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442
Mycologia,
95(3), 2003, pp. 442–456.
q2003 by The Mycological Society of America, Lawrence, KS 66044-8897
Phylogeny and taxonomy of
Macrolepiota
(Agaricaceae)
Else C. Vellinga
1
Department of Plant & Microbial Biology, University
of California at Berkeley, 111 Koshland Hall, No.
3102, Berkeley, California 94720-3102
Rogier P. J. de Kok
Centre for Plant Biodiversity Research, Australian
National Herbarium, CSIRO Plant Industry, G.P.O.
Box 1600, Canberra, Australian Capital Territor y
2601, Australia
Thomas D. Bruns
Department of Plant and Microbial Biology, University
of California at Berkeley, 111 Koshland Hall, No.
3102, Berkeley, California 94720-3102
Abstract:
The position and composition of
Macro-
lepiota
within the Agaricaceae and its phylogenetic
relationships with other members of the family were
investigated, using both molecular (ITS and LSU
rDNA sequences) and morphological characters. The
molecular data separate the genus into two clades.
The first clade comprises
M. procera
,
M. mastoidea
,
M. clelandii
and allies and is a sister group of
Leucoa-
garicus
and
Leucocoprinus
. The second, more diverse,
clade, with
M. rachodes
and allies,
M. globosa
,
Chlo-
rophyllum molybdites
,
Leucoagaricus hortensis
and
En-
doptychum agaricoides
, is a sister group of
Agaricus
.
The separation of the two clades is supported by mor-
phological characters, such as the structure of the
pileus covering, the stipitipellis and the shape of the
germ pore and the spore apex. The two clades are
regarded as genera for which the names
Macrolepiota
and
Chlorophyllum
are proposed.
Macrolepiota nym-
pharum
does not belong to either clade but is as-
signed to the genus
Leucoagaricus
, close to
L. leucoth-
ites
.
Endoptychum depressum
is transferred to the ge-
nus
Agaricus
as
A. inapertus
.
Key words:
Agaricaceae,
Chlorophyllum
,
Endopty-
chum
, ITS and LSU rDNA, phylogeny
INTRODUCTION
Macrolepiota
Singer belongs to the family Agaricaceae
(Fr.) Chev., one of the most diverse families in the
Accepted for publication September 30, 2002.
1
Corresponding author. E-mail: vellinga@uclink.berkeley.edu
Agaricales. Spore print color in the family varies from
white to brown, green and blue; the hymenophoral
trama is regular or trabecular; and the structure of
the pileus covering (of velar or pileipellicular origin)
ranges from epithelioid to trichodermal or cuticular.
Despite this morphological diversity, the family was
shown to be monophyletic, based on studies of nu-
clear ribosomal ITS, LSU, and mitochondrial SSU
( Johnson and Vilgalys 1998, Johnson 1999, Moncalvo
et al 2000, 2002). The molecular data support Sing-
er’s morphological concept of the family (1986), al-
though tribus Cystodermatae was excluded and the
family Lycoperdaceae was included (Kirk et al 2001,
Moncalvo et al 2002). A separate family for the white-
spored taxa, Lepiotaceae Over., as advocated by some
authors, was ruled out by the molecular data. How-
ever, relationships within the family Agaricaceae re-
mained largely unresolved, due to the small sample
of this diverse family, conservatively estimated to
comprise more than 900 species (Kirk et al 2001).
In the studies of Johnson (1999) and Johnson and
Vilgalys (1998),
Macrolepiota
comprising five species,
is polyphyletic but taxonomic conclusions were not
drawn. Morphologically,
Macrolepiota
has been rec-
ognized by these characters: big and fleshy basidi-
ocarps; hymenidermal or trichodermal universal veil
that splits up into coarse-to-fine squamules on the
pileus; an often complicated, double annulus, and
white-to-pink, thick-walled spores with a germ pore
that are dextrinoid, metachromatic in Cresyl blue
and congophilous. The lamella trama is trabecular
(Buller 1924, Heinemann 1989, Cle´menc¸ on 1997), a
character shared with
Leucoagaricus
Singer,
Leucoco-
prinus
Pat., and
Chlorophyllum
Mass.
Several genera are very similar to
Macrolepiota
.
Chlorophyllum
differs only in the distinctly green or
ochre spores; these colors obscure the staining reac-
tions of the spores. The genus
Chlorolepiota
Sathe &
Deshpande (1979) hypothetically occupies an inter-
mediate position between
Chlorophyllum
and
Macro-
lepiota
because the spore print is primrose yellow and
the spores are provided with a germ pore but are not
truncate; clamp connections are said to be absent.
Volvolepiota
Singer also closely resembles
Macrolepi-
ota
; a volva is present, the pileus covering is trichod-
ermal, clamp connections are present (in the context
of the stipe) and the spores have a germ pore (Hei-
nemann and De Meijer 1996).
443V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
The position of
Leucoagaricus hortensis
(Murrill)
Pegler is anomalous and has been the subject of re-
cent discussion (Akers and Sundberg 1997, Johnson
1999). This species was placed in the genus
Leucoa-
garicus
by Pegler (1983) because the spores lack a
germ pore; yet clamp connections are present, a
character state that does not occur elsewhere in the
genus
Leucoagaricus
. The structure of the veil resem-
bles that found in
Macrolepiota rachodes
(Vittad.)
Singer.
The secotioid genus
Endoptychum
was considered
a member of the Agaricaceae by some authors (e.g.,
Moser 1983). Singer (1986), on the other hand, re-
garded it as a Gasteromycete and as such an ancestor
of the Agaricales.
Two competing infrageneric classifications of the
genus
Macrolepiota
have been proposed: i) a division
into two sections based on the presence or absence
of clamp connections in which section
Macrolepiota
has clamp connections and section
Macrosporae
(Singer) Bon is lacking clamp connections in the tra-
ma (Singer 1986, Candusso and Lanzoni 1990, Bal-
lero and Contu 1991, though the last authors treated
Macrolepiota
and
Lepiota
as belonging to one genus);
ii) a classification with three sections,
Macrolepiota
,
Macrosporae
and
Laevistipedes
(Pa´zma´ny) Bon (Bon
1993), based on diverse macroscopical and micro-
scopical characters, such as the shape of the spore
and the germ pore, the structure of the annulus and
the covering of the stipe. According to Singer’s
(1986) classification,
M. rachodes
and
M. procera
be-
long to the same section, though according to the
molecular analyses of Johnson (1999), and our own,
these taxa belong to different lineages.
Several toxic species are known within
Macrolepiota
and
Chlorophyllum
, and a natural question is whether
these species form one clade, setting them apart
from the edible species, sharing characters that facil-
itate recognition and medical treatment.
Chlorophyl-
lum molybdites
(G. Meyer : Fr.) Mass. causes gastro-in-
testinal problems and is a particular threat to chil-
dren, because it frequents lawns and other man-made
habitats in tropical, subtropical areas and other pla-
ces with humid summers. It is ver y common in urban
and suburban areas in the eastern and southern parts
of the United States. An extensive list of references
to this species and its toxicity can be found in Reid
and Eicker (1991). The toxic component of this spe-
cies is unknown (Lehmann and Khazan 1992).
En-
doptychum agaricoides
Czern. caused hemolytic an-
emia in a dog, which had eaten a mature basidiocarp
(DE Desjardin pers obs).
Macrolepiota neomastoidea
(Hongo) Hongo and
M. venenata
Bon are reportedly
toxic (in the case of the former, by Yokoyama and
Yamaji 1981; in the case of the latter, by Bon et al
1979, Mazzolai 1989), whereas many other species,
especially
M. rachodes
and
M. procera
, are considered
excellent edibles. Some species are commercially
grown or sold in cultivation kits. Arora (1986) sus-
pected that
Leucoagaricus hortensis
(as
Lepiota humei
)
might be very toxic, but this species is sold on the
market in Bolivia (E Boa pers obs; identification by
senior author).
Some
Macrolepiota
species are widespread, and re-
ported from all over the world, others, such as
M.
excoriata
(Schaeff. : Fr) Wasser, and
M. phaeodisca
Bellu`, are restricted in their area of distribution
(Courtecuisse and Duhem 1994, Nauta and Vellinga
1995; resp. Candusso and Lanzoni 1990). Many spe-
cies occur in man-made habitats, such as gardens,
lawns, compost-heaps; others occur in grasslands or
open places in woods. There are striking disparities
in the number of species in different areas. These
differences are genuine and are not artifacts of the
extent to which they have been studied or of taxo-
nomic perspective. Western Europe is rich, with 11
to 19 species recorded, depending on the author. In
North America, on the other hand, only two species
generally are recorded, although the true number
might be seven (senior author pers obs). Despite the
fact that many species form big, conspicuous basidi-
ocarps, several species are still undescribed. It is im-
portant to note that European names often have
been applied prematurely to similar-looking species
in other parts of the world, complicating clear un-
derstanding of this group. For example, the name
M.
procer
a has been misapplied to a North American en-
tity (designated
M
. spec. nov. 5 in this study) and to
the Australian
M. clelandii
Grgur. (Grgurinovic
1997).
This study has three themes. First, it focuses on
phylogeny of
Macrolepiota
as inferred from ribosomal
DNA data and how this phylogeny relates to the clas-
sification, based on morphology; second, on the
placement of the taxa within the family Agaricaceae;
and third, on the relationship of the secotioid genus
Endoptychum
Czern. to the Agaricaceae. In the course
of the study several new taxa were discovered; they
will be described separately. Many collections from a
large area (including representatives from Africa,
and Australia) were examined, and ITS and LSU se-
quences, and morphological characters were used.
The choice deliberately was made to use a high num-
ber of taxa and samples instead of data from other
genes from a small sample (e.g., Greybeal 1998).
MATERIAL AND METHODS
Material.
Initially 92 collections of taxa belonging to
Ma-
crolepiota
(sensu Singer 1986) or to putative species in this
444 M
YCOLOGIA
group were used for molecular screening, along with an
additional seven sequences, which were accessed through
Genbank. These 99 collections represent at least 24 taxa or
monophyletic species complexes, and one sample of each
(i.e., 24 in total) was arbitrarily chosen from each set of
sequences with up to five differences in base pairs and used
in these analyses; in a few cases the ITS sequence was taken
from a different specimen than the LSU-sequence. Material
of 28 species representing the genera
Agaricus
,
Allopsalliota
,
Endoptychum
,
Lepiota
,
Leucoagaricus
,
Leucocoprinus
, and
Micropsalliota
was used for comparison (see T
ABLE
I for col-
lections and species analyzed).
Molecular identification.
DNA was extracted from fresh
and herbarium material; the internal transcribed spacer
(ITS) and part of the Large SubUnit (LSU) of the nuclear
ribosomal repeat were amplified by the polymerase chain
reaction (PCR) with the fungal specific primers ITS1F and
ITS4 (Gardes and Bruns 1993) for the ITS region, and
primers LR0R, LR3R, LR7, and LR16 for LSU (see http://
www.biology.duke.edu/fungi/mycolab/primers.htm). Se-
quencing of both strands was performed with an ABI model
377 sequencer (Applied Biosystems, Foster City, CA, U.S.A.)
using a Thermo SequenaseyDye terminator Cycle Se-
quencing Pre-Mix Kit (Amersham Pharmacia Biotech, Pis-
cataway, NJ, U.S.A.) or a BigDyeyTerminator Cycle Se-
quencing Ready Reaction Kit (Applied Biosystems Co.); the
primers ITS1 or ITS5, ITS2, ITS3, and ITS4 were used for
the ITS region, and primers LR0R, LR3R, LR7, and LR16
for the LSU-region. Raw data were processed with DNA Se-
quencing Analysis v.2.1.2 and Sequence Navigator v.1.0.1
(also of Applied Biosystems). The sequences thus obtained
were aligned in ClustalW (Thompson et al 1994), with these
settings: opening gap penalty 10, end gap penalty 10, ex-
tending gap and separation gap penalty both 0.05. Visual
alignment was the next step. The alignments have been de-
posited in TreeBASE under number S782. Sequences have
been deposited in Genbank, and the accession numbers are
given in T
ABLE
I.
Morphological data.
Nine morphological character states
were scored for all taxa and added to the databases (T
ABLE
II).
Phylogenetic analysis.
Maximum-parsimony analyses were
performed with PAUP* version 4.0b8 (Swofford 2001). One
hundred heuristic searches were conducted with random
sequence addition and tree bisection-reconnection (TBR)
branch-swapping algorithms, collapsing zero-length branch-
es and saving all minimal-length trees (MulTrees) on dif-
ferent datasets (ITS data, LSU sequences, and ITS and LSU
combined). Three members of tribus Lepioteae,
L. brun-
neoincarnata
,
L. cristata
and
L. rufipes
, were chosen as out-
group. Noninformative characters and part of the ITS1 re-
gion (190 base pairs) that was not unambiguously alignable
were excluded from the analyses. Gaps were treated as miss-
ing data. To measure relative support for the resulting
clades, 500 bootstrap replications (Felsenstein 1985) were
performed with the same parameters as for the parsimony
analyses.
To test alternative phylogenetic relationships, the Kishi-
no-Hasegawa maximum-likelihood ratio test (Kishino and
Hasegawa 1989) was performed, as implemented in PAUP*
with default settings, under the model developed by Hase-
gawa et al (1985). The hypotheses tested are enumerated
in T
ABLE
III under results.
RESULTS
Analyses of ITS dataset.
The aligned data set in-
cludes a total of 906 base pairs (including gaps); 273
parsimony-informative characters were analyzed (a
stretch of 190 base pairs in ITS1 was excluded from
the analyses). A total of 90 most-parsimonious trees
was recovered, with these characteristics: l
5
1353, CI
5
0.3836, CR
5
0.2449.
Four lineages were recovered (F
IG
. 1): (i) Lineage
1, composed of
Agaricus
,
Micropsalliota
, and
Endo-
ptychum depressum
. (ii) Lineage 2 included
M. rach-
odes
and allies,
M. neomastoidea
,
Chlorophyllum mol-
ybdites
,
M. globosa
,
Endoptychum agaricoides
and
Leu-
coagaricus hortensis
. This lineage shows relatively long
branches. Lineages 1 and 2 are sister groups of each
other. (iii) Lineage 3 to which
Leucoagaricus
and
Leu-
cocoprinus
species belonged. (iv) Lineage 4 compris-
ing
M. procera
,
M. mastoidea
,
M. excoriata
, and allies;
the branches within this lineage in general are short.
All four lineages are present in the same topology
on the consensus tree (not shown). However, Line-
age 3, the
Leucoagaricus-Leucocoprinus
assemblage, is
not bootstrap supported. Lineage 1 has 100% boot-
strap support, whereas Lineage 2 has a bootstrap sup-
port of 76%, and Lineage 4 has 73% (however, Lin-
eage 4 is 100% bootstrap supported when
Macrole-
piota
spec. nov. 1 is excluded (data not shown)).
The topological constraint enforcing
Macrolepiota
sensu Singer (1986), i.e., without
Chlorophyllum mol-
ybdites
and
M. globosa
, without
Leucoagaricus horten-
sis
, and without
Endoptychum agaricoides
, does not
yield a less likely tree (T
ABLE
III). But, enforcing
Leu-
coagaricus hortensis
with Lineage 3 has to be rejected
(P
,
0.05). Significantly less likely than the best tree
are the options in which
Endoptychum agaricoides
is
excluded from Lineage 2,
Macrolepiota nympharum
is
included in
Macrolepiota
,or
E. depressum
and
E. agar-
icoides
form a monophyletic group. A position of
Ma-
crolepiota
(lineages 2 and 4 together) together with
Lineage 3 (
Leucocoprinus
and
Leucoagaricus
), is sig-
nificantly worse than the option of a monophyletic
group of lineages 1, 2 and 4, which is the most likely
topology recovered.
Analyses of LSU dataset.
The aligned dataset shows
a total of 934 characters, of which 82 are parsimony
informative. One hundred most-parsimonious trees,
occurring in four islands, were recovered (l
5
225).
445V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
Only a few distinct lineages were recognized in all
four:
Agaricus
and
Endoptychum depressum
; the group
of
M. rachodes
;
M. procera
plus two closely related
species; all other species are paraphyletic throughout
the phylogram, in different topologies (the tree with
the highest
2
ln likelihood value is given in F
IG
. 2).
The consensus tree also is highly unresolved, and
only the lineages of
Micropsalliota
and
Allopsalliota
,
Leucoagaricus meleagris
and
L. americanus
, and
L. leu-
cothites
and
M. nympharum
respectively have a boot-
strap support higher than 70% (consensus tree not
shown; bootstrap values given in F
IG
. 2).
Most topological constraints do not yield signifi-
cantly less likely trees than the unconstrained tree
(T
ABLE
III under LSU data). However, enforcing
M.
nympharum
with
Macrolepiota
sensu Singer (1986),
enforcing the two
Endoptychum
species together and
enforcing a monophyletic clade containing all
Ma-
crolepiota
,
Chlorophyllum
, and
Agaricus
species are sig-
nificantly less likely.
Analyses of the combined ITS and LSU dataset.
The
combined dataset was tested for incongruence with
the partition homogeneity test, as implemented in
PAUP* 4.0b8, as advocated by Cunningham (1997).
The original ITS-dataset was incongruent with the
LSU-dataset (P
5
0.0200), because of the placement
of
Allopsalliota geesterani
. This species is a sister spe-
cies of
Micropsalliota
spec. in the phylogram, based
on the LSU-data set, but appears in the
Leucoagari-
cus-Leucocoprinus
lineage in the analyses of the ITS-
data set. The position of this species based on mor-
phological characters is also rather enigmatic; it
shares characters with
Agaricus
and
Micropsalliota
(Nauta 1999) and has some chemical reactions in
common with
Leucoagaricus americanus
and its allies.
A similar situation was found in the analyses of sev-
eral molecular datasets illuminating the phylogeny of
the tribus Triticeae within the Gramineae (Mason-
Gamer and Kellogg 1996), where the position of
Trit-
icum monococcum
caused incongruence. After remov-
al of
A. geesterani
in these datasets, the two datasets
were congruent (P
5
0.4100) and were used for fur-
ther analysis.
The combined dataset of ITS and LSU sequences
yields 287 parsimony-informative characters (out of
1883); 20 most-parsimonious trees could be recov-
ered, with a length of 988 (CI
5
0.4615, CR
5
0.2969), and all show the same four lineages as de-
rived from the ITS-data set (the strict consensus tree
is given in F
IG
. 3). These phylograms differ from the
ones based on ITS data only in the fact that lineages
3 and 4 are now sister groups.
Leucoagaricus hortensis
is a sister taxon of the
Endoptychum agaricoides
-
Chlo-
rophyllum
clade in half of the phylograms, and a sister
group to the
M. rachodes
clade in the other 10 phy-
lograms. The bootstrap support for Lineage 1 is
100%, 75% for Lineage 2, and 95% for Lineage 4;
again Lineage 3 gets very low bootstrap support (
,
50
%). All four lineages are present in the consensus
tree (F
IG
. 3) in the same topology as in the individual
most parsimonious trees.
The most likely tree recovered is the one in which
lineages 1, 2 and 4 form a monophyletic group, fol-
lowed by the unconstrained tree (lineages 1 and 2
monophyletic and sister groups, Lineage 4 mono-
phyletic and a sister group to Lineage 3) (T
ABLE
III).
The topological constraint enforcing lineages 2 and
4 together does yield an acceptable tree (P
5
0.2),
but all the other topological constraints yield much
less likely trees (T
ABLE
III). The hypothesis that lin-
eages 2, 3 and 4 form a monophyletic group (i.e.,
Macrolepiota
in a wide sense is a sister group to the
Leucoagaricus
-
Leucocoprinus
assemblage) cannot be
rejected, based on these data.
Adding morphological data to the data set or treat-
ing the gaps as fifth characters does not change the
topology of the trees (data not shown), although
bootstrap values might change slightly.
DISCUSSION
Concept of
Macrolepiota
.
The traditional concept of
Macrolepiota
, as an agaricoid genus comprising white
to pink-spored species with a germ pore in the
spores, cannot be maintained, based on the results
of the analyses of LSU data and combined ITS-LSU
data.
Leucoagaricus hortensis
whose spores are not
provided with a germ pore,
Chlorophyllum molybdites
and
M. globosa
with greenish spores, and the seco-
tioid
Endoptychum agaricoides
appear to belong to the
same lineage as some
Macrolepiota
taxa.
Macrolepiota
nympharum
, on the other hand, is a sister taxon of
Leucoagaricus leucothites
, according to these analyses.
The phylograms based on the ITS sequences, and
ITS-LSU-data (F
IGS
. 1 and 3), show two distinct clades
within
Macrolepiota
, viz. lineages 2 and 4, which do
not form a monophyletic group. However, topologi-
cal constraints forcing the two lineages together
(analyses of all three data sets) that result in one
monophyletic ‘‘big’’
Macrolepiota
clade is neither sig-
nificantly better, nor worse, than the two-clade op-
tion. These alternatives cannot be discriminated on
the present molecular grounds alone.
Morphological considerations.
The number of mor-
phological characters of mushrooms available for a
phylogenetic analysis on the generic level is unfor-
tunately very small in general. Nevertheless, several
morphological characters do support recognition of
446 M
YCOLOGIA
T
ABLE
I. Overview of species and collections and the GenBank accession numbers for the ITS and LSU sequences
Species
Collection &
herbarium
Location & date
of collection
ITS
GenBank
accession #
LSU
GenBank
accession #
Agaricus arvensis
Schaeff. GenBank AF161015 U11910
Agaricus bisporus
( J.E.
Lange) Imbach GenBank AF161014 U11911
Agaricus bitorquis
(Que´l.)
Sacc. E.C. Vellinga 2462 (UCB) U.S.A., Michigan, Washtenaw Co., Ann Arbor, 10-VIII-2000 AF482829
Agaricus californicus
Peck E.C. Vellinga 2319 (UCB) U.S.A., California, Alameda Co., Berkeley, U.C. Berkeley cam-
pus, 29-I-1999
AF482830
E.C. Vellinga 2337 (UCB) U.S.A., California, Alameda Co., Berkeley, U.C. Berkeley cam-
pus, 3-VIII-1999
AF482876
Agaricus campestris
L.: Fr. GenBank U85307 U85273
Agaricus diminutives
Peck E.C. Vellinga 2360 (UCB) U.S.A., Washington, Olympic Peninsula, Grays Harbor Co.,
north side of Quinault Lake, July Creek Campground, 16-
X-1999
AF482831 AF482877
Agaricus subrutilescens
(Kauffman) Hotson &
Stuntz
E.C. Vellinga 2418 (UCB) U.S.A., California, Sonoma Co., Salt Point State Park, 15-XII-
1999
AF482832
Agaricus
spec. N.L. Bougher H6271
(CSIRO-Wembley)
Australia, West Australia, 86.2 km east of Broome along the
Great Northern Hig, 10-II-1993
AF482833
Allopsalliota geesterani
(Bas
& Heinem.) Nauta
E.C. Vellinga 2263 (L) Netherlands, prov. Noord-Holland, Amsterdam, Amsterdamse
Bos, 23-IX-1998
AF482857 AF482888
Chlorophyllum molybdites
(G.
Meyer: Fr.) Massee J. States AEF1097 (MICH) U.S.A., Arizona, Maricopa Co., Peoria, 4-IX-1994 AF482836
GenBank U85274
Endoptychum agaricoides
Czern. R. Brotzu (herb. Brotzu) Italy, Sardinia, Nuoro, X-1990 AF482837 AF482885
Endoptychum depressum
Sing-
er & A.H. Sm.
E.C. Vellinga 2339 (UCB) U.S.A., California, Sierra Nevada, Fresno Co., Dinkey Creek
Rd, 6-VIII-1999
AF482834 AF482878
Lepiota brunneoincarnata
Chodat & Martin
E.C. Vellinga 2260 (L) Netherlands, prov. Noord-Holland, Amsterdam, Amsterdamse
Bos, 23-IX-1998
AF482875
M. Enderle (L) Germany, Baden-Wu¨rttemberg, ‘‘Ho¨rnle’’ close to Ulm-Grim-
melfingen, 29-VIII-1996
AF482896
Lepiota
‘‘
carmineobasidia
’’
Sundberg (1967)
E.C. Vellinga 2596 (UCB) U.S.A., California, San Mateo Co., San Francisco watershed,
8-XII-2000
AF482860
Lepiota cristata
(Bolt.: Fr.)
Kummer E.C. Vellinga 1445 (L) Luxembourg, Hollenfels, 28-IX-1988 AF391027
GenBank U85292
Lepiota rufipes
Morgan sensu
European authors H.A. Huijser (L) Netherlands, prov. Limburg, Bemelen, 9-X-1991 AF391066 AF482897
447V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
T
ABLE
I. Continued
Species
Collection &
herbarium
Location & date
of collection
ITS
GenBank
accession #
LSU
GenBank
accession #
Lepiota viriditincta
(Berk. &
Broome) Sacc. R.P.J. de Kok F 61 (L) Indonesia, East Kalimantan, Wanariset, 1-III-1993 AF482873
Leucoagaricus americanus
(Peck) Vellinga
S.J.W. Verduin (L) U.S.A., North Carolina, Durham, Duke University campus,
Duke Medical Center, 29-VII-1998
AF295928
E.C. Vellinga 2454 (UCB) U.S.A., Michigan, Washtenaw Co., Ann Arbor campus, Uni-
versity of Michigan, 6-VIII-2000
AF482891
Leucoagaricus barssii
(Zeller)
Vellinga E.C. Vellinga 2342 (UCB) U.S.A., California, Alameda Co., San Leandro, 21-IX-1999 AF295931
E.C. Vellinga 2268 (L) Netherlands, prov. Noord-Holland, Texel, Loosmansduinen,
24-IX-1998
AF482894
Leucoagaricus croceovelutinus
(Bon & Boiffard) Bon &
Boiffard
E.C. Vellinga 2243 (L) Netherlands, prov. Limburg, Elsloo-Geulle, Bunderbos, 19-IX-
1998
AF482862 AF482889
Leucoagaricus crystallifer
Vel-
linga
H.A. Huijser (L) Germany, Baden-Wu¨rttemberg, Gottenheim, Wasenweiler
Wald, 3-IX-1998
AF482863
Leucoagaricus hortensis
(Mur-
rill) Pegler D.E. Hemmes 1365 (SFSU) U.S.A., Hawai‘i Islands, Hawai‘i, Waipio Valley, 10-XII-1996 AF482843
GenBank U85284
Leucoagaricus leucothites
(Vit-
tad.) Wasser
E.C. Vellinga 2050 (L) Netherlands, prov. Noord-Holland, Amsterdamse Waterlei-
dingduinen, Van Lennepkanaal, 15-X-1996
AF482865
GenBank U85280
Leucoagaricus marriagei
D.A.
Reid
E.C. Vellinga 2005 (L) Netherlands, prov. Limburg, Elsloo-Geulle, Bunderbos, 9-IX-
1996
AF482866
Leucoagaricus meleagris
(Sow.) Singer
E.C. Vellinga 2095 (L) Netherlands, prov. Zuid-Holland, Capelle aan den IJssel,
18&19-VIII-1997
AF482867 AF482890
E.C. Vellinga 1990 (L) Netherlands, prov. Noord-Holland, Amsterdam, Zorgvlied, 30-
VII-1996
Macrolepiota nympharum
(Kalchbr.) Wasser C. Bas 9269 (L) Germany, Rheinland-Pfalz, Eifel, near Gerolstein, 21-IX-1990 AF482868
R.P.J. de Kok 3 (L) Germany, Rheinland-Pfalz, Eifel, near Dohm, 24-IX-1990 AF482895
Leucoagaricus purpureolilaci-
nus
Huijsman
E.C. Vellinga 2291 (L) Netherlands, prov. Zeeland, Schouwen-Duiveland, near
Haamstede, 6-XI-1998
AF482869
Leucoagaricus serenus
(Fr.)
Bon & Boiffard
E.C. Vellinga 1930 (L) Belgium, prov. Lie`ge, Tilff, Vallon de la Chavresse, 11-IX-
1995
AF482871 AF482893
Leucoagaricus sericifer
(Locq.) Vellinga
E.C. Vellinga 2116 (L) Netherlands, prov. Utrecht, Breukelen, Nijenrode, 6-X-1997 AF482872
Leucoagaricus wichanskyi
(Pi-
la´t) Bon & Boiffard
H.A. Huijser (L) Netherlands, prov. Limburg, Neercanne, Cannerbos, IX/X-
1987
AF482874
448 M
YCOLOGIA
T
ABLE
I. Continued
Species
Collection &
herbarium
Location & date
of collection
ITS
GenBank
accession #
LSU
GenBank
accession #
Leucoagaricus
spec. E.C. Vellinga 2777 (UCB) U.S.A., California, Alameda Co., Berkeley, U.C. Berkeley cam-
pus, 6-XII-2001
AF482858
Leucocoprinus brebissonii
(Godey) Locq. E.C. Vellinga 1784 (L) France, dept Pas-de-Calais, Foreˆt de Boulogne, 13-X-1991 AF482859
Leucocoprinus cretaceus
(Bull.: Fr.) Locq.
T. Læssøe 6171 (C) Malaysia, Sabah, Tabin Wildlife Reserve, Mud Volcano trails 3
& 4, 9-II-1999
AF482861
J. Engelen (L) Netherlands, prov. Gelderland, Apeldoorn, 9-IX-1997 AF482892
Leucocoprinus heinemannii
Migl.
E.C. Vellinga 2101 (L) Netherlands, prov. Zuid-Holland, Leiden, Hortus Botanicus,
29-VIII-1997
AF482864
Leucocoprinus straminellus
(Bagl.) Narducci & Caroti
E.C. Vellinga 2080 (L) Netherlands, prov. Zuid-Holland, Leiden, Hortus Botanicus,
16-VI-1997
AF482870
Macrolepiota clelandii
Grguri-
novic
K.R. Thiele 2650 (MEL) Australia, Victoria, 52 km north of Orhost on the Bonang
Road, Martins Creek, 17-V-2000
AF482838 AF482882
Macrolepiota colombiana
Franco-Molano GenBank U85311 U85276
Macrolepiota dolichaula
(Berk.) Pegler & Rayner
E.M. Canning 6603 (CANB) Australia, New South Wales, between Blakney Creek and Be-
vendale, just off road inside entrance to ‘Kunama Cottage,’
4-IV-1989
AF482839
R.P.J. de Kok 901 (CANB) Australia, ACT, Canberra, Black Mountain, CSIRO site, 14-IV-
2000
AF482883
Macrolepiota excoriata
(Schaeff.: Fr) Wasser R. Chrispijn (L) Netherlands, prov. Groningen, Dollarddijk, 11-IX-1997 AF482840
Macrolepiota fuliginosa
(Bar-
la) Bon
E.C. Vellinga 2275 (L) Netherlands, prov. Limburg, Elsloo, Bunderbos, 29-IX-1998 AF482841
Macrolepiota globosa
Mossebo H. Neda N421 (TFM) Nigeria, Edo State, Benin City, University of Benin campus,
24-V-2000
AF482842
Macrolepiota mastoidea
(Fr.:
Fr.) Singer
E.C. Vellinga 1685 (L) Germany, Rheinland-Pfalz, Eifel, west of Wiesbaum, 18-IX-
1990
AF482844
GenBank U85279
Macrolepiota neomastoidea
(Hongo) Hongo
E. Nagasawa (TMI 14182) Japan, Shimane Pref., Naka-gun, Yasaka-son, Nishikawauchi,
23-IX-1990
AF482845
Macrolepiota olivieri
(Barla)
Wasser
E.C. Vellinga 2230 (L) Netherlands, prov. Limburg, Elsloo-Geulle, Bunderbos, 19-IX-
1998
AF482846 AF482887
Macrolepiota phaeodisca
Bellu` P. Roux 994 (L) France, Corsica, Porticcio, 6-XI-1990 AF482847
Macrolepiota procera
(Scop.:
Fr.) Singer
E.C. Vellinga 2293 (L) Netherlands, prov. Zeeland, Schouwen-Duiveland, Haamste-
de, 7-XI-1998
AF482848
R.P.J. de Kok s.n. (L) Netherlands, prov. Groningen, Ter Apel, Ter Haar, 18-X-1990 AF482880
Macrolepiota rachodes
(Vit-
tad.) Singer E.C. Vellinga 2106 (L) Netherlands, prov. Zuid-Holland, Leiden, 24-IX-1997 AF482849
449V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
T
ABLE
I. Continued
Species
Collection &
herbarium
Location & date
of collection
ITS
GenBank
accession #
LSU
GenBank
accession #
GenBank U85277
Macrolepiota
spec. nov. 1 Z.L. Yang 2286 (HKAS) China, Sichuan Prov., Xianghcheng, Co., in the vicinity of
Shagong, 11-VII-1998
AF482850
Macrolepiota
spec. nov. 2 O. Oku (herb. Oku) Japan, Ibaraki Pref., Tsukuba, X-2000 AF482851 AF482884
Macrolepiota
spec. nov. 3 Z.L. Yang 2172 (HKAS) China, Yunnan Prov., Jinghong, Damenglon, 14-VIII-1995 AF482853
Macrolepiota
spec. nov. 4 B. Bayliss, E4505 (CSIRO-
Wembley)
Australia, Queensland, Mareeba, Davies Creek Road, 3-II-
1992
AF482854
(ITS1)
Macrolepiota
spec. nov. 5 R.E. Tulloss (UCB) U.S.A., Connecticut, Tolland Co., Gay City State Park, 23-IX-
2000
AF482852
R.E. Tulloss RET 9-25-99-H
(UCB)
U.S.A., Connecticut, Middlesex Co, Meshomasic State Forest,
Cobalt, 25-IX-1999
AF482881
Macrolepiota
spec. nov. A H. Lepp 1142 (CANB) Australia, Australian Capital Territory, Striling, 8 km SW of
Capital Hill, Canberra, 19-I-1995
AF482855
Macrolepiota
spec. nov. B E.C. Vellinga 2317 (UCB) U.S.A., California, Alameda Co, Berkeley, UC-Berkeley cam-
pus, 25-I-1999
AF482856
E.C. Vellinga 2361 (UCB) U.S.A., Oregon, Multnomah Co., Portland, S.E. Locust Ave,
20-X-1999
AF482886
Micropsalliota
spec. T. Læssøe 6025 (C) Malaysia, Sabah, Danum Valley, Field Centre, Main East Trail,
30-I-1999
AF482835 AF482879
450 M
YCOLOGIA
T
ABLE
II. Overview of morphological characters
Clamp-connections absent present
Spore colour white to pink green brown
Germ pore absent present
Cap over germ pore absent present missing
Basidia 2-spored 4-spored
Hymenophoral trama regular trabecular
Spore apex truncate rounded
Habit agaricoid secotioid
Causing gastrointesti-
nal problems no yes
lineages 2 and 4 at genus level. (i) The veil structure
is hymenidermal in Lineage 2 (with clavate to lagen-
iform terminal elements) versus trichodermal (all el-
ements cylindrical and elongate) in Lineage 4; (ii) a
stipe covering is absent in Lineage 2, although pre-
sent in all taxa of Lineage 4, giving the stipes of the
species a striking, banded appearance; (iii) the
spores of the taxa of Lineage 2 lack either a germ
pore, or have a truncate to rounded apex, without a
hyaline cap covering the germ pore; the pore is
caused by a depression of the episporium (Mele´ndez-
Howell 1967); the spores of the Lineage 4 taxa always
have a rounded apex, with a germ pore, covered by
a hyaline cap; the germ pore in this case is caused by
an interruption of the episporium, and it is filled by
a refringent plug (Mele´ndez-Howell 1967).
The data suggest that representatives of Lineage 2
are more thermophilic or thermotolerant than those
of Lineage 4. Lineage 2 includes species with a wide
distribution in the tropics, or with a preference for
compost heaps.
All agaricoid taxa of lineages 2 and 4 share the
general
Macrolepiota
features: The basidiocarps are
relatively big and fleshy (though the two undescribed
taxa basal to Lineage 4 form relatively small basidi-
ocarps), with a more or less complex annulus on the
stipe, made up of both universal and partial veil rem-
nants. The spores of all taxa are ellipsoid to amyg-
daloid-ellipsoid, and relatively large (8.0–22
m
m
long).
Presence or absence of clamp connections previ-
ously was considered a good character to distinguish
sections within the genus
Macrolepiota
(Singer 1986,
Pa´zma´ny 1985). In our observations, clamp connec-
tions were found, at least at the base of the basidia,
in almost all investigated taxa of Lineage 4 (they are
absent to rare in collections belonging to the com-
plex of
M. mastoidea
(Vellinga 2001)). Clamp con-
nections are absent or rare in some taxa of Lineage
2, e.g.,
Chlorophyllum molybdites
(Sundberg 1971),
and absent in others (
M. venenata
,
E. agaricoides
, and
M
. spec. nov. A (senior author pers obs). In short,
clamp connections are present and absent in both
lineages, as indicated in F
IG
. 3, and clearly this char-
acter cannot be used to distinguish the two lineages.
Taxonomic implications.
Morphological characters,
particularly features of the covering layers, and the
spore apex, provide the decisive factor to rule out
the ‘‘big’’
Macrolepiota
clade alternative and to re-
gard the two lineages as separate genera.
Lineage 4 comprises the type species of
Macrole-
piota
,
M. procera
, and keeps the name
Macrolepiota
.
Macrolepiota
in this restricted sense comprises section
Macrolepiota
and section
Macrosporae
with subsec-
tions
Excoriatae
Bon and
Microsquamatae
(Pa´zma´ny)
Bellu` & Lanzoni of the infrageneric classification ad-
vocated by Bon (1993).
Lineage 2 is more diverse, with relatively long-
branch lengths in the phylograms based on the se-
quence data, and the species morphologically are
more diverse than those in Lineage 4. Lineage 2 is
made up of
Macrolepiota
section
Laevistipedes
(Pa´z-
ma´ny) Bon,
M. globosa
, the genus
Chlorophyllum
,
Leu-
coagaricus hortensis
, and
Endoptychum agaricoides.
Macrolepiota neomastoidea
is basal to this lineage. Mo-
reno et al (1995) concluded on morphological
grounds that
Chlorophyllum
and
Macrolepiota
should
be merged, the only differences between the two gen-
era being the different spore color. The green pig-
ments in
Chlorophyllum
spores obscure the staining
reactions that nevertheless are similar to those of
spores of
Macrolepiota
species. Moreno et al (1995)
used the name
Macrolepiota
for the combined group,
though the genus name
Chlorophyllum
(Massee 1898)
predates
Macrolepiota
(Singer 1948) by 50 years.
Endoptychum agaricoides
is the type species of the
genus
Endoptychum
(Czernajew 1845), which means
that Lineage 2 should be called
Endoptychum
, be-
cause it is the oldest generic name available. How-
ever, it would be infelicitous to call Lineage 2
Endop-
tychum
, because this name is associated in usage and
etymology with the secotioid habit of only one taxon
in Lineage 2. Furthermore, the name is not well
known and has been used for species now shown to
belong to several genera (discussed further below).
Vellinga and De Kok (2002) officially proposed to
conserve
Chlorophyllum
against
Endoptychum
.
Akers and Sundberg (1997) placed the species
Leu-
coagaricus hortensis
in the genus
Leucoagaricus
on ac-
count of the absence of the germ pore, despite the
presence of clamp connections. In all analyses, that
position is significantly worse than including
L. hor-
tensis
within Lineage 2, the
M. rachodes
-
Chlorophyl-
lum
-
Endoptychum
clade.
Macrolepiota nympharum
is shown to belong to nei-
451V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
T
ABLE
III. Results from Kishino-Hasegawa tests for the three data sets.
P
is the probability of getting a more extreme T-value under the null hypothesis of no difference
between the two trees (two-tailed test)
Constraints
ITS data
Tree
length
No.
trees 2ln L
P
LSU data
Tree
length
No.
trees 2ln L
P
ITS1LSU data
Tree
length
No.
trees 2ln L
P
Unconstrained 1353 90 6167.58989 0.5992 225 100 1049.61794 (best) 988 20 4401.49163 0.5343
Macrolepiota
including
Chlorophyl-
lum
species,
Endoptychum agari-
coides
, and
Leucoagaricus horten-
sis
, but excluding
L. nympharum
,
is monophyletic 1359 84 6171.74242 0.1287 229 306 1064.59536 0.1187 994 12 4413.93499 0.1863
Macrolepiota
sensu Singer (1986) is
monophyletic, i.e., excluding
Chlorophyllum
,
Endoptychum agar-
icoides
, and
Leucoagaricus horten-
sis
(but also excluding
L. nym-
pharum
) 1369 120 6194.14041 0.0681 231 204 1068.66069 0.0823 1001 9 4439.71153 0.0012*
Macrolepiota
including
Chlorophyl-
lum
, but excluding
E. agaricoides
and
L. hortensis
is monophyletic 1373 120 6199.17163 0.0363* 231 165 1069.43603 0.0544 1012 28 4443.77166 0.0002*
Macrolepiota
including
E. agaricoides
and
Chlorophyllum
, excluding
Leucoagaricus hortensis
is mono-
phyletic 1387 58 6200.62042 0.0304* 229 66 1064.14571 0.1132 1004 20 4441.08814 0.0102*
Leucoagaricus nympharum
is not a
sister taxon of
L. leucothites
but
belongs to
Macrolepiota
(lineages
2 and 4) 1357 1170 6230.88738 0.0042* 239 48 1106.47892 ,0.0001* 1060 2 4607.29662 ,0.0001*
Macrolepiota
(lineages 2 and 4)
form one clade with
Agaricus
1357 120 6159.82350 (best) 245 408 1088.52286 0.0034* 991 6 4398.41866 (best)
Macrolepiota
(lineages 2 and 4) is a
sister group to the
Leucoagaricus-
Leucocoprinus
assemblage 1383 669 6234.02119 0.0030* 230 237 1060.50652 0.2053 994 3 4410.9952 0.2396
Endoptychum
forms a monophyletic
group, comprising
E. depressum
and
E. agaricoides
1427 180 6345.85121 ,0.0001* 237 116 1075.09145 0.0242* 1068 6 4582.82402 ,0.0001*
Leucoagaricus hortensis
belongs in
the
Leucoagaricus-Leucocoprinus
assemblage 1394 62 6263.64613 ,0.0001* 228 117 1055.32346 0.4337 1006 10 4444.83291 0.0060*
*
P
,0.05.
452 M
YCOLOGIA
F
IG
. 1. Maximum-parsimony analysis of ITS data, one of 90 most parsimonious trees. Bootstrap values over 70% are
indicated in bold below the branches.
ther of the two
Macrolepiota
lineages, but to be a sis-
ter taxon of
Leucoagaricus leucothites
. Migliozzi and
Bizzi (1994) suggested on morphological grounds
that
M. nympharum
(as
M. puellaris
) was an inter-
mediate between
Macrolepiota
and
Leucoagaricus
.
The structure of the pileus covering is much looser
than in
Macrolepiota
species; the shape of the cheil-
ocystidia and, in particular, the absence of clamp con-
nections support placement in
Leucoagaricus
(see
also Vellinga 2001).
The position of
Endoptychum
.
It is not a new obser-
vation that secotioid fungi do not form a monophy-
letic group but are either mutants of normally agar-
icoid taxa (Hibbett et al 1994) or recently derived
from diverse agaricoid and boletoid ancestors.
En-
doptychum depressum
and
E. agaricoides
both recently
are derived from representatives of Agaricales, it ap-
pears. In such cases, taxonomic conclusions have
been drawn in different ways. Kretzer and Bruns
(1997) combined the genera
Gastrosuillus
Thiers and
453V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
F
IG
. 2. Best (2ln Likelihood) of 100 equally most-par-
simonious trees, based on LSU data. Bootstrap values over
70% are indicated in bold below the branches.
Suillus
Gray, but Redhead et al (2001) kept
Montag-
nea
Fr. and
Coprinus
Pers. s. str. as separate genera
within the Agaricaceae.
Endoptychum depressum
in
Lineage 1 is morphologically an
Agaricus
species in
which the lamellae are not exposed and the basidia
do not actively discharge spores. It is closely related
to
Agaricus arvensis
. The name
Agaricus depressus
Le´veille´ , used for a small marasmioid species from
Java, Indonesia, predates a new combination by at
least 150 years, so the new name
Agaricus inapertus
Vellinga is proposed; basionym:
Endoptychum depres-
sum
Singer & A.H. Sm. in Brittonia 10: 216. 1958.
Endoptychum agaricoides
is a sister taxon of
Macro-
lepiota globosa
and
Chlorophyllum molybdites
. Conard
(1915) had presumed a close relationship with
Agar-
icus campestris
, because of similarities in develop-
ment, although this opinion was disputed strongly by
Lohwag (1924), who interpreted the agaricoid fungi
as derived from secotioid forms. A relationship with
Chlorophyllum
already was suggested by Singer and
Smith (1958), on account of similar spore colors. The
spores of
E. agaricoides
, the secotioid member of Lin-
eage 2, are greenish to yellowish brown under the
microscope, not gray-green like the spores of
Chl.
molybdites
. A germ pore is lacking or vaguely visible.
Young, uncolored spores have all the characteristic
staining reactions: red in Congo Red, blue in Cotton
Blue, red-brown in Melzer’s reagent, and with a pink
inner wall in Cresyl Blue. Clamp connections are ab-
sent at the base of the basidia and on hyphal septa,
and the hymenophoral trama hyphae are inflated,
character states shared with
Chl. molybdites
.
Singer and Smith (1958) listed two more
Endopty-
chum
species:
E. melanosporum
(Berk.) Singer & A.H.
Sm. with black spores, and
E. arizonicum
(Shear &
Griffiths) Singer & A.H. Sm. with white, globose
spores, and clamp connections in the trama (senior
author pers obs). These taxa await sequencing for
correct taxonomic placement.
Taxa other than the ones studied here putatively
belonging to the
Chlorophyllum
lineage are
M. abrub-
tibulba
(Heim) Heinem.,
M. brunnea
(Farlow & Burt)
S. Wasser,
M. venenata
,
M. subrhacodes
Murrill, and
probably
Chlorolepiota mahabaleshwarensis
Sathe &
Deshpande.
Classification above genus level.
Current molecular
data also show that recognition of two families, Le-
piotaceae for the non-brown-spored taxa and Agari-
caceae for brown-spored taxa, cannot be justified, just
as shown in earlier papers (e.g., Johnson 1999, Mon-
calvo et al 2000). In fact, Lineage 2 appears to be
more closely related to the brown-spored genus
Agar-
icus
than to the white-spored genera
Leucoagaricus
and
Leucocoprinus
, despite discordant morphological
data, such as the structure of the trama (regular in
Agaricus
versus trabecular in
Chlorophyllum
,
Macrole-
piota
,
Leucoagaricus
and
Leucocoprinus
) and the struc-
ture of the pileus coverings.
Classification of the family Agaricaceae into three
tribus has to be reconsidered. Division into two tri-
bus, Lepioteae and Agariceae, seems to be better sup-
ported by the current molecular data, but analysis of
data in the family as a whole is better suited to re-
solving the intrafamilial classification.
Coprinus com-
atus
(O.F. Mu¨ll. : Fr.) Pers. and
C. sterquilinus
(Fr. :
Fr.) Fr., and their secotioid allies
Montagnea arenarius
(DC.) Zeller, and
Podaxis
Desv. should be included
in these analyses (Hopple 1994, Johnson 1999) and
so should gasteroid taxa in the Lycoperdaceae (Hib-
bett et al 1997, Kru¨ger et al 2001, Moncalvo et al
2002).
454 M
YCOLOGIA
F
IG
. 3. Strict-consensus tree, based on maximum-parsimony analysis of the combined data set. Presence or absence of
clamp connections is indicated. Bootstrap values over 70% are given below the branches. The # sign indicates that the species
causes gastro-intestinal problems.
Molecular and morphological characters in Line-
age 3, the
Leucoagaricus-Leucocoprinus
assemblage,
are highly diverse, and our datasets do not warrant
any conclusions on this lineage because a relatively
small number of representatives have been chosen
for a comparison with
Macrolepiota
and
Chlorophyl-
lum
.
In conclusion.
The three questions raised in the in-
troduction now can be answered. The genus
Macro-
lepiota
has to be emended and now is restricted to
species with trichodermal pileus covering, a stipe cov-
ering made up of hymeni-trichodermal patches, and
spores with a rounded apex with a covered germ
pore.
Macrolepiota procera
,
M. dolichaula
,
M. masto-
455V
ELLINGA ET AL
:
M
ACROLEPIOTA PHYLOGENY AND TAXONOMY
idea
and allies belong here. A second genus com-
prises
Endoptychum agaricoides
,
Chlorophyllum molyb-
dites
,
M. globosa
,
Leucoagaricus hortensis
and the
group of
M. rachodes
. The agaricoid members are
characterized by a hymenidermal pileus covering, a
smooth stipe, and often truncate spores with an un-
covered germ pore, or the germ pore might be ab-
sent. We suggest that the generic name
Chlorophyllum
is appropriate for this lineage, pending the outcome
of a proposal submitted to Taxon. Toxic species
group together in the latter genus, Although they do
not constitute a separate clade. It also is shown that
secotioid taxa, formerly in the genus
Endoptychum
,
belong to different genera:
E. depressum
in the genus
Agaricus
, and
E. agaricoides
in
Chlorophyllum
(pro-
posal pending).
ACKNOWLEDGMENTS
The authors are grateful to the curators of the herbaria C,
CANB, KUN, L, MEL, MICH, the Mycology Herbarium,
CSIRO Forestry and Forest Products, Wembley, Western
Australia, Australia, and these people for material: Frances-
co Bellu` (Italy), Martin I. Bidartondo (California), Eric Boa
(Great Britain), Renato Brotzu (Italy), Rob Chrispijn (the
Netherlands), Manfred Enderle (Germany), Lisa C. Gru-
bisha (California), Jacques Guinberteau (France), Don E.
Hemmes (Hawaii), Henk A. Huijser (the Netherlands), Piet
H. Kelderman (the Netherlands), Richard W. Kerrigan
(Pennsylvania), Thomas Læssøe (Denmark), K. Maruyama
( Japan), Buck McAdoo (Washington), Dominique C. Mos-
sebo (Cameroon), Eiji Nagasawa ( Japan), Mr Oku (Japan),
Clark L. Ovrebo (Oklahoma), Pierre Roux (France), Clive
Shirley (New Zealand), Tamara Spillis (Ohio), R.K. Thiele
(Australia), Richard Tofts (Great Britain), Steve Trudell
(Washington), and Rod Tulloss (New Jersey). Partial fi-
nancing of the lab work by the ‘‘Rijksherbariumfonds Dr.
E. Kits van Waveren’’ is acknowledged (ECV). The MSA Al-
exander H. and Helen V. Smith award to ECV enabled her
to visit the herbarium of the University of Michigan at Ann
Arbor in August 2000. Dennis E. Desjardin, Richard W. Ker-
rigan and an anonymous reviewer gave insightful comments
on the manuscript. John Lennie provided linguistic and ed-
itorial advice.
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