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Phytotaxa 306 (3): 223–233
http://www.mapress.com/j/pt/
Copyright © 2017 Magnolia Press Article PHYTOTAXA
ISSN 1179-3155 (print edition)
ISSN 1179-3163 (online edition)
Accepted by Genevieve Gates: 18 Apr. 2017; published: 12 May 2017
https://doi.org/10.11646/phytotaxa.306.3.5
223
A new species and a new combination of Rhodophana (Entolomataceae,
Agaricales) from Africa
PABLO P. DANIËLS1, TIMOTHY J. BARONI2*, OUMAROU HAMA3, KERRI KLUTING4, SARAH BERGEMANN5,
FELIX INFANTE GARCÍA-PANTALEÓN1, MOUSSA BARAGE6 & DAHIRATOU IBRAHIM7
1Department of Botany, Ecology and Plant Physiology, University of Cordoba, Ed. Celestino Mutis, Campus Rabanales, Cordoba
14071, Spain.
2Department of Biological Sciences, State University of New York, College at Cortland, PO Box 2000, Cortland,New York 13045, USA.
3Department of Plant Production and Irrigation, Faculty of Agronomy, University of Tahoua, Tahoua BP-255, Niger.
4Department of Evolutionary Biology, Evolutionary Biology Centre, Uppsala University, Norbyvägen 18D, SE-752 36 Uppsala, Sweden.
5Biology Department, Middle Tennessee State University, PO Box 60, Murfreesboro, Tennessee 37132, USA.
6Department of Plant Production, Faculty of Agronomy, University Abdou Moumouni, Niamey BP-10960, Niger.
7Life Sciences and Earth Department, High School of Education, University Abdou Moumouni, Niamey BP-10963, Niger.
* Corresponding author: tim.baroni@cortland.edu
Abstract
Multiple collections of a new member of the Entolomataceae were found in W National Park of Niger during a survey of
macrofungi. This new species with a dark reddish brown scaly pileus surface and a yellow stipe belongs in the genus Rhodo-
phana of the Rhodocybe-Clitopilus clade in the Entolomataceae. Using a three-gene analysis of the phylogenetic position of
Rhodophana flavipes it is most closely related to the recently described Rhodophana squamulosa from India and is a sister
taxon to Rhodophana nitellina and R. melleopallens. Micromorphological examination of the type of Rhodocybe fibulata,
another African species with a scaly cap, confirms that it belongs in Rhodophana, thus a new combination Rhodophana
fibulata is proposed.
Key words: atp6, rpb2, tef1, molecular phylogeny
Introduction
The Entolomataceae Kotl. & Pouzar consists of two major clades, an Entoloma clade and a Rhodocybe-Clitopilus
clade (Co-David et al. 2009, Baroni et al. 2011, Baroni & Matheny 2011, Kluting et al. 2014) with the Rhodocybe-
Clitopilus clade containing fewer taxa globally (300 species in the Rhodocybe-Clitopilus clade vs. ca. 1200 species for
the Entoloma clade). Co-David et al. (2009) performed a phylogenetic analysis with a limited sample of representative
species and proposed the recognition of two genera (Entoloma Fr. ex P. Kumm. s.l. (1871:23) and Clitopilus (Fr.
ex Rabenh.) P. Kumm (1871:23)) in the Entolomataceae. The generic concepts proposed by Co-David et al. (2009)
were based on the conclusion that the morphological concepts used to recognize most of the segregate genera in the
Entoloma clade were unsupported and a similar situation existed in the Rhodocybe-Clitopilus clade. Baroni & Matheny
(2011) conducted a phylogenetic analysis of the Entolomataceae using an increased sample size with the same three
partial genes employed by Co-David et al. (2009) and demonstrated that four well-supported groups in the Rhodocybe-
Clitopilus clade exist. Kluting et al. (2014) expanded the sample size significantly for the Rhodocybe-Clitopilus clade
and analyzed three different partial protein-coding genes (atp6, tef1, and rpb2) resolving five, statistically supported
clades which can be recognized as distinct genera based on morphological characters: Rhodocybe Maire (1926:298) in
a redefined sense, Rhodophana Kühner (in Kühner & Lamoure 1971:23), Clitopilopsis Maire (1937:113), Clitopilus
(Fr.ex Rabenh.) P. Kumm. (1871:23), and a newly recognized genus Clitocella Kluting, T. J. Baroni & Bergemann
(2014:1135) that is a sister group to Clitopilus and Rhodocybe. We follow here the generic concepts proposed by
Kluting et al. (2014).
There are a limited number of publications describing species of the Rhodocybe-Clitopilus clade from Africa
(Baroni 1999, Beeli 1928, Daniëls et al. 2015, Malençon & Bertault 1970, 1975, Pegler 1977) and only 17 taxa of
Rhodocybe and/or Clitopilus have been documented in these works. We discuss here two taxa of Rhodophana now
known for Africa, one of which is a new species.
DANIËLS ET AL.
224 • Phytotaxa 306 (3) © 2017 Magnolia Press
A recent project entitled “Edible and cultivable Macromycetes of Niger”, carried out jointly by the University of
Córdoba, the Spanish Agency for International Development Cooperation, with the Abdou-Moumouni University of
Niamey, from 2010 to 2013 had as one of its aims a project to inventory Niger’s macrofungal biodiversity (Daniëls
et al. 2015). The field work led to the discovery of an undescribed species of the Rhodocybe-Clitopilus clade in the
genus Rhodophana. This distinctive new species of Rhodophana is common in the W National Park of Niger (Fig.
1), especially within the West Sudanian savanna that is located in a transition region between savanna and woodlands
with three habitats: The woody savanna, the riparian gallery forests and the bushy savanna. The rainy season is from
July to September with an annual rainfall of 600–1000 mm stimulating the production of macrofungi. This new species
appears to be a saprotroph in these environments and is morphologically recognized as belonging in Rhodophana
by the elongate, strongly bumpy basidiospores with minute angles in polar view, cyanophilic spore walls, and large
obvious clamp connections on most hyphae.
We present here morphological and phylogenetic data to support recognition of this new species of Rhodophana
and provide a new combination into Rhodophana of another African species originally described in Rhodocybe.
FIGURE 1. Map of the W National Park showing the collection sites of Rhodophana flavipes. 1) General Seyni Kountché, 2) Mékrou
River on the border with Benin.
Materials and Methods
Phylogenetic analysis:—Kluting et al. (2014) extracted DNA from Hama 434 (COFC-F 5029) and produced partial
sequences of the nuclear RNA polymerase subunit II (rpb2) and the nuclear translation elongation factor 1-α (tef1)
gene regions for a three-locus phylogenetic analysis of the Rhodocybe and Clitopilus species. Aligned sequence data
for 34 ingroup and five outgroup taxa (Panellus stipticus (Bull.) P. Karst. (1879:96), Mycena aff. pura (Pers.) P.Kumm.
(1871:107), Catathelasma imperiale (Quél.) Singer (1940:9), Tricholoma flavovirens (Pers.) S. Lundell (in Lundell
& Nannfeldt 1942:1102) [= T. equestre (L.) P. Kumm. (1871:130)], and Tricholoma aurantium (Schaeff.) Ricken
(1915:332); Table 1) were obtained from the three-locus alignment (with the intron regions of tef1 excluded) used
in the analysis by Kluting et al. (2014). Geneious 6.1.8 (http://www.geneious.com, Kearse et al. 2012) was used to
generate multiple sequence alignments of the atp6, rpb2, and tef1 gene regions (471, 906, and 898 bp, respectively),
and a phylogenetic analysis of this subset is presented here. A concatenated matrix of the three loci (with six apt6 and
four tef1 sequences coded as missing, Table 1) is available on TreeBASE (http://purl.org/phylo/treebase/phylows/study/
TB2:S16270). Matrices for the individual gene regions were partitioned across codon position, and the concatenated
dataset was partitioned across gene region and codon position for the phylogenetic analyses.
Maximum Likelihood analyses were conducted using RAxML ver. 8.0.24 (Stamatakis 2006, Stamatakis 2014) on
the CIPRES gateway (Miller et al. 2010). For all analyses, 1000 bootstrap (MLBS) pseudoreplications were conducted
using the rapid bootstrap algorithm (Stamatakis et al. 2008) and a GTR+Γ model of nucleotide substitution was applied
to all partitions. All trees were visualized using FigTree v1.4.0 (Rambaut 2012). Individual gene regions were first
A NEW SPECIES AND A NEW COMBINATION OF RHODOPHANA Phytotaxa 306 (3) © 2017 Magnolia Press • 225
analyzed separately. The program compat.py (Kauff & Lutzoni 2003) was used to detect topological incongruence
(conflicting topologies with BS support values of 70 or greater) between the three resulting phylograms. In the absence
of topological incongruence, an analysis was conducted on the concatenated dataset of the three loci.
TABLE 1. GenBank accession numbers for sequences used in phylogenetic analyses.
Species Collection Identifier Herbarium Accession
No. GenBank No.
atp6 rpb2 tef1
Clitocella fallax 52/85 O-F88953 KC816767 KC816936 KC816845
Clitocella fallax 25668OKM 25668OKM KC816768 KC816937 KC816846
Clitocella mundula 7161 TJB 7161 TJB KC816782 KC816952 KC816862
Clitocella popinalis K(M): 167017 KC816797 KC816972 KC816879
Clitocella popinalis 6378 TJB 6378 TJB KC816801 KC816976 KC816882
Clitopilopsis hirneola 8490 R.E. Halling REH8490 KC816904 KC816820
Clitopilopsis hirneola PM 247-08 Artsobs. 1376857 KC816977 KC816883
Clitopilus apalus 26394 Watling WAT26394 KC816738 KC816906 KC816822
Clitopilus crispus 9982 TJB 9982 TJB KC816742 KC816910 KC816826
Clitopilus hobsonii 5967 TJB 5967 TJB KC816748 KC816917
Clitopilus paxilloides 5809 TJB 5809 TJB KC816750 KC816919 KC816832
Clitopilus peri 10041 TJB 10041 TJB KC816753 KC816922 KC816835
Clitopilus prunulus 6805 TJB 6805 TJB KC816755 KC816924 KC816837
Clitopilus venososulcatus 8111 TJB 8111 TJB KC816761 KC816930
Rhodocybe alutacea 5726 TJB 5726 TJB KC816762 KC816931 KC816842
Rhodocybe caelata 6919 TJB 6919 TJB KC816764 KC816933 KC816843
Rhodocybe caelata J. Parkin J. Parkin KC816765 KC816934
Rhodocybe collybioides 10417 TJB 10417 TJB KC816766 KC816935 KC816844
Rhodocybe fuliginea E537 Gates E537 KC816770 KC816940 KC816850
Rhodocybe hondensis 6103 TJB 6103 TJB KC816771 KC816941 KC816851
Rhodocybe mellea 6883 TJB 6883 TJB KC816774 KC816944 KC816854
Rhodocybe reticulata E2183 Gates E2183 KC816804 KC816980 KC816887
Rhodocybe rhizogena 5551 TJB 5551 TJB KC816805 KC816981 KC816888
Rhodocybe roseiavellanea 8130 TJB 8130 TJB KC816806 KC816982 KC816889
Rhodocybe stipitata 5523 TJB 5523 TJB KC816815 KC816993
Rhodophana flavipes Hama 434 COFC-F 5029 KC816984 KC816891
Rhodophana melleopallens K(M): 143160 KC816775 KC816945 KC816855
Rhodophana melleopallens 415/83 O-F172919 KC816776 KC816946 KC816856
Rhodophana nitellina K(M): 132700 KC816960 KC816867
Rhodophana nitellina Artsobs. 1541959 KC816790 KC816961 KC816868
Rhodophana nitellina 6740 TJB 6740 TJB KC816964 KC816871
Rhodophana nitellina 7861 TJB 7861 TJB KC816789 KC816959 KC816866
Rhodophana nitellina HH74/10 O-F293352 KC816788 KC816958 KC816865
Rhodophana stangliana 2073 T. Læssøe 2073TL KC816992 KC816899
Catathelasma imperiale 11CA01A 11CA01A KC816816 KC816994 KC816900
Mycena aff. pura 11CA007 11CA007 KC816817 KC816995 KC816901
Panellus stipticus 11CA052 11CA052 KC816818 KC816996 KC816902
Tricholoma flavovirens 11CA038 11CA038 KC816819 KC816997 KC816903
Tricholoma aurantium LCG2308 JN019434 JN019705 JN019386
Bayesian analyses of the four datasets were conducted using MrBayes ver. 3.2.2 (Ronquist et al. 2012). The
best fitting model of sequence evolution for each gene region was first determined using the model test function
in TOPALi ver. 2.5 (Milne et al. 2008) based on the Akaike Information Criterion. For these tests, MrBayes was
selected as the method for tree estimation. The SYM+Γ model was selected for the rpb2 and tef1 sequence data, and
the GTR+Γ model was selected for the atp6 sequence data. Since MrBayes does not allow for multiple outgroup
taxa, only Panellus stipticus was specified. Eleven chains (one cold and ten heated) with default temperatures were
used for two concurrent runs of 1,000,000 generations, sampling every 100 generations with three swaps per chain
per generation for each analysis. The substitution rates, transition/transversion rate ratios, character state (stationary
nucleotide frequencies) and the alpha shape parameter were unlinked to estimate these parameters independently for
each partition. For all four analyses, the two runs converged on the same topology as indicated by standard deviation
DANIËLS ET AL.
226 • Phytotaxa 306 (3) © 2017 Magnolia Press
split frequencies below 0.01. Scatterplots were generated to determine when the runs had become stationary. The first
30,000 generations were discarded as the burn-in for each analysis before parameter values were summarized, and
Bayesian Posterior probabilities (BPP) and branch lengths were calculated for a 50% majority rule consensus tree.
Morphological analysis:—Collections were obtained using routine sampling methods (Eyi-Ndong et al. 2011)
and photographed in the field with a digital camera. Notes on morphological characters of basidiomata were taken
before drying the samples with warm air using a Bunsen burner attached to a folding portable dryer (De Kesel 2001).
Once dried the preserved specimens were placed inside minigrip bags to eliminate ambient moisture. Color references
were coded following Kornerup & Wanscher (1981) from the fresh specimens and additional information on color
and surface morphology was taken from the color digital images used in preparing the description for the protologue.
Herbarium abbreviations follow Thiers (2017, continuously updated).
For the study of anatomical features, dried tissues were revived in 95% ETOH and then dH2O before mounting
and observing materials in 3% KOH or Congo Red in a 10% ammonia solution. Dry tissue samples of thin sections
of the pileus and stipe surfaces and lamella edges were also mounted directly in 3% KOH or Melzer’s Reagent to
observe hyphal structure or amyloid color reactions respectively. To determine cyanophilic reactions, small dry lamella
pieces were placed in Cotton Blue in lactic acid and heated for about 30 seconds over an alcohol lamp flame until the
mountant began to vaporize. The blue stained tissue was then moved into a drop of clear lactic acid on the same slide
to destain the sample for approximately 1 minute. The tissue was finally mounted in a drop of clear lactic acid on a new
clean slide and gently squashed under the coverslip to separate cells for clear observation.
SEM images of basidiospores were obtained from rehydrated, then dehydrated and critical point dried samples of
lamella pieces (Baroni et al. 2014) on an ISI DS130c scanning electron microscope. Images were obtained on a digital
camera using DIC optics of an Olympus BX50 light microscope.
FIGURE 2. Phylogeny of the Rhodocybe-Clitopilus clade showing the position of Rhodophana flavipes in a ML tree for the concatenated
dataset of the atp6, rpb2, and tef1 gene regions with MLBS ≥ 70 and BPP values ≥ 0.95 displayed on the branches.
A NEW SPECIES AND A NEW COMBINATION OF RHODOPHANA Phytotaxa 306 (3) © 2017 Magnolia Press • 227
Results
Phylogenetic analysis:—No conflict was detected between the three gene regions analyzed separately, and the ML tree
for the concatenated dataset is presented here with MLBS and BPP values of ≥ 70 and ≥ 0.95, respectively, displayed
on the branches (Fig. 2). The five genera described by Kluting et al. (2014) are well resolved and supported: Clitopilus
(MLBS = 89, BPP = 1.0), Clitocella (MLBS = 100, BPP = 1.0), Clitopilopsis (MLBS = 100, BPP = 1.0), Rhodocybe
(MLBS = 99, BPP = 1.0), and Rhodophana (MLBS = 100, BPP = 1.0). The results of this analysis demonstrate the
phylogenetic placement of Hama 434 within Rhodophana (MLBS = 100, BPP = 1.0), but separate from all collections
of R. nitellina (Fr.) Kühner (in Kühner & Lamoure 1971:23), R. melleopallens (P.D. Orton) Kluting, T.J. Baroni &
Bergemann (2014:1138) and R. stangliana (Bresinsky & Pfaff) Vizzini (2014:1) (MLBS = 80, BPP = 0.98).
The sequences generated from the new species, KC816984 (rpb2) and KC816891 (tef1), are deposited in
GenBank.
Morphological analysis:—The morphological analysis is in agreement with the phylogenetic analysis supporting
recognition of a new species of Rhodophana in the Rhodocybe-Clitopilus clade of the Entolomataceae. We cite seven
collections of this taxon, all from the W National Park of Niger.
Taxonomy
Rhodophana flavipes T.J. Baroni, Daniëls & Hama sp. nov. (Figs. 3–13)
Mycobank: MB# 811694
Diagnosis:—Characterized as a species of Rhodophana by the strongly undulate-pustulate subamygdaliform
basidiospores with cyanophilic spore walls that are angular by 7–11 facets in polar view and the abundant clamp
connections present on most hyphae of the basidiomata, differing from other species in Rhodophana by the erect
pyramidal reddish brown squamules on the pileus surface when young, the pileus becoming orange-brown and strongly
sulcate with age and the yellow stipe that produces thick cord like rhizoids at the base which are covered with acicular
crystals, and by the distinctive rbp2 (KC816984) and tef1 (KC816891) DNA sequences.
Etymology:—“flavipes” refers to the yellow color of the stipe.
Holotype:—NIGER. Tillabéri Region: Department of Say, Tamou Commune, W National Park, General Seyni
Kountché I, elev. 236 m, 12°26’52”N, 02°26’09”E, 14 September 2009, in bushy savanna with Combretum micranthum
G.Don, C. nigricans Leprieur ex Guill. & Perr., C. glutinosum Perr. ex DC. and Guiera senegalensis J.F.Gmel, in leaf
litter and woody debris, leg. O. Hama, M. Barage & D. Ibrahim, Hama 259 (COFC-F 5050).
Isotype:—CORT and Université Abdou Moumouni, Niamey (Niger).
Description:—Pileus dark reddish (9C5—Dull Red to 9D5—Rosewood) at first when moist, becoming more
brownish (7D7–8—Burnt Sienna or Brick Red or 7E6–8—Agate) with loss of moisture, especially over the disc and
with margin becoming a much paler buff orange (5B4—Grayish Orange to 5A3—Pale Orange) or eventually light
yellow or cream color (4A3–4) in dry conditions, slightly hygrophanous with reddish brown fibrils and squamules
contrasting with the paler ground color, 30–80 mm broad, at first convex, then becoming plano-convex with the disc
broadly depressed, surface uniformly covered with dark brownish silky appressed fibrils and squamules, squamules
often erect, pyramidal and/or squarrose, obvious, especially over the disc, surface opaque, smooth at first, becoming
subsulcate or strongly sulcate with expansion and age, margin incurved becoming decurved, fleshy. Flesh white or
pale peach (5A2—Orange White), 2 mm thick, unchanging when exposed. Lamellae at first pale yellowish (4A2–
3—Cream), but soon light orange or pale peach or orange buff (5–6A2—Orange White or 5–6A3—Pale Orange),
becoming darker and more flesh colored (7A–B3—Pale Red or Greyish Red), eventually darker brownish orange
(7C5–6) with age, adnexed, broadly adnexed or somewhat sinuate, seceding, subdistant, thick, broad (3–4 mm deep),
edge concolorous, even becoming eroded. Stipe pale yellow (4A3—Cream) or light yellow (4A4—Light Yellow),
becoming darker (5B4—Greyish Orange) and also from handling (reddish or reddish brown bruising of fibrils on
the stipe), 4–10 mm broad, 40–65 mm long, equal, cylindrical, straight or sometimes curving, surface smooth but
innately fibrillose splitting, hollow, with thick white rhizomorphs (0.5 mm in diam.) at base. Odor herbaceous. Taste
not distinctive.
DANIËLS ET AL.
228 • Phytotaxa 306 (3) © 2017 Magnolia Press
FIGURES 3−6. Basidiomata of Rhodophana flavipes. Figs. 3−4 are young fresh samples of Hama 259 (Holotype). Figs. 5−6 represent the
older mature stages of the basidiomata with faded colors of the pileus, Hama 434. Scale bar = 10 mm. Photos by O. Hama.
Basidiospores (6.4–) 7.2–10.5 (–11.3) × (4.8–) 5.6–6.4 (–7.2) µm (x = 8.2 ± 1.16 × 5.97 ± 0.47 µm, Q = 1.13–1.83,
Qm = 1.4 ± 0.18: n=61/3), subamygdaliform or ellipsoid in profile view, ellipsoid or obovate in face view, minutely
angular in polar view with 7−11 facets, strongly undulate-pustulate all over, walls evenly cyanophilic, inamyloid.
Basidia mostly 4-sterigmate, some 2-sterigmate, narrowly clavate, 26−31.6 × 8.8−10 µm. Hymenial cystidia absent.
Lamella trama stramineous in 3% KOH, composed of parallel, cylindrical hyphae, 6−14 µm in diam and mostly short
in length, 20−50 µm, also with repository hyphae filled with shiny yellow or golden pigments and scattered through the
trama. Pileipellis yellowish ochre or yellowish orange in 3% KOH, composed of repent, cylindrical hyphae, 2.4−7.2
µm in diam, not encrusted, hardly differentiated from context except by the narrower width of the hyphae, also with
scattered groups of erect fascicles of agglutinated cylindrical hyphae that are slightly broader than those of the rest of the
pellis, 8−14 µm in diam. Pileus context hyaline, composed of inflated, interwoven hyphae (4−)8−18 µm in diam, also
undulating refractive repository hyphae filled with diffuse yellow pigment, 3−12 µm in diam, occasionally branched,
A NEW SPECIES AND A NEW COMBINATION OF RHODOPHANA Phytotaxa 306 (3) © 2017 Magnolia Press • 229
scattered through the context. Stipitipellis near the apex of the stipe a hyaline or pale stramineous layer of repent,
cylindrical hyphae, 2.4−4.8 µm in diam, producing scattered clusters of erect fascicles of agglutinated cylindrical
hyphae similar to those of the pileipellis. Clamp connections present and obvious in all tissues. Rhizomorphs composed
of cylindrical interwoven hyphae, 3−16 µm diam, external layer of hyphal walls covered with easily dislodged long,
acicular crystals that are often fractured into smaller blunt end pieces, individual crystals 2−10−60 × 0.5−2.0 µm,
small bacilliform crystals also present. Repository hyphae scattered or abundant in the pileus context and lamella
trama, yellow or golden yellow in 3% KOH, mostly cylindrical, occasionally irregularly swollen, 3−18 µm in diam,
sometimes septate, sometimes branched, without clamps.
FIGURES 7−13. Basidiospores of Rhodophana flavipes (Hama 259, Holotype). Fig. 7: Various views of basidiospores and strongly
developed ornamentation. Fig. 8: Polar view of a single basidiospore. Fig. 9: SEM image of a single spore in profile view. Fig. 10:
Microscopic image of a pyramidal scale of the pileipellis. Fig. 11: Clamp connection on hyphae of stipe basal cords. Fig. 12-13: Crystals
on the hyphae of the stipe basal cords. Scale bars = 10 µm except Fig. 9 bar = 1 µm. Photos by T. J. Baroni.
DANIËLS ET AL.
230 • Phytotaxa 306 (3) © 2017 Magnolia Press
Habit, habitat and fruiting period:—Single or gregarious in small numbers, saprotrophic on rich organic litter
areas, notably in the gallery forests with Cola laurifolia Mast., Mitragyna inermis (Willd.) K.Schum. and Diospyros
mespiliformis Hochst. ex DC. present.
Additional specimens examined:—NIGER. Tillabéri Region: Department of Say, Tamou Commune, W National
Park, General Seyni Kountché II, elev. 254 m, 12°27’06,3”N, 02°25’38,9”E, 28 August 2008, in open savanna with
plant debris under Isoberlinia doka Craib & Stapf and Acacia ataxacantha DC., leg. O. Hama, M. Barage & D.
Ibrahim; Hama 105 (COFC-F 5431). Tillabéri Region, Department of Say, Tamou Commune, W National Park, close
to the Mékrou River on the border with Benin, elev. 216 m, 12°15’17”N, 02°23’24”E, 6 August 2010, riparian forest
with Cola laurifolia and Mitragyna inermis, in leaf litter and woody debris, leg. O. Hama, P. Daniëls, M. Barage, D.
Ibrahim & M. Rosas; Hama 315 (COFC-F 5030); same location, 8 September 2012, leg. O. Hama; Hama 591 (COFC-
F 5554). Tillabéri Region, Department of Say, Tamou Commune, W National Park, close to the Mékrou River on the
border with Benin, elev. 332 m, 12°15’16’’N, 02°23’22’’E, 29 August 2010, riparian forest, on rotten wood, leg. O.
Hama, M. Barage & D. Ibrahim; Hama 434 (COFC-F 5029). Tillabéri Region, Department of Say, Tamou Commune,
W National Park, close to the Mékrou River on the border with Benin, elev. 218 m, 12°15’18’’N, 02°23’25’’E, 15
August 2012, riparian forest with Kigelia africana (Lam.) Benth. and Mitragyna inermis, in clay-sandy soil with leaf
litter and woody debris, leg. O. Hama, Hama 526 (COFC-F 5552); same location, Hama 527 (COFC-F 5553).
Discussion
Rhodophana flavipes is distinguished by the dark reddish or reddish brown pileus with pyramidal or squarrose
squamules (Fig. 3) covering the medium (30 mm in diam) to moderately large (80 mm in diam) pileus that contrasts
sharply with the cream or light yellow stipe that can darken to reddish brown from handling or age. The lamellae are
also thick, fleshy-rigid and pale yellow becoming pale orange or orange-brown with age (Fig. 4−5). The colors of this
species when fresh are attractive. With age, the colors become muted with the pileus turning orange buff or even light
yellow or cream color from drying out (Fig. 6); also the gills reflect a varying amount of color depending of the angle
of vision, from yellowish to pink. Under the microscope the obvious bumpy-pustulate ornamented, ellipsoid or slightly
almond shaped basidiospores that are minutely angled in polar view (Fig. 7−9), and the prominent clamp connections
on the hyphae of the basidiomata (Fig. 11) are typical features of the genus Rhodophana and separate these species
morphologically from all other members of the Rhodocybe-Clitopilus clade. The well-developed tapered pyramidal
scales of the pileus composed of agglutinated cylindrical hyphae (Fig. 10) combined with the large size and yellow
stipe are characteristic for this new species of Rhodophana. The acicular crystals on the hyphae of the rhizomorphs
(Fig. 12−13) are also distinctive and have not been reported for any other species of Rhodophana or other members
of the Rhodocybe-Clitopilus clade.
The pileus colors are reminiscent of, and yet differ sharply from, the orange-brown and smaller species,
Rhodophana nitellina and R. melleopallens that have glabrous smooth pileus surfaces with concolorous stipes and
pilei. Although Rhodocybe nitellina has been recorded from Africa (Pegler 1977, Morris 1990) neither R. nitellina nor
R. melleopallens were found during this study.
There are two other species of Rhodophana that have a squamulose pileus surface, Rhodocybe fibulata Pegler
(1977:526), described from Uganda and Tanzania in east Africa and Rhodophana squamulosa K.P.D.Latha & Manim.
(in Raj et al. 2016:91) from India. Rhodocybe fibulata is a smaller species than R. flavipes, with pileus size 20−35 mm
broad, and is characterized by its ferruginous to dark brown finely squamulose pileus, ferruginous to pale brown stipe,
pale pinkish brown lamellae, ovoid to broadly ellipsoid undulate bumpy basidiospores [6−8(−9) × 4.3−6 µm—Pegler
1977], and inflated subglobose or pyriform terminal cells in the erect squamules of the pileus. The basidiospores, the
abundant clamp connections on the hyphae and the pigmentation and general habit of this taxon indicate it belongs in
Rhodophana. A request to borrow paratype material of Rhodocybe fibulata Pegler from Kew (K) in order to extract
DNA to produce molecular profiles has not been granted as of this writing. However, one of us (TJB) has previously
performed a micromorphological examination of the type of R. fibulata, and confirms that morphologically it belongs
in Rhodophana, thus the following combination is in order.
Rhodophana fibulata (Pegler) T. J. Baroni, Kluting & Daniëls, comb. nov.
Basionym: Rhodocybe fibulata Pegler, Kew Bull. Additional Series VI. p. 526. 1977.
Mycobank: MB# 819907
A NEW SPECIES AND A NEW COMBINATION OF RHODOPHANA Phytotaxa 306 (3) © 2017 Magnolia Press • 231
Rhodophana squamulosa is readily distinguished from R. flavipes by numerous features, but most significantly by
its much smaller size, different colors of the basidiomata, the smaller and differently shaped basidiospores and by
the 5% difference in rpb2 sequences (Genbank KT180331, Raj et al. 2016) when compared with BLAST search in
the GenBank database (www.ncbi.nlm.nih.gov). The pileus of R squamulosa is 2−19 mm broad, centrally depressed
becoming infundibuliform, uniformly dark brown in color, not hygrophanous and the margin is not striate. The stipe is
2−3 mm broad and 5−20 mm long, brown with grayish orange towards the base, while the basidiospores are (5.5−) 6−8
× 4−5.5 µm, which are not only significantly smaller than those of R. flavipes, but of different shape being lacrymoid
or subglobose (Raj et al. 2016).
Conclusions
The rhizomorphs and the basal mycelium are frequently overlooked in fungal descriptions, as are many other characters
such as chlamydospores, skeletal hyphae, dendroid hyphae, repository (secretory) hyphae, acanthocytes and oxalate
crystals which have proved to be of taxonomic and/or functional value in fungi (Christan 2008, Clémençon 2003,
Horner et al. 1995, Hutchison et al. 1996, Luo et al. 2004, 2006, Zamora et al. 2013). Some of these structures appear
to be formed in order to cause a mechanical injury to soil nematodes to obtain an extra amount of nutrients (Luo et al.
2004, 2006) or for protection purposes (Whitney & Arnott 1986, Krisai & Mrazec 1986). Since no previous references
about mycelial structures of Rhodocybe s.l. or Rhodophana were known, this new species persuades us to look for such
structures more carefully within the Entolomataceae.
The fungal diversity of the African savanna and Niger are still poorly known. Rhodophana flavipes may also be
present in other west African countries with similar habitats that border the W National Park of Niger (e.g., Benin and
Burkina Faso) and possibly other countries such as Nigeria.
Acknowledgments
This study was part of the “Edible and cultivable Macromycetes of Niger (Ethnomycology)” project, financed by the
Spanish Agency for International Cooperation and Development (AECID-C/023163/09; D/031488/10; A1/039675/11).
The authors are grateful to the curators of the COFC-F herbarium for the exsiccata loan management.
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