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The genus Macrolepiota (Agaricaceae, Basidiomycota)
in China
Z. W. Ge & Zhu L. Yang & Else C. Vellinga
Received: 21 June 2010 / Accepted: 13 September 2010 / Published online: 30 September 2010
#
The Author(s) 2010. This article is published with open access at Springerlink.com
Abstract Species of the genus Macrolepiota (Agaricaceae)
in China were investigated on the basis of morphology and
DNA sequences data. Six species, i.e., M. detersa, M.
dolichaula, M. mastoidea, M. orientiexcoria ta, M. procera,
and M. velosa are recognized, of which M. detersa and M.
orientiexcoriata are new species. All of them are described
and illustrated with line drawings, and a key is provided to
those recognized species. The taxonomic uncertainty of M.
crustosa, originally described from China, is also discussed.
ITS sequences were used to support the new species
delimitations and to test the conspecificity between the
Chinese specimens and their relatives from other conti-
nents. Phylogenetic analyses identify three clades within
Macrolepiota: /macrolepiota, /macrosporae, and /volvatae
clade. /macrolepiota clade and /macrosporae clade respec-
tively correspond to section Macrolepiota and section
Macrosporae in Bon’s infrageneric classification. Section
Volvatae is proposed to accommodate species with a volva
but without clamp connections within Macrolepiota.
Keywords Agaricales
.
Chlorophyllum
.
Morphology
.
Systematics
.
Taxonomy
Introduction
The genus Macrolepiota (Agaricaceae, Agaricales, Basi-
diomycota) was established by Singer (1948). Macroscopi-
cally, basidiomata of species in this genus are typically big,
fleshy, and often with squamules on the pileus; lamellae are
white to cream ; a prominent annulus is usually present
which is often movable. Microscopically, clamp connec-
tions are present on the septa of the hyphae in lamellae;
basidiospores are thick-walled, relat ively big, white to
cream when accumulated, and the inner spore-wall is
metachromatic in cresyl blue (Singer 1948).
Originally, Macrolepiota only accommodated non-
volvate species. Species with a well-formed cup-like volva
were placed in a separate genus, namely Volvolepiota
Singer (Singer 1959). A recent study indicated that a volva
at the base of the stipe does not warrant a separate genus,
and thus, Volvolepiota is synonymous with Macrolepiota
(Vellinga and Yang 2003). Accordingly, the genus Macro-
lepiota in the current sense also contains species with a
cup-like volva.
Based on morphological and molecular data, Johnson
(1999) investigated the traditional classification of the light-
spored Lepiota s.l., and found Macrolepiota
is not
monophyletic. Later on, Vellinga et al. (2003) evaluated
the generic level of Macrolepiota, which was shown to be a
monophyletic genus after transferring species with pileal
squamules made up of a hymenidermal layer, spores with
truncated germ pore or without a germ pore, and a smooth
stipe to Chlorophyllum Massee. Consequently, representa-
tives of Macrolepiota in the present sense are characterized
by the combination of the following characters: pileal
squamules of a trichodermal layer made up of long
subcylindric elements, spores with a germ pore caused by
an interruption of the episporium covered by a hyalinous
Z. W. Ge
:
Z. L. Yang (*)
Key Laboratory of Biodiversity and Biogeography,
Kunming Institute of Botany, Chinese Academy of Sciences,
Kunming 650204, People’s Republic of China
e-mail: fungi@mail.kib.ac.cn
Z. W. Ge
Harvard University herbaria,
Cambridge 02138 MA, USA
E. C. Vellinga
Department of Plant and Microbial Biology,
University of California,
Berkeley, CA 94720-310, USA
Fungal Diversity (2010) 45:81–98
DOI 10.1007/s13225-010-0062-0
cap, and the presence of stipe squamules, often visible as
colored bands in the full-grown specimens (Vellinga 2003
Vellinga et al. 2003).
Currently, there are about 30 species recognized world
wide (Kirk et al. 2008). Although the genus contains some
edible species, which have been an interest to cultivate by
researchers (e.g. Ding and Huang 2003), knowledge of this
genus in East Asia is poor and fragmentary. Although a few
species of Macrolepiota were recorded from China (Shao
and Xiang 1981; Zang et al. 1996; Bi et al. 1997; Mao
2000; Teng 1996; Vellinga and Yang 2003), literature on
some of these records has very limited information in the
descriptions, and information on voucher specimens is
lacking (e.g. Mao 2000, 2009 ; Teng 1996). Based on
extensive morphological examination and molecular phy-
logenetic analyses, species diversity of Macrolepiota in
China and the affinities to other species of Macrolepiota are
presented. In addition, the infra-generic classification of
Macrolepiota is also discussed.
Materials and methods
Morphological studies
The examined materials were collected in China, and
deposited in KUN (with HKAS numbers), HMAS, GDGM,
BPI and HMJAU. Herbarium codes used follow Thiers
(2010). Color notations indicated in the descriptions are from
Kornerup and Wanscher (1978), and Color codes are
according to the Online Auction Color Chart™,indicated
by ‘oac’ before a number. The descriptions of species are in
alphabetical order by species epithet. In the description,
macromorphology is based on the field notes and color slides
of the material; micromorphology is based on observation of
the material under microscope. Melzer’sreagentwasusedto
test the amyloidy of spores. Other structures (e.g. pileal
structure, cheilocystidia and basidia) were observed in 5–10
%KOHandwithCongo–red before making line drawings.
The abbreviation [n/m/p] shall mean n basidiospores
measured from m fruit bodies of p collections in 5–10 %
KOH solution. At least 20 basidiospores were meas ured for
each collection. Dimensions for basidiospores are given as
(a-) b-c (-d). The range b-c contains a minimum of 90% of
the measured values. Extreme values (a and d) are given in
parentheses. Q is used to mean “length/width ratio” of a
spore in side view; avQ means average Q of all basidio-
spores ± sample standard deviation.
DNA isolation and amplification
Genomic DNA was extracted from dried material.
Small parts of the pileus tissue were ground in an
eppendorf tube using a pestle. DNA was isolated with a
modified Cetyltrimethylammonium bromide (CTAB)
procedure of Doyle and Doyle (19 8 7). ITS/5.8S rDNA
were amplified using primers ITS1F and ITS4 (White et
al. 1990; Gardes and Bruns 1993). PCR was performed in
a total volume of 25 μl containing 1 U Taq DNA
polymerase, 2.5 μl of 10×Taq polymerase reaction buffer,
1 μl of 25 mM magnesium chloride (QIAGEN Inc.,
Valencia, California, USA), 5 nmol of each dNTP, 0.6 μl
of 10 μM each of the two pr im ers and 1 μl o f the DNA
extract. PCR reactions were performed with 4 min initial
denaturationat95°C,followedby34cyclesof50sat
94°C, 40 s at 53°C, 50 s at 72°C, and a final extension of
7 min at 72°C followed the last cycle. PCR products were
purified using a QIAquick PCR purification kit (QIAGEN
Inc., Valencia, California, USA). Sequencing was per-
formed using a Bigdye terminator cycle sequencing kit
(Applied Biosystems, Foster City, California, USA)
following the manufacturer ’s protocol. Sequencing pri-
mers for the ITS regions were ITS1F and ITS4. Sequenc-
ing react ions were purified using Pellet Paint (Novagen,
Madison, Wisconsin, USA) and were run on an Applied
Biosystems 377 XL automated DNA sequencer. Sequence
chromatograms were c ompiled with Sequencher 4 .1
software (GeneCodes Corporation, Ann Arbor, Michigan,
USA).
Phylogenetic analyses
Sequences were aligned using CLUSTAL X 1.81 followed
by manual inspection and correction in MacClade
(Thompson et al. 1997; Maddison and Maddison 2000).
The resulting ITS data set was evaluated using two tree-
building methodologies: the maximum parsimony (MP)
criterion in PAUP* and the Bayesian criterion. Gaps were
treated as missing data in all analyses.
Maximum Parsimony ana lysis was pe rformed using
PAUP* 4.0b10 (Swofford 2004). One thousand heuristic
searches were conducted with random sequence addition
and tree bisection-reconnection (TBR) branch-swapping
algorithms, collapsing zero-length branches and saving all
minimal-length trees (MulTrees). To measure relative
support for the resulting clades, 500 bootstrap replications
were p erformed with the same parameters as for the
parsimony analyses (Felsenstein 1985).
To test alternative p hylogenetic relationships, the Bayes-
ian analysis were performed using MCMC with Mr. Bayes
V3.0b3 (Ronquist and Huelsenbeck 2003). Bayesian
analyses were repeated 4.2 million generations and sampled
every 100. The first 25% of generations were discarded as
burn-in, and Bayesian posterior probabilities (PP) were then
calculated from the posterior distribution of the retained
Bayesian trees.
82 Fungal Diversity (2010) 45:81–98
Results
Morphological observations
115 putative Macrolepiota specimens were examined, and
87 specimens of Macrolepiota are cited in this paper. These
examined specimens represent six Macrolepiota species of
which two are new to science. The six recognized species
are Macrolepiota detersa, M. dolichaula, M. mastoidea, M.
orientiexcoriata, M. procera and M. velosa, and they will
be described in detail in the taxonomy part. Some of the
previous records of M. dolichaula and M. procera are
misidentified in the literature and these will be addressed
under the material examined part of each species.
Molecular phylogenetic results
Sequences generated in this study were deposited in
GenBank with accession numbers from HM125507 through
HM125532, and the GenBank accession numbers for ITS
sequences are given with the lists of examined collections
and in the phylogenetic tree (Fig. 1). The final alignment
was deposi ted in TreeBASE (Study Accession URL: http://
purl.org/phylo/treebase/phylows/study/TB2:S10499). The
alignment comprises of 72 Macrolepiota sequences, plus
2 species of Leucoagaricus Locq. ex Singer. Leucoagaricus
barssii (Zeller) Vellinga and L. meleagris (Sowerby) Singer
were designated as outgroup based on a more inclusive
analysis of sequences of Agaricaceae (unpublished person-
al data). The aligned data set included 752 base pairs, of
which 22 bases were ambiguous and were excluded in the
analyses. Among the analyzed 730 base pairs, 482 are
constant, 48 are variable parsimony-uninformative charac-
ters, and 200 variable parsimony informative characters
were used to reconstruct the phylogeny. Maximum parsi-
mony analysis resulted in 9 equally parsimonious trees with
a tree length of 448 steps, CI=0.730, RI=0.947, HI=0.270.
Figure 1 shows one of the most parsimoni ous trees .
Bayesian analysis resulted in similar results, except the
clade formed by Macrolepiota clelandii changed to be basal
instead of the clade formed by Macrolepiota eucharis and
Macrolepiota velosa. However, basal status of M. clelandii
does not get statistical support.
In order to distinguish clade names from traditional
taxonomic names, clade names are written in lower cases,
never italicized, and preceded with the symbol “/”.As
shown in Fig. 1, Macrolepiota forms a well supported
monophyletic group and got strong bootstrap (100%) and
bayesian PP supports (1.00). Within Macrolepiota, three
clades were recovered.
Clade 1, here referred to as /volvatae clade, includes two
volvate species, M. eucharis and
M. velosa, this clade got
98 % bootstrap support and 1.00 bayesian PP support.
Macrolepiota velosa , described from southern China, is
sister to M. eucharis, a species described from Australia.
Clade 2, here referred to as /macrosporae clade, includes
M. excoriata, M. mastoidea, M. orientiexcoriata, M.
phaeodisca, M. konradii, M. psammophila, and M. sub-
squarrosa. T his clade got 100% bootstrap a nd 1.00
bayesian PP support. Within this clade, collections of M.
mastoidea from China clustered with collections from other
areas; M. orientiexcoriata collections from China clustered
together and got 64% bootstrap support.
Clade 3, here referr ed to as /macrolepiota clade, includes
the generic type M. procera, and its related allies such as M.
colombiana, M. detersa, M. dolichaula, M. fuliginosa, M.
rhodosperma, and an undescribed species from North
America. Macrolepiota clelandii, a species described from
Australia which may represent an independent clade (with
100% bootstrap support), formed a sister clade of the core /
macrolepiota clade (excluding M. clelandii) and got 51%
bootstrap support. For now, we tentatively include it in the /
macrolepiota clade.
Wi thin this Clade 3, the core /macrolepiota c lade
received 98% bootstrap support and 1.00 Bayesian poste-
rior probabilities support. Collections of M. procera from
China, clustered together w ith a Japanese collection,
forming an East Asian clade. This clade got 80% bootstrap
support and 0.99 Bayesian PP support and turns out to be
sister to European M. procera. Collections of M. dolichaula
from China and Australia form a monophyletic clade and
got 100% bootstrap support and 1.00 bayesian PP support.
Macrolepiota detersa, a novel species described in the
present paper, clustered with 3 collections of M. sp. from
Japan and 100 % bootstrap support and 1.00 bayesian PP
support.
Taxonomy
Macrolepiota detersa Z. W. Ge, Zhu. L. Yang & Vellinga
sp. nov. Fig. 2
MycoBank: MB 518349
Pileus 8–12 cm diametro, primo ovoideus vel hemi-
sphaericus, dein convexus vel plano- convexus, albus vel
albidus, squamuli s crustatis, griseolis-aurantiacis vel pal-
lide brunneis. Lamellae liberae, albae, confertae. Stipes
13.0–15.0×1.8–2.4 cm, subcylindricus, minutus sursum,
albidus, basim incrassatus. Annulus superus, albidus,
membranaceus. Caro alba; sapor mitis. Basidia 30–38×
11–15 μm, clavata, hyalina, 4-sporigera, raro 2-sporigera.
Basidiosporae 14.0
–16.0 (18.0)×(9.0) 9.5 – 10.5 (11.0) μm,
ellipsoideae, glabrae, hyalinae, dextrinoideae. Pleurocysti-
dia absentia. Cheilocystidia clavata, lato-clavata vel
pyriformia, raro subfusiformia, hyalina, 18–38×7–15 μm.
Squamulae pilei trichoderma, apicalis hyphis erectibus,
Fungal Diversity (2010) 45:81–98 83
84 Fungal Diversity (2010) 45:81–98
luteis vel luteo-brunneis, subcyl indricis compositae. Fibu-
lae praesentes. Habitatio: terrestris.
Holotypus: C. L. Hou 603 (HKAS 55306), 2 Oct. 2007,
Jingde County, Anhui Province, China.
Etymology: “detersa” refers to the easily detachable
squamules on the pileus.
Basidiomata (Fig. 2a) medium-sized to large. Pileus 8–
12 cm in diam., ovoid to hemispherical when young,
becoming c onvex to plano-convex with age, white to
whitish, covered with scattered, greyish orange (5B5-5B6,
oac688 or oac729) to light brown (6C7-6D7, oac777)
patch- or crust-like squamules which are easily detachable
from the pileus; disc smooth, light brown (6C7-6D7,
oac777). Lamellae free, moderately crowded, white when
young, white to cream colored when mature, up to 1 cm in
height, thin, with lamellulae, sometimes with brown spots
on the lamellae. Stipe whitish, subcylindrical, 13.0–15.0×
1.8–2.4 cm, attenuating upwards, with tiny brownish to
brown (oac721 ) squamules, hollow. Annulus ascending,
whitish, membranous, complex, big, with brownish patchy
squamules on the underside; movable when mature.
Context white to whitish, spongy, unchanging when cut,
odorless. Taste mild or indistinct.
Basidiospores (Fig. 2c)[48/2/1] 14.0–16.0 (18.0)×(9.0)
9.5–10.5 (11.0) μm, Q=(1.40) 1.43–1.67 (1.71), avQ=
1.53±0.07, ellipsoid to ovoid in side view, ellipsoid in front
view, thick-walled, smooth, hyaline, dextrinoid, congophi-
lous, metachrom atic in cresyl blue, with a germ pore caused
by an interruption in the episp orium on the rounded apex,
covered with a hyalinous cap in KOH; apiculus about 1 μm
long. Basidia (Fig. 2d)30–38×11–15 μm, clavate, thin-
walled, hyaline, 4-spored, rarely 2-spored . Cheilocystidia
(Fig. 2e)18–38×7–15 μm, clavate to broadly clavate to
pyriform, rarely subfusiform, colorless and hyaline, thin-
walled. Pleurocystidia absent. Squamules on pile us
(Fig. 2b) a palisade of vertically arranged subcylindric,
clampless hyphae [18–40 (55) μminlength,7
–13 (15) μm
in diam.], fr equently septate , rarely branch ed, with
terminal elements slightly attenuate toward the tip, with
Fig. 2 Macrolepiota detersa (HKAS 55306) a. Basidiomata; b. Squamules on pileus; c. Basidiospores; d. Basidia; e. Cheilocystidia
Fig. 1 One of the 9 equally parsimonious trees (L=448, CI=0.730,
RI=0.947, HI=0.270,) obtained in parsimony analysis of ITS
sequence data. Terminal taxa represent individual specimens with
GenBank accession number, and branch lengths are proportional to
the number of steps (character changes) along the branch. Bootstrap
support (≥50%) is shown above the branches and clade with posterior
probabilities greater than 0.90 is indicated with thick branches. Strict
consensus tree resulted in the same topology. New sequences
generated in this paper are marked with asterisks (*), and other
sequences are mainly from Vellinga et al. (2003) and Johnson (1999 )
Fungal Diversity (2010) 45:81–98 85
yellowish to brownish vacuolar pigment, slightly thick-
walled. Clamp connections common at the base of
basidia and cheilocystidia.
Habitat and known distribution in China: Terrestrial
and saprotrophic, solitary to scattered. Distributed in
eastern China.
Materials examined: Anhui Province: Jingde County,
Zaoyuan, bamboo forest, 2 Oct. 2007, C. L. Hou 603
(HKAS 55306, holotype).
Comments: Macrolepiota detersa is a good edible
species. It is a striking species, characterized by the
combination of scattered, reflexed, patch- or crust-like,
easily detachable, brown squamules on the white pileal
background, a relatively big membranous annulus, and
clavate to broadly clavate to pyriform cheilocystidia.
Macrolepiota detersa is very similar to M. procera in
morphology. However, M. procera has smaller plate-like
squamules on pileus which are more closely attached to the
pileus, and the stipitipellis of M. procera has conspicuous
contrasting dark brown squamules compared with those of
M. detersa. Microscopically, the cheilocystidia of M.
procera are mainly clavate to utriform, and hyphal seg-
ments in the squamules on pileus of M. procera are longer
(25–90×7–14 μm) than those of M. detersa (15–25×7–
11 μm). Ph ylogenetically, a close relationship with M.
dolichaula, not with M. procera, was suggested based on
ITS sequences data set. Morphologically, M. detersa can
easily be separated from M. dolichaula by forming plate-
like pileus squamules, and the squamules, made up of short,
rarely branched filamentous hyphae. Macrolepiota detersa
is also known from Japan based on DNA sequence data
(Fig. 1), and probably occurs in other East Asian countries.
Macrolepiota prominens (Viv.: Fr.) M.M. Moser (in the
M. mastoidea complex), originally described from Europe,
comes close but differs in a protruding umbo on the pileus,
a simple broad annulus, and lamellae edges which become
black with age (Wasser 1993).
Macrolepiota dolichaula (Berk. & Broome) Pegler &
Rayner in Kew Bull. 23: 365. 1969.
Agaricus dolichaulus Berk. & Broome in Trans. Linn.
Soc. London. 27: 150. 1871 (‘ 1870’).
Lepiota dolichaula (Berk. & Broome) Sacc., Syll. Fung.
5: 32. 1887.
Leucocoprinus dolichaulus (Berk. & Broome) Pat. in
Bull. trimest. Soc. mycol. Fr. 29: 215. 1913.
Leucocoprinus dolichaulus
(Berk. & Broome) Boedijn
in Sydowia 5: 221. 1951.
Leucocoprinus dolichaulus var. cryptocyclus Pat. in
Bull. trimest. Soc. mycol. Fr. 29: 215. 1913.
Agaricus beckleri Berk. in J. linn. Soc. 13: 156. 1872.
Lepiota beckleri (B erk.) Sacc., Syll. Fung. 5: 56. 1887.
Agaricus stenophyllus Cooke & Massee in Grevillea 15:
98. 1887.
Lepiota stenophylla (Cooke & Massee) Sacc. in Syll.
Fung. 9: 4. 1891.
Basidiomata (Fig. 3a) medium-sized to large. Pileus 6–
16 cm in diam., fleshy, campanulate when young, become
convex to plano- convex with age, with a low umbo at disc,
white to whitish, covere d with yellow brownish to
brownish granular squamules, which become minute and
sparse toward margin; disc smooth, yellow brown to
brown; margin down-reflexed, appendiculate, sometimes
inconspicuously short striate. Lamellae free, crowded, with
short lamellulae, white when young, white to cream colored
when mature, off white to cream when dried, at times hay
colored after years of deposit. Stipe white to whitish,
subcylindrical, 7– 24×0.8–2.5 cm, attenuating upwards,
with minute farinose granules; base slightly enlarged;
hollow. Annulus ascending, simple, whitish, membranous.
Context whitish, sometimes becoming orange at the base
of the stipe when cut. Taste mild.
Basidiospores (Fig. 3c)[69/3/3] (10.0) 12.5–16.0×(6.5)
8.0–10.5 (12.0) μm (x=13.95±1.23×9.26±0.99 μm), Q=
(1.29) 1.30–1.67 (1.94), avQ=1.51±0.13, ovoid to ellipsoid
in side view, ellipsoid in front view, thick-walled (about
0.5 μm), smooth, hyaline, dextrinoid, congophilous, meta-
chromatic in cresyl blue, with a germ pore caused by an
interruption in the episporium on the rounded apex, covered
with a hyalinous cap in KOH; apiculus 1–1.5 μm long.
Basidia (Fig. 3d)28–33×12–16 μm, clavate, thin-walled,
hyaline, 4-spored; sterigmata up to 4.5 μ m long. Cheilo-
cystidia (Fig. 3e)20–33×11–15
μm, clavate to broadly
clavate, hyaline, thin-walled. Pleurocystidia absent. Squa-
mules on pileus (Fig. 3b) a palisade of short, frequently
branched, subcylindric, clampless hyphae with terminal
elements subcylindric to subfusiform, 6–15 μm in diam.,
hyaline or with yellowish vacuolar pigment, thin-walled to
slightly thick-walled. Clamp connecti ons common at the
base of basidia and cheilocystidia, but rare elsewhere.
Habitat and known distribution in China: Terrestrial
and saprophytic, solitary to scattered on the ground in
mixed forests or on road sides. Distributed in southern and
southwestern China.
Materials examined: Fujian Province: Fuzhou City,
Apr. 1934, S. Q. Deng 2473 (BPI 752291). Guangdong
Province: Yangchun County, alt. 400 m, 19 May 1987, Z.
S. Bi 11703 (GDGM 11703); Nan’ao County, Huanghua
Mt., alt. 150–200 m, 12 Sept. 1986, Z. S. Bi and G. Y.
Zheng 10789 (GDGM 10789); Boluo County, Luofu Mt.,
alt. 140 m, G. Li 11957 (GDGM 11957, as M. procera in
Bi et al . 1994). Hainan Province: Ledong County,
Jianfenglin, alt. 201 m, 4 Aug. 1999, P. Q. Sun 4277
[HKAS 34692, as M. rhacodes (Vittad.) Singer, synonym
of Chlorophyllum rachodres (Vittad.) Vellinga, in Yuan and
Sun 2007 ]; Ledong County, Fanyangang, 11 June 1936, X.
X. Liu 28414 (HMAS 24977); Ledong County, 12
86 Fungal Diversity (2010) 45:81–98
June1936, X. X. Liu 28415 [HMAS 22675 (M)]; Yeda
Tropical Crops Research Institute, 26 May 1960, J. H. Yu
and R. Liu 1402 [HMAS 29852 (M)]; Yeda Tropical Crops
Research Institute, 28 May 1960, J. H. Yu and R. Liu 1461
[HMAS 29851 (M)]; Qiongzhong County, Limu Mt., 6 July
1960, J. H. Yu and R. Liu 1761 [HMAS 28817 (S)];
Lingshui County, Diaoluo Mt., 28 Oct. 1987, GDGM
14161 [as Macrolepiota procera (Scop.: Fr.) Singer in Bi et
al. 1997]; Lingshu i County, Diaoluo Mt., 27 Mar. 1989,
GDGM 15514 (as M. procera in Bi et al. 1997). Sichuan
Province: Xichang City, 4 July1971, X. L. Mao and Q. M.
Ma 129 [HMAS 36880 (S), as M. gracilenta (Krombh.)
Wasser in Ying et al. 1994,asLepiota gracilenta (Krombh.)
Quél. in Ying and Zang 1994 ]; Kangding County, Gongga
Mt., alt. 2800 m, under Picea and Betula, 17 July 1982, Y.
Xuan (HKAS 9751); Miyi County, 27 July 1986, M. S.
Yuan 1186 (HKAS 18396, as M. procera in Yuan and Sun
2007). Tibet (Xizang Autonomous Region): Mêdog
(Motuo) County, alt. 850 m, 2 Aug. 1983, X. L. Mao
M1160 [HMAS 52719 (S), as M. procera in Mao 1995];
Fig. 3 Macrolepiota dolichaula (HKAS 43813, Basidiomata from HKAS 38718) a. Basidiomata; b. Squamules on pileus; c. Basidiospores; d.
Basidia; e. Cheilocystidia
Fungal Diversity (2010) 45:81–98 87
Mêdog (Motuo), 3 Aug. 1983, X. L. Mao M1166 [HMAS
54142, as Leucoagaricus excoriatus (Schaeff.) Singer in Li
et al 1995]. Yunnan Province: Dongshan, alt. 2000 m.,
Sept. 1982, W. K. Zheng 828 (HKAS 10342); Kunming
City, 29 June 1942, W. F. Chiu [HMAS 12189 (S)];
Kunming Institute of Botany, Oct. 2000, X. H. Wang 1201
(HKAS 38171); Kunming City, Heilongtan, 15 Aug. 1974,
M. Zang 954 (HKAS 954); Kunming City, Heilongtang, 18
Aug. 1975, X. J. Li 2608 (HKAS 40470); Kunming City,
Heilongtan, 14 July 1976, M. Zang 2716 (HKAS 40455);
Kunming City, Changchong Mt., 12 July 1984, L. S. Wang
1 (HKAS 13115); Kunming City, Heilongtan, 11 July 1986,
L. S. Wang 31594 (HKAS 3365); Kunming City, Heilong-
tan, 20 Aug. 1987, Y. Xuan 1375 (HKAS 18311); Kunming
City, Kunming Institute of Botany, 25 July 1990, Z. L.
Yang 1019 (HKAS 22693); Kunming City, 20 June 1973,
L. W. Xu and Y. C. Zong and Q. M. Ma 209 [HMAS 36287
(S), as Lepiota excoriata (Schaeff.) P. Kumm. in Ying et al.
1994]; Kunming City, Heilong tan, alt. 1980 m., 15 Oct.
2001, Z. L. Yang 3214 (HKAS 38718); Kunming City,
Heilong tan, 17 Sept. 2001, Z. L. Yang 3203 (HKAS
38462); Fuming County, under Pinus yunnanensis, 27 July
1998, Z. J. Li and M. Zang 12977 (HKAS 34016);
Songming County, Liangwang Mt., 17 Sept. 1979, G. M.
Feng 1 (HKAS 4632); Songming County, Baiyi Xiang, 22
July 1998, X. H. Wang 412 (HKAS 35957); Songming
County, Aziying, 29 July 1998, M. Zang 12979 (HKAS
34018); Yiliang County, 1 Sept. 1999, Z. L. Yang 2622
(HKAS 34066); Yuxi City, 20 July 1991, X. X. Liu 3a
(HKAS 23404a); Gejiu City, Datun, 15 Sept. 1986, K. K.
Chen 157 (HKAS 18200); Lüchun County, 11 Oct. 1973,
M. Zang 325 (HKAS 325); Lufeng County, Yipinglang, alt.
1800 m, 27 June 1978, 86048 (HKAS 4493); Guangnan
County, 29 June 1959, Q. Z. Wang 747 [HMAS 25146
(M)]; Qiubei County, 15 July 1959, Q. Z. Wang 787
[HMAS 25143 (M), as M. gracilenta in Ying et al. 1994];
Jinghong City, 30 Oct. 1958, S. J. Han and L. Y. Chen 5327
[HMAS 26225 (M)], Menglun County, 14 Sept. 1974, M.
Zang 1522 (HKAS 1522); Menglun County, Botanical
Garden, 23 Aug. 1988, Z. L. Yang 582 (HKAS 21810); 23
Aug. 1988, Z. L. Yang 582 (HKAS 21810); Menglun
County, 6 Aug. 1988, Z. L. Yang 279 (HKAS 21809);
Jingdong County, Ailao Mt., 18 July 2006, Z. L. Yang 4660
(HKAS 50457); Luxi County, 3 July 1977, X. J. Li 86
(HKAS 2915, as M. procera in Zang et al. 1994); Ruili
City, alt. 1000 m, 25 July 1979, W. K. Zheng 79069
(HKAS 4839); Genma County, 23 Aug. 1980, M. Zang
6647 (HKAS 6647); Lijiang City, Yulong mt., alt. 2600 m.,
14 Aug. 1982, J. X. Xi 333 (HKAS 10029); Lijiang City,
Xiangshan, 1 Aug. 1985, M. Zang 10194 [HKAS 15093, as
Macrolepiota permixta (Barla) Pacioni in Zang et al. 1996];
Lijiang City, near Jinsha river, alt. 1800 m, 6 July 2004, Z.
W. Ge 61 (HKAS 45862); Malong County, 1 Aug. 1992, Y.
Xiang 3 (HKAS 25481); Pu’er (Simao) City, Caiyanghe,
Heilongtan , alt. 1450 m, 16 June 2000, M. Zang 13339
(HKAS 36104); Xiaguang City, 21 Aug. 1938, C. I. Wei
8238 [HMAS 04238 (S)]; Tengchong County, Qushi, 9 Oct.
2002, H. C. Wang 247 (HKAS 42006); Longlin County,
Longjiang Xiang, alt. 2100 m, 4 Sept. 2002, Z. L. Yang
3437 (HKAS 41506); Yingjiang County, 14 Aug. 1980, M.
Zang 6635 (HKAS 6635); Yingjiang County, Tongbiguang
Xiang, alt. 1450 m, 12 July 2003, L. Wang 73 (HKAS
43169); Jianchuan County, Shibao Mt., alt. 2500 m, 14
Aug. 2003, Z. W. Ge 1 (HKAS 43813).
Comments: Macroscopically, M. dolichaula differs from
the other species of Macrolepiota by its relatively big,
umbonate pileus with minute, pallid squamules and long
slender stipe which sometimes becomes orange at the base
when cut. Microscopically, it differs from other species by
its clavate to broadly clavate cheilocystidia, and squamules
made up of a palisade of short, more branched, subcylin-
dric, clampless hyphae.
Macrolepiota dolichaula was originally described from
Sri Lanka and later also found in China (Chiu 1948), east
Africa (Pegler 1977), Australia (Grgurinovic 1997), and
Vietnam (Yang 2000), and northern Thailand (pers. obs.). It
is considered an edible mushroom in China. Macrolepiota
dolichaula is the most frequently found species in southern
and southwestern China, but often misiden tified as M.
procera, M. mastoidea,
M. perm ixta (Barla) Pacioni, or
Chl. rachodes (Vittad.) Vellinga. In fact, M. procera is
much browner, has a stipe with brown squamules, a pileus
with plate-like squamules made up of a trichodermal layer
of yellowish-brown walled hyphae which seldom branch,
and larger spores; M. mastoidea usually has relatively small
basidiomata, irregularly patchy or sometimes star-shaped
pileal squamules, a subtle banded pattern covering of the
stipe, and the rare presence of clamp connections on the
base of the basidia. Macrolepiota permixta, regard ed as a
variety of M. procera by some authors, differs from M.
dolichaula b y big, plate-like squamules on the pileus, a
stipe context that turns wine-red to orange-red when
scratched or cut (Breitenba ch and Kränzlin 1995). It might
be a color variant of M. procera. Macrolepiota rachodes,
presently placed in the genus Chlorophyllum , has plate-like
pileal squamules made up of a hymeniform layer and
truncated spores whose gem pore is caused by a depression
in the episporium and its germ pore without a cap (Vellinga
2001; Ge and Yang 2006).
Macrolepiota mastoidea (Fr. : Fr.) Singer in Lilloa 22:
417. 1951 (‘1949’).
Agaricus mastoideus Fr. : Fr., Syst. mycol. 1: 20. 1821.
Lepiota mastoidea (Fr. : Fr.) P. Kumm., Führ. Pilzk.:
135. 1871.
Lepiotophyllum mastoideum (Fr. : Fr.) Locq. in Bull.
mens. Soc. linn. Lyon 11: 40. 1942.
88 Fungal Diversity (2010) 45:81–98
Leucocoprinus mastoideus (Fr. : Fr.) Locq. in Bull.
mens. Soc. linn. Lyon 14: 46. 1945.
Basidiomata (Fig. 4a) medium-sized to large. Pileus 5–
11 cm in diam., fleshy, ovoid when young, becoming
convex to plano-convex when mature, with a distinct umbo
at disc, white to off-white, covered with grey-brownish
furfuraceous squamules, which are at first smooth and
continuous, then gradually break up into irregular patches,
and become minute and sparse toward margin; margin
slightly appendiculate. Lamellae free, crowded, white to
greyish white, with lamellulae of 2–3 lengths. Stipe
subcylindrical, 6– 15×0.5–1.0 cm, attenuating upwards,
whitish, covered with tiny furfuraceous brownish squa-
mules, especially above the annulus; base slightly enlarged.
Annulus ascending, simple, whitish, membranous. Context
whitish, not changing color when cut. Taste mild.
Basidiospores (Fig. 4c)[41/2/2](11.0)12.0–14.0
(15.0) ×8.0– 9.5 (10.0) μ m, x = 12.95 ±0.84 ×8.69±
0.60 μm, Q=(1.33) 1.38–1.63 (1.65), avQ=1.49±0.09,
ellipsoid to ovoid in side view, ellipsoid in front view,
thick-walled, smooth, hyaline, dextrinoid, congophilous,
metachromatic in cresyl blue, with a germ pore caused by
an interruption in the episporium on the rounded apex,
covered with a hyalinous cap in KOH; apiculus 1–1.5 μm
Fig. 4 Macrolepiota mastoidea (HKAS 11084) a. Basidiomata; b. Squamules on pileus; c. Basidiospores; d. Basidia; e. Cheilocystidia
Fungal Diversity (2010) 45:81–98 89
long. Basidia (Fig. 4d)32–44×12.0–14.0 μm, clavate,
thin-walled, hyaline, 4-spored. Cheilocystidia (Fig. 4e)
(10) 15–20×7 – 10 μm, clavate, hyaline, thin-walled, in
bunches forming a sterile edge. Pleurocystidia absent.
Squamules on pileus (Fig. 4b) a palisade of subcyl indric,
clampless hyphae (6–12 μm in diam.), with terminal
elements slightly attenuate toward the tip, with yellowish
to brownish vacuolar pigment, slightly thick -walled.
Clamp connections occasionally observed at the base of
basidia.
Habitat and known distribution in China: Terrestrial
and saprotrophic, solitary to scattered in open meado ws or
in mixed forests. Distributed in northeastern and south-
western China.
Materials examined: Heilongjiang Province: Yichun
City, Beishan, alt. 400 m, 8 Aug. 2000, M. S. Yuan 4646
(HKAS 37384); Huma County, 29 July 2000, X.L. Mao, H.
A. Wen and S.X. Sun 120 (HMAS 76557, determined as
Macrolepiota crustosa L.P. Shao & C.T. Xiang by Mao).
Jilin Province: Antu County, Baima town, alt. 740 m, 17
Aug. 2004, L. F. Zhang 605 (HKAS 11084); Antu County,
Jinyuetan Park, alt. 220 m, 29 Sept. 2004, L. F. Zhang 628
(HKAS 11207); Dunhua City, Huangnihe, 5 Sept. 2006, X.
H. Wang 2016 (HKAS 50914). Sichuan Province:
Chengdu City, 30 Sept. 2006, Z. W. Ge 938 (HKAS
51950).
Comments: Macrolepiota mastoidea is an edible spe-
cies. Macroscopically, it differs from the other Chinese
species of Macrolepiota by its distinctive umbonate pileus
covered with grey-brownish velvet squamules which are
irregularly arranged or star-shaped, and its slender stipe
covered with brownish squamules. Microscopically, it is
characterized by the combination of its clavate cheilocys-
tidia, and pileal squamules made up of a palisade of rarely
branched, subcylindric, clampless hyphae.
Macrolepiota mastoidea is very close to M. excoriata
(Schaeff.) Wasser, but the latter has a smooth stipe and
more common clamp connections on the septa of the
basidia (Wasser 1993; Vellinga 2001).
Macrolepiota mastoidea is a complex of species with
different morphologies, but with very small differences in
ITS (Fig. 1). Now it is shown to be present in Asia as well.
Macrolepiot a mastoidea was previously recorded in
China, but re-examination confirmed that some collections
were misidentified. e.g. HMAS 28232 was cited as M.
mastoidea (M. gracilenta) (Ying et al. 1994), but is actually
Lepiota clypeolaria (Bull.) P. Kumm.
Macrolepiota orientiexcoriata Z. W. Ge, Zhu. L. Yang
& Vellinga, sp. nov. Fig. 5
MycoBank: MB 518350
Pileus 8–12 cm diametro, convexus vel applanatus,
albus vel albidus, squamulis furfuraceis, luteo-brunneis
vel brunneis-au ranti acis, obtus e umbona tus. Lamella e
liberae, albae, angustae. Stipes 9.0–11.0 × 1.0–2.0 cm,
subcylindricus, minutus sursum, albidus, basim incrassa-
tus, non-discolorans. Annulus superus, albidus, membra-
naceus. Caro alba; sapor mitis. Basidia 35–52×13–16 μm,
clavata, hyalina, 4-sporigera, raro 2-sporigera. Basidio-
sporae (12.0) 13.0–15.0 (16.0)×(7.5) 8.5–10.0 (10.5) μm,
ellipsoideae, glabrae, hyalinae, dextrinoideae. Pleurocysti-
dia absentia. Cheilocystidia obtusifusiformia vel subcla-
vata, raro subcylindrica vel vesiculosa, hyalina, 20–43×
9–15 μm. Squamulae pilei trichoderma, apicalis hyphus
erectibus, subhyalinus vel luteo-brunneis, subcylindricis
compositae. Fibulae praesentes. Habitatio: terrestris.
Holotypus: Z. W. Ge 96 (HKAS 45863), 12 July 2004,
Xiangcheng County, Sichuan Province, China.
Etymology: “orienti- ” refers to the locality of the type
specimens collected; “excoriata” refers to the squamules of
the pileus.
Basidiomata (Fig. 5a) medium to large-sized. Pileus 8–
12 cm in diam., white to whitish, convex to plano-convex,
sometimes with slightly reflexed margin, with a low umbo
at disc, covered with brownish yellow (5C8, oac687)
furfuraceous squamules, which become minute and sparse
toward margin, revealing white flesh between them; disc
smooth, brownish orange (6C7, oac719). Lamellae free,
moderately crowded, white when young, white to cream
colored when mature, sometimes slightly with pinkish
tinge, thin, with lamellulae. Stipe whitish, subcylindrical,
9–11×1.0–2.0 cm, gradually attenuating upwards, glabrous
or with shiny hairs, hollow; base slightly enlarged to
subglobose, 3.5– 4.0 cm wide. Annulus ascending, whitish,
membranous, slightly complex, with brownish p atchy
squamules on the underside. Context white to whitish,
spongy, unchanging color when cut, but at edge of stipe
slightly with wine red tinge, odorless. Taste mild or
indistinct.
Basidiospores (Fig. 5c )[136/8/6 ] (12.0) 13.0–15.0
(16.0)×(7.5) 8.5–10.0 (10.5)
μm, Q=(1 .38) 1.40–1.63
(1.67), avQ=1.50±0.08, ellipsoid to ovoid in side view,
ellipsoid in front view, thick-walled, smooth, hyaline,
dextrinoid, congophilous, metachromatic in cresyl blue,
withagermporecausedbyaninterruptioninthe
episporium on the rounded apex, covered with a hyalinous
cap in KOH; apiculus not distinctive, about 1 μm long.
Basidia (Fig. 5d)35–52×13–16 μm, clavate, thin-walled,
hyaline, 4-spored rarely 2-spored. Cheilocystidia (Fig. 5e)
20–43×9.5–15 μm, obtusely fusiform to subclavate in most
cases, occasionally subcylindric to vesiculose, hyaline, thin-
walled, in bunches forming a sterile edge. Pleurocystidia
absent. Squa mules on pileus (Fig. 5b) a palisade of
subcylindric, clampless hyphae (20–65 μm in length, 5 –
10 μm in diam.), seldomly branched, with terminal
elements slightly attenuate toward the tip, with yellowish
to brownish vacuolar pigment, slightly thick -walled.
90 Fungal Diversity (2010) 45:81–98
Clamp connections common at the base of basidia and
cheilocystidia.
Habitat and known distribution in China: Terrestrial
and saprotrophic, solitary to scattered in open meado ws or
under bushes. Distributed in northern and southwestern
China (Inner Mongolia and Sichuan).
Materials examined: Inner Mongolia Autonomous
Region: Wulanchabu (Ulanqab) City, alt. 1590 m, 1 Aug.
1990, P. G. Liu 623 (HKAS 23040); Aug. 1994, X. L. Mao
8111 [HMAS 63157 (M); Aug. 1994, X. L. Mao 8116
[HMAS 73304 (M); Sichuan Province: Xiangcheng
County, alt. 3000 m, 12 July 2004, Z. W. Ge 96 (HKAS
45863, holotype); Xiangcheng Coun ty, shago ng, alt.
3000 m, 11 July 1998, Z. L. Yang 2286 (HKAS 32153);
on the way from Jiulong County to Mianning County, 16
July 2005, Z. W. Ge 505 (HKAS 49001).
Comments: Macrolepiota orientiexcoriata is character-
ized by the combination of brownish to reddish-brown
furfuraceous squamules composed of a palisade of sub-
cylindric, seldomly branched, clampless hyphae, and the
obtusely fusiform to subclavate cheilocystidia.
Macrolepiota orientiexcoriata is very close to M.
excoriata (Schaeff.) Wasser both morphologically and
molecularly. However, M. excoriata has a pileus with an
indistinct umbo, the pileal squamules are more often star-
shaped, and the stipe is smooth (Candusso and Lanzoni
1990; Breitenbach and Kränzlin 1995; Vellinga 2001).
Microscopically, cheilocystidia of M. excoriata are more
often narrowly clavate to subcylindric (Wasser 1993;
Vellinga 2001). The ITS data separate the two taxa, with
M. orientiexcoriata in its own clade separate from M.
excoriata (Fig. 1).
Macrolepiota phaeodisca Bellù, which is sister to M.
orientiexcoriata in the phylogenetic tree, originally de-
scribed from the Mediterranean region (Sardinia, Italy),
grows in sandy environment, differs in the dark squamules-
fibrillose pileus, and lack of clamp connections (Bellù
1984). Macrolepiota orientiexcoriata is also very similar to
Fig. 5 Macrolepiota orientiexcoriata (HKAS45863) a. Basidiomata; b. Squamules on pileus; c. Basidiospores; d. Basidia; e. Cheilocystidia
Fungal Diversity (2010) 45:81–98 91
M. mastoidea. However, the latter has a distinctive
umbonate pileus covered with grey-brownish velvet squa-
mules which are irregularly arranged or star-shaped, and its
slender stipe covered with pale brownish squamules .
Chlorophyllum neomastoideum (Hongo) Vellinga, orig-
inally described from Japan, is somewhat similar, but it
differs from M. orientiexcoriata by the reddening of the
flesh when cut, vesicular to clavate cheilocystidia, smaller
(7–8.5×4.5–6 μm) and truncate spores (Hongo 1970).
Macrolepiota procera (Scop. : Fr.) Singer in Papers
Mich. Acad. Sci., Arts Letters 32: 141. 1948 (‘1946’).
Agaricus procerus Scop., Fl. Carn. 2: 418. 1772.
Agaricus procerus Scop. : Fr., Syst. Mycol. 1: 20. 1821.
Lepiota procera (Scop. : Fr.) S.F. Gray, Nat. Arr. Brit. Pl.
1: 601. 1821.
Mastocephalus procerus (Scop. : Fr.) O. Kuntze, Rev.
Gen. Pl. 2.: 1860. 1891.
Leucocoprinus procerus (Scop. : Fr.) Pat., Essai Taxon.
Hymen.: 171. 1900.
Lepiotophyllum procerum (Scop. : Fr.) Locq. in Bull.
mens. Soc. linn. Lyon 11: 40. 1942.
Basidiomata (Fig. 6a) medium to large-sized. Pileus 7–
25 cm in diam., ovoid to drum stick shaped when young,
becoming convex to plano-convex with age, with an obtuse
umbo at disc, white to whitish, covered with brown, dark
brown to grayish brown plate-like squamules; disc smooth,
brown; covering disrupting into small plate-like squamules
which are irregularly arranged toward margin on the dirty
white background. Lamellae free, densely crowded, thin,
white when young, white to cream colored when mature,
with lamellulae in 2-3 lengths. Stipe whitish, subcylindri-
cal, 18.0–34×1.0–2.2 cm, attenuating upwards, at base
enlarged (3.5–4.0 cm), covered with brown to dark brown
velvet squamules sometimes in irregular bands, hollow or
fibrous-stuffed. Annulus superior, about 5 cm below stipe
apex, dirty white above, underside brownish, membranous,
complex, moveable. Context spongy, white to cream at the
pileus, grayish red to purplish brown at the stipe; not
changing color. Smell not recorded. Taste mild.
Basidiospores (Fig. 6c)[64/4/ 4](12.0)13.0–16.0
(19.0)×8.0–10.0 (12.0) μm, Q=(1.35) 1.40–1.63 (1.65),
avQ=1.50±0.07, ellipsoid to ovoid in side view, ellipsoid
in front view, thick-walled, smooth, hyaline, dextrinoi d,
congophilous, metachromatic in cresyl blue, with a germ
pore caused by an interruption in the episporium on the
rounded apex, covered with a hyalinous cap in KOH;
apiculus not distinctive. Basidia (Fig. 6d)30–43×12–
17 μm, clavate, thin-walled, hyaline, 4-spored. Cheilocys-
tidia (Fig. 6e)20–39×10–23 μm, clavate to utriform to
irregularly clavate, hyaline, thin-walled, in bunches forming
a sterile edge. Pleurocystidia absent. Squamules on pileus
(Fig. 6b) a palisade of subcylindric, slightly thick-walled,
clampless hyphae which are 7–11 (14) μm in diam., seldom
branched, with terminal elements slightly attenuate toward
the tip, with yellowish brown vacuolar pigment, slightly
thick-walled. Clamp connections common at the base of
basidia and cheilocystidia.
Habitat and known distribution in China: Terrestrial
and saprotrophic; solitary to scattered on edge of the forest
or in the forest dominated by coniferous and Fagaceous
trees. Distributed in northeastern and eastern China (Hei-
longjiang, Jilin, Shangdong, Jiangsu and Guangdong).
Specimens examined: Guangdong Province: Chang-
jiang Cou nty, Bawangling, GDGM 11851; Heilongjiang
Province: Hulin City, Dongfanghong natural reserve, 19
Sept. 2004, Tolgor 2702 (HMJAU 2702). Jilin Province:
Fusong County, Songji anghe, alt. 1300 m, 12 Aug. 2000,
M. S. Yuan 4659 (HK AS 37383); Yanbian Chosenzu
Zizhizhou, Baihe, alt. 840 m, 15 Aug. 2004, L. F. Zhang
517 (HKAS 8108); Fusong County, Lushuihe, alt. 625 m,
11 Aug. 2004, L. F. Zhang 381 (HKAS 5722). Shangdong
Province: 26 Aug. 1980, H. A. Wen and Y. C. Zong 10
[HMAS 42757 (M)]. Jiangsu Province: Nanjing City, 21
June 1931, S. Q. Teng 490 (BPI 75231).
Comments: Macrolepiota procera is an edible species.
Morphologically, it is characterized by the big, fles hy
basidiomata, the stipe covered with zig-zag banded squa-
mulae, and the squamules on pileus composed of a palisade
of subcylindric, slightly thich-walled, clample ss brown
hyphae.
Macrolepiota fuliginosa (Barla) M. Bon and M. permixta
(Barla) Pacioni are two close ly related species. But M.
fuliginosa has grayish brown basidiomata, and M. permixta
red-brown basidiomata (Bon 1996; Candusso and Lanzoni
1990; Vellinga 2001).
According to the ITS tree, the East Asian collections
differ from those of Europe; t his may in dicate that
collections from East Asia and those from Europe represent
different phylogenetic species. As we have n ot found
discernable morphological characters to separate them, we
continue to recognize the East Asian collections as M.
procera.
Macrolepiota velosa Vellinga & Zhu L. Yang in
Mycotaxon 85: 184. 2003.
Basidiomata (Fig.
7a) medium to large-sized. Pileus 7–
9 cm in diam., plano-convex, with a wide indistinct umbo,
purplish to pale brownish or grey with purplish tinge,
fibrillose, covered with brown to dark brown furfuraceous
squamules; disc smooth, dark brown. Sometimes with
white to dirty white membranous volval remnants as
patches on the surface. Lamellae free, remote from the
stipe, moderately crowded, white to cream color ed, with
whitish cystidiose edge. Stipe brownish to purplish brown,
cylindrical, 10–17×0.4–1.0 cm, attenuating and paler
upwards, with fine fibril s or squamules, hollow; base
slightly enlarged up to 1.3 cm. Annulus ascending, whitish
92 Fungal Diversity (2010) 45:81–98
on upperside with brown rim, and brownish underside,
membranous. Volva limbate, white, membranous. Context
white, with pinkish to brownish tinge both in pileus and
stipe, odorless. Smell indistinct. Taste mild or indistinct.
Basidiospores (Fig. 7c)[145/6/6] (8.0) 9.0–11.0 (11.5)×
(5.5) 6.0–7.5 (8.0) μm, Q=(1.2)1.36– 1.5 (1.62), avQ=
1.42±0.06, amygdaloid-ellipsoid in side view, ellipsoid in
front view, thick-walled, smooth, hyaline, dextrinoid,
congophilous, metachromatic in cresyl blue, with a germ
pore caused by an interruption in the episporium on the
rounded apex, covered with a hyalinous cap in KOH,
apiculus not distinctive, about 1 μm long. Basidia (Fig. 7d)
25–30×9.5–11.5 μm, clavate, 4-spored, without clamp
connections. Cheilocystidia (Fig. 7e)44–68×4.5–7.5 μm,
cylindrical, some slightly widened at apex, with rounded
apex, with grayish granular contents, and refractive patch at
apex, thin-walled, forming a sterile edge. Pleurocystidia
absent. Squa mules on pileus (Fig. 7b) a palisade of
ellipsoid to subglobose, clampless elements (20–65 μmin
length, 5–10 μm in diam.) in chains, rarely branched, with
clavate to narrowly clavate terminal elements (up to 100×
25 μm), slightly thick-walled, brownish, interspersed with
some cylindrical hyphae 5 – 10 μm wide. Velar patches
made up of hyaline, non-colored, cylindrical narrow hyphae
about 2–4 μm. Clamp connections not observed at the
base of basidia, cheilocystidia.
Habitat and known distribution in China: Terrestrial
and saprotrophic, solitary to scattered on the ground in
mixed forest. So far only found in Yunnan and Hainan.
Materials examined: Yunnan Province: Jinghong City,
Damenglong, alt. 650 m, 14 Aug. 1995, Z. L. Yang 2172
(HKAS 29487); Mengla County, Menglun Natural Reserve,
alt. 700–800 m, 2 Sept. 1990, Z. L. Yang 1271 (HKAS
23312); Mengla County, Menglun Nature Reserve, alt.
580 m, 12 Aug. 1988, Z. L. Yang 381 (HKAS 21808);
Mengla County, Menglun, Botanical Garden, alt. 580 m,
12 Oct. 1989, Z. L. Yang 767 (HKAS 22131). Hainan
Province:
Changjiang County, Bawangling Nature Re-
Fig. 6 Macrolepiota procera (HKAS 8108) a. Basidiomata; b. Squamules on pileus; c. Basidiospores; d. Basidia; e. Cheilocystidia
Fungal Diversity (2010) 45:81–98 93
serve, alt. 680 m, 19 Aug. 2009, N. K. Zeng 518 (HKAS
58050); same locality, alt. 693 m, 23 Aug. 2009, N. K.
Zeng 562 (HKAS 58051).
Comments: The distinctive characters of M. velosa are
the basidiomata with a volva at the base of the stipe,
sometimes with whi te to whitish volval remnant patches on
the pileus; small basidiospores and the squamules made up
of ellipsoid to subglobose brown-walled elements in chains
interspersed with some brown filamentous hyphae.
Comparing to other Macrolepiota species with a volva,
M. pulchella de Meijer & Vellinga from Braz il differs from
M. velosa b y longer basidiospores (10.0–14.5×6.0–
Fig. 7 Macrolepiota velosa (HKAS 29487, Basidioma from HKAS 58051) a. Basidiomata; b. Squamules on pileus; c. Basidiospores; d. Basidia;
e. Cheilocystidia
94 Fungal Diversity (2010) 45:81–98
7.5 μm), shorter cheilocystidia (23– 42 μm), and the
squamules made up of clavate elements and long, colorless
emerging hyphae; M. eucharis Vellinga & Halling from
Australia differs in larger basidiospores (10.8–15.5×7.0–
9.0 μm), wider and shorter cheilocystidia (25–53×5.0–
12.0 μm), and squamules lacking ellipsoid to globose or
clavate elements. Macrolepiota brunnescens Vellinga, also
from South America, has velar patches on the pileus, but
becomes brown in all part. Macrolepiota clelandii Grgur.
superficially resembles M. velosa, but differs from the latter
by the absence of a volva at the base of the stipe, the
predominantly 2-spored basidia, and much bigger spores up
to 28.5×15.5 μm (Vellinga 2003).
Macrolepiota velosa is also known from northern Thai-
land; its edibility remains unknown.
Doubtful species and taxa recorded from China
but with uncertainty
Macrolepiota crustosa L.P. Shao & C.T. Xiang in Journal
of North-eastern Forestry Institute 8 (4): 36. 1980.
The original description reads:
“Fructificatio solitarius vel gregarius; pileus 6–13, cm.
latus, carnosus, mollis e globoso demum explanatus vel
depressus, centro mamillis, crustis albo-cinerus vestitus,
centro demum fuligineus, lobo frastoso, peripheria facile
exutus, exer albo-caro, polygonalibus, fibrillosis; caro alba,
fracta demum lutescens, inodora, sapore grato; stipes cavus,
levis, albo-griseus, bulbo amplisssimo, 17–22 cm. longus, 8–
11 mm. crassus; annulus mobolis, fibroso-lacerus; lamellae
distantes, latae albae, fractae incarnatae; sporae ovoideo-
ellipsoideae, chlorino-hyalinae, 11–14.5×6– 8 μ; basidia
cylindraceo-clavata, 34–44×10–11.9 μ.—Esculenta.
Hab: Heilongjiang, Dai-ling, ad terram in pinelis, 18,
VIII, 1974, Shao Li-ping, Siang Cun-ti, no. 74210 (Typus)
Obs: Species Macrolepiotae procerae et Macrolepiotae
rachodese affinis, a priore stipes minute squamis differt, a
posteriore carne aere rubesc ente et in centro pileo emamil-
lae, sapore parum grato facile dignoscendo differ.”
Comment: According to the description and the
habitdepictedinFig.2 in Shao et Siang (1980), M.
crustosa is more similar to M. mastoidea than to M.
procera,but“the smooth stipe and the white context
changes yellow” remindsusofaChlorophyllum species.
HMAS 76557, collected from Huma in Heilongjiang
province and determained by X. L . Mao as M. crustosa,
turned out to be a misidentification of M. mastoidea.
Because the type of M. crustosa is lost, its taxonomic
uncertainty remains.
Macrolepiota prominens (Viv. : Fr.) M.M. Moser, in
Gams, Kleine Kryptogamenflora, Edn 3 (Stuttgart) 2b/2:
184. 1967
Macrolepiota prominens, clearly belongs to the M.
mastoidea group, is characterized by a conspicuous protrud-
ing umbo on the pileus, a simple broad annulus, and lamellae
edges which become black with age (Wasser 1993). Teng
(1996)andMao(2000)recordedthisspeciesforChina.As
there is no specimens cited, and no specimens examined fit
with characters mentioned above, voucher specimens for this
species still need to be found or collected.
Key to the recognized species of Macrolepiota from China
1 Basidiomata with a volva at the base of the stipe
M. velosa
1* Basidiomata without a volva at the base of the stipe
2 Pileus surface with brown plate-like squamules; annulus complex; clamp connections common at the base of the
basidia
3 Stipe surface with conspicuous fine brown squamules on whitish background; pileus squamules made up of
yellowish-brown walled long hyphal segments, mainly 25– 90×7– 11 (14) μm
M. procera
3* Stipe surface with fine brown squamules on whitish backgro und; pileus squamules made up of yellowish-brown
walled short hyphal segments, mainly 15–25×7–11 μ m
M. detersa
2* Pileus surface with pale ochraceous to brown fine squamules; annulus simple, or only slightly thicker near the
edge; clamp connections absent or present
4 Stipe surface with brown squamules; usually without clamps at the base of basidia
M. mastoidea
4* Stipe surface smooth; usually with clamps at the base of basidia
Fungal Diversity (2010) 45:81–98 95
Discussion
New species within Macrolepiota and species diversity
in China
As shown in Fig. 1, M. detersa is phylogenetically closely
related to, but distinct from M. dolichaula and M. procera
based on the ITS data. Similarly, M. orientiexcoriata is
phylogenetically closely related to M. excoriata, M.
mastoidea, and M. phaeodisca, but forms a clade of its
own. As both M. detersa and M. orientiexcoriata have
discrete characters to tell them apart from the currently
described species, we described them as new species in this
paper.
In addition, the result that M. detersa clustered with 3
collections of M. sp. from Japan, whi ch as a whole gets
strong statistical supports, 100% of bootstrap and 1.00
bayesian PP support respectively, indicates that the three
Japanese collections are M. detersa (Fig. 1).
By far, Europe is the species richest region of Macro-
lepiota, with 11 speci es in the current sense recorded
(Candusso and Lanzoni 1990; Vellinga 2001; but numbers
depend on species concepts), then followed by Asia with 9
species recorded (Manjula 1983; Pegler 1986; Shao and
Xiang 1981; Teng 1996; Vellinga and Yang 2003), and 4
species in east Africa (Pegler 1977), and 3 species in
Australia (Grgurinovic 1997; Vellinga 2003). Based on our
present results, at least 6 morphological species were found
in China, with representatives belonging to three different
phylogenetic clades recovered by the analyses of the ITS
data set.
Morphological features of the major monophyletic clades
within Macrolepiota
The monophyly of Macrolepiota shown here is in accor-
dance with earlier findings based on ITS (Vellinga et al.
2003). Vellinga et al. (2003) detected similar major clades
(Fig. 1 in their paper), however, only one of the clades
containing M. excoriata, M. mastoidea, M. “spec. nov. 1”
(which is M. orientiexcoriata) and
M. phaeodisca got
bootstrap support. In our present study, two of the three
clades recovered by the ITS data set got strong bootstrap
and Bayesian post probability supports. The separation of
the three clades is supported by morphological characters
and will be discussed as following:
/volvatae clade (Clade 1) is characterized by species
having a volva at the base of the stipe, finely
squamulous stipe surfaces, relatively small (usually
less than 15 μm) amygdaliform-ellipsoid spores, and
no clamp connections at the base of the cheilocystidia
and basidia. Species of this clade so far are mainly
distributed in tropical regions (Vellinga 2003; Vellinga
and Yan g 2003).
/macrosporae clade (Clade 2) is characterized by a
smooth stipe, a simple annulus and rare clamp
connections. In contrast to those in /macrolepiota
clade, species within this clade do not have big plate-
like squamules on pileus, but furfuraceous fine
squamules composed of a single layer with rarely
branched, pale brownish and thin-walled cylindrical
hyphae.
/macrolepiota clade (Clade 3) is characterized by
having a complex annulus, relatively big (usually 14–
20 μm) ovoid-el lipsoid spores, with a common
presence of clamp connections at the base of the
cheilocystidia and basidia, stipe usually 2-3 time the
pileus diameter (Bon 1996), and the cheilocystidia are
mainly broadly clavate. The stipes usually ha ve fine
brown squamules, but M. dolichaula and M. clela ndii
have farinose stipe surfaces. The pileus covering of
species within this clade forms big-plate like squamules,
and the squamules are composed of two layers with the
terminal layer composed of seldom branched brownish
and thick-walled cylindrical hyphae arising from a layer
which is composed of thin-walled, often branched
hyphae (but M. dolichaula is the exception here as well).
Infrageneric classification and systematic position
of species with volva in Macrolepiota
In traditional taxonomic classifications, Singer partitioned
Macrolepiota into two groups (section Macrolepiota and
section Macrosporae) based on the presence or absence of
clamp connections (Singer 1986). Bon (1996) divided the
genus Macrolepiota into three sections by adding sect.
Laevistipedes (Pázmány) Bon. Vellinga (2003) transferred
5 Stipe base sometimes becomes orange when cut, pileus squamules composed of more frequently branched
hyphae, cheilocystidia mainly clavate to broadly clavate
M. dolichaula
5* Stipe base n ot changing color when cut, pileus squamules composed of seldom branched hyphae, cheilocystidia
mainly obtusely fusiform to clavate
M. orientiexcoriata
96 Fungal Diversity (2010) 45:81–98
the section Laevistipedes to the genus Chlorophyllum, and
Vellinga and Yang (2003 ) synonymized Volvolepiota with
Macrolepiota without discussion of the taxonomic positions
of those species with a volva within the genus. In this study,
our molecular phylogenetic analysis recovered three major
clades with strong statistical support. This result and the
corresponding morphological characters of each clade
indicate that Macrolepiota could be divided into three
sections. /macrolepiota clade and /macrosporae clade
mainly correspond to the curren t infra-generic classification
proposed by Bon (1996). Considering the species with a
volva form a well-supported /volvatae clade (Clade 1), we
propose a new section to accommodate the species with a
volva within Macrolepiota.
Macrolepiota sect. Volvatae Z. W. Ge, Zhu L. Yang &
Vellinga, sect. nov.
MycoBank: MB 518351
Stipes basi marginatus-bulbosus, volvatus, Basidiospora
parvula, 15.5 μm minus. Fibulae absentes.
Stipe with a volva at the base, annulus simple or only
thickening at the edge of the annulus or only somewhat
reflexed near the annulus margin, basidiospores less than
15.5 μm in length, clamp connections absent.
Type species: Macrolepiota velosa Vellinga & Zhu. L.
Yang in Mycotaxon 85: 184 (2003).
Other species included in this section are Macrolepiota
pulchella de Meijer & Vellinga, M. eucharis Vellinga &
Halling and M. brunnes cens Vellinga.
Acknowledgements Z. W. Ge would like to thank Dr. D. S. Hibbett
(Clark University, USA) for allowing him to generate some sequences
in his lab, and Prof. D. H. Pfister for support during his stay in the
Harvard University Herbaria. The authors are very grateful to Dr. C. L.
Hou for sending the type material and image of Macrolepiota detersa.
Thanks are also due to Dr. T.H. Li, Guangdong Institute of
Microbiology (GDGM), and Dr. Y. J. Yao, Institute of Microbiology,
Chinese Academy of Sciences (HMAS) for allowing us access to the
relevant specimens in their herbaria. This study was supported by the
National Natural Science Foundation of China (grants No. 30800004),
the Natural Science Foundation of Yunnan Province (No. 2008CD164),
the Ministry of Science and Technology of China (2008FY110300), the
Joint Funds of the National Natural Science Foundation of China and
Yunnan Provincial Government (No. U0836604) the Hundred Talents
Program of the Chinese Academy of Sciences, and the National Key
Technology R & D Program (No. 2008BADA1B00).
Open Access This article is distributed under the terms of the
Creative Commons Attribution Noncommercial License which per-
mits any noncommercial use, distribution, and reproduction in any
medium, provided the original author(s) and source are credited.
References
Bellù F (1984) Contributo al genere Macrolepiota Singer-2. Bollettino
Gruppo Micologico G. Bresadola 27(1–2):5 –20
Bi ZS, Zheng GY, Li TH (1994) Macrofungus Flora of Guangdong
Province. Guangdong Science and Technology, Guangdong,
p 879, in Chinese
Bi ZS, Li TH, Zhang WM, Song B (1997) A preliminary agaric flora
of Hainan Province. Guangdong Higher Education, Guangzhou,
p 388, in Chinese
Bon M (1996) Die Großpilzflora von Europa 3. Lepiotaceae (übersetzt
und bearbeitet von F. Medjebeur-Thrun F., Thrun WU). Eching:
IHW-Verlag
Breitenbach J, Kränzlin F (1995) Fungi of Switzerland, Vol. 4.
Agarics 2nd part. Verlag Mykologia, Lucerne (Switzerland)
Candusso M, Lanzoni G (1990) Fungi Europaei 4. Lepiota s. l.
Saronno, Giovanna Biella
Chiu WF (1948) The Amanitaceae of Yunnan. Sci. Rept. Natl. Tsing
Hua Univ. Ser. B., Biol. and Psychol. Sci 3(3):165–178
Ding ZQ, Huang SZ (2003) Characteristics and high-yield culture
technique of Macrolepiota procea. Edible Fungi 4:33, in Chinese
Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small
quantities of fresh leaf material. Phytochem Bull 19:11–15
Felsenstein J (1985) Confidence limits on phylogenies: an approach
using the bootstrap. Evolution 39:783–791
Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for
basidiomycetes—application to the identification of mycorrhizae
and rusts. Mol Ecol 2:113–118
Ge ZW, Yang ZL (2006) The genus Chlorophyllum (Basidiomycetes)
in China. Mycotaxon 96:181– 191
Grgurinovic CA (1997) Larger fungi of South Australia. Botanic
Gardens of Adelaide and State Herbarium and Flora and Fauna of
South Australia Handbooks Committee, Adelaide
Hongo T (1970) Notulae mycologicae 9. Memoirs of the Shiga
University. Nat Sci 20:49–54
Johnson J (1999) Phylogenetic relationship s within Lepiota sensu lato
based on morphological and molecular data. Mycologia 91:443–458
Kirk PM, Cannon PF, Minter DW, Stalpers JA (2008) Dictionary of
the fungi, 10th edn. CABI, Wallingford
Kornerup A, Wanscher JH (1978) Methuen handbook of color, 3rd
edn. Eyre Methuen Ltd., London
Maddison DR, Maddison WP (2000) MacClade 4: analysis of
phylogeny and character evolution. Sinauer Associates, Sunderland
Manjula B (1983) A revised list of the agaricoid and boletoid
basidiomycetes from India and Nepal. Proc Indian Acad Sci
(Plant Sciences) 92(2):81–213
Mao XL (1995) Macrofungal flora of the Mt. Namjagbarwa Region. In:
Li BS, Mao XL, Wang ZW (eds) Biota of the Mt. Namjagbarwa
Region. Science, Beijing, p 118, in Chinese
Mao XL (2000) The mac rofungi in China . Henan Science and
Technology, Zhengzhou, p 719, in Chinese
Mao XL (2009) The macromycetes of China. Science, Beijing, p 816,
in Chinese
Pegler DN (1977) A preliminary Agaric Flora of East Africa. Kew
Bulletin Additional Series 6: 1– 615. London, HMSO
Pegler DN (1986) Agaric Flora of Sri Lanka. Kew Bulletin Additional
Series 12:1–519
Ronquist F, Huelsenbeck JP (2003) MrBayes 3: Bayesian phylogenetic
inference under mixed models. Bioinformatics 19:1572–1574
Shao LP, Xiang CT (1981) (
‘1980’) The study on the Macrolepiota
spp. in China. Journal of Northeastern Forestry Institute 4:35–38
Singer R (1948) (‘1946’) New and interesting Species of Basidiomycetes.
Papers Michigan Academy of Science, Arts and Letters 32:103–150
Singer R (1959) Dos generos de hongos nuevos para Argentina. Bol
Soc Argent Bot 8:9–13
Singer R (1986) The Agaricales in modern taxonomy, 4th edn. Koeltz
Scientific Books, Koenigstein
Swofford DL (2004) PAUP*. Phylogenetic analysis using parsimony
(* and othe r methods), Version 4.01. Sinauer, Sunderland,
Massachusetts
Fungal Diversity (2010) 45:81–98 97
Teng SC (1996) In: Korf RP (ed) Fungi of China, Mycotaxon Ltd,
Ithaca, New York, pp 453–456
Thiers B (2010) [continuously updated]. Index Herbariorum: A global
directory of public herbaria and associated staff. New York
Botanical Garden’s Virtual Herbarium. http://sweetgum.nybg.org/
ih/]
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG
(1997) The CLUSTAL_X windows interface: flexible strategies
for multiple sequence alignment aided by quality analysis tools.
Nucleic Acids Res 25:4876–4882
Vellinga EC (2001) Macrolepiota. In: Noordeloos ME, Kuyper TW,
Vellinga EC (eds) Flora Agaricina Neerlandica, vol. 5. A. A.
Balkema Publishers, Lisse (Netherlands)
Vellinga EC (2003) Chlorophyllum and Macrolepiota (Agaricaceae) in
Australia. Austr Syst Bot 16:361–370
Vellinga EC, Yang ZL (2003) Volvolepiota and Macrolepiota—Macro-
lepiota velosa, a new species from China. Mycotaxon 85:183–186
Vellinga EC, De Kok RPJ, Bruns TD (2003) Phylogeny and taxonomy
of Macrolepiota (Agaricaceae). Mycologia 95(3):442–456
Wasser SP (1993) Tribes Cystodermateae Sing. and Leucocoprineae
Sing. of the CIS and Baltic States. Libri botanici 9. Eching:
IHW-Verlag
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct
sequencing of fungal ribosomal RNA genes for phylogenies. In:
Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR
protocols: a guide to methods and applications. Academic Press,
San Diego, pp 315–322
Yang ZL (2000) Type studies on agarics described by N. Patouillard
(and his co-authors) from Vietnam. Mycotaxon 75:431–476
Ying JZ, Zang M (eds.) (1994) Economic macrofungi from south-
western China. Beijing: Science Press. 399 pp. (in Chinese)
Ying JZ, Wen HA, Zong YC (1994) The Economic macromycetes
from western Sichuan. Science, Beijing, in Chinese
Yuan MS, Sun PQ (2007) Atlas of Chinese mushrooms. Sichuan
Publishing House of Science and Technology, Chengdu, p 552,
in Chinese
Zang M, Li B, Xi JX (1996) Fungi of Hengduan mountains. Science,
Beijing, in Chinese
98 Fungal Diversity (2010) 45:81–98
