Studies in African thelephoroid fungi: 1. Tomentella capitata and Tomentella brunneocystidia, two new species from Benin (West Africa) with capitate cystidia

Abstract

This paper presents Tomentella capitata and Tomentella brunneocystidia as new species based on molecular data and anatomical features. Both T. capitata and T. brunneocystidia form sister species with Tomentella pilosa. All three taxa are well supported by bootstrap values. Anatomically, T. capitata and T. brunneocystidia are very close and are similar in shape, size, ornamentation of basidiospores, and size and colour of subicular hyphae. Monomitic
rhizomorphs sometimes covered by irregularly shaped thin hyphae are present in both species. Shape and pigmentation of the
cystidia are the most discriminating features between T. capitata and T. brunneocystidia. The cystidia of T. capitata are maximum 35μm long, show a distinctive globose apex and are sometimes covered with dark brown pigmentation and/or encrustation,
whereas cystidia of T. brunneocystidia are bigger, up to 55μm long, with a sub-capitate shape and dark blue to dark green contents all over their length. The differences
to species, already described as having capitate and clavate cystidia, are discussed. A key for the identification of cystidioid
Tomentella species is given.

Full text

ORIGINAL ARTICLE
Studies in African thelephoroid fungi: 1. Tomentella capitata
and Tomentella brunneocystidia, two new species
from Benin (West Africa) with capitate cystidia
Nourou Soulemane Yorou &Urmas Kõljalg &
Brice Sinsin &Reinhard Agerer
Received: 2 June 2006 /Revised: 30 October 2006 / Accepted: 3 November 2006 / Published online: 22 December 2006
#German Mycological Society and Springer-Verlag 2006
Abstract This paper presents Tomentella capitata and
Tomentella brunneocystidia as new species based on
molecular data and anatomical features. Both T. capitata
and T. brunneocystidia form sister species with Tomentella
pilosa. All three taxa are well supported by bootstrap
values. Anatomically, T. capitata and T. brunneocystidia are
very close and are similar in shape, size, ornamentation of
basidiospores, and size and colour of subicular hyphae.
Monomitic rhizomorphs sometimes covered by irregularly
shaped thin hyphae are present in both species. Shape and
pigmentation of the cystidia are the most discriminating
features between T. capitata and T. brunneocystidia. The
cystidia of T. capitata are maximum 35 μm long, show a
distinctive globose apex and are sometimes covered with
dark brown pigmentation and/or encrustation, whereas
cystidia of T. brunneocystidia are bigger, up to 55 μm
long, with a sub-capitate shape and dark blue to dark green
contents all over their length. The differences to species,
already described as having capitate and clavate cystidia,
are discussed. A key for the identification of cystidioid
Tomentella species is given.
Introduction
Donk (1964) defined corticioid fungi as an assemblage of
extremely diverse organisms. The corticioid fungal group
includes indeed many paraphyletic genera (Parmasto 1995)
and has been considered as the outcome of a regressive
evolution of erect basidiomata (Corner 1954), whilst Heim
(1948) regarded them as primitive forms of the current
erected basidiomes. The last assumption is supported by
recent phylogenetic analysis (Larsson et al. 2004)as
corticioid fungi have proven to occupy basal clades within
almost all major identified lineages, among them the
thelephoroid one.
Thelephoroid fungi are characterised by the presence
of thelephoric acid (Bresinsky and Rennschmid 1971;
Gill and Steglich 1987) and the strongly ornamented, often
dark brown basidiospores. The group is rather diverse
regarding shape of the basidiomes, and includes species
with either poroid, irpicoid, lamellate, hydnoid or odontoid
hymenophores (Stalpers 1993).
Tomentella Persoon ex. Pat. is a member of Thelephorales
(Stalpers 1993). It has been previously described under se-
veral generic names including Caldesiella Sacc., Hypochnus
Fr. per Fr., Odontia Pers., Corticium subg. Tomentel l a Pers.,
and Thelephora Pers. with Thelephora ferruginea (Pers.)
Pers. as type species. The generic name Toment e l l a is
conserved and Tomentella ferruginea (Pers.) Pat. is quoted
as type species (Larsen 1974).
Microscopically, the genus Tomentella comprises species
with either monomitic or dimitic hyphal systems, clamped
or simple septate hyphae (Stalpers 1993; Kõljalg 1996),
Mycol Progress (2007) 6:7–18
DOI 10.1007/s11557-006-0519-4
Taxonomical novelties:Tomentella capitata Yorou & Agerer,
Tomentella brunneocystidia Yorou & Agerer.
N. S. Yorou (*):R. Agerer
Department Biology I and Geo-Bio Centre, LMU,
Biodiversity Research Group: Mycology,
Menzinger Str. 67,
80638 München, Germany
e-mail: n.s.yorou@gmail.com
N. S. Yorou :B. Sinsin
Laboratoire d´Ecologie Appliquée, Faculté des Sciences
Agronomiques, Université d´Abomey-Calavi,
01 BP 526 Cotonou, Benin
U. Kõljalg
Institute of Botany and Ecology, University of Tartu,
40 Lai St.,
51005 Tartu, Estonia

basidia and cystidia (if present) of various shape and for
some species relatively complex rhizomorphs. The most
recent monograph of this genus (Kõljalg 1996) based the
delimitation of Tomentella from Tomentellopsis Hjortstam
and Pseudotomentella Svrček on the size of basidiospores,
their ornamentation and their colour in 3% KOH and in
water. In this context, members of the genus Tomentellopsis
possess simple septate hyphae, colourless subicular and
sub-hymenial hyphae, and colourless echinulate basidio-
spores not exceeding 8 μm in size. The species of
Pseudotomentella hold colourless, yellowish to brownish
(in 3% KOH) spores with bi- or tri-furcate ornamentations,
whereas Tomentella covers species with spores different
from those of Tomentellopsis and Pseudotomentella. Within
the genus Tomentella, Larsen (1974); Stalpers (1993) and
Kõljalg (1996) based the species delimitation on the
presence or absence of rhizomorphs and cystidia but also
on the size, shape and ornamentation of spores.
If present, cystidia are either hyphoid, acuminate, clavate
or capitate (Kõljalg 1996) and with or without encrustation
and pigmentation.
In this study, we describe T. capitata and T. brunneocys-
tidia as new species. Both species have capitate cystidia that
do not resemble cystidia of already described Tom e n t ella
species.
There are many papers which include one to few species
of resupinate thelephoroid fungi found in Africa (Wakefield
1966; Malençon 1952,1954; Patouillard 1897; Martini and
Hentic 2002). This paper is a first of a series exclusively
devoted to tropical African resupinate thelephoroid species
and their ectomycorrhizae with native trees.
Materials and methods
Collections and specimens
The study is based on specimens collected during the rainy
season of 2003, 2004 and 2005 in woodlands and dry
forests of the Soudanean Endemism Centre (White 1983).
Specimens are collected under native trees and dried using
a propane gas heated dryer (De Kesel 2001). Preliminary
morphological descriptions are made using both fresh and
dried material. Colour patterns are given according to
Kornerup and Wanscher (1978). All specimens used for
descriptions and the holotypes are deposited in M
(Holmgren et al. 1990). Isotypes of both new species are
deposited in TU (Holmgren et al. 1990).
Light microscopy and drawings
Microscopic structures are described using dried specimens.
Fine sections through the basidiocarp were made using a
razor blade under a stereomicroscope (WILD Heerbrugg
M5, Switzerland) and mounted in water and afterwards in
2.5% KOH. Microscopic studies were performed using a
light microscope Leica DM LB2. Measurements were made
at a magnification of ×1,000. Measurements of basidia and
basidiospores do not include sterigmata, apiculus and orna-
mentation. Line drawings were made at magnification ×1,000
using a drawing tube. Observations were also made in
Cotton blue, Congo Red and Melzer’s reagent (Kreisel and
Schauer 1987). The format for descriptions follows
Kõljalg (1996).
Molecular methods and analyses
DNA from fungal fruit bodies were extracted and ITS
regions sequenced following methods described in Kõljalg
et al. (2000). Sequences are kept in Genbank NCBI with
accession numbers DQ848610–DQ848613. ITS sequences
of two specimens of Tomentella capitata and two of
T. brunneocystidia were obtained and aligned manually
against ITS data matrix used in Kõljalg et al. (2001). This
paper (Kõljalg et al. 2001) studied Tomentella species
which form cystidia and these data are therefore vital in
molecular recognition of new cystidioid taxa. In addition,
two unpublished ITS sequences of Tomentella pilosa (Burt)
Bourdot & Galzin from Costa Rica and USA and one
T. pilosa sequence from ectomycorrhizae (GenBank no
AY874385) were included into data matrix.
Distance neighbour joining analyses with Hasegawa–
Kishino–Yano (HKY85) substitution model, pairwise base
differences and bootstrap support were calculated by using
the beta version 4.0d81 of PAUP (Swofford 2003).
Results
Sequence analyses
Neighbour-joining tree demonstrating the position of T.
capitata and T. brunneocystidia sequences is shown on
Fig. 1. The two specimens of T. capitata have identical
sequences and cluster together. Specimens of T. brunneo-
cystidia cluster together too. Both T. capitata and T.
brunneocystidia are close to T. pilosa with which they form
a monophyletic group with a high bootstrap value. Sequence
differences between all three species are given in Table 1.
T. capitata Yorou & Agerer, sp. nov.
Description
Basidiocarpis resupinatis, separabilibus, pelliculosis, con-
tinuis. Hymenio ochraceo-brunneo usque ad atro-brunneo,
8 Mycol Progress (2007) 6:7–18

interdum viride-brunneo, granuloso vel farinoso, subiculo
concolorato. Marginibus indeterminatis.
Rhizomorphae in subiculo et in margine, atro-brunneae
usque ad atro-umbrinae, monomiticae, leves, interdum
cystidiis capitiformibus; cystidiae brunneae usque ad atro-
brunneae in 2.5% KOH et in aqua, nec congophilae, nec
cyanophilae, nec amyloideae; hyphis in superficie tenuis,
1–2μm in diametro, defibulatis, irregulariter ramificatis;
hyphae internae fibuligerae, 0.5–1μm crassitunicatae,
brunneae usque ad atro-brunneae in 2.5% KOH et in
aqua, nec congophilae, nec cyanophilae, nec amyloideae;
Tab l e 1 Pairwise base difference between Tomentella capitata,
Tomentella brunneocystidia and Tomentella pilosa
Tomentella
capitata (%)
Tomentella
brunneocystidia (%)
Tomentella pilosa 4.3–5.7 6.5–7.0
Tomentella
brunneocystidia
7.5
Fig. 1 The neighbour-joining
tree showing the placement of
Tomentella capitata and
T. brunneocystidia among
Tomentella and Thelephora
species. Bootstrap values are
shown above the branches.
The GenBank, EMBL or
UNITE codes of samples are
indicated after the species name
Mycol Progress (2007) 6:7–18 9

hyphae sub superficio rhizomorpharum 4–6μm in diame-
tro, hyphae in centro rhizomorpharum 4–7μm in diametro.
Hyphae subiculi fibuligerae, septa simplicia deficientia,
(3.5)4–5.5(6)μm in diametro, 0.5–1μm crassitunicatae,
non incrustatae, pallide brunneae in 2.5% KOH et in aqua,
nec congophilae, nec cyanophilae, muribus in parte
amyloideis.
Hyphae subhymenii fibuligerae, 3–5μm in diametro,
tenuitunicatae usque ad 0.5 μm crassitunicatae, cellulis
brevibus, inflatis, incoloratae usque ad pallide brunneae in
2.5% KOH et in aqua, nec congophilae, nec cyanophilae,
nec amyloideae, hyphae juveniles hymenio et subhymenio
in parte muribus hyphis subamyloideis.
Cystidia rhizomorpharum capitiformia, base fibuligera,
20–48(50) μm longa, apice 4–6μm in diametro, base
3–4.5 μm, incolorata in 2.5% KOH et in aqua, rare
incrustata; cystidia hymenii late capitiformia, 25–35 μm
longa, apice 9–14(16) μm in diametro, base 4–6.5 μm, non
septata, interdum flavo-brunnea, plerumque incolorata vel
capitibus incrustatis atro-brunneis.
Basidia 25–45 μm longa, 8–12 μm in diametro, base
6–8μm, basibus fibuligeris, suburniformia, non stipitata,
rare sinuosa, septa transversa deficientia, 4-sterigmatica,
sterigmatibus 5–7μm longis, basaliter 1–1.2 μm latis,
incolorata in 2.5% KOH et in aqua, cyanophilae et
congophilae.
Basidiosporae 7–9(9.5)×7–9(9.5) μm in aspectu fron-
tali, (7.5)8–9(9.5)×7–8(9) μm in aspectu laterali, lobatae
in aspectu frontali, ellipsoideae in aspectu laterali, echinu-
latae, aculeis usque ad 2 μm longis, guttulis frequentibus,
pallide brunneae usque ad brunneae in 2.5% KOH et in
aqua, nec cyanophilae, nec congophilae, nec amyloideae.
Chlamydosporae absentes
Basidiocarp—resupinate, separable from the substrate,
pelliculose, is continuous. Hymenium is light brown (6D3),
brown (6F4) to dark brown (7F4), granulose/farinose,
concolorous with subiculum; sterile margin indeterminate.
Rhizomorphs—present in subiculum and at the margins,
dark brown to dark umber under a dissection microscope,
dark brown in water and in 2.5% KOH, monomitic, young
rhizomorphs (thinner than 30 μm) smooth, rarely with
irregularly shaped thin hyphae on the surface (Fig. 2), older
rhizomorphs (thicker than 30 μm), patchily covered by
irregularly shaped thin hyphae (Fig. 3), sometimes with
cystidia emerging from the surface (Fig. 3), cystidia colour-
less in 2.5% KOH and in water, sometimes with encrusta-
tion, neither congophilous nor cyanophilous, nor amyloid;
superficial thin hyphae 1–2μm diameter, emerging from
thicker hyphae (Fig. 4), frequently branched, exceptionally
with clamps, frequently with simple septa, some without
septa and recalling then binding hyphae of polypores;
internal hyphae (hyphae below surface plus central hyphae)
clamped, thick-walled (0.5–1μm thick), brown to dark
brown in 2.5% KOH and in water, not congophilous, not
cyanophilous, not amyloid; hyphae below surface 4–6μm,
few hyphae repeatedly simple septate; central hyphae wider,
4–7μm, simple septa infrequent.
Subicular hyphae—consistently broadly clamped
(Fig. 5), simple septa lacking, (3.5)4–5.5(6) μm, thick-
walled (0.5–1μm), without encrustation, pale brown in
2.5% KOH and water, not congophilous, not cyanophilous,
walls amyloid at patches.
Subhymenial hyphae—clamped, 3.5–5μm diameter,
thin- to thick-walled (0.5 μm), cells not short and inflated,
colourless to pale brown in water and in 2.5% KOH, neither
congophilous nor cyanophilous, walls of young hyphae of
hymenium and subhymenium slightly amyloid.
Cystidia—from rhizomorphs capitate, clamped at base,
aseptate along their length, 20–48(50) μm long, 4–6μmat
apex and 3–4.5 μm at base, colourless in 2.5% KOH and in
water, rarely with encrustation, neither congophilous nor
cyanophilous, nor amyloid, cystidia of the hymenium
broadly capitate, clamped at base, aseptate along their
length, 25–35 μm long, 9–14 (16) μm at apex and 4–
6.5 μm at base, sometimes with yellow–brown contents,
often colourless but then with dark brown pigment and/or
encrustation covering the inflated apex.
Basidia—25–45 μm long, 8–12 μm at apex and 6–8μm
at base, clamped at base, suburniform, not stalked, rarely
sinuous, without transverse septa, colourless in 2.5% KOH
and in water, cyanophilous and congophilous, 4-sterigmate,
sterigmata 5–7μm long and 1–1.2 μm at base.
Basidiospores—7–9(9.5)×7–9(9.5) μm in frontal face
and (7.5)8–9(9.5)×7–8(9) μm in lateral face, lobed in
Fig. 2 Young rhizomorph of
Tomentella capitata Yorou &
Agerer
10 Mycol Progress (2007) 6:7–18

frontal view, ellipsoid in lateral view, echinulate, aculei
relatively long, 1–2μm, oil drops common, pale brown to
brown in 2.5% KOH and in water, not cyanophilous, not
congophilous, not amyloid.
Chlamydospores—absent.
Type material
Benin, central part, Borgou Province, reserved forest of
Wari–Maro, Wari–Maro site, 08° 49′19.0″N, 002° 16′
32.4″E, on dead bark and log, leg. N. S. Yorou,
05.08.2005, herb SYN 860. Holotype in M; isotype in
TU. Genbank NCBI, accession number DQ848612.
Etymology
The epithet is proposed in reference to the distinctly
capitate cystidia.
Material studied
Benin, central part, Borgou Province, Sinendé region, forest
close to Fo–Bouko Village, 10° 8′46.6″N, 002° 15′6.00″
E, on dead bark and log, leg. R. Agerer, 22.08.2003, herb
RA 13781 (M). Benin, central part, Borgou Province,
reserved forest of Wari–Maro, Agbassa area, 08° 53′31.5″
N, 002° 19′08.8" E, on dead bark and log, leg. N. S. Yorou,
14.07.2004, herb SYN 631 (M). Benin, central part, Borgou
Province, reserved forest of Wari–Maro, Wari–Maro area,
09°00′47.1″N, 002° 01′36.9″E, on dead bark and logs of
native trees, leg. N. S. Yorou, 05.08.2005, herb SYN 862
(M), SYN 856 (M), SYN 846 (M), SYN 848 (M), SYN 841
(M), SYN 844 (M), and SYN 855 (M).
Habitat
In woodlands and dry forests of the Soudanean Endemism
Centre (White 1983) dominated by Ceasalpiniaceae,
Fig. 3 Rhizomorphs of Tomen-
tella capitata,acapitate
cystidium with irregularly
shaped thin hyphae and
boptical section through the
rhizomorph
Fig. 4 Surface view of rhizomorph of T. capitata showing the
connection between irregularly shaped thin hyphae and generative
ones
Mycol Progress (2007) 6:7–18 11

Euphorbiaceae and Dipterocarpaceae. On dead bark and
log, under native trees such as Afzelia africana Smith,
Isoberlina doka Craib & Stapf, Isoberlina tomentosa
(Harms) Craib & Stapf, Monothes kerstingii Gilg, Uapaca
togoensis Pax, Burkea africana Hook. F. and Detarium
microcarpum Guill. & Poir. Sporophores growing on dead
bark and logs of various trees, always at the interface
between soil and logs.
Tomentella brunneocystidia Yorou & Agerer, sp. nov
Description
Basidiocarpis resupinatis, separabilibus, arachnoideis, con-
tinuis. Hymenio aureo-brunneo usque ad atro-brunneo,
granuloso, subiculo concolorato. Marginibus indeterminatis.
Rhizomorphae in subiculo et in margine, frequentes,
frequenter ramificantes, atro-brunneae, monomiticae, leves,
cystidiis deficientibus; hyphis in superficie tenuis infre-
quentibus, 1–2μm in diametro, defibulatis, irregulariter
ramificatis; hyphae internae fibuligerae, 0.5–1μm crassi-
tunicatae, subbrunneae usque ad brunneae in 2.5% KOH et
in aqua, nec congophilae, nec cyanophilae, nec amyloi-
deae; hyphae sub superficio rhizomorpharum 3–4μmin
diametro, frequentibus hyphis multis septis simplicibus,
hyphae in centro rhizomorpharum 3–6μm in diametro,
septis simplicibus raris vel deficientibus.
Hyphae subiculi fibuligerae, septa simplicia deficientia,
(3)4–6(6.5) μm in diametro, 0.5–1μm crassitunicatae,
incrustatae granulis brunneis, pallide flavae vel flavae in
2.5% KOH et in aqua, nec congophilae, nec cyanophilae,
nec amyloideae.
Hyphae subhymenii fibuligerae, 4–7μm in diametro,
tenuitunicatae usque ad 0.5 μm crassitunicatae, cellulis
brevibus, inflatibus deficientibus, flavae in 2.5% KOH et in
aqua, nec congophilae, nec cyanophilae, nec amyloideae.
Fig. 5 Tomentella
capitata Yorou & Agerer,
abasidiospores in frontal view,
bbasidiospores in lateral
view, and csection through
the basidiocarp
12 Mycol Progress (2007) 6:7–18

Cystidia ab hyphae subiculae formantes, rhizomorphis
cystidiis deficientibus, subcapitiformia, base fibuligera, (22)
25–50(55) μm longa, apice 7–10 μm in diametro, base 5–
7μm, non septata, non incrustata, frequenter contento atro-
brunneo, in 2.5% KOH brunneo-atro, interdum incolorata.
Basidia (20)22–38(40) μmlonga,6–10 μmindiametro,
base 5–8μm, basibus fibuligeris, clavata, non stipitata,
frequenter sinuosa, septa transversa deficientia, 4-sterigma-
tica, sterigmatibus 5–8μm longis, basaliter 1–1.2 μmlatis,
frequenter incolorata in 2.5% KOH et in aqua sed interdum
subflava, cyanophilae et congophilae, non amyloidea.
Basidiosporae 7.5–8×7–8μm in aspectu frontali, 6.5–7
(8) μm in aspectu laterali, triangulares vel lobatae in aspectu
frontali, ellipsoideae in aspectu laterali, echinulatae, aculeis
usque ad 1 μm longis, guttulis frequentibus, pallide brunneae
usque ad subflavae in 2.5% KOH et in aqua, nec cyanophilae,
nec congophilae, nec amyloideae .
Chlamydosporae absentes
Basidiocarp—resupinate, separable from the substrate,
arachnoid, continuous. Hymenium is golden brown (5D7)
to dark brown (6F5), granulose, and concolorous to the
subiculum. Sterile margin is indeterminate.
Rhizomorphs—present in the subiculum and at margin, in
great numbers and repeatedly branched, dark under a dissection
microscope, pale brown to brown in 2.5% KOH and in water,
without cystidia, monomitic; young rhizomorphs (thinner than
40 μm) smooth (Fig. 6), sometimes peripheral clamped hyphae
with many simple septa (Fig. 7); older rhizomorphs (thicker
than 40 μm) commonly patchily covered with dense
irregularly shaped thin hyphae (Fig. 8); superficial thin hyphae
1–2μm diameter, emerging from commonly simple septate
hyphae (Fig. 9), multiply and densely branched, clampless,
some without septa then similar to binding hyphae of
polypores, sometimes growing along and around the thicker
hyphae (Fig. 10); internal hyphae (hyphae below surface plus
central hyphae) clamped, thick-walled (0.5–1μm), yellow in
2.5% KOH and in water, not congophilous, not cyanophilous,
not amyloid; hyphae below surface 3–4μm, many hyphae
with several simple septa; central hyphae wider, 3–6μm,
simple septa infrequent.
Subicular hyphae—clamped (Fig. 11), (3)4–6(6.5) μm;
thick-walled (0.5–1μm), simple septa lacking, pale yellow
to yellow in 2.5% KOH and in water, not cyanophilous, not
congophilous, not amyloid, with red–brown encrustation
that are observable in water and Melzer’s reagent, rapidly
dissolving in KOH.
Subhymenial hyphae—clamped, 4–7μm in wide, cells
not short and inflated, thin to thick-walled (0.5 μm), yellow
in water and in 2.5% KOH, not congophilous, not
cyanophilous, not amyloid.
Cystidia—sub-capitate, arising from subicular hyphae,
clamped at base, aseptate along their length, (22)25–50(55)
μm long, 7–10 μm at apex and 5–7μm at base, frequently
with dark green or dark blue contents all over their length,
without encrustation, sometimes colourless.
Basidia—(20)22–38(40) μm long, 6–10 μm at apex and
5–8μm at base, clamped at the base, clavate, not stalked,
often sinuous, without transverse septa, often colourless but
sometimes light yellow in 2.5% KOH and in water,
cyanophilous and congophilous, not amyloid, 4-sterigmate,
sterigmata 5–8μm long and 1,5–2μm at base.
Basidiospores—7.5–8×7–8μm frontal face and 6.5–7
(8) μm lateral face, triangular to lobed as seen in frontal
view and ellipsoid in lateral view, echinulate, aculei short,
not exceeding 1 μm, oil drops common, pale brown to pale
yellow in 2.5% KOH and in water, not cyanophilous, not
congophilous, not amyloid.
Chlamydospores—absent.
Fig. 6 Optical section through
young rhizomorph of
T. brunneocystidia Yorou
& Agerer
Fig. 7 Optical section of young rhizomorph of T. brunneocystidia showing mostly simple septate peripheral hyphae
Mycol Progress (2007) 6:7–18 13

Type material
Benin, central part, Borgou Province, reserved forest of Wari–
Maro, Wari–Maro area, Wari–Maro, 09° 00′47.3" N, 002°
52′36.9" E, on dead barks and logs of native trees, leg. N. S.
Yorou, 05.08.2005, herb. SYN 839. Holotype in M; isotype in
TU. Genbank NCBI, accession number DQ848613.
Etymology
The epithet is proposed in reference to the brown to dark
green content of the cystidia.
Material studied
Benin, central part, Borgou Province, reserved forest of
Ouémé Supérieur, Arboretum of Kpessou–Samari, 09° 00′
47.3″N, 002° 52′36.9″E, on dead bark and logs of native
trees, leg. R. Agerer, 23.08.2003, herb. RA 13815 (M), RA
13779 (M), RA 13823 (M). Benin, central part, Borgou
Province, reserved forest of Wari–Maro, Wari–Maro area,
9°00′7.7″N, 002° 00′18.8″E, on dead bark and logs, leg
N. S. Yorou, 05.08.2005, herb. SYN 859 (M), SYN 863
(M). Benin, central part, Borgou Province, forest of Wari–
Maro, Wari–Maro area, Wari–Maro, 08° 12′25.6″N, 002°
47′31.8″E, leg. N. S. Yorou, 06.08.2005, herb SYN 865
(M), SYN 868 (M), SYN 883 (M), SYN 885 (M), SYN 889
(M), SYN 894 (M), SYN 897 (M).
Habitat
Collected in woodlands and dry forests of various native
trees such as Isoberlinia doka,I. tomentosa,U. togoensis
and A. africana. On dead bark and logs, either at the
interface between soil and bark or on free bark.
Discussion
Both T. capitata and T. brunneocystidia are comparatively
different regarding the ITS sequences (4.3–5.7 and 6.5–
7.0% respectively) with T. pilosa. All three species form a
monophyletic group with T. atroarenicolor Nikol. as a
sister species. The last taxon is also clearly deviating from
T. capitata and T. brunneocystidia by 5.9 and 7.2%,
respectively.
T. capitata and T. brunneocystidia are anatomically very
similar. Both species are separable from the substrate, form
granulose to farinose hymenophores and present dark
rhizomorphs under a dissection microscope. Microscopic
similarities of these species include the thick-walled (0.5–
1μm) subicular hyphae of 4–6μm(rarely7–8μm), the dark
Fig. 8 Optical section through an old rhizomorph of T. brunneocystidia
Fig. 9 Surface view of the rhizomorph of T. brunneocystidia showing
the connection between surface hyphae and generative ones
14 Mycol Progress (2007) 6:7–18

brown (in water and in 2.5% KOH) rhizomorphs that form
irregularly shaped thin hyphae on their surface and the
triangular to lobed basidiospores in frontal view and ellipsoid
in lateral view. The size of the basidiospores is slightly
dissimilar, ranging from 7–9.5 μminT. capitata and 6.5–
8μminT. brunneocystidia. However, the aculei of the
basidiospores of T. capitata are distinctly longer (1–2μm)
than those of the basidiospores of T. brunneocystidia that are
at most 1 μm. The species also differ in shape, colour, and to
some extent, the contents of cystidia. The cystidia in T.
capitata are 25–35 μm and have a distinctly capitate apex
9–14 (16) μm in diameter. They commonly present a dark
brown pigmentation and/or encrustation covering the inflated
head (Fig. 5). The cystidia in T. brunneocystidia are longer,
ranging from 25 to 50 (55) μm have a subcapitate apex of
7–10 μm diameter (Fig. 11) and brown, dark blue to dark
green contents all over their length. However, colourless
cystidia are sometimes present in T. brunneocystidia.Other
differences include hyphal dimension, presence of cystidia
on rhizomorph and the colour of subhymenial hyphae in
water, 2.5% KOH and Melzer’s reagent. In T. capitata,
hyphae are pale brown to brown (in water and in 2.5%
KOH) slightly amyloid in patches. An amyloid reaction is
also present in young hyphae of the hymenium and the
subhymenium. In T. brunneocystidia, subicular and subhy-
menial hyphae are pale yellow to yellow (in water and in
2.5% KOH) and no positive reaction is observed with
Melzer’s reagent. The subicular hyphae of T. brunneocysti-
dia have encrustation that quickly dissolves in KOH. No
encrustations are observed on subicular hyphae of T.
capitata. Subhymenial hyphae of T. capitata are 3.5–5μm
wide while those of T. brunneocystidia range from 4 to 7 μm
diameter. In T. capitata, old rhizomorphs have capitate
cystidia that emerge from the surface. No cystidia are
observed on rhizomorphs of T. brunneocystidia.
Among the described Tomentella species, T. pilosa and
Tomentella viridula Bourdot & Galzin have capitate
cystidia (Kõljalg 1996; Melo et al. 1998). The cystidia of
T. pilosa are 50–120 μm and of 6.5–13 μm diameter at
apex. Those of T. viridula are 40–80 μm long and have a
diameter of 5–8 (12) at apex. Furthermore, T. virudula
lacks rhizomorphs and the rhizomorphs in T. pilosa are
dimitic with distinctive skeletal hyphae of 2–3μm (Kõljalg
1996). Both T. capitata and T. brunneocystidia present
brown to dark brown rhizomorphs that are monomitic and
show on their surface rather irregularly shaped and
repeatedly branched thin hyphae of 1–2μm diameter.
Distinctive skeletal hyphae have not been observed.
Clavate cystidia within the genus Toment e l l a are known
from Tomentella muricata (Ellis & Everh.) Wakel., Tomen-
tella clavigera Litsch. and T. subclavigera Litsch. (Kõljalg
1996). In T. clavigera, cystidia are 80–140 μmlongand
6–8μm wide at apex (Kõljalg 1996). Those of T.
subclavigera are 105–145 μmlongand7–11 μmatapex
(Kõljalg 1996;Meloetal.1998), and cystidia in T. muricata
are 50–105 μm long and 6.5–8.5 μmdiameteratapex.
Cystidia in all three species are therefore longer and
narrower than cystidia of T. brunneocystidia. Neither T.
clavigera nor T. subclavigera has rhizomorphs in contrast to
T. brunneocystidia.
The presence of irregularly shaped thin hyphae on the
surface of rhizomorphs has only rarely been reported for
thelephoroid fungi. Raidl and Müller (1996) and Raidl (1997)
mentioned the presence of repeatedly branched thin hyphae
on the surface of rhizomorphs of ectomycorrhizae formed by
T. ferruginea (Pers.) Pat. on Fagus sylvatica L. Similar
narrow surface hyphae have been shown for rhizomorphs of
ectomycorrhizae formed by T. subtestacea Bourdot & Galzin
on Populus alba (Jakucs and Agerer 2001) where they are
described as highly specialised, tortuous, repeatedly rami-
fied, densely entwined, glued and of dark yellow to brown
colour. In both, T. capitata and T. brunneocystidia, the thin
hyphae on the rhizomorphs are tortuous, too, densely and
repeatedly branched, mostly clampless but they are brown to
dark brown in water and in 2.5% KOH. According to Raidl
Fig. 10 Surface view of rhizomorph of T. brunneocystidia showing
thin hyphae growing along and around generative hyphae
Mycol Progress (2007) 6:7–18 15

(1997), the surface narrow hyphae occur at a late stage of the
rhizomorphs’ontogeny. We agree with this statement
because young rhizomorphs (under 35 μm diameter) of our
specimens commonly lack such hyphae.
All studied specimens are collected in woodlands and dry
forests of the Soudanean Endemism Centre (White 1983).
Woodlands and dry forests in Benin are dominated by
Ceasalpiniaceae (mainly I.doka and I. tomentosa,B.
Fig. 11 Tomentella brunneocys-
tidia Yorou & Agerer,
abasidiospores in frontal view,
bbasidiospores in lateral
view, and csection through the
basidiocarp
Fig. 12 Shape of cystidia within Tomentella species
16 Mycol Progress (2007) 6:7–18

africana,D. microcarpum, few specimens of A. africana are
also recorded); Euphorbiaceae (U. togoensis,Uapaca
guinneensis,) Fabaceae (Pterocarpus erinaceus) and one
single member of Dipterocarpaceae (Monotes kerstingii).
All mentioned species are putative ectomycorrhizae formers
(Sanon et al. 1997;Ducoussoetal.2002). Sporophores of T.
capitata were always found at the underside of logs or bark.
Sporophores of T. brunneocystidia have also been collected
on dead trunks that had no contact with the soil. In temperate
and boreal forests, members of resupinate telephoroid fungi
(including Tomentel l a ) are proved to be ectomycorrhizal
symbionts (Agerer 1996; Agerer and Bougher 2001;Jakucs
and Agerer 1999;Jakucsetal.2005; Kõljalg et al. 2000,
2001). It is very likely that both Tome n t e lla species form
ectomycorrhizal symbiosis with native trees.
Key for the identification of cystidioid Tomentella species
(Shapes of cystidia are illustrated in Fig. 12)
1. Cystidia subcapitate to capitate
2. Rhizomorphs absent, cystidia capitate, 40–80 μm long,
5–9 (12) μm at apex, colourless, often with encrusta-
tion around the head. T. viridula
2*. Rhizomorphs present
3. Rhizomorphs dimitic, with distinctive skeletal hyphae,
cystidia capitate, up to 130 μm long. T. pilosa
3*. Rhizomorphs monomitic, covered by irregularly
shaped, tortuous and repeatedly ramified narrow hy-
phae similar to binding hyphae of Polyporaceae
4. Cystidia with distinctive distal apex, 25–35 μmlong
and 9–14 μm at apex, old rhizomorphs sometimes
with distinctive capitate cystidia of 20–48(50) μm
long and 4–6μm at apex, aculei 1–2μm long.
T. capitata
4*. Cystidia subcapitate, 22(25)–50(55) long and
7–10 μm at apex, often with dark brown to blue–
green content over their length, aculei 0.5–1μm long.
T. brunneocystidia
1*. Cystidia of other shape
5. Cystidia clavate
6. Rhizomorphs present, cystidia 50–105 μm long. T.
muricata
6*. Rhizomorphs absent
7. Cystidia 80–140 μm long, basidia sometimes greenish
in KOH, basidiospores triangular to lobed in frontal
view. T. clavigera
7*. Cystidia 105–145 μm long, basidia never greenish,
basidiospores ellipsoid to globose in frontal view. T.
subclavigera
5*. Cystidia acuminate or hyphoid
8. Cystidia acuminate, with brown deposits at apices,
rhizomorphs absent,
9. Hymenophore greenish, basidiospores 7–8.5 μm long
in frontal and lateral faces, light brown in KOH. T.
galzinii
9*. Hymenophore brownish, basidiospores 6.5–8.5 μm
long in frontal and lateral faces, light brown to reddish–
brown in KOH. T. subtestacea
8*. Cystidia hyphoid, rhizomorphs present. T. atroarenicolor
Acknowledgement Financial supports were provided by the Ger-
man Academic Exchange Service (DAAD) through the grant no. A/
03/15106. Field works and equipment were financially supported
by the African Forests Research Network (AFORNET) with the
grant no. 02/2005 and the International Foundation for Science
(IFS) under grant no. D/4033-1. Leho Tedersoo and Triin Suvi are
also much thanked for their daily assistance during the visit of the
first author at the Institute of Botany and Ecology, University of
Tartu in Estonia.
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