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Molecular phylogeny and taxonomy of the genus
Veloporphyrellus
Yan-Chun Li
Key Laboratory for Plant Diversity and Biogeography of
East Asia, Kunming Institute of Botany, Chinese
Academy of Sciences, Kunming 650201, China
Beatriz Ortiz-Santana
US Forest Service, Northern Research Station, Center for
Forest Mycology Research, One Gifford Pinchot Drive,
Madison, Wisconsin 53726-2398
Nian-Kai Zeng
Department of Pharmacy, Hainan Medical University,
Haikou 571199, China
Bang Feng
Zhu L. Yang
1
Key Laboratory for Plant Diversity and Biogeography of
East Asia, Kunming Institute of Botany, Chinese
Academy of Sciences, Kunming 650201, China
Abstract
:
Veloporphyrellus
is a genus known from
North and Central America, southeastern Asia, and
Africa. Because species of this genus are phenotypi-
cally similar to some taxa in several genera, such as
Boletellus
,
Leccinum
,
Strobilomyces
,
Suillus
and
Tylopi-
lus
s.l. belonging to Boletales, its phylogenetic
disposition has never been addressed. We analyzed
four DNA regions, the nuclear ribosomal LSU and
tef-
1a
, and the mitochondrial mtSSU and
atp
6 genes, to
investigate the phylogenetic disposition of
Velopor-
phyrellus
. Although the monophyly of the genus and
its systematic placement within the Boletaceae was
well supported, its relationship to other genera was
not resolved. Morphologically
Veloporphyrellus
is
distinguished from other boletoid genera by the
combination of the pinkish or grayish pink hymeno-
phore, the membranous veil hanging on the pilea
margin, the trichoderm-like pileus covering and the
smooth basidiospores. Five species, including two new
species and two new combinations, are described and
illustrated. A key to the species of
Veloporphyrellus
also
is provided.
Key words:
Boletaceae, mycorrhiza, new taxa,
phylogeny, taxonomy
INTRODUCTION
Veloporphyrellus
L.D. Go´mez & Singer, based on
V.
pantoleucus
L.D. Go´mez & Singer, originally was
described for a species from Costa Rica (Go´ mez and
Singer 1984). Watling and Turnbull (1993) contrib-
uted another species to this genus (i.e.
V. africanus
Watling). It is notable that the character of the veil
extending from the pilea margin and embracing the
stipe in younger basidiomata has been shared by
some members of related genera: for
Austroboletus
(Corner) Wolfe as in
A. dictyotus
(Boedijn) Wolfe and
A. fusisporus
(Kawam. ex Imazeki & Hongo) Wolfe;
for
Boletellus
Murrill as in
B. ananas
(M.A. Curtis)
Murrill,
B. emodensis
(Berk.) Singer,
B. ananiceps
(Berk.) Singer, and
B. projectellus
(Murrill) Singer; for
Leccinum
Gray as in
L. atrostipitatum
A.H. Sm., Thiers
& Watling; for
Strobilomyces
Berk. as in
S. floccopus
(Vahl.) P. Karst.; for
Suillus
Gray as in
S. luteus
(L.)
Roussel. In addition, the characters of the pale
pinkish to light pinkish or grayish pink hymenophore
and spore print, also are shared with species in
Australopilus
Halling & Fechner,
Austroboletus
,
Fistu-
linella
Henn.,
Harrya
Halling, Nuhn & Osmundson,
Tylopilus
P. Karst.,
Zangia
Yan C. Li & Zhu L. Yang
and some species in
Porphyrellus
E.J. Gilbert. Thus, a
few species in
Veloporphyrellus
have caused a number
of important systematic and evolutionary questions in
the past, but until now, the systematic position of
Veloporphyrellus
based on molecular data has not been
investigated.
In our study of boletes that have the characters of a
distinct membranous veil or a projecting appendicu-
late margin, a pink to pinkish hymenophore and
smooth basidiospores, we found four additional
species that morphologically agree with the generic
concept of
Veloporphyrellus
. We used both morpho-
logical data and molecular sequences of four genes,
together with ecological data to (i) compare the
morphological features among
Veloporphyrellus
and
other similar genera such as
Australopilus
,
Austrobo-
letus
,
Boletellus
,
Fistulinella
,
Harrya
,
Leccinum
,
Strobi-
lomyces
,
Suillus
,
Tylopilus
and
Zangia
; (ii) investigate
the phylogenetic position and sister groups of
Veloporphyrellus
; (iii) evaluate the relationships
among
Veloporphyrellus
species; and (iv) provide
insights about the distribution and mycorrhizal hosts
of the genus.
MATERIALS AND METHODS
Morphological studies.—
Macroscopic descriptions are based
on detailed field notes made on fresh basidiocarps.
Microscopic structures were revived in 5%KOH. Sections
of the pileus covering were cut radially, vertically and
halfway between center and margin of the pileus. All
Submitted 20 Apr 2013; accepted for publication 20 Sep 2013.
1
Corresponding author. E-mail: fungi@mail.kib.ac.cn
Mycologia,
106(2), 2014, pp. 291–306. DOI: 10.3852/106.2.291
#2014 by The Mycological Society of America, Lawrence, KS 66044-8897
291
microscopic features were drawn by hand (Zeng et al. 2012,
2013; Hosen et al. 2013). For explanations of spore data
see Li et al. (2009). Color codes are from Kornerup and
Wanscher (1981). Methods for scanning electron micros-
copy (SEM) followed Xiang et al. (2010). Briefly, basidio-
spores were scraped from the dried hymenophore, pasted
onto an SEM stub with double-sided tape, coated with gold-
palladium and photographed with an AMRAY 1000B SEM.
Specimens examined are deposited in CFMR, KUN and F
(herbaria codes according to Thiers 2011).
DNA extraction, PCR and DNA sequencing.—
Protocols for
DNA extraction, PCR, cloning, sequencing and sequence
alignment followed those in Li et al. (2011), Xiang et al.
(2012), Du et al. (2012) and references therein. The primer
pair used for amplifying the nrLSU region was LROR and
LR7 (Vilgalys and Hester 1990). However, for sample
Go´ mez 21232 deposited at herbarium F (the type of
V.
pantoleucus
), we failed to amplify the nrLSU region using
the above primer pair. Thus, the primer pairs LROR and
LR3 and LR3R and LR5 (Vilgalys and Hester 1990) were
used. To amplify the translation elongation factor 1a(
tef-
1a
), the primers EF1-a-F and EF1-a-R were used (Mikheyev
et al. 2006). For amplifying the mitochondrial small subunit
gene fragment (mtSSU), we used the primer pair MS1 and
MS2 (White et al. 1990). To amplify the mitochondrial atp6
gene fragment (
atp
6), the primer pair
atp
6-5 and
atp
6-R was
used (Li et al. 2011).
Phylogenetic analysis.—
The phylogenetic analyses were
based on four genes, nrLSU,
tef-1a
, mtSSU and
atp
6. Two
datasets were analyzed: the single-locus dataset for the
nrLSU gene and the concatenated multilocus dataset of
nuclear genes (nrLSU and
tef-1a
) and mitochondrial genes
(mtSSU and
atp
6). To test for phylogenetic conflict among
the four genes, the partition homogeneity (PH) or
incongruence length difference (ILD) test was performed
with 1000 randomized replicates, using heuristic searches
with simple addition of sequences in PAUP* 4.0b10
(Swofford 2002). The result of the partition homogeneity
test showed that the phylogenetic signals present in the
different gene fragments were not in conflict (
P
50.76)
and could therefore be combined.
Although the genus
Veloporphyrellus
originally was placed
in the Boletaceae, it also shares some features, such as the
extended veil on the pilea margin, the pored hymeno-
phore, and the elongate and smooth basidiospores, with a
few species in
Suillus
. Thus, sequences from samples
Suillus cavipes
(Opat.) A.H. Sm. & Thiers,
S. granulatus
(L.) Roussel and
S. luteus
of the Suillaceae were
downloaded from GenBank and included for outgroup
rooting in both analyses. The scientific names, origin,
collection information and GenBank accession numbers
for the sequences used in the combined nuclear and
mitochondrial DNA datasets are presented (TABLE I). DNA
sequences were edited and aligned with BioEdit and
Clustal X and manually checked and adjusted where
necessary (alignment deposited in TreeBASE with acces-
sion no. S14517).
Both datasets were analyzed with maximum parsimony
(MP), randomized accelerated maximum likelihood
(RAxML) and Bayesian Inference (BI) methods respective-
ly. MP analysis was estimated in PAUP*4.0b10 (Swofford
2002) with these settings: gaps as missing data; multistate
taxa interpreted as uncertainty; starting tree(s) obtained via
stepwise addition; 1000 random addition sequences; one
tree held at each step during stepwise addition; tree-
bisection-reconnection (TBR) branch swapping; steepest
descent and MULTREES options not in effect. One hundred
MP bootstrap replicates were completed using heuristic
searches with the same search parameters as above. All
parameters in the ML analysis used the default setting, and
statistical support values were obtained using nonparamet-
ric bootstrapping with 100 replicates. All datasets were
analyzed further with a Bayesian approach (metropolis-
coupled Monte Carlo) using MrBayes 3.1 (Huelsenbeck and
Ronquist 2005). The parameter model was selected by the
Akaike information criterion (AIC) as the best-fit likelihood
model with ModelTest 3.7 (Posada and Buckley 2004). The
models employed for each of the four partitions were:
TRN+I+Gfor nrLSU, GTR+I+Gfor
tef-1a
and mtSSU, and
TVM+I+Gfor
atp
6. Posterior probabilities (PP) were deter-
mined twice by running one cold and three heated chains
in parallel mode, saving trees every 1000 generations. Runs
were terminated once the average standard deviation of
split frequencies fell below 0.01. The trees during burn-in
were excluded, and the 50%majority-rule consensus tree of
the remaining trees was calculated by PAUP* to determine
Bayesian posterity probability of each clade.
RESULTS
Molecular data.—
Forty-two sequences, 12 nrLSU, 7
tef-1a
, 11 mtSSU and 12
atp
6, were newly generated
for this study (TABLE I). For the nrLSU dataset,
sequences representing most of the genera in the
Boletaceae were downloaded from GenBank (Castellano
et al. 1992; Binder and Fischer 1997; Binder and Besl
2000; Binder and Bresinsky 2002; Peintner et al. 2003;
Binder and Hibbett 2007; Halling et al. 2007; Halling
et al. 2008; Desjardin et al. 2008, 2009; Dentinger et al.
2010; Orihara et al. 2010; Li et al. 2011; Halling et al.
2012). The final nrLSU dataset included 53 nrLSU
sequences, and the alignment contained 976 nucleotide
sites (273 were parsimony informative). Parsimony
analysis resulted in six most parsimonious trees of 1123
steps, with consistency index (CI) 50.438 and retention
index (RI) 50.646. The combined nrLSU,
tef-1a
, mtSSU
and
atp
6 dataset consisted of 2517 nucleotides (583 sites
were parsimony informative). Parsimony analysis resulted
in 109 parsimonious trees of 1548 steps, with CI 50.673,
RI 50.755.
In our phylogenetic analyses on both datasets
using ML, MP and BI approach, very similar
estimates of tree topologies were produced. The
analyses differed in that ML and BI yielded greater
resolution within and among clades. The most
significant finding was that
Veloporphyrellus
typified
292 MYCOLOGIA
by
V. pantoleucus
nested within the Boletaceae clade,
and clustered with
V. velatus
(5
Tylopilus velatus
[Rostr.] F.L. Tai),
V. conica
(5
Fistulinella conica
[Ravenel] Pegler & T.W.K. Young),
V. alpinus
and
V. pseudovelatus
, with moderate to high support
values on single-locus and multilocus datasets in all
analyses (FIGS. 1, 2). Three main clades with signif-
icant statistical support were recovered within
Veloporphyrellus
. Three samples of
V. conica
formed
a well supported clade with 100%ML, MP bootstrap
support values and high PP values for both datasets
(clade I). Two sequences from the type specimen of
V. pantoleucus
and one sequence from
V. velatus
were clustered together with 100%ML, MP bootstrap
support values and high PP values on both datasets
(clade II). In addition, four samples of
V. pseudove-
latus
clustered with two samples of
V. alpinus
with
100%ML, MP bootstrap support values and high PP
values on both datasets (clade III).
Morphological observations.—
In this study 16 speci-
mens, representing five species (see descriptions
below) phenetically similar to the type species of
Veloporphyrellus
, were examined.
Veloporphyrellus
is
characterized by the pinkish or grayish pink hymeno-
phore, the projecting membranous veil, the smooth
basidiospores and the trichodermial pileus covering.
TAXONOMY
Veloporphyrellus alpinus Yan C. Li & Zhu L. Yang,
sp. nov. FIGS. 3A–C, 4A, 5
MycoBank MB801791
Etymology:
Named because of its subalpine to alpine
distribution.
Diagnosis:
This species is distinguished from others
in
Veloporphyrellus
by the sharp umbonate pileus, large
basidiospores, the subalpine to alpine distribution and
the mycorrhizal association with species of
Abies
.
TABLE I. Specimens used in molecular phylogenetic studies and their GenBank accession numbers
Taxon Voucher Locality
GenBank accession no.
nrLSU
atp6
mtSSU
tef-1a
Boletellus projectellus
— — AY684158 DQ534604 — AY879116
Boletinellus merulioides
— — AY684153 DQ534601 — DQ056287
*
Boletus edulis
KUN (HKAS 55836) Marburg, Germany HQ326927 HQ326839 HQ326903 HQ326860
Chamonixia caespitosa
— — AF336245 AF114444 AF213145 —
*
Leccinum holopus
KUN (HKAS 53417) Hunan, central China HQ326928 — HQ326904 HQ326861
*
L. manzanitae
KUN (HKAS 51277) Tibet, SW China HQ326929 — HQ326905 HQ326862
Paxillus involutus
— — AY612815 AF114447 AY615912 —
Phylloporus rhodoxanthus
— — DQ534631 AF114443 M91013 —
Strobilomyces floccopus
— — DQ534626 DQ534607 AY615918 AY883428
Suillus cavipes
— — AF071535 — M91016 —
S. granulatus
— — AB284479 AF002137 AY615920 —
S. luteus
— — AY612825 AF002135 — —
*
T. felleus
a
KUN (HKAS 54926) Marburg, Germany HQ326933 HQ326843 HQ326909 HQ326866
*
T. felleus
b
KUN (HKAS 55832) Jilin, NE China HQ326934 HQ326844 HQ326910 HQ326867
*
Veloporphyrellus alpinus
a
KUN (HKAS 57490) Yunnan, SW China JX984537 JX984514 JX984526 JX984549
*
V. alpinus
b
KUN (HKAS 68301) Yunnan, SW China JX984538 JX984515 JX984527 JX984550
*
V. conicus
a
CFMR (BZ 1670) Cayo District, Belize JX984543 JX984520 JX984532 JX984555
*
V. conicus
b
CFMR (BZ 1705) Cayo District, Belize JX984544 JX984521 — —
*
V. conicus
c
CFMR (BZ 2408) Cayo District, Belize JX984545 JX984522 JX984533 —
*
V. pantoleucus
a
F (Go´ mez 21232-1) Cartago, Costa Rica JX984548 JX984525 JX984536 —
*
V. pantoleucus
b
F (Go´ mez 21232-2) Cartago, Costa Rica JX984547 JX984524 JX984535 —
*
V. velatus
KUN (HKAS 63668) Hainan, south China JX984546 JX984523 JX984534 JX984554
*
V. pseudovelatus
a
KUN (HKAS 59444) Yunnan, SW China JX984542 JX984519 JX984531 JX984553
*
V .pseudovelatus
b
KUN (HKAS 52673) Yunnan, SW China JX984541 JX984518 JX984530 JX984552
*
V. pseudovelatus
c
KUN (HKAS 52244) Yunnan, SW China JX984539 JX984516 JX984528 —
*
V. pseudovelatus
d
KUN (HKAS 52258) Yunnan, SW China JX984540 JX984517 JX984529 JX984551
*
Zangia citrina
KUN (HKAS 52677) Fujian, SE China HQ326940 HQ326849 HQ326916 HQ326871
*
Z. olivacea
KUN (HKAS 55830) Yunnan, SW China HQ326946 HQ326855 HQ326922 HQ326874
*
Z. olivaceobrunnea
KUN (HKAS 52275) Yunnan, SW China HQ326947 HQ326856 HQ326923 HQ326875
*
Sequences obtained in this study and form study by Li et al. (2011). Others were from GenBank. SW 5southwestern, NE 5
northeastern, SE 5southeastern. Superscripts (a, b, c and d) relate individual collections of the same taxon to their
corresponding sequence data (FIGS. 1, 2).
LIETAL.:
V
ELOPORPHYRELLUS
PHYLOGENETICS 293
FIG. 1. Cladogram resulting from the nrLSU rDNA dataset using RAxML. ML and MP BS support values .50%are
indicated above or below the branches as ML BS/MP BS. In Bayesian analysis, PP .0.95 are indicated with thick branches.
GenBank accession numbers are provided after the species name.
294 MYCOLOGIA
Basidiomata 1.8–3.5 cm wide, small to medium-
sized. Pileus conical to applanate, always with a sharp
umbo, densely covered with radially arranged brown
(7D7–8) to cocoa brown (8D7–8) or chestnut brown to
dark reddish brown (8C7–8) squamules, dry, not viscid
when wet; margin extended, forming membranous veil
which is concolorous with pileus; the veil attached to
apex of stipe in younger basidioma but broken into
pieces and hanging on pilea margin in aged ones.
Hymenophore adnate to depressed around apex of
stipe, pallid (1B1) to pale pinkish (11B3–5) or pinkish
(13A2) to pink (12A3–4); tubes and pores concolor-
ous, tubes up to 0.6 cm long and pores up to 0.1 cm
wide, color unchanged when bruised. Stipe 5.5–6.5 3
0.4–0.7 cm, clavate, sometimes enlarged downward,
glabrous, yellowish orange (5A4–5) to grayish orange
(5B5–6) upward and brown (7D7–8) to reddish brown
(8C7–8) downward; basal mycelia white (1A1). Con-
text of pileus and stipe solid, white (1A1) to bright
white, color unchanged when bruised.
Basidia 34–39 38.5–12 mm, clavate, hyaline to light
yellowish in KOH, thin-walled, four-spored, occasion-
ally two-spored. Basidiospores [60/3/3] (15.5–)
16–19(–19.5) 3(4.5–)5–6(–6.5), (Q 5[2.77–]2.82–
3.56[–3.60], Q
m
53.13 60.18), boletoid, slightly
thick-walled (up to 0.5 mm thick), subhyaline to light
olivaceous in KOH and yellowish brown in Melzer’s
reagent, smooth under SEM (FIG. 4A). Pleurocystidia
47–69 35.5–9 mm, fusiform to subfusiform or
subfusoid-mucronate to ventricose-mucronate with a
short pedicel, sometimes narrowly mucronate, ros-
trate, thin-walled, hyaline in KOH and yellowish to
yellow in Melzer’s reagent. Cheilocystidia 33–81 36–
9mm, broadly clavate to subfusiform or ventricose,
thin-walled, some with 1–2 septa or secondary septa,
hyaline in KOH and yellowish to yellow in Melzer’s
reagent. Hymenophoral trama bilateral, composed of
broad hyaline hyphae up to 10 mm wide. Pileus
covering (squamules on pileus) composed of more or
less vertically to almost radially arranged to loosely
interwoven, colorless to yellowish, filamentous hyphae
4.5–7 mm diam and often with yellow to yellowish
brown vacuolar pigment soon dissolved in KOH
solution; terminal cells 27–69 34.5–6.5 mm, sub-
cylindrical. Pilea trama composed of hyphae 5–10 mm
diam, colorless to yellowish in KOH. Clamp connec-
tions absent in all tissues.
Habitat:
Solitary on the ground in mixed forest of
Abies delavayi
,
Castanea henryi
and
Fargesia
spp. or in
mixed forest of
Abies kawakamii
and
Fargesia
spp.
Distribution:
Currently known from subalpine to
alpine regions, 3100–3600 m, in China (Yunnan
province, Taiwan).
Holotype
: CHINA. YUNNAN PROVINCE: Lijiang
County, Shitou, 3100 m, 2 Sep 2009,
B. Feng 761-
HKAS 57490
(KUN).
Additional specimens examined:
CHINA. YUNNAN PROV-
INCE: Dali County, Canshan National Forest Park, 3600 m,
12 Aug 2010,
X.T. Zhu 125-HKAS 68301
(KUN). TAIWAN:
Hehuan Mountain, 3200 m, 15 Sep 2012,
B. Feng 1266-
HKAS 63669
(KUN).
Notes: Veloporphyrellus alpinus
is characterized by
the conical to applanate but sharply umbonate pileus,
the dense tomentose, brown to cocoa brown or
chestnut brown to dark reddish brown squamules
on the pileus, the white membranous veil remnants,
the pinkish to flesh-colored hymenophore, the
grayish orange to brown or brownish stipe surface,
large basidiospores, the mainly mycorrhizal associa-
tion with species in
Abies
. It is easy to separate this
species from the remaining taxa in
Veloporphyrellus
.
Phylogenetically,
V. alpinus
was clustered with
V.
pseudovelatus
, with high support values based on both
single-locus and multilocus sequence datasets using
ML, MP and Bayesian approach (FIGS. 1, 2). But they
FIG. 2. Cladogram resulting from the combined nuclear
(nrLSU and
tef-1a
) and mitochondrial (mtSSU and
atp6
)
DNA dataset using RAxML. ML and MP BS support values
.50%are indicated above or below the branches as ML
BS/MP BS. In Bayesian analysis, PP .0.95 are indicated
with thick branch. GenBank accession numbers for the four
genes of each species are provided (TABLE I).
LIETAL.:
V
ELOPORPHYRELLUS
PHYLOGENETICS 295
differ in the morphology of pileus, size of the
basidiospores and mycorrhizal associations (see our
brief description of
V. pseudovelatus
below).
Veloporphyrellus conicus (Ravenel) B. Ortiz, Yan C.
Li & Zhu L. Yang, comb. nov. FIGS. 3D–F, 4B, 6
MycoBank MB801792
FIG. 3. Habitat of
Veloporphyrellus
species. A–C.
V. alpinus
(A–B. holotype,
B. Feng 761-HKAS 57490
, KUN. C.
B
.
Feng
1266-HKAS 63669
, KUN). D–F.
V. conicus
(D.
B. Ortiz-Santana 321-BZ 1670
, CFMR. E. B.
Ortiz-Santana 356-BZ 1705
, CFMR.
F.
B. Ortiz-Santana 480-BZ 2408
, CFMR). G–I.
V. pseudovelatus
(G.
L.P. Tang 1212-HKAS 63032
, KUN. H. holotype,
Z.L.
Yang 4941-HKAS 52258
, KUN. I.
Y.C. Li 2815-HKAS 63670
, KUN). J–L.
V. velatus
(J–L.
N.K. Zeng 763-HKAS 63668
, KUN).
296 MYCOLOGIA
Boletus conicus
Ravenel, Ann. Mag. Nat. Hist. 12:430. 1853
(Basionym).
Tylopilus conicus
(Ravenel) Beardslee, Mycologia 26:253.
1934.
Mucilopilus conicus
(Ravenel) Wolfe, Mycotaxon 10:119.
1979.
Fistulinella conica
(Ravenel) Pegler & T.W.K. Young,
Trans Br Mycol Soc 76:140. 1981.
This species is characterized by the hemispheri-
cal, convex or broadly conical pileus, decurved and
appendiculate pileus margin, appressed or pulvi-
nate yellow to brownish yellow or greenish brown
squamules on the pileus and the adnate to
depressed hymenophore (Beardslee 1934, Wolfe
1979, Pegler and Young 1981, Ortiz-Santana et al.
2007). For the comparison of species in
Velopor-
phyrellus
, the micromorphological characters of
this species were described and illustrated bellow
based on three collections from Belize (Central
America).
Basidia 28–33 311–12.5 mm, broadly clavate to
clavate, hyaline to light yellowish in KOH, thin-walled,
four-spored, occasionally two-spored. Basidiospores
[60/3/3] (14.5–)15–17 34.5–5(–5.5) mm, (Q 5
[2.82–]2.9–3.44[–3.56], Q
m
53.22 60.16), boletoid,
slightly thick-walled (up to 0.5 mm thick), subhyaline to
light olivaceous in KOH and yellowish brown in
Melzer’s reagent, smooth under SEM (FIG.4B).
Pleurocystidia 25–51 37–11 mm, fusiform to subfusi-
form or subfusoid-mucronate to ventricose-mucronate
with a short pedicel, sometimes narrowly mucronate,
rostrate, thin-walled, hyaline in KOH and yellowish
to yellow in Melzer’s reagent. Cheilocystidia 27–34 3
7–9 mm, broadly clavate to subfusiform or ventricose,
thin-walled, some with 1–2 septa or secondary septa,
hyaline in KOH and yellowish to yellow in Melzer’s
reagent. Hymenophoral trama bilateral, composed
of broad hyaline hyphae up to 8 mm wide. Pileus
covering composed of more or less vertically to
almost radially arranged to loosely interwoven,
colorless to yellowish, filamentous hyphae 4–7 mm
diam and often with yellow to yellowish brown
vacuolar pigment but soon dissolved in KOH
solution; terminal cells 15–65 33–8 mm, subcylind-
rical. Pilea trama made up of hyphae 7–10.5 mm
diam, colorless to yellowish in KOH. Clamp connec-
tions absent in all tissues.
Habitat:
Gregarious on soil under
Pinus caribaea
and
Quercus
spp.
Distribution:
Currently known from North America
(from North Carolina south to Florida, Mexico) and
Central America (Belize).
Specimens examined:
BELIZE. CAYO DISTRICT: Moun-
tain Pine Ridge Forest Reserve, Douglas da Silva, swamp
near British Military Camp, 450 m, 4 Oct 2002,
B. Ortiz-
Santana 321-BZ 1670
(CFMR). Douglas da Silva, Forestry
Station campground, 456 m, 13 Oct 2002,
B. Ortiz-Santana
356-BZ 1705
(CFMR). Five Sister Lodge, near lodge, 387 m,
30 Nov 2002,
B. Ortiz-Santana 480-BZ 2408
(CFMR).
Notes: Veloporphyrellus conicus
originally was de-
scribed as
Boletus conicus
Ravenel (in Berkeley and
Curtis 1853) based on a collection from South
FIG. 4. Basidiopores of
Veloporphyrellus
under SEM. A.
V. alpinus
(holotype,
B. Feng 761-HKAS 57490
, KUN). B.
V. conicus
(
B. Ortiz-Santana 321-BZ 1670
, CFMR). C.
V. pantoleucus
(type,
Go´ mez 21232
, F). D.
V. pseudovelatus
(holotype.
Z.L. Yang
4941-HKAS 52258
, KUN). E.
V. velatus
(
N.K. Zeng 763-HKAS 63668
, KUN).
LIETAL.:
V
ELOPORPHYRELLUS
PHYLOGENETICS 297
Carolina, eastern North America. This species also has
been reported from southern North America and
Central America (Beardslee 1934, Ortiz-Santana et al.
2007). It would be better to study type or authentic
materials of this species from the type locality to
determine whether materials from these areas repre-
sent a single species, although North America-Central
America distribution pattern of boletes do occur
(Halling and Mueller 2005, Dengtinger et al. 2010,
Feng et al. 2012). Unfortunately DNA from the type
specimen of
B. conicus
may be unattainable based on
the age of the collection. Further field investigations,
careful morphological observations and molecular
analyses using multiple genes based on additional and
authentic materials might help us better understand
the delimitation of this species. However, our three
collections from Belize (CFMR: BZ 1670, CFMR: BZ
1705, CFMR: BZ 2408) generally have morphological
characters similar to
B. conicus
according to recent
descriptions (Wolfe 1979, Pegler and Young 1981,
Ortiz-Santana et al. 2007). Consequently the new
combination is proposed.
Veloporphyrellus conicus
has been placed in several
genera. Beardslee (1934) transferred it to the genus
Tylopilus
based on the pink spores and flesh-colored
hymenophore. Wolfe (1979) regarded it as a member
of
Mucilopilus
due to the ‘‘ixotrichodermial’’ pileus
covering. Pegler and Young (1981) transferred
species in the genus
Mucilopilus
to
Fistulinella
and
transferred
Boletus conicus
to
Fistulinella
. From the
protolog of this species (Ravenel in Berkeley and
Curtis 1853), the pileus of
V. conicus
is described as
FIG.5.
Veloporphyrellus alpinus
(holotype,
B. Feng 761-HKAS 57490
, KUN). a. Basidiospores. b. Basidia and
pleurocystidium. c. Pleurocytidia. d. Cheilocystidia. e. Pileus covering. Bars: a–e 520 mm.
298 MYCOLOGIA
‘‘pulvinate’’. Thus, it is preferable to regard the pileus
covering as a trichoderm rather than an ixotricho-
derm. In our molecular phylogenetic study, it turned
out that collections from Belize clustered together
with species in
Veloporphyrellus
,ratherthanin
Tylopilus
and
Fistulinella
, with high support values
on both datasets using ML, MP and BI approach
(FIGS. 1, 2).
Veloporphyrellus pantoleucus L.D. Go´ mez & Singer,
Brenesia 22:293, FIG. 1, 1984. FIGS. 4C, 7
Veloporphyrellus pantoleucus
is characterized by the
white, tomentose pileus, the white to whitish squa-
mules, the white membranous veil, the pale pinkish to
light pinkish or grayish pink hymenophore, the
ocher-purple spore print, the white pubescent to
subglabrous stipe, the white basal mycelia on the base
of stipe, the trichodermial pileus covering and the
clavate to cylindrical cheilocystidia with 1–2 septa or
secondary septa.
Basidia 28–42 37.5–10 mm, clavate, hyaline in
KOH and yellowish in Melzer’s reagent, four-spored.
FIG.6.
Veloporphyrellus conicus
(
B. Ortiz-Santana 321-BZ 1670
, CFMR). a. Basidiospores. b. Basidia and pleurocystidium.
c. Pleurocytidia. d. Cheilocystidia. e. Pileus covering. Bars: a–e 520 mm.
LIETAL.:
V
ELOPORPHYRELLUS
PHYLOGENETICS 299
Basidiospores (40/1/1) (11.5–)12–14(–14.5) 34–
5mm, (Q 5[2.4–]2.56–3.25(–3. 5), Q 52.91 6
0.19), boletoid, smooth, slightly thick walled (up to
0.5 mm thick), subhyaline to light olivaceous in KOH
and yellowish brown in Melzer’s reagent, smooth
under SEM (FIG. 4, C). Pleurocystidia 51–57 36–
9mm, subfusoid-mucronate or ventricose-mucronate
with a long pedicel, thin-walled, hyaline in KOH and
yellowish to yellow in Melzer’s reagent. Cheilocystidia
70–92 37–10 mm, abundant, most clavate to cylindri-
cal, or attenuate upwards, some with 1–2 septa or
secondary septa, thin-walled, hyaline in KOH and
yellowish to yellow in Melzer’s reagent. Pileus covering
composed of trichoderm, non-viscid, colorless to
yellowish hyphae 4.5–8 mm diam; terminal cells
35.5–56 34–6.5 mm, subcylindrical. Pilea trama hyphae
6–11 mm diam, hyaline to yellowish brown in KOH.
Clamp connections absent.
Habitat:
Solitary on the ground in mixed forest
of
Quercus
-
Magnolia
forest at about 1800–2000 m
altitude.
Distribution
: Currently known only from Costa Rica.
Specimen examined:
COSTA RICA. CARTAGA: San
Cristo´ bal Jul 1983,
Go´ mez 21232
(Type: F).
Notes:
The type of
V. pantoleucus
consists of one
immature (
Go´mez 21232-1
)andahalfmature(
Go´mez
21232-2
) basidiocarp in good condition. In our
phylogenetic study,
V. pantoleucus
clustered together
with
V. velatus
with high support values on both
datasets, but they differ in the color of the pileus and
FIG.7.
Veloporphyrellus pantoleucus
(type,
Go´ mez 21232
, F). a. Basidiospores. b. Basidia and pleurocystidia. c. Pleurocytidia.
d. Cheilocystidia. e. Pileus covering. Bars: a–e 520 mm.
300 MYCOLOGIA
stipe,thesizeofthebasidiospores,themycorrhizalhost
association and geographical distribution (Go´mez and
Singer 1984; see our description of
V. velatus
below).
Veloporphyrellus pseudovelatus Yan C. Li & Zhu L.
Yang, sp. nov. (FIGS. 3G–I, 4D, 8)
MycoBank MB801793
Etymology:
Named because of its similarity to
V. velatus
.
Diagnosis:
This species differs from the other
species in
Veloporphyrellus
by the hemispherical to
subconical coca brown to chestnut brown pileus, the
chestnut brown surface of the stipe, the mycorrhizal
association with species of
Keteleeria
and
Pinus
.
Pileus 2–5 cm diam, convex to hemispherical, dry,
not viscid, densely covered tomentose, cocoa brown
to chestnut brown squamules on the white back-
ground; margin projecting, forming membranous veil
which is concolorous with pileus, the veil attached to
apex of stipe in younger basidioma but broken into
pieces and hanging on pilea margin. Hymenophore
FIG.8.
Veloporphyrellus pseudovelatus
(holotype,
Z.L. Yang 4941-HKAS 52258
, KUN). a. Basidiospores. b. Basidia and
pleurocystidium. c. Pleurocytidia. d. Cheilocystidia. e. Pileus covering. Bars: a–e 520 mm.
LIETAL.:
V
ELOPORPHYRELLUS
PHYLOGENETICS 301
free to subfree or sinuate around apex of the stipe,
white initially, pinkish to flesh-colored when mature;
tubes and pores concolorous; tubes 3–6 mm deep,
with rust brown stains here and there when old or
touched; pores small, about 0.3–1 mm diam. Stipe 3–
730.5–0.8 cm, subcylindrical or slightly attenuate
upward, base sometimes enlarged, surface of stipe
pale chestnut brown, smooth and macroscopically not
scabrous or reticulate. Mycelia on base of stipe white
to pale white. Context of pileus and stipe cream to
white, unchanging when injured. Flavor and odor not
distinctive.
Basidia 23–30 38–11 mm, clavate, hyaline in KOH
and yellowish in Melzer’s reagent, four-spored, some-
times two-spored. Basidiospores (200/10/8) (12)12.5–
15(16) 34–5(5.5) mm, (Q 5[2.45]2.6–3.38[3.63],
Q52.94 60.18), boletoid, smooth, light olivaceous
to pale melleous in KOH and yellowish brown in
Melzer’s reagent, smooth under SEM (FIG.4D).
Pleurocystidia 50–69 36–9 mm, subfusoid-mucronate,
or ventricose-mucronate with a long pedicel, thin-
walled, hyaline in KOH and yellowish to yellow in
Melzer’s reagent. Cheilocystidia 41–68 36–10 mm,
abundant, most clavate to cylindrical, or attenuate
upward, some with 1–2 septa or secondary septa, thin-
walled, hyaline in KOH and yellowish to yellow in
Melzer’s reagent. Pileus covering composed of more or
less vertically to almost radially arranged to loosely
interwoven, colorless to yellowish, filamentous hyphae
3–6 mm diam and often with yellow to yellowish brown
vacuolar pigment but dissolved quickly in KOH
solution; terminal cells 26–61 34–6 mm, subcylindrical.
Pilea trama composed of hyphae 7–15 mm diam,
colorless to yellowish in KOH. Clamp connections
absent in all tissues.
Habitat:
Solitary to scattered, in the forest of
Keteleeria
spp. or in mixed forest of
Keteleeria
spp.
and
Pinus yunnanensis
.
Distribution:
KnownfromsouthwesternChina
(Yunnan province).
FIG.9.
Veloporphyrellus velatus
(
N.K. Zeng 763-HKAS 63668
, KUN). a. Basidiospores. b. Basidia and pleurocystidia.
c. Pleurocytidia. d. Cheilocystidia. e. Pileus covering. Bars: a–e 520 mm.
302 MYCOLOGIA
Holotype:
CHINA. YUNNAN PROVINCE: Kunming,
Jindian, 2000 m, 1 Aug 2007,
Z.L. Yang 4941-HKAS
52258
(KUN).
Additional specimens examined:
CHINA. YUNNAN PROV-
INCE: Chuxiong, Nanhua wild mushroom market, 25 Aug
2007,
Z.L. Yang 4927-HKAS 52244
(KUN). Kunming,
Xishan, 2050 m, 10 Aug 2007,
Y.C. Li 986-HKAS 52673
(KUN). Dehong, Yingjiang City, on route from Tengchong
to Longlin, 2010 m, 19 Jul 2009,
Y.C. Li 1697-HKAS 59444
(KUN). Chuxiong, Nanhua wild mushroom market, 2 Aug
2009,
Y.C. Li 1947-HKAS 59695
(KUN). Baoshan, Dax-
ishan, 1900 m, 9 Aug 2010,
L.P. Tang 1212-HKAS 63032
(KUN); same location, 10 Aug 2010,
L.P. Tang 1219-HKAS
63039
(KUN). Kunming, Yeyahu, 1950 m, 18 Aug 2012,
Y.C. Li 2815-HKAS 63670
(KUN).
Notes: Veloporphyrellus pseudovelatus
is character-
ized by the cocoa brown to chestnut brown tomentose
pileus, the concolorous membranous veil remnants
on the pilea margin, the pinkish to flesh colored
hymenophore, and the chestnut brown surface of
the stipe. This species is phenotypically similar to
V. alpinus
, and phylogenetically, they were clustered
together. However,
V. alpinus
has a yellowish orange
to grayish orange or brown to reddish brown stipe
surface, relatively larger basidiospores, a subalpine to
alpine distribution, 3100–3600 m, and a mycorrhizal
association with
Abies
species.
Veloporphyrellus velatus (Rostr.) Yan C. Li & Zhu L.
Yang, comb. nov. FIGS. 3J–L, 4E 9
MycoBank MB801794
;
Suillus velatus
Rostr. Bot. Tidsskr. 24:207. 1902
(Basionym).
;
Boletus velatus
(Rostr.) Sacc. & D. Sacc., Syll. fung.
(Abellini) 17:97. 1905.
;
Tylopilus velatus
(Rostr.) F.L. Tai, Syll. fung. Sini-
corum. 1165. 1979.
Basidiomata 4 cm wide, small to medium-sized.
Pileus subhemispherical, densely covered with brown
(7D7–8) to cocoa brown (8D7–8) or chestnut brown
to dark reddish brown (8C7–8) squamules on the
white background, dry, not viscid when wet; margin
extended, with white (1A1) membranous veil rem-
nants. Hymenophore depressed around apex of the
stipe, pallid (1B1) to pale pinkish (11B3–5) or
pinkish (13A2) to pink (12A3–4); tubes and pores
concolorous, tubes up to 0.5 cm long and pores up to
0.5 mm wide. Stipe 7.2 30.6–0.8 cm, clavate,
glabrous, white (1A1) to bright white; basal mycelia
white (1A1). Context of pileus and stipe solid, white
(1A1) to bright white, unchanging when bruised.
Basidia 25–32.5 310–12.5 mm, broadly clavate to
clavate, hyaline to light yellowish in KOH, thin-walled,
four-spored, occasionally two-spored. Basidiospores
[60/1/1] 11–12.5 (13) 3(4.0)4.5–5 mm, (Q 5
[2.40]2.44–2.67[2.75], Q
m
52.56 60.12), ellipsoid,
oblong or fusiform to subfusiform, slightly thick-
walled (up to 0.5 mm thick), subhyaline to light
olivaceous in KOH and yellowish brown in Melzer’s
reagent, smooth under SEM (FIG. 4E). Pleurocystidia
61–75 38.5–12 mm, fusiform to subfusiform or
subfusoid-mucronate to ventricose-mucronate with a
short pedicel, sometimes narrowly mucronate, ros-
trate, thin-walled, hyaline in KOH and yellowish to
yellow in Melzer’s reagent. Cheilocystidia 25–38 37–
11 mm, broadly clavate to subfusiform or ventricose,
thin-walled, some with 1–2 septa or secondary septa,
hyaline in KOH and yellowish to yellow in Melzer’s
reagent. Hymenophoral trama bilateral, composed
of broad hyaline hyphae up to 9.5 mm wide. Pileus
covering composed of more or less vertically to
almost radially arranged to loosely interwoven,
colorless to yellowish, filamentous hyphae 3–7 mm
diam and often with yellow to yellowish brown
vacuolar pigment but soon dissolved in KOH
solution; terminal cells 15–57.5 33–5.5 mm, sub-
cylindrical. Pilea trama composed of hyphae 5–11 mm
diam, colorless to yellowish in KOH. Clamp connec-
tions absent in all tissues.
Habitat:
Solitary on the ground in mixed forest of
Lithocarpus
spp and
Pinus fenzeliana
.
Distribution:
Currently known from Thailand and
southern China (Hainan province).
Specimen examined:
CHINA. HAINAN PROVINCE: Wuz-
hishan county, Wuzhishan National Nature Reserve, 1200 m,
31 Jul 2010,
N.K. Zeng 763-HKAS 63668
(KUN).
Notes: Veloporphyrellus velatus
originally was de-
scribed from Thailand as a member of the genus
Suillus
(Rostrup 1902), then it was transferred to the
genus
Boletus
(Saccardo 1905). However, considering
that the colors of the hymenophore and the spore
print were different from those in
Boletus
, Tai (1979)
transferred it to the genus
Tylopilus
, yet the combina-
tion is invalid because of the shortage of literature
citation of the basionym (McNeill et al. 2012). To
understand the species concept of
V. velatus
, it would
be ideal to study the type or authentic materials, even
only morphologically. Unfortunately, the type of
Suillus velatus
studied by Rostrup could not be traced
(Corner 1972). In addition this species has not been
collected for the second time since its original
description from Siam. However, our collections
(HKAS 63668) made from Hainan, China, share
several characters with Rostrup’s species, such as the
dense tomentose, brown to cocoa brown or chestnut
brown to dark reddish brown squamules on the pileus,
the veil remnants on the pilea margin, the depressed
hymenophore at apex of the stipe, the glabrous stipe
and the tropical distribution. Thus, a new combination
is proposed.
Veloporphyrellus velatus
is characterized by
the dense tomentose, brown to cocoa brown or
LIETAL.:
V
ELOPORPHYRELLUS
PHYLOGENETICS 303
chestnut brown to dark reddish brown squamules on
the pileus, the white membranous veil remnants, the
pinkish to flesh-colored hymenophore and the white
surface of the stipe. Phylogenetically
V. velatus
was
clustered with
V. pantoleucus
with high statistical
support based on both single-locus and multilocus
sequence datasets using ML, MP and Bayesian ap-
proach (FIGS. 1, 2). However,
V. pantoleucus
has a
white, tomentose pileus, a concolorous subtomentose
veil and relatively bigger spores (Go´mez and Singer
1984; see
V. pantoleucus
above).
For the convenience of identification, a key to the
species is given below. Noticeably,
V. africanus
is the
only species in this genus with blue reaction when
bruised and olivaceous hymenophore. Such traits are
shared by some species in
Boletellus
,
Leccinum
and
Boletus
L., and thus it is doubtful whether it is a true
Veloporphyrellus
species. Unfortunately, the type spec-
imen of this species was unavailable for the present
phylogenetic study. Moreover, no additional materials
of this species have been reported since its original
description. The taxonomic position of this species
should await further molecular and morphological
data.
KEY TO THE SPECIES IN
V
ELOPORPHYRELLUS
1. Context without bluish color reaction when
bruised; known from southeastern Asia or North
and Central America . . . ................... 2
1. Context with bluish reaction when bruised; known
from Africa . ....................
V. africanus
2. Pileus with brown, chestnut brown, cocoa
brown or reddish brown squamules; known
from Asia .......................... 3
2. Pileus with white to whitish, or yellow to
brownish yellow or greenish brown squamules;
known from North or Central America . . . . 5
3. Stipe surface orange yellow to orange brown or
reddish brown; distributed at a high altitude
(2000–3600 m) .......................... 4
3. Stipe surface white; distributed at a low elevation
(#1500 m) .....................
V. velatus
4. Basidiospores 15.5–19.5 34.5–6.5 mm; distrib-
uted at higher elevation (3100–3600 m);
associated with species of
Abies
....
V. alpinus
4. Basidiospores 12–16 34–5.5 mm; distributed
at a relatively lower elevation (approx.
2000 m); associated with species of
Keteleeria
or
Pinus
................
V. pseudovelatus
5. Pileus surface covered with yellow to brownish
yellow or greenish brown squamules, margin
decurved and appendiculate; distributed in North
America and Central America ........
V. conicus
5. Pileus surface covered white to whitish squamules,
margin with white membranous veil; distributed in
Central America ................
V. pantoleucus
DISCUSSION
Four species were newly delimitated in
Veloporphyrellus
,
including two new species (i.e.
V. alpinus
and
V.
pseudovelatus
) and two new combinations (i.e.
V.
conicus
and
V. velatus
). The type species of
Velopor-
phyrellus
,
V. pantoleucus
, is poorly known, and its
molecular data of the type specimen has not been
investigated. Accordingly, the taxonomic boundaries
of
Veloporphyrellus
were unknown before this study.
Some species in
Veloporphyrellus
have caused a number
of important systematic and evolutionary questions in
the past. Specifically,
V. conicus
was placed in several
genera of the family Boletaceae, such as
Boletus
L.,
Tylopilus
,
Mucilopilus
,
Fistulinella
(Ravenel 1853,
Beardslee 1934, Wolfe 1979, Pegler and Young 1981)
based on single or inadequate morphological charac-
ters. Likewise,
V. velatus
originally was placed in the
genus
Suillus
Gray in Suillaceae (Rostrup 1902).
Subsequently, it was transferred to different genera
in Boletaceae. such as
Boletus
and
Tylopilus
(Sac-
cardo 1905, Tai 1979). In this study the combined
morphological and molecular data help us better
resolve the systematic position of these species and
their evolutionary relationships.
Veloporphyrellus
species generally share the charac-
ters of the membranous veil remnants hanging on the
pilea margin with some species in
Austroboletus
,
Boletellus
,
Leccinum
,
Strobilomyces
and
Suillus
. Howev-
er, species in
Boletellus
,
Leccinum
and
Suillus
all lack
the characters of the pale pinkish to light pinkish or
grayish pink hymenophore and pink to ocher-purple
spore print that distinguishes
Veloporphyrellus
species.
Species in
Austroboletus
and
Strobilomyces
have orna-
mented basidiospores, while those in
Veloporphyrellus
have smooth basidiospores (FIG. 4).
Veloporphyrellus
species also shared the same hymenophore and
spore-print color with species in
Australopilus
,
Aus-
troboletus
,
Fistulinella
,
Harrya
,
Tylopilus
,
Zangia
and
some species in
Porphyrellus
. However, species in
Austroboletus
have ornamented basidiospores, and
the species in the other six genera do not present
membranous veil remnants on the pilea margin. Thus,
Veloporphyrellus
can be distinguished from other
boletoid genera by the feature combination of the
pale pinkish to light pinkish or grayish pink hymeno-
phore and spore print, the extending membranous
veil remnants on the pilea margin, the trichoderm-like
pileus covering and the smooth basidiospores.
Molecular phylogenetic relationships at the generic
level within the Boletaceae have been investigated in
several studies (e.g. Binder and Besl 2000; Binder and
Bresinsky 2002; Binder and Hibbett 2007; Halling et al.
2007, 2008, 2012; Desjardin et al. 2008, 2009; Li et al.
2011; Lebel et al. 2012; Zeng et al. 2012, 2013). However,
304 MYCOLOGIA
sequences of the genus
Veloporphyrellus
were not
included. In our phylogenetic analyses based on single-
locus and multilocus datasets,
Veloporphyrellus
was nested
into the Boletaceae clade and clustered as a monophy-
letic group in all analyses using both datasets, but its
relationships to other genera in the family remains
unresolved (FIGS.1,2).Within
Veloporphyrellus
,cladesI,
II and III all had significant statistical support for
monophyly and morphological features indicated clear
distinctions. However, in our single-locus dataset analysis,
clades II and III clustered together with moderate ML
bootstrap support (59%,F
IG. 1); while in our multilocus
dataset analysis, clades I and III formed a monophyletic
group with moderate ML, MP bootstrap support values
(75%,70%respectively) and PP values (FIG.2).Thus,
the infrageneric relationships of species among the three
clades were not well inferred in this study. Future studies
with more additional materials and more molecular
sequences would help to resolve their relationships.
Veloporphyrellus
is a genus with species from North
and Central America, southeastern Asia and Africa.
Most species are distributed in subtropical to tropical
regions, except
V. alpinus
, which is restrict to the
subalpine to alpine areas in Yunnan and Taiwan.
Species in this genus can form putatively ectomycor-
rhizal associations mainly with plants of Caesalpiniaceae,
Dipterocaceae, Fagaceae and Pinaceae.
Veloporphyrellus
africanus
is undoubtedly connected with either
Brachys-
tegia
spp. (Caesalpiniaceae) or
Marquesia
spp. (Dipter-
ocarpceae) in Zambia (Watling and Turnbull 1993).
Veloporphyrellus alpinus
is found in the forests of
Abies
(Pinaceae) in Yunnan and Taiwan.
Veloporphyrellus
conicus
originally was found in the forest of
Pinus
(Pinaceae) in Carolina (Berkeley and Curtis 1853). For
the generic type
V. pantoleucus
, it is found in mixed
forest of
Quercus
spp. (Fagaceae) and
Magnolia
spp.
(Magnoliaceae) in Costa Rica (Go´mez and Singer
1984).
Veloporphyrellus pseudovelatus
is clearly associated
with
Keteleeria
(Pinaceae) and
Pinus
at about 2000 m in
southwestern China. For
V. velatus
, no mycorrhizal
hosts were indicated in its original description (Rostrup
1902.). However, our field investigation indicates that it
may be associated with trees of
Lithocarpus
(Fagaceae)
and
Pinus
in tropical China.
ACKNOWLEDGMENTS
The authors thank Dr Robert Lu¨cking (F) for providing type
specimen of
V. pantoleucus
on loan and granting permission
to extract DNA from the type specimens. The authors are very
grateful for the help of Dr M. Binder and two anonymous
reviewers for the critical review and helpful comments on the
manuscript. We also thank Dr L.P. Tang and Mr X.T. Zhu
(KUN) for providing collections. This study was supported by
the National Natural Science Foundation of China
(Nos. 31000012, 31210103919, 31370001), the Independent
Research Program of the Chinese Academy of Sciences
(KSCX2-EW-J-24), the Natural Science Foundation of
Yunnan Province (2013FB066) and the CAS/SAFEA Inter-
national Partnership Program for Creative Research Teams.
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306 MYCOLOGIA
