Fungal Diversity

, Volume 58, Issue 1, pp 281–298

Molecular evidence for novel Cantharellus (Cantharellales, Basidiomycota) from tropical African miombo woodland and a key to all tropical African chanterelles

Authors

    • Département Systématique et évolution, UMR7205Muséum National d’Histoire Naturelle
  • Frank Kauff
    • Fachbereich Biologie, Abt. Molekulare PhylogenetikTechnische Universität Kaiserslautern
  • Corinne Cruaud
    • Centre national de séquençageGenoscope
  • Valérie Hofstetter
    • Department of plant protectionAgroscope Changins-Wädenswil Research Station ACW
Article

DOI: 10.1007/s13225-012-0215-4

Cite this article as:
Buyck, B., Kauff, F., Cruaud, C. et al. Fungal Diversity (2013) 58: 281. doi:10.1007/s13225-012-0215-4

Abstract

The authors present a combined morphological and molecular approach of the genus Cantharellus in Africa. Morphological descriptions and detailed illustrations are provided for five new species from the Zambezian savannah woodlands in tropical Africa: C. afrocibarius, C. gracilis, C. humidicolus, C. miomboensis and C. tanzanicus. A maximum likelihood analysis of tef-1 sequences obtained for 83 collections of Cantharellus that are representative of all major groups in world wide Cantharellus, places a total of 13 African chanterelles, including the five newly described taxa. The recognition of a separate genus Afrocantharellus is rejected. An identification key based on the re-examination of all existing type material is provided for all presently known African Cantharellus.

Keywords

AfrocantharellusBiodiversityGuineocongolian rain forestIdentification keyPhylogenyTanzaniaZambezian woodlands

Introduction

Chanterelles are among the most common edible mushrooms for the local peoples all over tropical Africa, particularly in the Zambezian savannah woodlands, also often named ‘miombo’ woodlands, referring to the Swahili name for Brachystegia, one of the dominant tree genera in these woodlands. During the rainy season, many different species are offered for sale on nearly every market and along road sides, sometimes in impressive quantities (Buyck 1994). Yet, many African Cantharellus—even among the very commonly marketed ones—still remain poorly known or undescribed. This paper presents several new taxa from the African miombo woodlands. This area constitutes the most important vegetation type in the Zambezian region and is characterized by a high degree of endemism (54 % of the 8,500 species of phanerogames). The miombo woodlands cover 2.6 million km2 south of the equator and are distributed over eight countries (the major part of Angola, Southern Shaba in the Democratic Republic of Congo, the south-eastern and southern parts of Malawi and Burundi and a the larger part of Mozambique, Zambia and Zimbabwe).

There have been several earlier taxonomic studies dealing with Cantharellus in the African miombo. The first important revision of the miombo chanterelles was by Heinemann (1966), who described many new taxa based on specimens gathered by resident Belgian expatriates in the Southern Shaba area of the Democratic Republic of Congo and sent to the Botanical Garden in Brussels for study. That revision had clearly demonstrated the very different fungal assemblage that inhabits these savannah woodlands as compared to the neighbouring rain forest area (Heinemann 1958; De Kesel et al. 2011; Eyi Ndong et al. 2011). Since then, several new taxa were described from the miombo woodlands in Burundi (Buyck 1994), Tanzania (Tibuhwa et al. 2008; Buyck et al. 2000), Zimbabwe (Eyssartier et al. 2002) and Zambia (Eyssartier and Buyck 1998) resulting in a total of 22 known Cantharellus for the area, all solely based on morphological features. The limits of the few morphological features available for the recognition of well-defined taxa in Cantharellus were nevertheless clearly demonstrated by Buyck et al. (2000).

In a very recent molecular paper, Tibuhwa et al. (2012) recognized a new genus, Afrocantharellus, for some African Cantharellus with well-developed gill folds based on a phylogenetic analysis of rDNA LSU and concatenated LSU/5.8-ITS2/ATP6 data. The present paper is the only other molecular study on African Cantharellus, with five new species based on their morphology and a phylogenetic analysis of the translational elongation factor alpha-1 (tef-1).

This regional revision supplements the recently published papers on African (De Kesel et al. 2011; Buyck 2012; Buyck et al. 2012) and American chanterelles (Buyck et al. 2010, 2011; Buyck and Hofstetter 2011) in preparation of the forthcoming multigene phylogeny of worldwide Cantharellus, in which the systematic arrangement of Cantharellus will be discussed in more detail.

Materials and methods

Morphological data

All studied collections (Table 1) were gathered by the senior author and collaborators over the past 15 years and specimens are deposited at the mycological herbarium of the Paris Natural History Museum (PC). Color notations follow Kornerup and Wanscher (1978). Spacement of the hymenophore elements (veins or gills) distinguishes between ‘L’ (for radial elements running from cap margin all the way to the stipe surface) and ‘l’ (for shorter radial elements near the cap margin that never reach the stipe); their density can be expressed for either of these types separately, or for the total of all radial elements near the cap margin (as L + l/cm). Original drawings for all elements of the hymenium or pellis were made at a magnification of 2,400 × by B. Buyck with the aid of a camera lucida. All microscopic observations and measurements were made in ammoniacal Congo red after pretreatment with a 5 % aqueous KOH solution to improve tissue dissociation and matrix dissolution. Measurements of basidiospores cite length, width and length/width ratio (Q) in this format: (minimum–)mean minus standard deviation– mean value–mean plus standard deviation(− maximum measured); spore measurements are based on “n” spores. References to infrageneric placements follow the most recent morphology-based classification of the genus by Eyssartier and Buyck (2001b). Author names of sequenced taxa are listed in Table 1.
Table 1

Voucher table

Taxon

Voucher extraction nr/collector nr.

PC Herb. accession nr.

Origin

tef-1 Genbank accession nr

Genus Cantharellus

C. addaiensis

495/BB 98.033 neotype

PC0084717

Tanzania

JX192992

C. addaiensis Henn. 1898

267/BB 98.057

PC0084718

Tanzania

JX192976

C. afrocibarius

497/BB 96.236

PC0084125

Zambia

JX192994

C. afrocibarius Buyck & V.Hofst. 2012

496/BB 96.235 holotype

PC0084124

Zambia

JX192993

C. altipes

344/BB 07.162

PC0084090

USA

GQ914945

C. altipes Buyck & V.Hofst.2011

318/BB 07.019 holotype

PC0084085

USA

GQ914939

C. amethysteus

352/BB 07.309

PC0084071

Slovakia

GQ914954

C. amethysteus (Quél.) Quél.1888

349/BB 07.284

PC0084070

Slovakia

GQ914953

C. cibarius

479/GE 07.025

PC0084088

France

GQ914949

C. cibarius Fr. :Fr. 1821

351/BB 07.300

PC0084077

Slovakia

GQ914950

C. cinnabarinus

326/BB 07.053

PC0084093

USA

GQ914984

C. cinnabarinus (Schwein.) Schwein. 1834

312/BB 07.001 neotype

PC0084094

USA

GQ914985

C. congolensis

512/BB 98.058

PC0084776

Tanzania

JX192996

C. congolensis aff.

66/BB 06.176

PC0084078

Madagascar

JX192967

C. congolensis aff.

19/BB 06.166

PC0084979

Madagascar

JX192998

C. congolensis aff.

69/BB 06.197

PC0084076

Madagascar

GQ914982

C. congolensis Beeli 1928

511/BB 98.039

PC0084123

Tanzania

JX193015

C. decolorans aff.

57/BB 06.146

PC0084757

Madagascar

JX192964

C. decolorans aff.

466/BB 08.243

PC0084733

Madagascar

JX192987

C. decolorans aff.

56/BB 06.145

PC0084111

Madagascar

JX193003

C. decolorans aff.

20/BB 06.168

PC0124633

Madagascar

JX192999

C. decolorans aff.

471/BB 08.290

PC0084806

Madagascar

JX193001

C. decolorans aff.

472/BB 08.291

PC0084807

Madagascar

JX193002

C. decolorans aff.

467/BB 08.260

PC0084808

Madagascar

JX193000

C. decolorans aff.

61/BB 06.151

PC0124634

Madagascar

JX193004

C. decolorans Eyssart. & Buyck 1999

469/BB 08.278

PC0084098

Madagascar

GQ914968

C. densifolius Heinem. 1958

510/BB 98.013

PC0084126

Tanzania

JX193014

C. ferruginascens P.D.Orton 1969

348/BB 07.283

PC0084099

Slovakia

GQ914952

C. fistulosus Tibuhwa & Buyck 2008

517/DT 43 holotype

PC0084738

Tanzania

JX192997

C. gracilis Buyck & V.Hofst. 2012

251/BB 98.234 holotype

PC0084737

Tanzania

JX192970

C. humidicolus

493/BB 98.036-2

PC0084724

Tanzania

JX193006

C. humidicolus Buyck & V.Hofst. 2012

250/BB 98.036 holotype

PC0084724

Tanzania

JX193005

C. isabellinus var. parvisporus

256/BB 98.020 holotype

PC0084753

Tanzania

JX192972

C. isabellinus var. parvisporus Eyssart. & Buyck 2000

249/BB 98.037

PC0084100

Tanzania

GQ914966

C. lateritius

330/BB 07.058

PC0084105

USA

GQ914959

C. lateritius (Berk.) Singer 1949

320/BB 07.025 neotype

PC0084103

USA

GQ914957

C. lewisii

314/BB 07.003 holotype

PC0084074

USA

GQ914962

C. lewisii Buyck & V.Hofst. 2011

301/BB 02.197

PC0084073

USA

GQ914961

C. lilacinopruinatus Hermitte, Eyssart. & Poumarat 2005

347/BB 07.221

PC0084106

Slovakia

GQ914951

C. minor

329/BB 07.057

PC0084721

USA

JX192979

C. minor Peck 1872

313/BB 07.002

PC0084747

USA

JX192978

C. miomboensis Buyck & V. Hofst. 2012

255/BB 98.021 holotype

PC0084748

Tanzania

JX192971

C. platyphyllus

262/BB 98.126 epitype

PC0084723

Tanzania

JX192975

C. platyphyllus Heinem. 1966

259/BB 98.012

PC0084108

Tanzania

GQ914969

C. platyphyllus subsp. bojeriensis

459/BB 08.160

PC0084740

Madagascar

JX192984

C. platyphyllus subsp. bojeriensis

461/BB 08.164

PC0084804

Madagascar

JX193007

C. platyphyllus subsp. bojeriensis Eyssart. & Buyck 1999

458/BB 08.158

PC0084741

Madagascar

JX192983

C. pseudominimus Eyssart. & Buyck 1999

477/JV 00.663

PC0084725

Portugal

JX192991

C. quercophilus Buyck, Lewis, Eyssart. & V.Hofst. 2010

455 = 636/BB 07.097 holotype

PC0084726

USA

JX192981

C. sp. ined.

505/DT 62

s.n.

Tanzania

JX193012

C. sp. ined.

506/DT 72

s.n.

Tanzania

JX193013

C. sp. ined. 1

464/BB 08.210

PC0085130

Madagascar

JX193008

C. sp. ined. 2

462/BB 08.196 holotype

PC0084109

Madagascar

GQ914980

C. sp. ined. 2

463/BB 08.203

PC0084722

Madagascar

JX192985

C. sp. ined. 3

475/BB 08.336 holotype

PC0084754

Madagascar

JX192989

C. sp. ined. 4

460/BB 08.162 holotype

PC0084079

Madagascar

GQ914981

C. sp. ined. 4

465/BB 08.234

PC0084736

Madagascar

JX192986

C. sp. ined. 5

58/BB 06.147

PC0084110

Madagascar

JX193016

C. sp. ined. 5

67/BB 06.179 holotype

PC0084129

Madagascar

JX192968

C. sp. ined. 5

68/BB 06.180

PC0084130

Madagascar

JX192969

C. sp. ined. 5

70/BB 06.247

PC0085151

Madagascar

JX193017

C. sp. ined. 6

457/BB 08.070 holotype

PC0084754

Madagascar

JX192982

C. sp. ined. 7

474/BB 08.320 holotype

PC0084729

Madagascar

JX192988

C. subcyanoxanthus aff.

257/BB 98.014

PC0084745

Tanzania

JX192973

C. subcyanoxanthus Buyck, Randrianjohany & Essart 2012.

476/BB 00.1173 holotype

PC0084746

Madagascar

JX192990

C. subincarnatus

55/BB 06.096

PC0084755

Madagascar

JX192963

C. subincarnatus Eyssart. & Buyck 2001 aff.

13/BB 06.080

PC0084727

Madagascar

JX192962

C. symoensii

261/BB 98.113 epitype

PC0084756

Tanzania

JX192974

C. symoensii Heinem. 1966

260/BB 98.011

PC0084113

Tanzania

GQ914970

C. tabernensis

340/BB 07.119

PC0084116

USA

GQ914976

C. tabernensis Feibelman & Cibula 1996

328/BB 07.056 epitype

PC0084115

USA

GQ914974

C. tanzanicus aff.

17/BB 06.153

PC0084112

Madagascar

JX193009

C. tanzanicus aff.

59/BB 06.148

PC0084127

Madagascar

JX192965

C. tanzanicus aff.

60/BB 06.149

PC0084128

Madagascar

JX192966

C. tanzanicus aff.

62/BB 06.159

PC0085037

Madagascar

JX193010

C. tanzanicus aff.

63/BB 06.165

PC0084789

Madagascar

JX193011

C. tanzanicus Buyck & V.Hofst. 2012

268/BB 98.040 holotype

PC0084728

Tanzania

JX192977

C. tenuithrix

343/BB 07.125 holotype

PC0084084

USA

GQ914947

C. tenuithrix Buyck & V,Hofst. 2011

322/BB 07.035

PC0084087

USA

GQ914946

C. texensis

341/BB 07.120

PC0084096

USA

GQ914987

C. texensis Buyck & V.Hofst. 2011

317/BB 07.018 holotype

PC0084097

USA

GQ914988

C. tomentosus

500/BB 98.060 holotype

PC0084732

Tanzania

JX192995

C. tomentosus Eyssart. & Buyck 2000

248/BB 98.038

PC0084121

Tanzania

GQ914965

Genus Craterellus

Cr. tubaeformis

350/BB 07/293

PC0084122

Slovakia

GQ914989

Genus Hydnum

H. repandum

356/BB 07.341/MTS3757

PC0084749

Slovakia

JX192980

Taxon sampling, molecular data and phylogenetic analyses

The taxon sampling includes 85 collections (Table 1) comprising 83 Cantharellus and two outgroups (Craterellus and Hydnum). This dataset is representative of all major clades that were recognized in the molecular phylogeny of the genus Cantharellus based on a multilocus analysis (Buyck and Hofstetter 2008); it is also representative of all subgenera recognized in the latest morphological classification by Eyssartier and Buyck (2001b). DNA was isolated from fresh material stored in CTAB buffer or from dried fruit bodies following the protocol described in Hofstetter et al. (2002). Tef-1 amplification was conducted with the primers and conditions by Morehouse et al. (2003). When amplification of tef-1 gene generated multiple bands, PCR products were cloned with pSTBlue-1 AccepTor VectorTM Kit (Novagen). Amplification of four clones was sufficient to recover at least one PCR product of the expected 850–900 bp length for the partial tef-1 gene, secondary bands being much shorter (500–650 bp). Sequencing with the primers for amplification was performed by A. Couloux at the Genoscope or “Consortium National de Recherche en Génomique”. His co-authorship is part of the agreement No. 2005/67 on the project Macrophylogeny of Life (G. Lecointre, director) between the Genoscope and the service of molecular systematicss of the Muséum National d’Histoire Naturelle (CNRS IFR 101). With the exception of 29 tef-1 sequences already deposited in GenBank for previous papers on American chanterelles (Buyck et al. 2011), sequences were newly produced for this study. Sequences were assembled and edited with the software package Sequencher 3.0 (Gene Codes Corp., USA). A maximum likelihood (ML) analysis was conducted on our tef1, 85-specimen dataset in RAxML v. 7.0.3 (Stamatakis 2006), using a GTRmodel with a gamma distribution. Bootstrap proportions (MLbs) (Felsenstein 1985; Stamatakis et al. 2008) were estimated with 1,000 replicates, and frequencies >70 % were considered significant. We also conducted Bayesian analyses in MrBayes v3.1.1 (Huelsenbeck and Ronquist 2001), sampling every 500th tree for 10,000,000 generations, and implementing a GTR model of nucleotide substitution with an estimated proportion of invariable sites and a gamma distribution approximated by four categories. We conducted three independent runs to verify that all runs converged to the same log-likelihood stationary level. After discarding the burn-in, the last 10,000 trees of each run were pooled, and the resulting 30,000 trees were used to calculate a 50 %-majority rule consensus tree in order to obtain Bayesian posterior probabilities (PP).

Results

Molecular analyses

The alignment of the tef-1 sequences for 85 taxa, is available as online supplementary data (DATA SUPPLEMENT 1). The full alignment includes 1,025 characters. After exclusion of ambiguous regions and four spliceosomal introns, the dataset accounts for 633 bp. The ML tree with the highest likelihood value (lnL −5353.986075) searches is shown in Fig. 1. Tef-1 locus provided low support for basal relationships except for the monophyly of genus Cantharellus that received significant support (MLbs = 76 %). The tef-1 gene did, however, better resolve terminal relationships for the species newly described here (see gray boxes in Fig. 1).
https://static-content.springer.com/image/art%3A10.1007%2Fs13225-012-0215-4/MediaObjects/13225_2012_215_Fig1_HTML.gif
Fig. 1

ML best tree inferred from the analysis of tef-1 locus for 85 specimens. Branches that received bootstrap support ≥70 and PP ≥95 % are in bold. Only BS values are reported along the branches except for the branch in bold grey which is supported only by Bayesian analysis (MLbs/PP). Some branches are extremely short due to the high degree of sequence similarity. The specimens collected in the African miombo woodlands, the subject of this paper, are in bold red font, with gray boxes indicating the species newly described in this paper. Malagasy collections are indicated in black, including 7 still undescribed taxa. North American and European chanterelles are in green and blue respectively

The most basal clade in our phylogeny consists of Malagasy collections that most likely represent C. subincarnatus (MLbs = 100 %), a species still only known from the original description based on a few Central African collections. C. miomboensis sp. nov. and C. tanzanicus sp. nov. are part of the next basal clade which received weak support (MLbs = 64 %; PP < 95 %). Within this clade, C. miomboensis sp. nov. occupies the most basal position within a subclade that clusters, although without support, this new species with a still undescribed Malagasy species (C. ined.7) and three African species, C. addaiensis, C. isabellinus var. parvisporus and C. tomentosus. This subclade forms an unsupported monophyletic and sister relationship with a second subclade that clusters C. tanzanicus sp.nov. with three still undescribed Malagasy species (C. ined.4,5 and 6), the African C. densifolius and one other African species (C. sp. ined. which is not further discussed here because the specimens molded completely after collecting and have become useless for morphological examination and typification). C. tanzanicus sp.nov. is monophyletic with several Malagasy collections, labeled here as C. cf. tanzanicus, with full support (MLbs = 100 %). ML best tree did not resolve C. cf. tanzanicus collections 62 and 63 from C. tanzanicus sp. nov. but ML bootstrap analyses and Bayesian posterior probabilities both supported the monophyly of the five C. cf. tanzanicus collections (MLbs = 83 %, PP = 99 %) and consequently support the distinction between C. tanzanicus and C. cf. tanzanicus.

C. gracilis sp. nov. and C. humidicolus sp. nov. are both part of a significantly supported clade (MPbs = 81 %). Within this clade two still undescribed Malagasy species (C. ined. 3 & 4) are basal, while the new species C. gracilis, C. humidicolus and the Malagasy C ined. 2 are monophyletic (MLbs = 81 %) and more derived. C. gracilis is clearly separated (MLbs = 100 %) from the sister species C. humidicolus and C. ined. 2 (MLbs = 100 %).

The last novel species described in this study, C. afrocibarius sp. nov., belongs to a more derived subclade and is suggested to be monophyletic, however without support (MLbs = 64 %; PP < 95 %), and sister to C. fistulosus in our tree (MLbs = 100 %).

Morphological analyses

The new species are described below in alphabetical order.

C. afrocibarius Buyck & V.Hofst. sp. nov. — Mycobank MB 800901; Figs. 2–4
https://static-content.springer.com/image/art%3A10.1007%2Fs13225-012-0215-4/MediaObjects/13225_2012_215_Fig2_HTML.gif
Figs. 2–4

Cantharellus afrocibarius. 2. Spores. 3. Elements of the hymenium. 4. Terminal elements of the pileipellis. Scale bar = 5 μm for spores, 10 μm for the other elements. (drawings B.Buyck, from holotype)

Diagnosis: Apart from the very unusual caespitose habit, this species differs from C. cibarius in the absence of strongly thickened terminal cells at the cap surface.

Holotypus: Zambia. Copperbelt province, near Chibuli in Brachystegia woodland, 31 Jan. 1996, Buyck & Eyssartier leg., in Buyck 96.235 (PC0084124)

Fruit body very fleshy and firm, up to 180 mm diam. and more than 100 mm high, but often buried for an additional 50–70 mm in the soil, usually multipileate and when young entirely covered by a chalk-white bloom. Cap smooth, dull, with the centre remaining convex to plane, compared to C. cibarius of a darker, more brownish yellow (between 2A4 and 3A4–5) or toward corn yellow, amber yellow (4B5–6) when young, often remaining whitish for a long time in the cap center and near the extreme cap margin (when humid, the white color seems to be subcutaneous as from air-filled spaces); margin often irregular from the very beginning, rapidly strongly lobed and undulating, smooth, remaining inflexed at the extreme end, either remaining convex or becoming sometimes uplifted outside the centre but never infundibuliform. Hymenophore very strongly decurrent, often up to the very stipe base, composed of anastomosing veins and low, forking gill-folds, off-white to cream (concolorous with the stipe at first), then deeper yellow with ageing. Stipe thick and often with a larger part buried in the soil, often very large compared to a single cap, confluent and splitting upward giving rise to several fruit bodies, whitish, compact and hard. Context whitish, but in section marbled with a pale, brownish-gray, yellow underneath the pileipellis. Taste mild. Smell fruity. Spore print not obtained (probably yellow).

Spores ellipsoid to slightly reniform in side view, (7.9)8.5–9.2–9.8(10.2) × (5.0)5.2–5.5–5.9(6.5) μm, Q = (1.5)1.6–1.7–1.75(1.8), smooth. Basidia long and slender, mostly 70–90 × 6–7.5 μm, very narrowly clavulate to almost subcylindrical, irregularly undulate or constricted, (5)6–spored. Subhymenium filamentous, composed of narrow and long cells measuring 3–5 μm diam. Cystidia not differentiated. Pileipellis composed of quite regular and slender, thin- to slightly thick-walled and then refringent hyphal extremities, ca. 3–5 μm diam.; terminal cell often quite short, measuring (25)30–50(75) × 3–5(7) μm, subcylindrical or often slightly constricted subapically. Clamp connections abundant everywhere.

Other examined collection: Zambia. Copperbelt province, near Chibuli in Brachystegia woodland, 31 Jan. 1996, Buyck & Eyssartier leg., in Buyck 96.236 (PC0084125, paratypus).

Commentary: This appears to be a rare species, so far only known from both sequenced collections. Apart from the very unusual caespitose field aspect, this yellow species is characterized by omnipresence of clamps, rather long basidia, ellipsoid, slightly reniform spores and absence of strongly thick-walled extremities in the pileipellis. Other yellow chanterelles from the woodland area include C. defibulatus (Heinem.) Eyssart. & Buyck, which is easily separated by the absence of clamp connections, and some ill-known taxa that were described as varieties of C. cibarius (Heinemann 1966), lacking the caespitose habit.

C. fistulosus, which occupies an unsupported sister-relationship with our new species (Fig. 1), shares with the latter the presence of clamps, but is a very small, thin-fleshed, brownish yellow species with well-developed gill folds.

C. gracilis Buyck & V.Hofst., sp. nov. — Mycobank MB 800911; Figs. 5–7
https://static-content.springer.com/image/art%3A10.1007%2Fs13225-012-0215-4/MediaObjects/13225_2012_215_Fig3_HTML.gif
Figs. 5–7

Cantharellus gracilis. 5. Spores. 6. Elements of the hymenium. 7. Terminal elements of the pileipellis. Scale bar = 5 μm for spores, 10 μm for the other elements. (drawings B.Buyck, from holotype)

Diagnosis: This species differs from C. microcibarius by the absence of clamp connections, the longer stipe and the well-developed, not-anastomosing gill folds.

Holotypus: Tanzania. Dar-es-salaam, on a hill slope near the university campus, in a highly secundary patch of woodland dominated by Brachystegia utilis, B. microphylla and Combretum molle, with a grass layer dominated by Rottboellia exaltata, Themeda triandra and Panicum trichocladum, on sandy clay near termite mound, 27 April 1998, Buyck 98.034 (holotypus PC0084737, isotypus UDSM).

Cap 10–20 mm diam., subplane to plano-convex with slightly inflexed margin when young, soon depressed at the center to strongly infundibuliform with an often incised margin; surface smooth, dry, vivid yellow (3A6–7), with some amber yellow to orange yellow tinges, especially at the disc (4A6–7), paler towards the margin. Hymenophore of well-developed relatively spaced gill-folds, strongly decurrent, pale cream yellow to almost concolorous with cap and stipe, forked towards the cap margin, not veined-anastomosed in between. Stipe slender and usually nearly twice as long as the cap diam., 20–35 × (1)2–3 mm, subcylindrical or slightly widened at the base, concolorous to the cap but often becoming paler to almost whitish toward the stipe base, smooth. Context fragile, pale yellowish. Taste mild. Smell weak. Spore print not obtained.

Spores short ellipsoid, (6.2)6.5–6.9–7.3(7.7) × (4.6)4.7–5.0–5.3(5.6) μm, Q = 1.3–1.4–1.5, n = 20, ovoid to almost subglobose in face view, smooth, unpigmented or very slightly yellowish. Basidia (4)5(6)-spored, not very long, mostly 50–65 × 6–8 μm, slightly clavate to subcylindrical. Cystidia not differentiated. Subhymenium composed of cylindrical cells of approx. the same diam. as the basidia and basidiola, not remarkably slender. Pileipellis a loose trichocutis composed of long, slender extremities of 4–6(10) μm diam., with spaced septa that are remarkably rounded resulting in less large contact between cells, thin-walled or with slightly thickened and then refringent walls, not densely branched; the terminal cells mostly 25–50(−80) μm long and often very slightly constricted subapically. Clamp connections absent.

Commentary: This is the only collection known so far. This species was described as C. cf microcibarius by Buyck et al. (2000), a taxon for which the original material has been lost (fide Eyssartier 2001). Based on the original description of the latter (Heinemann 1966), our species differs morphologically from the latter by the absence of clamp connections, the longer stipe and the well-developed, not anastomosing gill folds. Other yellowish, equally small chanterelles from the African miombo woodlands include C. schmitzii Heinem. and C. fistulosus Tibuhwa & Buyck, both easily separated by their abundant clamp connections. However, in our phylogenetic tree (Fig. 1), our new species clusters with significant support with two bright orange chanterelles C. humidicolus (see below) and a still undescribed Malagasy species (C. ined. 2). Both orange species share a similar microscopy but differ essentially in size and form of their spores.

C. humidicolus Buyck & V.Hofst., sp. nov. — Mycobank MB 800912; Figs. 8–10
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Figs. 8–10

Cantharellus humidicolus. 8. Spores. 9. Elements of the hymenium. 10. Terminal elements of the pileipellis. Scale bar = 5 μm for spores, 10 μm for the other elements. (drawings B.Buyck, from holotype)

Diagnosis: This species differs from other small to medium-sized, orange chanterelles in Africa by the combination of medium-sized caps (up to 65 mm diam.), the more elongated spores (Qmean = 2.4) and lack of clamp connections.

Holotypus: Tanzania. Morogoro district, Kinguruwira Sisal Estate, Uluguru Mts., 300 m. behind Sokoine Agricultural University campus, mixed woodland dominated by Pteleopsis myrtifolia, Brachystegia spiciformis, Combretum molle and Markhamia obtusifolia, on a vertical slope of a small riverbed, 28 April 1998, Buyck 98.036 (PC0084724)

Cap (14)30–65 mm diam., sometimes convex at first, but soon depressed in the centre, with the margin becoming irregularly and strongly lobed-undulate, but not radially striate; the surface smooth, slightly greasy, of a beautiful, deep orange, mandarin orange (6AB8) in a quite homogeneous fashion, becoming rapidly paler when drying out. Hymenophore strongly decurrent, very variable, ranging from rather well-developed gill folds to a barely ridged-veined surface, particularly in smaller specimens; gills equal and becoming increasingly forked near the extreme cap margin, in other cases not forked and distinctly unequal with regular lamellulae of different length, close to moderately spaced (4–5 L/cm and 1 l/mm), up to 2 mm high, sometimes also strongly anastomosed-veined in between, much paler than the cap and off-white to pale yellowish. Stipe slender and subcylindrical in smaller fruit bodies, mostly firm and fleshy, often narrowing downward in larger ones, usually not exceeding the cap diam., (13)25–50 x (3)12–22 mm, often split to produce two caps or 2–3 stipes aggregated at the base, often laterally compressed, subconcolorous to the cap to off-white, solid. Context 1–2(4) mm thick, firm, whitish. Taste mild. Smell distinct, very agreeable of typical Cantharellus. Spore print whitish to very pale cream.

Spores elongate, (7.1)7.6–8.1–8.799.20 x (2.9)3.2–3.4–3.7(3.8) μm, Q = (2.0) 2.2–2.4–2.6 (2.9), n = 20, smooth. Basidia moderately long, mostly 50–65 × 5–6.5 μm, subcylindrical to slightly clavate, as basidiola sinuous and irregular in outline, 5(6)-spored. Subhymenium more or less filamentous, of slender to short cells that measure 3–5 μm diam., often only slightly narrower than the terminal basidia and basidiola. Cystidia not differentiated. Pileipellis composed of slender and mostly quite regular, thin-walled to slightly thickened and then refringent hyphae with distant septa, (3)4–5.5(7) μm diam., the terminal cells of similar diam. and often long, mostly measuring 50–100 μm long, slightly constricted subapically. Clamp connections absent.

Other material examined: Tanzania. Kisarawe district, Vigama, near Kazimzumbwi forest, ca 10 km from Kisarawe, 12 May 1998, Buyck 98.124 (PC0085052, paratypus).

Commentary: This species grows in large groups under Brachystegia, particularly in more humid areas without a well-developed herbaceous layer. The small and bright orange chanterelles are a complex and probably very diverse species group in Africa (as well as in Madagascar). Their identification is mostly based on a combination of spore size and form as well as presence/absence of clamp-connections. C. humidicolus is characterized by the combination of the clearly elongated spores (Qmean = 2.4) and lack of clamp-connections. Based on the modern type revisions of Eyssartier (2001), there are at least 2 other small, orange African chanterelles that lack clamp connections: C. pseudocibarius Henn. produces fruit bodies of almost the same size, has comparable but somewhat longer basidia, but possesses much rounder spores (Qmean = 1.4) and a more orange hymenophore. C. pseudofriesii Heinem. is a quite smaller species (max 3 cm diam.) with larger hyphae in the pileipellis (5–10(15) μm diam.), small basidia (35 μm long) and ellipsoid spores (Qmean = 1.7) that are smooth (contrary to the original description, fide Eyssartier 2001) and has remarkably crowded gills. It was originally described from the rain forest area.

C. miniatescens Heinem., another taxon originally described from rain forest, is of similar size and the detailed original description (Heinemann 1958) corresponds quite well to our C. humidicolus but has a tomentose cap with clear reddish and pinkish tints and, under the microscope, ellipsoid spores and wider hyphae in the pileipellis, as well as abundant clamp connections. It is therefore unrelated to our species. Unfortunately, as Eyssartier (2001) reported, the type specimen of C. miniatescens does not correspond to the original description and is very small (cap < 15 mm diam.).

C. miomboensis Buyck & V.Hofst., sp. nov. — Mycobank MB 800900; Figs. 11–13
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Figs. 11–13

Cantharellus miomboensis. 11. Spores. 12. Elements of the hymenium. 13. Terminal elements of the pileipellis. Scale bar = 5 μm for spores, 10 μm for the other elements. (drawings B.Buyck, from holotype)

Diagnosis: This species differs from C. rufopunctatus in the thin-walled elements at the cap surface that are < 7.5 μm diam. and never so short that they become almost subglobose.

Holotypus; Tanzania. Dar-es-salaam, in loamy clay soil on a hill slope with degraded woodland near University campus, 350–400 m alt., 28 April 1998, Buyck 98.021 (PC0084748 holotype)

Fruit bodies dispersed or fasciculated, usually in groups of many individuals. Pileus up to 150 mm diam., very fleshy and firm, first convex, becoming plano-convex and often irregularly lobed when mature; margin first very regular, thick and strongly incurved, later more wavy and retaining mostly a narrow, incurved border, never striated but at the extreme margin often shortly wavy to almost crenulate from the lamellulae pattern underneath; general color variable, ranging from a warm ochre or orange yellow or greenish yellow to a grayish brown or even almost entirely whitish gray; the whole surface areolate-squamulose from a strongly adhering, felty, brownish to whitish tomentum that quickly disintegrates with cap expansion into flattened and appressed squamulae towards the cap margin, exposing the paler context in between, showing yellowish-brown discolorations where touched or injured. Hymenophore composed of well-developed, decurrent, well-spaced and unequal gill folds, especially near the extreme cap margin with many very short lamellulae or also often forked, compared with the cap and stipe surface of a brighter, egg-yolk yellow, sometimes with slight olivaceous to grayish hues. Stipe very thick and firm, subcylindrical or narrowing downwards, concolorous to the cap or slightly paler, and covered with a similar felty-tomentose layer that typically disrupts horizontally into appressed to even recurved squamae, solid. Context fleshy, firm, off-white, often marbled pale gray when fresh (from water imbibition ?), pale yellow underneath the pileipellis, weakly and slowly yellowing with age or when touched. Odor indistinct. Taste pleasant. Spore print insufficient for correct appreciation.

Spores ellipsoid to elongate, usually slightly reniform, (8.9)9.2–10.1–10.9(11.67) x 4.8–5.3–5.7(6.0) μm, Q = (1.5)1.7–1.9–2.1(2.2), n = 20, smooth. Basidia medium long, mostly 55–65 × 7–9 μm, clavulate to almost subcylindrical, 5(6)–spored. Subhymenium composed of cells that are nearly similar in diam. as the base of the hymenial elements. Cystidia not differentiated. Pileipellis composed of thin-walled, quite densely septate and frequently ramifying hyphal extremities, 4–7 μm diam., the terminal cell usually subcylindrical, often subapically slightly constricted and subcapitate, often irregularly sinuous-undulate, mostly (25)30–50(80) μm long. Clamp connections absent.

Other material examined: Tanzania. Morogoro district, Kinguruwira Sisal Estate, Uluguru mountain slope behind the Sokoine Agricultural University campus, 300 m alt., in a Brachystegia specifiormis woodland with Pteleopsis myrtifolia, Dalbergia nitidula, Combretum molle, Markhamia obtusifolia and exotic Eucalyptus camadulensis, on loamy clay, 2 April 2004, Buyck 04.129 (PC0085156); ibid. 4 April 2004, Buyck 04.148 (PC0084858). Zambia. Along road from Kabwe to Kiposhi, in Brachystegia miombo woodland, 14 Jan. 1996, Buyck & Eyssartier leg., in Buyck 96.007 (PC0085088); Along road from Mpongwe to Lusaka, in Brachystegia woodland near Mikati, Buyck & Eyssartier leg., in Buyck 96.117 (PC0085087); near Miputu, in Brachystegia woodland, Buyck & Eyssartier leg., in Buyck 96.154 (PC0085082); at Lake Kashyba, in Brachystegia miombo woodland, 24 Jan. 1996, Buyck & Eyssartier leg., in Buyck 96.182 (PC0085083); near Chibuli, in Brachystegia miombo woodland, 31 Jan. 1996, Buyck & Eyssartier leg., in Buyck 96.237 (PC0085086), 96.238 (PC0085085); bought from villagers along road from Lusaka to Kapiri-Mposhi, 4 Feb. 1996, Buyck 96.315 (PC0085084).

Commentary: C. miomboensis is characterized by the large, extremely firm and fleshy fruit bodies that find their equal only in the bright orange-red C. splendens Buyck 1994. All cited collections form a very homogeneous morphological group under the microscope. The general color of the collections varies from off-white to yellow to olivaceous brown or relatively dark grayish brown. The group of squamulose chanterelles around C. rufopunctatus is a very divers and complex group in Africa, involving probably a number of macroscopically similar, but genetically different taxa, some of which occur in the rain forest area, whereas others are probably typical inhabitants of the savannah woodlands, and perhaps even of exotic eucalypt plantations. It is a group in great need of further study.

C. miomboensis is one of the most common chanterelles of the African miombo woodlands. It is gathered in Zambia in enormous quantities during the rainy season and is even exported under the European name C. cibarius from Lusaka-based companies to e.g. Japan and Germany (Buyck unpubl.). This species has been referred to as C. rufopunctatus var. ochraceus Heinem. in the past (Buyck 1994; Buyck et al. 2000; Eyssartier 2001) and both species indeed share nearly identical features of hymenium and spores. The type specimens of C. rufopunctatus and its variety ochraceus have been re-examined by Eyssartier (2001). These type studies confirm the elongated shape of the spores of the type collections for both African varieties, as well as medium-long basidia (50–80 μm) and an overall strong similarity not only with C. miomboensis but also with C. tanzanicus (see below). The main difference with C. miomboensis lies in the much larger hyphal extremities in the pileipellis of Heinemann’s taxa (short, sometimes nearly subglobose terminal cells measuring 7.5–18 μm diam. or (5)10–15(20) μm according to Eyssartier’s findings) that are frequently thick-walled (overlooked in the original description), whereas our species possesses thin-walled elements that are <7.5 μm diam. and never so short that they become almost subglobose.

Since its original description, C. rufopunctatus has never been reported again from rain forest, until very recently by Eyi Ndong et al. (2011). These authors identified as C. rufopunctatus a number of specimens collected in Gabon, Cameroon and Benin and in its original habitat, the Guineo-congolian rain forest. Unfortunately, the latter publication contributed considerable confusion. Their description of a vividly orange species (Methuen 7A5) with pale orange gills (5A4), white flesh, and rarely forked gills correspond to the type description, but differs completely from the color pictures and watercolors illustrating these recent collections, showing a yellow (Methuen 3A series) species with concolorous gills that are frequently forked and has yellowish flesh. The original description (Heinemann 1958) and later illustrations (Heinemann 1959), however, depict a species that is 6–15 cm diam. (versus 3–6(8) cm in Eyi Ndong et al.) that has concolorous squamulae (distinctly darker and brown squamulae in Eyi Ndong et al.). The few microscopic features mentioned for these recent collections of C. rufopunctatus by Eyi Ndong et al. (2011) are also in strong contrast with those of the type which has elongated spores (versus ellipsoid, Qmean = 1.38 in Eyi Ndong et al.). Whereas the watercolors accompanying the descriptions of C. rufopunctatus and C. luteopunctatus (Beeli) Heinem. in Eyi Ndong et al. may very well belong to one and the same taxon (gill spacement is described as L + l = 13/cm and as L + l = 18/cm for these recent collections of C. rufopunctatus and C. luteopunctatus respectively, which is very different from the L + l = 35–40/cm mentioned in the original description of the latter), but the color slides do not. In both cases, the identifications are most likely erroneous. The macroscopic descriptions, on the other hand, seem to have been copying (a large) part of the features from the original publication and stand in contrast with what is illustrated in both cases. Unfortunately, as none of the illustrations specifies what specimen was illustrated or used for measurements, it is impossible to unravel this mixture. The microscopic features cited in Eyi Ndong et al. (2011) seem to be original and almost superimposable between the two species (C. rufopunctatus and C. luteopunctatus), although incomplete (lacking data on the pileipellis composition for instance) as well as imprecise (all illustrated basidia for all described Cantharellus are 4–spored, but not so in the descriptions, and the size of basidia for C. rufopunctatus is given as 36–44 μm long, but illustrated with basidia that exceed 50 μm long [from the type?]).

C. tanzanicus Buyck & V.Hofst., sp. nov. — Mycobank MB 800897; Figs. 14–16
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Figs. 14–16

Cantharellus tanzanicus. 14. Spores. 15. Elements of the hymenium. 16. Terminal elements of the pileipellis. Scale bar = 5 μm for spores, 10 μm for the other elements. (drawings B.Buyck, from holotype)

Diagnosis: This species differs from C. miomboensis in the much smaller and fragile stature and the presence of thick-walled hyphal extremities of the cap.

Holotypus: Tanzania. Morogoro district, Kinguruwira Sisal Estate, Uluguru mountain slope behind the Sokoine Agricultural University campus, 300 m alt., in a Brachystegia specifiormis woodland with Pteleopsis myrtifolia, Dalbergia nitidula, Combretum molle, Markhamia obtusifolia and exotic Eucalyptus camadulensis, on loamy clay, 28 April 1998, Buyck 98.040 (PC0084728)

Cap 27–40 mm diam., thin-fleshed, slightly undulate-lobed, near the margin with distinctly sulcate appearance, covered with a grayish-greenish brown tomentum, but soon more or less concentrically squamose-areolate, leaving a yellowish layer underneath exposed, hygrophanous and almost whitish grey or pale brown when drying out. Hymenophore of well-differentiated, 2–3(5) mm high gill folds that are very fragile and frequently fissured vertically from gill edge to dorsal attachment, rarely forked, strongly unequal especially close to the extreme cap margin, yellowish orange (4A5–6 or slightly more orange, but not 5). Stipe slender, 24–37 × 3–6 mm, more or less equal to the cap diam., often bent in its lower part, subcylindrical or a little bit broader at the tip, whitish with a pale yellowish hue, densely squamulose, solid. Context quite fragile, whitish, marbled with pale grayish-brownish veins when fresh. Taste mild. Smell typical of Cantharellus. Spore print distinctly cream (near 4A3).

Spores ellipsoid to elongate, usually slightly reniform, (7.3)7.9–8.4–8.9(9.2) x (3.7)3.8–4.1–4.4(4.6) μm, Q = (1.7)1.8–2.0–2.2(2.3), n = 20, smooth. Basidia medium long, mostly 45–60 × 6–7 μm, clavulate to almost subcylindrical, 5(6)-spored. Subhymenium composed of cells that are larger than in the hymenium and not narrowly filamentous. Cystidia not differentiated. Pileipellis composed of distinctly thick-walled, quite densely septate hyphal extremities, 5–8(10) μm diam., the terminal cell usually narrowing upward, rarely fusiform, often constricted subapically and subcapitate, sometimes irregularly sinuose-curved, mostly 30–50(60) μm long. Clamp connections absent.

Commentary: The presence of Eucalyptus near this collection is not without importance. Preliminary sequence data put this species close to other collections from Madagascar (C. cf. tanzanicus in our Fig. 1) that were gathered in pure Eucalyptus stands. However, our recent revision of Australian Cantharellus has revealed no species that could be a possible close relative (Eyssartier and Buyck 2001a).

This species shares with C. miomboensis, described above, the squamulose surface as well as identical spores, but differs in the smaller and much more fragile stature, the less bright coloration as well as in the presence of thick–walled hyphal extremities of the cap.

Discussion

Even though all of the here presented new taxa are not affected by the newly described Afrocantharellus and are all part of Cantharellus s.s. in Tibuhwa et al. (2012), we can not adhere to the views expressed in the latter publication. The present ML analysis of tef-1 data adds no support to the recognition of two separate genera (see Fig. 1) and presents at least one incongruence as to the position of C. fistulosus Tibuhwa & Buyck in the Tibuhwa et al. paper. The latter species, for which we sequenced the type specimen (see Table 1), is part of Afrocantharellus in Tibuhwa et al. (l.c.). It is morphologically completely different from the remainder of Afrocantharellus, but much more similar to the here newly described C. afrocibarius, which seems its closest relative (see Fig. 1). In our experience, part of the explanation for the results obtained by Tibuhwa et al. (l.c.) may reside in problems of ambiguous alignment as the entire ITS region is completely unalignable in Cantharellus, whereas it is well-established that compositional bias in DNA sequences can adversely affect phylogenetic analyses based on those sequences (Foster and Hickey 1999), in particular in cases such as the AT-rich ATP6 gene. Furthermore, the morphological argumentation supporting the distinction between Afrocantharellus from Cantharellus is entirely false. Afrocantharellus species are by all means typical Cantharellus: the absence of clamps characterizes the large majority of African Cantharellus and also the widely spaced and well-differentiated gill-folds are typical of so many other tropical species in the genus (Eyssartier 2001). Even the argument that all of the Afrocantharellus produce fruit bodies with variegated colors (as opposed to uniformely coloured fruit bodies in Cantharellus) is entirely false: apart from C. platyphyllus, none of the African Afrocantharellus have variegated colors (Buyck 1994) and variegated colours are typical for many Cantharellus. For all these reasons, the recognition of a separate Afrocantharellus is here entirely rejected. Furthermore, we have also examined the type of Goossensia cibarioides Heinem.. The microscopical features of this species, that has a smooth hymenophore, are by all means those of typical Cantharellus (Buyck unpubl.) and we have, therefore, integrated this species in the identification key provided below.

It can be concluded from this first molecular approach of African Cantharellus, that this genus is extremely diverse in tropical Africa, much more so than on other continents. The already high number of described species may still be far from the true level of diversity. One of the most urgent aspects that need attention is the recollection and redescription of the earlier taxa, especially those described more than half a century ago from the rain forest area. Care should be taken to supply more precise and fully illustrated data on morphology, and preferably also include a multigene molecular characterization in view of the selection and designation of fully documented epitypes. Especially the rain forest taxa are in need of redescription and epitypification. With the exception of C. afrocibarius, all newly described species are characterized by the absence of clamp connections in their tissues, a feature so far unknown from northern hemisphere chanterelles, but typical for the majority of the African Cantharellus.

For the moment, the key below, which is based on morphological re-examination of all existing type material, may allow for a more precise identification of newly collected specimens. As we think that the general climate may be very important for the distribution of the various chanterelles, we mentioned the general habitat, distinguishing open woodlands versus dense humid forest. The latter forest type, however, comprises not only the equatorial rain forest area in tropical Africa, but extends also into the many gallery forests that border the rivers and streams that ingress the surrounding woodlands. Therefore, care should be taken when interpreting geographical data that lack any ecological precision.

Identification key for tropical African CANTHARELLUS

  1. 1a.

    Hymenophore mouse gray and densely veined, becoming black with ageing or when handled or injured. Originally described for the rain forest area, but frequently reported from the surrounding woodlands...............................Cantharellus congolensis

     
  2. b.

    Hymenophore neither gray nor black and fruit body not blackening with age or when handled...................2

     
  3. 2a.

    Cap and sometimes also stipe surface exhibiting, at least partly, distinct lilac-bluish-violaceous tints; fruit bodies generally distinctly yellowing with age............3

     
  4. b.

    Cap and stipe without such distinct lilac-bluish-violaceous tints............................................................7

     
  5. 3a.

    Fruit body thin-fleshed and fragile; cap strongly squamulose; hymenophore densely veined and anastomosing, whitish. Only known from the woodland type collection..............................Cantharellus conspicuus

     
  6. b.

    Fruit body firm and fleshy, at least in the cap center and stipe; cap often with some appressed squamulae; hymenophore usually not strongly anastomosing and with well-developed, unequal gill-folds, young very pale with yellowish or pinkish tints, then gradually yellowing.....................................................................4

     
  7. 4a.

    Spores narrowly ellipsoid, 5.5–6.72–8 × 3–3.97–5 μm, Q = 1.37–1.71–2.0. Cap very dark violaceous when young; context with faint pinkish hue. Only known from equatorial rain forest....Cantharellus goossensiae

     
  8. b.

    Spores more elongate with mean length-width ratio usually >2....................................................................5

     
  9. 5a.

    Mean spore width <4 μm and mean length <9 μm. Woodland species.......Cantharellus subcyanoxanthus

     
  10. b.

    Spores distinctly larger with mean spore width equal or >4 μm and mean spore length usually >9 μm. Equatorial rain forest species.....................................................6

     
  11. 6a.

    Cap completely smooth or only wrinkled-striate or crenulate near the extreme margin, lacking vividly yellow colors and more pinkish, lilac to cream. Spores narrow (ca 4 μm wide).........Cantharellus longisporus

     
  12. b.

    Cap radially wrinkled or striate outside the center, partly with vividly yellow colors; spores larger (ca 5 μm wide). Equatorial rain forest, often misapplied to woodland collections...........................Cantharellus cyanoxanthus

     
  13. 7a.

    Cap and/or stipe at least partly with distinct red, bright orange or pink colors...................................................8

     
  14. b.

    Cap and stipe lacking such colors entirely, either yellow, brown or gray or a mixture of these colors.........23

     
  15. 8a.

    Fruit bodies medium-sized to large, with very well-developed unequal gills that hardly fork but may have a much lower anastomosing network in between, the whole mushroom reminding strongly of the genus Cuphophyllus (Hygrophoraceae). Clamp connections entirely absent.............................................................9

     
  16. b.

    Fruit bodies not reminiscent of Hygrophoraceae, medium-sized to (very) small; hymenophore reticulately veined or forming less well-developed gill-folds that usually fork...................................................................11

     
  17. 9a.

    Fruit bodies large (up to 18 cm diam.) with cap and stipe thick, firm and fleshy, of an intense, bright orange to reddish orange or blood red that sticks to the fingers upon handling; gills yellowish orange. Only known from woodland............Cantharellus splendens

     
  18. b.

    Fruit bodies usually distinctly smaller and with different colors..................................................................10

     
  19. 10a.

    Spores ellipsoid, Qmean = 1.6–2.0. Cap distinctly smaller and less fleshy than in C. splendens, typically blood red and contrasting with the much more yellowish orange gills and stipe. Only known from woodland.........................................Cantharellus symoensii

     
  20. b.

    Spores shortly ellipsoid to nearly subglobose, Qmean = 1.2–1.4. Cap versicolorous, usually a mixture of gray, vinaceous, grayish red and sometimes clear bluish tints (especially in the context). Gills cream-colored to pale yellow. Only known from woodland..........................Cantharellus platyphyllus

     
  21. 11a.

    Fruit bodies principally yellowish orange to brightly orange.......................................................................12

     
  22. b.

    Fruit bodies pink to red............................................17

     
  23. 12a.

    Clamp connections present. Cap very small (8–15 mm diam.), yellow-orange; gills yellowish.....................................Cantharellus microcibarius

     
  24. b.

    Clamp connections always absent. Not this combination of characters......................................................13

     
  25. 13a.

    Fruit bodies up to 14 cm diam. and fleshy, with numerous, concolorous squamulae on cap and stipe surface. Equatorial rain forest...........Cantharellus rufopunctatus var. rufopunctatus

     
  26. b.

    Fruit bodies much smaller, mostly 1–6 cm diam. and not squamulose........................................................14

     
  27. 14a.

    Gill folds very crowded (up to 30/cm at cap margin)...16

     
  28. b.

    Gill folds, if well-developed, more spaced (up to 15/cm at cap margin)...............................................15

     
  29. 15a.

    Cap up to 3.5 cm diam.; stipe quickly becoming fistulose; spores shortly ellipsoid, 6–6.73–7.5 x 4–4.85–6 μm. Q = 1.17–1.39–1.62. Originally described from equatorial rain forest, reported later from woodland................................Cantharellus pseudocibarius

     
  30. b.

    Cap up to 6 cm diam.; stipe solid; spores distinctly elongate, (8) 8.1–8.63–9.2 (10) x (3) 3.4–3.78–4.1 (4.7) μm, Q = (1.9) 2.1–2.3–2.5 (2.8). Only known from woodland...................Cantharellus humidicolus

     
  31. 16a.

    Cap up to 3.5 cm diam.; spores ellipsoid, 5.5–6.2–7 × 3–3.73–4.5 μm, Q = 1.33–1.67–2. Only known from equatorial rain forest.........Cantharellus pseudofriesii

     
  32. b.

    Cap very small, only up to 1.5 cm diam.; spores ellipsoid but distinctly larger, 6.5–7.64–9 × 4.5–5.14–6 μm, Q = 1.3–1.49–1.8. Only known from equatorial rain forest.....................................Cantharellus tenuis

     
  33. 17a.

    Clamp connections present; spores ellipsoid (Qmean = 1.2–1.4)....................................................................18

     
  34. b.

    Clamp connections absent; spores more variable in form, ranging from subglobose to elongate..............20

     
  35. 18a.

    Cap up to 8 cm diam., reddish pink, quickly paler when drying out; hymenophore pink and densely reticulate-anastomosed. Only known from woodland...........................................Cantharellus ruber

     
  36. b.

    Cap up to 10 cm diam., overall colors more brownish vinaceous to dull reddish, not becoming rapidly paler when drying out; hymenophore with distinct gill folds, but eventually veined-anastomosed in between. Equatorial rain forest...............................................19

     
  37. 19a.

    Cap and stipe squamulose-hirsute........................................Cantharellus rhodophyllus

     
  38. b.

    Cap and stipe smooth or nearly so. (microscopically identical to the preceding species)....................................Cantharellus subincarnatus

     
  39. 20a.

    The whole fruit body more or less concolorous.......21

     
  40. b.

    Hymenophore much paler, off-white to pale yellowish, strongly contrasting with the reddish surface of stipe and cap............................................................22

     
  41. 21a.

    Cap up to 6 cm diam.; spores elongate, Q > 2. Only known from woodland...Cantharellus heinemannianus

     
  42. b.

    Cap < 3 cm diam.; spores ellipsoid, Q < 1.6. Only known from woodland.........Cantharellus addaiensis

     
  43. 22a.

    Cap up to 3 cm diam., gill-folds crowded (L + l = 30/cm). Spores (type) nearly subglobose to shortly ellipsoid, 6–6.68–7 × 4–4.85–5.5 μm, Q = 1.2–1.38–1.55. Only known from equatorial rain forest, although frequently misapplied to woodland collections..............................................Cantharellus floridulus

     
  44. b.

    Cap up to 1.5 cm diam., gill-folds spaced. Spores (type) ellipsoid, 7–7.69–9 × 4–4.94–5.5 μm, Q = 1.4–1.56–2. Only known from equatorial rain forest.............................................Cantharellus alboroseus

     
  45. 23a.

    Hymenophore smooth or nearly so..........................24

     
  46. b.

    Hymenophore with well-developed anastomosing veins or gill-folds.....................................................25

     
  47. 24a.

    Clamp connections present. Basidia two-spored. Only known from equatorial rain forest................................................Cantharellus solidus

     
  48. b.

    Clamp connections absent. Basidia not two-spored. Only known from equatorial rain forest...............................................Goossensia cibarioides

     
  49. 25a.

    Cap and stipe remaining smooth and never distinctly squamulose-scaly at maturity. Cap surface principally yellow, although sometimes with more brownish center, especially when young.......................................26

     
  50. b.

    Cap and often also stipe distinctly squamulose or scaly or at least disrupting concentrically near the cap margin at maturity. Cap surface often pale brownish gray to brown or even olivaceous, at least when young, later becoming paler and more yellow through disruption of the tomentose surface layer...............30

     
  51. 26a.

    Clamp connections present......................................27

     
  52. b.

    Clamp connections absent.......................................29

     
  53. 27a.

    Fruit bodies fleshy, very compact and firm; cap yellow with paler, off-white areas, forming generally cespitose groups of up to 18 cm diam. Only known from woodland..................Cantharellus afrocibarius

     
  54. b.

    Cap <3 cm diam; stipe becoming fistulose at maturity................................................................28

     
  55. 28a.

    Cap with brown matted center and yellow margin and stipe. Hymenophore with pinkish hue producing a pale pink spore print. Spores ellipsoid, with Qmean = 1.7. Hyphal extremities of cap measuring 3–5 μm diam. Only known from the woodland type........................................Cantharellus fistulosus

     
  56. b.

    Very similar but with the hymenophore more yellow orange, lacking pinkish hues. Spores subglobose to shortly ellipsoid, with Qmean = 1.3. Hyphal extremities of cap measuring (8) 10–15 (20) μm diam. Only known from the woodland type collection.........................................Cantharellus schmitzii

     
  57. 29a.

    Fruit body fleshy and medium-sized, with usually robust stipe (strongly reminding of C. cibarius). Woodland............................Cantharellus defibulatus

     
  58. b.

    Fruit body very small and fragile, with slender stipe. Woodland..................................Cantharellus gracilis

     
  59. 30a

    Hymenophore composed of crowded gill folds (L + l = 20–40/cm)...............................................................31

     
  60. b.

    Hymenophore composed of distinctly more spaced gill folds...................................................................33

     
  61. 31a.

    Cap brown, not squamulose but sometimes disrupting concentrically near the margin. Gill folds brown at maturity. Only known from the woodland area............................................Cantharellus tomentosus

     
  62. b.

    Cap distinctly squamulose-scaly, pale lemon yellow, cream to grayish or brown. Gill folds very pale, brownish to yellowish gray, remaining so at maturity...........32

     
  63. 32a.

    Taste bitterish. Equatorial rain forest area.....................................Cantharellus luteopunctatus

     
  64. b.

    Taste mild. Originally described from the equatorial rain forest area, but often reported from humid habitats in the surrounding woodlands............Cantharellus densifolius

     
  65. 33a.

    Fruit bodies firm and fleshy, up to 15 cm diam or more.........................................................................34

     
  66. b.

    Fruit bodies neither particularly fleshy nor firm, rarely exceeding 5 cm diam...........................................35

     
  67. 34a.

    Cap surface having thin-walled elements that are <7.5 μm diam. and never so short that they become almost subglobose. Woodland.................Cantharellus miomboensis

     
  68. b.

    Cap surface under the microscope with locally strongly inflated, nearly subglobose terminal elements. Equatorial rain forest, frequently misapplied to woodland collections.....Cantharellus rufopunctatus var. ochraceus

     
  69. 35a.

    Basidia predominantly 5-6-spored. Only known from the woodlands..........................................................37

     
  70. b.

    Basidia 2-4-spored. Only known from equatorial rain forest........................................................................36

     
  71. 36a.

    Cap and stipe concolorous, very pale coloured. Basidia four-spored. Spores 8–8.78–10 × 5–5.82–6.5 μm. Q = 1.42–1.51–1.67. Often misapplied to woodland collections.........................................Cantharellus isabellinus

     
  72. b.

    Cap and stipe concolorous, a dirty grayish to brownish. Basidia two-spored. Hymenophore safran colored. Spores 7–8.36–10 × 5.5–6.10–7 μm, Q = 1.14–1.32–1.54...........................Cantharellus croceifolius

     
  73. 37a.

    Fruit bodies clearly squamulose to areolate on the surface of cap and stipe. Gill folds pale yellowish. Spores ellipsoid to elongate, usually slightly reniform, (7.3)7.9–8.4–8.9(9.2) x (3.7)3.8–4.1–4.4(4.6) μm, Q = (1.7)1.8–2.0–2.2(2.3). Woodlands with ingressin of eucalypts................................Cantharellus tanzanicus

     
  74. b.

    Fruit bodies with tomentose cap and smooth stipe. Gill folds dull colored, a pale grayish cream to pinkish gray. Spores a bit less elongate, (7)7.5–8.1–8.5(9) x (4)4.4–4.75–5 μm, Q = (1.5)1.6–1.7–1.8(1.9). Woodland area, likely represents a distinct species..................Cantharellus isabellinus var.parvisporus

     

Acknowledgments

We are grateful to the staff members of AMU in Dar-es-Salaam for logistic support and field assistance, and to L. Mwasumbi and F.M. Mbago for their botanical expertise. The field work in Tanzania was financed by SIDA/SAREC under the «Propagation and phytochemical studies of endangered or economically important plants and fungi of Tanzania » project. The sequencing for this study was performed by C. Cruaud at the Genoscope or “Consortium National de Recherche en Génomique” near Paris (France) as part of the agreement n°2005/67 on the project “Macrophylogeny of life” between the Genoscope and the “service de systématique moléculaire” (CNRS IFR 101) of the Muséum National d’Histoire Naturelle and receives continuing support from the ATM-project “Barcode of life” (Dirs. L. Legall and S. Samadi).

Supplementary material

13225_2012_215_MOESM1_ESM.nex (95 kb)
ESM 1(NEX 95 kb)

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© Mushroom Research Foundation 2012