic tree. A total of nine species from China nested into this well-supported clade, including the seven new species that are described above: C. aurantinus, C. austrosinensis, C. galbanus, C. luteolus, C. luteovirens, C. minioalbus, and C. sinominor, and two previously reported species C. albus and C. zangii. In the phylogenetic analyses of the tef 1 dataset, species in the subg. Parvocantharellus formed similar interspecific relationships in the multi-locus dataset, the seven new species were also well-supported, and further proved that the tef 1 gene is suitable to determine the interspecific relationships for most species in Cantharellus.
In this study, nine species were discovered from China, and they all belong to the C. section Flavobrunnei, which is characterized by the presence of medium-sized to extremely small basidiomata, a yellowish to brownish pileus, a long stipe, and abundant clamp connections. Phylogenetic analyses in our study all support the sect. Flavobrunnei as a monophyletic subclade in the subg. Parvocantharellus. In addition to the nine species that were introduced above, C. appalachensis and C. minor belonging to the subg. Parvocantharellus have also been reported in China [47,[54][55][56]. C. appalachensis was proposed to demonstrate a geographically disjunct distribution from southeastern North America to eastern Asia [47]. However, only the LSU sequences of C. appalachensis that were derived from the Chinese samples were available, and these were not included in the phylogenetic analyses owing to the markedly low levels of genetic variation and the challenges that were encountered in distinguishing them among most of the species [57]. Thus, to determine the distribution of C. appalachensis in China, additional useful gene sequences from more samples are warranted. C. minor is considered a broad species and is widely distributed in most parts of China [54][55][56], including northeast, central, southern, and southwest China. Unfortunately, there were no specimen vouchers that were available for these records. The name "Cantharellus minor" is a collective name and has been misapplied to almost any small yellow Cantharellus species in China. The four new species C. austrosinensis, C. galbanus, C. luteovirens, and C. sinominor are easily misidentified as C. minor based on their morphological characteristics. However, phylogenetic analyses indicated that they represented five distinct species and C. minor, originally reported from North America under oaks and other hardwoods, can be distinguished by the presence of its large basidiospores (8-11 × 5-7 µm) [51][52][53]. In the present study, no samples of C. minor from China were detected in the phylogenetic analyses, and further studies with extensive sampling are warranted to determine the distribution of the species in China. The distribution of C. minor may be similar to that of C. cibarius in China. A recent study has shown that the distribution of C. cibarius is limited to northeast China, and the popular edible mushroom that is marketed in Yunnan, Guizhou, and Sichuan Provinces is, in fact, the native species C. yunnanensis W.F. Chiu [18].
Geographically, the species that are in the C. subg. Parvocantharellus are mainly distributed in the northern hemisphere and are especially diverse in Asia [16,35,38]. Except for C. zangii which has been reported in the subalpine regions of China and India, the remaining eight species were all reported in the subtropical and tropical regions of southern China, revealing an unexpectedly large number of new Cantharellus species in China, with a considerable number of species remaining to be discovered. Remarkably, in the phylogenetic trees, specimens that were from the northern hemisphere were clustered together in the well-supported sect. Flavobrunnei of subg. Parvocantharellus, representing a distinct northern hemisphere distribution clade. Meanwhile, three species, namely C. avellaneus, C. congolensis, and C. subcyanoxanthus, from the southern hemisphere formed the basal and sub-basal branches in the subgenus. C. avellaneus and C. congolensis, which belong to the sect. Congolenses Heinem., formed an isolated sub-basal clade in the subgenus and were characterized by their strongly blackening context and strong reaction with most macrochemical reagents. These distinct morphological characteristics make the species in this section easily distinguishable from Flavobrunnei, demonstrating a unique tropical African geographic distribution clade. C. subcyanoxanthus, which is characterized by its strong blue-violet-lilac to vinaceous basidiomata and yellow context, formed a monospecific sect. Cyanomaculati Buyck & V. Hofstetter, was located in the basal clade of subg. Parvocantharellus, but without significant support in the multi-locus phylogenetic tree. Based on molecular correlations, Buyck et al.
[1] roughly divided the two sections in the subg. Parvocantharellus, but their morphological characteristics are not well-matched with the definition of the subg. Parvocantharellus. Thus, their distinct morphological characteristics and relatively independent phylogenetic positions may result in their assignment to a new subgenera level in the future as more related species continue to be discovered.  Data Availability Statement: In Publicly available datasets were analyzed in this study. This data can be found here: [https://www.ncbi.nlm.nih.gov/; https://www.mycobank.org/; https://www. treebase.org/treebase-web/home.html, accessed on 28 July 2021].