Hypocrea / Trichoderma species with pachybasium-like conidiophores: teleomorphs for T. minutisporum and T. polysporum and their newly discovered relatives

Abstract

We describe or redescribe species of Hypocrea/Trichoderma (Ascomycetes, Hypocreales) having hyaline ascospores and pachybasium-like conidiophores. Teleomorphs are reported for Trichoderma minutisporum (Hypocrea minutispora sp. nov.) and T. polysporum (H. pachybasioides). Hypocrea pilulifera/T. piluliferum is redescribed. Trichoderma croceum is synonymized with T. polysporum. The new species H. parapilulifera, H. stellata and H. lacuwombatensis are described. All of these species fall within the morphological concept of Trichoderma sect. Pachybasium and within the phylogenetic group pachybasium B5 of Kullnig-Gradinger et al (2002) Kullnig-Gradinger C, Szakacs G, Kubicek CP. 2002. Phylogeny and evolution of the genus Trichoderma: a multi-gene approach. Mycol Res 106:757–767. . Parsimony analysis of nucleotide sequences from three unlinked loci—ITS1 and 2, endochitinase (ech42) and translation elongation factor 1-alpha (tef1)—detects two distinct phylogenetic lineages within the group pachybasium B5. One comprises H. pachybasioides/T. polysporum, H. pilulifera/T. piluliferum, H. parapilulifera and H. stellata; this group, the “polysporum” lineage, is characterized by having conidia that are white in mass and is the only lineage within Hypocrea characterized by such conidia. The second group includes the green conidial T. minutisporum and H. lacuwombatensis. The partition homogeneity test reveals significant recombination within the “polysporum” lineage but not within the “minutisporum” lineage.

Full text

310
Mycologia,
96(2), 2004, pp. 310–342.
q2004 by The Mycological Society of America, Lawrence, KS 66044-8897
Hypocrea
/
Trichoderma
species with pachybasium-like conidiophores:
teleomorphs for
T. minutisporum
and
T. polysporum
and their newly
discovered relatives
Bingsheng Lu
1
Department of Plant Pathology, Agronomy College,
Shanxi Agricultural University, Taigu, Shanxi
030801, China
Irina S. Druzhinina
Section of Applied Biochemistry and Gene Technology,
Institute of Chemical Engineering, TU Wien,
Getreidemarkt 9-166.5, A-1060 Wien, Austria
Payam Fallah
2
Priscila Chaverri
3
Pennsylvania State University, Department of Plant
Pathology, 301 Buckhout Laboratory, University Park,
Pennsylvania 16802
Cornelia Gradinger
4
Christian P. Kubicek
Section of Applied Biochemistry and Gene Technology,
Institute of Chemical Engineering, TU Wien,
Getreidemarkt 9-166.5, A-1060 Wien, Austria
Gary J. Samuels
5
USDA-ARS, Systematic Botany and Mycology
Laboratory, Room 304, B-011A, BARC-West,
Beltsville, Maryland 20705-2350
Abstract:
We describe or redescribe species of
Hy-
pocrea
/
Trichoderma
(Ascomycetes, Hypocreales) hav-
ing hyaline ascospores and pachybasium-like conidi-
ophores. Teleomorphs are reported for
Trichoderma
minutisporum
(
Hypocrea minutispora
sp. nov.) and
T.
polysporum
(
H. pachybasioides
).
Hypocrea pilulifera
/
T.
piluliferum
is redescribed.
Trichoderma croceum
is syn-
onymized with
T. polysporum.
The new species
H. par-
apilulifera, H. stellata
and
H. lacuwombatensis
are de-
scribed. All of these species fall within the morpho-
logical concept of
Trichoderma
sect.
Pachybasium
and
within the phylogenetic group pachybasium B5 of
Accepted for publication June 16, 2003.
1
Current address: Agronomy College, Dept. of Plant Protection,
Zhongkai Agrotechnical College, Guangzhou 510225, China.
2
Current address: Environmental Microbiology Laboratory, Inc.,
10636 Scripps Summit Court Suite 103, San Diego, California
92131.
3
Current address: USDA-ARS, Systematic Botany and Mycology
Laboratory, Room 304, B-011A, BARC-West, Beltsville, Maryland
20705-2350.
4
Current address: Section of Industrial Microbiology, Institute of
Chemical Engineering, TU Wien, Getreidemarkt 9-172.5, A-1060
Wien, Austria.
5
Corresponding author. E-mail: gary@nt.ars-grin.gov
Kullnig-Gradinger et al (2002). Parsimony analysis of
nucleotide sequences from three unlinked loci—
ITS1 and 2, endochitinase (
ech42
) and translation
elongation factor 1-alpha (
tef1
)—detects two distinct
phylogenetic lineages within the group pachybasium
B5. One comprises
H. pachybasioides
/
T. polysporum,
H. pilulifera
/
T. piluliferum, H. parapilulifera
and
H.
stellata
; this group, the ‘‘polysporum’’ lineage, is
characterized by having conidia that are white in
mass and is the only lineage within
Hypocrea
charac-
terized by such conidia. The second group includes
the green conidial
T. minutisporum
and
H. lacuwom-
batensis.
The partition homogeneity test reveals sig-
nificant recombination within the ‘‘polysporum’’ lin-
eage but not within the ‘‘minutisporum’’ lineage.
Key words:
anamorph-teleomorph connection,
Ascomycetes, endochitinase gene, Hypocreaceae, Hy-
pocreales, ITS, molecular phylogenetics, systematics,
translation elongation factor
tef1
INTRODUCTION
Trichoderma
Pers. : Fr. Sect.
Pachybasium
(Sacc.) Bis-
sett, as characterized by Bissett (1991b) included
most of the described species of the genus. These
fungi were characterized by having relatively short
and broad phialides arising from, and often clustered
on, wide branches of the conidiophores. The section
included species that produce sterile or fertile elon-
gations of their conidiophores along with a few spe-
cies that lack any elongations of the conidiophores.
It also included species that have green conidia as
well as species in which conidia are white or yellow
in mass. Despite morphological similarity among the
species originally included in the section (Bissett
1991b), DNA sequence analysis has shown it to be
paraphyletic (Kindermann et al 1998, Kullnig-Gra-
dinger et al 2002) and separable into two major phy-
logenetic groups that have been termed clade A and
clade B (Kullnig-Gradinger et al 2002). Clade A,
which includes the neotype of the type species of
sect.
Pachybasium, T. hamatum
(Bon.) Bain., is de-
rived from within sect.
Trichoderma.
Most of the pach-
ybasium-like species are placed in clade B and form
a highly diverse but monophyletic sister group of
sect.
Trichoderma.
This large group of species has not
been given formal taxonomic status within
Hypocrea
/

311L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
Trichoderma
and is referred to here as pachybasium
B. The genetic diversity of
Trichoderma
sect.
Pachy-
basium
not withstanding, the morphological descrip-
tor ‘‘pachybasium-like’’ is informative and used
throughout the present work to describe a character-
istic type of conidiophore.
Several taxa belonging to pachybasium B are char-
acterized by green conidia and conidiophores from
which sterile or fertile elongations arise (Bissett
1991b). Chaverri et al (2003) recently reviewed the
taxonomy of these species. They described
Hypocrea
strictipilosa
Chaverri & Samuels as the teleomorph of
T. strictipile
Bissett and described or redescribed but
did not name pachybasium-like anamorphs for the
new and closely related species
H. cremea
Chaverri &
Samuels
, H. cuneispora
Chaverri & Samuels,
H. eston-
ica
Chaverri & Samuels and
H. surrotunda
Chaverri
& Samuels. All of those species were included in one
clade in pachybasium B.
Apart from the species mentioned above, other
taxa from pachybasium B have green conidia but lack
elongations of conidiophores. In addition, some spe-
cies have hyaline (white in mass) or yellow conidia;
depending on the species, their conidiophores may
have sterile elongations. Four of these species (
T. po-
lysporum
Rifai,
T. croceum
Bissett,
T. minutisporum
Bis-
sett and
H. pilulifera
Rifai & J. Webster) previously
have been shown to form one strongly supported
clade (subclade B5) in pachybasium B in a multigene
phylogenetic analysis. In this paper, we have investi-
gated this clade in detail. We synonymize
T. croceum
under
T. polysporum
Rifai and link that species to
Hy-
pocrea pachybasioides
Doi. We link
T. minutisporum
Bissett to the new
Hypocrea
species
H. minutispora.
We redescribe the teleomorph and anamorph of
H.
pilulifera
and describe the new species
Hypocrea par-
apilulifera, H. stellata
and
H. lacuwombatensis
and
their
Trichoderma
anamorphs.
MATERIALS AND METHODS
Isolates.
—The isolates used in this study are listed in T
ABLE
I. The authors isolated many of the cultures from
Hypocrea
collections; these are indicated as ‘‘G.J.S.’’, and represen-
tatives are deposited in Centraalbureau voor Schimmelcul-
tures, Utrecht, The Netherlands (CBS), Agriculture and
Agri-Food Canada, Eastern Cereals and Oilseeds Research
Centre, Ottawa, Canada (DAOM), the American Type Cul-
ture Collection, Manassas, Virginia (ATCC) and Institute of
Microbiology, Chinese Academy of Science, Beijing, China
(CGTCC). Other cultures were obtained from ATCC, CBS,
DAOM and the Tottori Mycological Institute, Tottori City,
Japan (TMI).
Single-ascospore isolations from fresh collections of
Hy-
pocrea
were made on CMD (Difco cornmeal agar 12%
dextrose 11% antibiotic solution [0.2% Sigma streptomy-
cin sulfate 10.2% Sigma neomycin sulfate 1distilled wa-
ter]) with the aid of a micromanipulator. These cultures are
maintained in CMA (Difco cornmeal agar) slant tubes at 9
C and in liquid nitrogen. The most frequently cited collec-
tor, G.J. Samuels, is abbreviated as G.J.S.
Growth and colony characterization.
—Growth trials were per-
formed to determine the growth rate and optimum tem-
perature for growth following the protocol of Samuels et al
(2002) on PDA (Difco potato-dextrose agar) and synthetic
low-nutrient agar (SNA, Nirenberg 1976). The isolates were
grown in the dark and the colony radius was measured at
24, 48, 72 and 96 h at 15, 20, 25, 30 and 35 C. Each growth-
rate experiment was repeated three times and the results
averaged for each isolate. The time of first appearance of
conidia, the presence of yellow pigmentation of young co-
nidia, the presence of diffusing pigment in the agar, odor
and colony appearance also were noted.
Morphological observations.
—Morphological observations of
the anamorph were taken from cultures grown on CMD in
9 cm diam vented plastic Petri plates in an incubator at 20
C with alternating 12 h fluorescent light and 12 h darkness
within 14 d. These standard characters were measured from
3% KOH or distilled water: width of phialide at the base,
phialide width at the widest point, phialide length, cells sup-
porting phialides, presence of chlamydospores and chla-
mydospore width. Measurements of continuous characters
were taken from images using the beta 4.0.2 version of Sci-
on Image (Scion Corp., Frederick, Maryland). Measure-
ments of asci, ascospores and anamorph characters in spe-
cies descriptions are reported as maxima and minima in
parentheses and the mean plus and minus the standard de-
viation of a minimum of 30 measurements. Four types of
compound microscopy were used, viz. bright field (BF),
phase contrast (PC), differential interference contrast
(DIC) and fluorescence (FL). Preparations studied for fluo-
rescence microscopy were prepared by flooding prepara-
tions that had been used for measurements with Calcofluor
(Sigma Fluorescent Brightener 28, C.I. 40622 Calcofluor
white M2 in a 2 molar phosphate buffer at pH 8.0). Colony
appearance was described from CMD at 20 C and PDA at
25 C with alternating 12 h fluorescent light and 12 h dark-
ness, including formation and shape of tufts or pustules.
The presence of chlamydospores was recorded by examin-
ing the reverse of a colony grown on CMD after ca 1 wk at
20 C under 12 h darkness and 12 h cool white fluorescent
light with 403objective of a compound microscope.
Illustrations of conidiophores and conidia were taken
from colonies grown on CMD at 20–21 C under 12 h dark-
ness/12 h cool white fluorescent light for 7–10 days, unless
otherwise noted.
The herbarium specimens of
Hypocrea
were rehydrated
briefly in 3% KOH. Rehydrated stromata were supported
by Tissue-Tek O.C.T. Compound 4583 (Miles Inc., Elkhart,
Indiana) and sectioned at a thickness of ca. 15 mm with a
freezing microtome. Permanent preparations of the sec-
tions were made following Volkmann-Kohlmeyer and Kohl-
meyer (1996). These teleomorph characteristics were eval-
uated: diameter, height, color and shape of the stroma; tex-
ture of surface of the stroma; perithecium shape, length

312 M
YCOLOGIA
T
ABLE
I. Strains of
Hypocrea/Trichoderma
studied
Morphological
species Isolate Identification
Geographic
origin Substratum
GenBank
ITS 1 ITS 2 ITS 1 12
ech42 EF-1
a
T. croceum
ex-type DAOM 167068
H. pachybasioides
/
T. polysporum
Canada (Quebec) soil, pine forest AF400259 AF400995 AF486002
H. pilulifera
/
T. pilulife-
rum
ex-type
CBS 814.68
H. pilulifera
UK
Betula
sp., decorti-
cated wood
Z48813 Z48813 AF486003
H. pilulifera
/
T. pilulife-
rum
CBS 341.93
H. pilulifera
not
studied mor-
phologically
Hypocrea
sp. G.J.S. 86-540
H. pachybasioides
/
T. polysporum
USA (New York) ?
Fagus,
decorticat-
ed wood
AY240839 AY24021 AY240845
Hypocrea
sp. G.J.S. 88-44
H. pachybasioides
/
T. polysporum
USA (North Caro-
lina)
bark
Hypocrea
sp. G.J.S. 88-59
H. pachybasioides
/
T. polysporum
USA (North Caro-
lina)
decorticated wood AY240840 AY240213 AY240846
Hypocrea
sp. G.J.S. 89-135
H. pachybasioides
/
T. polysporum
USA (North Caro-
lina)
bark of recently
dead hardwood
tree
AY240841 AY240214 AY24084
Hypocrea
sp. G.J.S. 90-28
H. pachybasioides
/
T. polysporum
Switzerland
Picea
sp., decorti-
cated wood
Hypocrea
sp. G.J.S. 90-63
H. pachybasioides
/
T. polysporum
USA (New York) bark AY240842 AY240215 AY240848
Hypocrea
sp. G.J.S. 90-116
H. pachybasioides
/
T. polysporum
USA (North Caro-
lina)
Aphyllophorales
on
Betula
sp.
AY24084 AY24021 AY240849
Hypocrea
sp. G.J.S. 90-126
H. pachybasioides
/
T. polysporum
USA (North Caro-
lina)
fungi on bark AY240844 AY240217 AY240850
Hypocrea
sp. G.J.S. 93-38
H. pachybasioides
/
T. polysporum
USA (New York) bark and decorti-
cated wood
Hypocrea
sp. G.J.S. 99-90
H. pachybasioides
/
T. polysporum
New Zealand
Nothofagus menzie-
sii,
bark
AY240167 AY240199 AY240218 AY240851
Hypocrea
sp. G.J.S. 99-155
H. pachybasioides
/
T. polysporum
Australia (New
South Wales)
bark AY240168
Hypocrea
sp. G.J.S. 99-159
H. pachybasioides
/
T. polysporum
Australia (New
South Wales)
bark AY240169
Hypocrea
sp. G.J.S. 99-207
H. pachybasioides
/
T. polysporum
New Zealand bark AY24017 AY240200 AY240219 AY24085
Hypocrea
sp. G.J.S. 99-219
H. pachybasioides
/
T. polysporum
Australia (Victo-
ria)
bark AY240171
Hypocrea
sp. G.J.S. 99-220
H. pachybasioides
/
T. polysporum
Australia (Victo-
ria)
Eucalyptus,
decor-
ticated wood
AY240172

313L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
T
ABLE
I. Continued
Morphological
species Isolate Identification
Geographic
origin Substratum
GenBank
ITS 1 ITS 2 ITS 1 12
ech42 EF-1
a
Hypocrea
sp. G.J.S. 99-221
H. pachybasioides
/
T. polysporum
Australia (Victo-
ria)
bark AY240173 AY240201 Ay240220 AY240853
Hypocrea
sp. G.J.S. 99-223
H. pachybasioides
/
T. polysporum
Australia (Victo-
ria)
Nothofagus
sp.,
bark
AY240174
Hypocrea
sp. G.J.S. 99-224
H. pachybasioides
/
T. polysporum
New Zealand branchlets of re-
cently dead tree
AY240175
Hypocrea
sp. G.J.S. 99-244
H. pachybasioides
/
T. polysporum
New Zealand
Nothofagus
sp.,
bark
T. polysporum
TMI 34109
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
AY240176
T. polysporum
TMI 60177
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
AY240177
T. polysporum
TMI 60466
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
AY240178
T. polysporum
TMI 60844
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
AY240179
T. polysporum
TMI 8084
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8215
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8217
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8217
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8218
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8220
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8221
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TMI 8258
H. pachybasioides
/
T. polysporum
Japan
Quercus
infected
with Shiitake
T. polysporum
TR 100 5
ATCC 18650
H. pachybasioides
/
T. polysporum
unknown unknown
T. polysporum
TR 54
H. pachybasioides
/
T. polysporum
USA (Oregon)
Pseudotsuga men-
ziesii,
roots in-
fected with
Phel-
linus weirii
T. polysporum
TR 80
H. pachybasioides
/
T. polysporum
USA (Wyoming) soil

314 M
YCOLOGIA
T
ABLE
I. Continued
Morphological
species Isolate Identification
Geographic
origin Substratum
GenBank
ITS 1 ITS 2 ITS 1 12
ech42 EF-1
a
Hypocrea
sp. G.J.S. 91-60
H. parapilulifera
USA (Virginia) decorticated wood AY241587 AY240221 AY240854
Hypocrea
sp. G.J.S. 99-188
H. parapilulifera
Australia (Victo-
ria)
Nothofagus,
bark AY240180
Hypocrea
sp. G.J.S. 99-222
H. stellata
New Zealand bark AY240181 AY24020 AY240222 AY240855
T. minutisporum
ex-type DAOM 167069
H. minutispora/T.
minutisporum
Canada (Quebec) maple forest soil AY240182
T. minutisporum
DAOM 175931
H. minutispora/T.
minutisporum
Canada (Ontario)
Betula papyrifera,
stump
AY240183 AY24020
T. minutisporum
DAOM 177713
H. minutispora/T.
minutisporum
Canada (Quebec)
Betula
sp., log AY240184
T. minutisporum
DAOM 178046
H. minutispora/T.
minutisporum
Canada (Ontario) wood AY240185 AY240204 AY240856
T. minutisporum
DAOM 179741
H. minutispora/T.
minutisporum
Canada (Ontario) ?
Betula
sp., wood AY240186
T. minutisporum
DAOM 179894
H. minutispora/T.
minutisporum
Canada (Ontario) wood AY240187
T. minutisporum
DAOM 180142
H. minutispora/T.
minutisporum
Canada (Ontario)
Betula
sp. AY240188
T. minutisporum
DAOM 212372
H. minutispora/T.
minutisporum
Canada (Alberta) soil under lodge-
pole pine
AY240189
T. minutisporum
CBS 901.72
H. minutispora/T.
minutisporum
Germany wood AY240190 AY240205 AY240223 AY240857
Hypocrea
sp. G.J.S. 90-80
H. minutispora/T.
minutisporum
USA (North Caro-
lina)
decorticated wood AY240191
Hypocrea
sp. Ex-type, G.J.S.
90-81 5
ATCC MYA-
2951
H. minutispora/T.
minutisporum
USA (Georgia) bark AY240192
Hypocrea
sp. G.J.S. 90-82
H. minutispora/T.
minutisporum
USA (Georgia) decorticated wood AY240193 AY240206 AY240224 AY240858
Hypocrea
sp. G.J.S. 90-112
H. minutispora/T.
minutisporum
USA (North Caro-
lina)
?
Acer,
bark AY240194 AY240207 AY240225 AY240859
Hypocrea
sp. G.J.S. 90-115
H. minutispora/T.
minutisporum
USA (North Caro-
lina)
Quercus,
bark AY240195 AY240208 AY240226 AY240860
Hypocrea
sp. G.J.S. 95-181
H. minutispora/T.
minutisporum
USA (Kentucky)
Stereum
sp. AY240196 AY240209 AY240861
T. koningii
ATCC 28012
H. minutispora/T.
minutisporum
USA (North Caro-
lina)
soil AY240197 AY240210 AY240227 AY240862
Hypocrea
sp. G.J.S. 99-198
H. lacuwombaten-
sis
New Zealand bark of recently
dead tree
AY240198 AY240211 AY240228 AY240863

315L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
T
ABLE
II. Results of the phylogenetic analyses of ITS,
tef1
and
ech42
Gene gaps
ITS 1 12
Missing Fifth base
tef1
Missing Fifth base
ech42
Missing Fifth base
Combined
Missing Fifth base
Number of trees
Total characters
Parsimony-informative
100
383
30
24
55
100
112
41
12
53
2
454
117
2
450
4
949
188
1
588
Polymorphic sites
Tree length
Homoplasy index
Consistency index
Retention index
115
0.35
0.65
0.62
233
0.35
0.65
0.63
196
0.43
0.57
0.71
263
0.41
0.58
0.71
396
0.46
0.54
0.61
1751
0.62
0.38
0.21
643
0.38
0.62
0.72
2104
0.54
0.46
0.43
and width; reaction to 3% KOH, color, width of peritheci-
um wall; ostiolar canal length; color and 3% KOH reaction
of stroma outer region; shape, diameter and wall thickness
of cells of the outer, middle (immediately below the outer
region) and inner region (below perithecia) of the stroma;
ascus length and width; distal and proximal part-ascospore
length and width. Measurements of continuous characters
were obtained with Scion Image beta 4.0.2. Statistical anal-
yses were performed using Systat 10.0 (SPSS Inc., Chicago,
Illinois).
DNA extraction, PCR amplifications and sequencing.
—DNA
was isolated from fresh mycelium as described previously
(Turner et al 1997). A region of nuclear rDNA, containing
the ITS1 and 2 and the 5.8S rRNA gene, was amplified by
PCR using the primer combinations SR6R and LR1 in 50
mL volumes (White et al 1990) in an automated tempera-
ture-cycling device (Biotron, Biometra, Go¨ttingen), using
the protocol described in Kullnig-Gradinger et al (2002). A
0.2 kb fragment of
tef1
was amplified by the primer pair
tef1fw and tef1rev. A 0.4 kb fragment of
ech42
was amplified
by the primer pair Chit42-1a and Chit42-2a (Kullnig-Gra-
dinger et al 2002). Template DNA (100 mL) was prepared
directly from PCR products by purifying it with a commer-
cial kit (Cleanmix; Talent s.r.l., Trieste) and sequenced with
the aid of a LI-COR 4000L automatic sequencing system,
using cycle-sequencing (Robocycler 40; Stratagene, La Jolla,
California) with the ThermoSequenasekit (Amersham Bio-
sciences Inc. Piscataway, New Jersey) as described in Kin-
dermann et al (1998).
The sequences of isolates submitted to GenBank, as in-
dicated in T
ABLE
I, are the sequences used in all phyloge-
netic analyses. Sequences not obtained during this work are
given by their respective GenBank accession numbers. The
nucleic acid matrix—ITS1 and 2,
tef1,
and
ech42
combined
tree F
IG
. 2—has TreeBase ID number SN1486.
RAPD analysis was performed as described earlier (Kull-
nig et al 2000), using M13 as a primer.
Molecular phylogenetic analysis.
—DNA sequences were
aligned first with ClustalX 1.81 (Thompson et al 1997) and
then visually adjusted, based on the algorithm of Waterman
(1986) using Gendoc 2.6.002 (Nicholas and Nicholas 1997).
Gaps were treated either as missing or as a fifth character.
Phylogenetic analyses were performed in PAUP* 4.0b10 us-
ing
Trichoderma hamatum
DAOM 167057 as outgroup. Par-
simony analysis was performed using a heuristic search, with
a starting tree obtained via stepwise addition, with random
addition of sequences with 1000 replicates, tree-bisection-
reconnection as the branch-swapping algorithm, Multrees
in effect. Stability of clades was assessed with 500 bootstrap
replications. The Partition Homogeneity Test (PHT) inte-
grated in PAUP was used to test the congruence among
different gene datasets (Cunningham 1997). For this test
parsimony-uninformative characters were excluded, gaps
were treated as missing, and 10 000 repetitions were per-
formed. A maximum of 100 trees were saved to conserve
memory.
RESULTS
Molecular phylogenetic analyses.
—Fifty-four isolates of
Trichoderma
/
Hypocrea
first were analyzed by sequenc-
ing ITS1 and 2, and RAPD analysis. Representatives
of each haplotype then were subjected to a parsi-
mony analysis using nucleotide sequences of a frag-
ment of
tef1
(encoding translation elongation factor
1-alpha, Kullnig-Gradinger et al 2002) and of
ech42
(encoding the 42-kDa endochitinase, Lieckfeldt et al
2000a). The ITS1 and 2 tree had the fewest parsi-
mony-informative polymorphic sites (30; correspond-
ing to 7.4%; T
ABLE
II). Both
tef1
and
ech42
produced
a high number of parsimony-informative characters
(41 and 117, corresponding to 36.2 and 24.3%). Phy-
logenetic analysis using gaps as a fifth base did not
give strongly different tree topologies but resulted in
higher bootstrap support for the clades. Both trees
obtained from the analysis of single genes only (F
IG
.
1a, b), as well as that from a combined analysis of
ITS1 and 2,
tef1
and
ech42
;F
IG
. 2), and using
T. ha-
matum
(sect.
Trichoderma
) as outgroup, grouped
these species in a strongly supported (98% when gaps
were treated as missing data) monophyletic group,
thus confirming the previously defined cluster B5 in
pachybasium B (Kullnig-Gradinger et al 2002).
Within pachybasium B5, the
Hypocrea
/
Trichoderma
isolates included in our study clustered in two main
clades, which we refer to as the ‘‘polysporum’’ and

316 M
YCOLOGIA
F
IG
. 1. One of the most parsimonious trees from
tef1
(A) and
ech42
(B). Numbers on branches represent bootstrap
coefficients; those above are gaps treated as missing information; those below as gaps treated as a fifth base. Ex-type strains
are shown in bold. GenBank accession numbers for
T. hamatum
DAOM 167057 are given in Kullnig-Gradinger et al (2002).
‘‘minutisporum’’ clades, respectively. Each major
clade included two or more subclades, or lineages,
that were more or less well supported. While most
lineages comprised only a single isolate, in some in-
stances ascospore isolates and cultures derived di-
rectly from nature (conidial) were almost identical
(T
ABLE
II). There were no strictly conidial lineages.
The minutisporum clade received high bootstrap
support both in the combined (ITS1
1
2,
ech42
and
tef1
) as well as the individual (
tef1, ech42
) trees and
includes the named species
T. minutisporum
ATCC
28012, which previously was misidentified (Kuhls et
al 1996) as
T. hamatum,
and several green-spored
Hy-
pocrea
species. A new
Hypocrea
species, G.J.S. 99-198
from New Zealand, described here as
H. lacuwom-
batensis,
always occurred basal to this cluster.
The polysporum clade was less strongly but still
reasonably well supported by bootstrap coefficients
and includes the described species
T. croceum, H. pi-
lulifera
/
T. piluliferum
and
T. polysporum.
The ex-type
culture of
T. croceum
and the epitype culture of
T.
polysporum
clustered in subclades within the larger
polysporum clade, but neither subclade received
strong support. Two other branches, each leading to
species doublets (viz. G.J.S. S 90-63 and G.J.S. 90-116;
G.J.S. 86-540 and G.J.S. 90-126) always received
strong support within this cluster. However, both are
morphologically indistinguishable from
H. pachyba-
sioides
(see below) and thus might be at the stage of
clonal isolation only.
Three sets of
Hypocrea
isolates (viz.
H. pilulifera,
isolate G.J.S. 91-60/G.J.S. 99-188, and isolate G.J.S.
99-222) always clustered at a basal position of the po-
lysporum clade. Low bootstrap support for their
branches indicates that these three sets represent sep-
arate lineages.
The partition homogeneity test (Huelsenbeck et al
1996) was used to examine the null hypothesis of
recombination (Koufoupanou et al 1997). The actual
summed tree length of 576 steps was exactly at the
lowest limit of that produced by any of the 10 000
artificial datasets (
P
-value 0.002; F
IG
. 3), and four
steps shorter than
.
95% of them, thus indicating in-
congruence among the different gene trees. Howev-
er, the
P
-value (gene partition incongruence value)
was 0.0549, which is slightly more than
P
5
0.05, be-
low which incongruence is considered significant. We
therefore have investigated whether this would be
due to recombination ocurring only within one of
the clusters in pachybasium B5. In addition, we re-
moved the ITS sequences from the analysis because
of the unequal and biased mutation rate of rDNA
genes (Maynard-Smith and Smith 1998). Thus the
partition homogeneity test was applied separately to
each of the two clades, with and without using ITS1
and 2 sequences (T
ABLE
IV). The corresponding data
(F
IG
.3,T
ABLE
IV) clearly show that recombination is
only apparent within the polysporum clade, whereas
it appears to be absent from the minutisporum clade.
Within the polysporum clade recombination only oc-

317L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
F
IG
. 2. One of the most parsimonious combined trees from ITS112,
tef1
and
ech42
Numbers on branches represent
bootstrap coefficients; those above are gaps treated as missing information; those below as gaps treated as a fifth base. Ex-
type strains are shown in bold. Vertical gray bars indicate species considered by Kullnig-Gradinger et al (2002) to belong to
subclades pachybasium B5 and pachybasium B4 respectively. GenBank accession numbers for
T. hamatum
DAOM 167057
and pachybasium B4—
T. harzianum
CBS 226.95,
T. tomentosum
CBS 349.93 and
T. virens
CBS 249.59—are given in Kullnig-
Gradinger et al (2002).
curs within
T. polysporum
/
T. croceum
/
H. pachybasioi-
des
but not between them and
H. stellata, H. parap-
ilulifera
and
H. pilulifera.
Biogeography.
—The strains shown in the phylograms
(F
IGS
. 1, 2) are representative. Strains that were iden-
tical to those shown in the phylograms in ITS 1
1
ITS 2 and having
.
80% similarity in RAPD profiles
are listed in T
ABLE
III. Within the polysporum clade
there is no geographic bias in favor of any of the
subclades. Most of the strains studied were identical
to the ex-type culture of
T. croceum
; these included a
wide geographic diversity including the eastern
U.S.A., Japan and Australia. The isolate G.J.S. 90-116
(North Carolina) was identical to isolates from Aus-
tralia. The isolate G.J.S. 91-60 (Virginia) was identical
to G.J.S. 99-188 (New Zealand).
Hypocrea
pilulifera
/
T. piluliferum
is unusual in that
it is limited to northern Europe. In this regard, it is
similar to the unrelated
H. aureoviridis
/
T. aureoviride
(Lieckfeldt et al 2001).
In contrast to the polysporum clade, where several
isolates were collected in Australia and New Zealand,
only one strain (G.J.S. 99-198) in the minutisporum
clade originated in Australasia. Most
T. minutisporum
isolates came from North America (eastern U.S.A.
and Canada), with only one isolate collected outside
of North America (CBS 901.72, Germany).
Phenotype analyses.
—All isolates included in the pre-
sent study have a pachybasium morphology as was
broadly defined by Bissett (1991b) in which phialides
tended to be short, wide and more or less densely
clustered on broad cells. The phialides of
T. pilulifer-
um
differs from the typical pachybasium morphology
in being more divaricate. The phialides typically were
slightly narrower than the cell from which they arose
and on average 1.5 times as wide. Each of the two
major clades, revealed by DNA sequence analysis, was
characterized by conidial color (viz. green in the min-
utisporum clade and white in the polysporum clade).
There were no green-conidial strains in the polyspo-
rum clade and no white-conidial strains in the min-
utisporum clade. The clades also differed in growth

318 M
YCOLOGIA
F
IG
. 3. Partition homogeneity test results for
H. pachybasioides
/
T. polysporum
(dark shade) and
H. minutispora
/
T. min-
utisporum
(light shade). For this test parsimony-uninformative characters were excluded, gaps were treated as missing, and
10 000 repetitions were performed.
T
ABLE
III.
Hypocrea
and
Trichoderma
strains appearing in phylograms and additional strains having identical ITS1 1ITS2
and .80% similar RAPD profiles
Species Strain in phylograms
1
Strains not in phylograms
2
H. pachybasioides
/
T. polysporum
ATCC 18650/CBS 820.68 5G.J.S. 99-90
H. pachybasioides
/
T. polysporum
G.J.S. 99-207 none found
H. pachybasioides
/
T. polysporum
DAOM 167068 (
T. croceum
ex-type) 5G.J.S. 88-44, G.J.S. 89-135, G.J.S. 99-219,
G.J.S. 99-220, G.J.S. 99-224, all TMI strains,
Tr 54, Tr 80
H. pachybasioides
/
T. polysporum
G.J.S. 90-116 5G.J.S. 88-59, G.J.S. 99-155, G.J.S. 99-159,
G.J.S. 99-223
H. pachybasioides
/
T. polysporum
G.J.S. 90-63 none found
H. pachybasioides
/
T. polysporum
G.J.S. 99-221 none found
H. pachybasioides
/
T. polysporum
G.J.S. 90-28 5Tr 100
H. pachybasioides
/
T. polysporum
G.J.S. 86-540 5G.J.S. 90-126, G.J.S. 93-38, G.J.S. 99-198,
G.J.S. 99-244
H. parapilulifera
G.J.S. 91-60 5G.J.S. 99-188
H. stellata
G.J.S. 99-222 none found
H. minutispora
/
T. minutisporum
DAOM 167069 5DAOM 175931, DAOM 177713, DAOM
179741, DAOM 180142, DAOM 213372,
DAOM 179894, DAOM 178046, CBS 901.72
H. minutispora
/
T. minutisporum
G.J.S. 90-82 5G.J.S. 90-80, G.J.S. 90-81, G.J.S. 95-181
H. minutispora
/
T. minutisporum
G.J.S. 90-115 none found
H. minutispora
/
T. minutisporum
G.J.S. 90-112 none found
H. minutispora
/
T. minutisporum
ATCC 28012 none found
H. lacuwombatensis
G.J.S. 99-198 none found
H. pilulifera
/
T. piluliferum
CBS 814.68 none found
H. pilulifera
/
T. piluliferum
CBS 341.93 none found
1
F
IGS
.1,2.
2
Provenance in T
ABLE
I.

319L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
T
ABLE
IV. Results of the partition homogeneity test
p
value
1
tef1,
ech42
ITS1 and
2,
tef1,
ech42
Polysporum clade
H. pachybasioides
/
T. polysporum
H. parapilulifera, H. pilulifera, H. stellata
Minutisporum clade
H. minutispora
/
T. minutisporum
Pachybasium B5
0.0186
0.0060
0.5681
0.3882
0.2030
0.0668
0.0075
0.0002
0.7088
0.5527
0.4486
0.0549
1
Gene partition incongruence is significant when
P
,
0.05.
rate. Although all the isolates grew slower on PDA at
25 C than is usual of
Trichoderma,
isolates in the po-
lysporum clade rarely reached a colony radius of 25
mm after 72 h at 25 C in in the dark, whereas the
isolates of the minutisporum clade reached a radius
of approximately 35 mm under those conditions.
Stromata of all collections studied were typical of
Hypocrea
in being lightly to brightly pigmented and
fleshy. Perithecia were completely immersed. Asco-
spores were bicellular but disarticulated early in their
development to give 16 unicellular part-ascospores in
each ascus. Part-ascospores of all collections were hy-
aline, dimorphic and finely spinulose. The stromata
of all collections tended to be less than 5 mm diam,
solitary and sharply delimited, circular to elliptic in
outline, with margins free from or united with the
substratum. Stromata of the Japanese specimens of
H. pachybasioides,
which were isolated from
Quercus
logs infected with shiitake (
Lentinula edodes,
T
ABLE
I), tended to be aggregated or gregarious and often
were effused and the margins were not free from the
substratum. In all cases the stromatal surface was
plane and the apices of individual perithecia were
not evident or barely rose above the stroma surface.
Each stroma contained many perithecia, the open-
ings of which were easily seen as dark dots against
the lighter surface of the stroma. The youngest, de-
veloping stromata tended to be vinaceous and to
have a lighter-colored margin, but as stromata aged
the tendency was to become a light brown or buff,
or to remain vinaceous. The color variation was not
associated with any of the subclades of either
T. min-
utisporum
or
T. polysporum.
There was no reaction to
3% KOH.
The stromata of all the
Hypocrea
specimens were
generalized to the extent that one could not predict
which anamorph any collection would give. There
were no single or combined gross morphological,
pigmentation or anatomical features of the teleo-
morph that characterized any of the clades. The only
exception to this is the stroma of
H. pilulifera,
the
type of which formed on an herbaceous substratum
(
Juncus effusus
). However, the ex-type culture of its
anamorph,
T. piluliferum,
was isolated from wood, as
were all the other ascospore collections (but not the
conidial isolates, which came from a diversity of
sources but mainly soil). Apparently there are no ex-
tant ascospore isolates of
T. piluliferum.
There were no anatomical differences in the stro-
mata or perithecia in any of the species included in
this study. The cells of the stroma surface were typi-
cally angular, 3–7
3
2.5–6.5
m
m, walls
,
1
m
m thick
and easily seen. All stromata were divided into three
regions—surface, subsurface and tissue below peri-
thecia. The surface region of the stroma was 20–35
m
m thick and was composed of pigmented cells
which, in face view, were 2.5–5.0
3
2–3
m
m in cross
section. In all cases the subsurface region was com-
posed of intertwined, ca. 3
m
m wide, septate, thin-
walled hyphae. The cells below the perithecia tended
to be hyphal or at least of textura intricata. Perithecia
were 200–300
m
m tall, 100–170
m
m wide and the os-
tiolar canal was 40–90
m
m long. The perithecial pa-
pilla only rarely protruded through the stroma sur-
face, and the cells of the perithecial apex were not
different from the surrounding cells of the stroma
surface.
The asci of all species were cylindrical and sessile;
ascospores were uniseriate or at most partially biser-
iate. The ascus apex of all collections was slightly
thickened and possessed an obscure pore. Part-asco-
spores were more or less dimorphic. The distal part-
ascospores were subglobose to ellipsoidal, rarely con-
ical. The proximal part-ascospores were oblong to
wedge-shaped, tapering slightly from the middle to
the end. The distal part-ascospores of members of
most subclades of the polysporum clade were 3.5–4.5
m
m long and ca. 3.5
m
m wide and the proximal part-
ascospores were 4.0–5.5(–6.0)
3
2.5–3.2
m
m. The
part ascospores of the subclade that includes G.J.S.
86-540 (USA: New York), G.J.S. 90-126 (USA: Virgin-
ia), and TMI 8084 ( Japan) were significantly smaller
than all others in all regards (distal part-ascospores:
3.3
6
0.4[2.5–4.6]
3
3.0
6
0.3[2.1–3.7]
m
m; proxi-
mal part-ascospores 3.8
6
0.5[2.5–5.2]
3
2.6
6
0.3[1.9–3.3]
m
m).
Cultures derived from ascospores could not be dis-
tinguished from conidial cultures of, respectively,
T.
polysporum
and
T. minutisporum.
The
Hypocrea
spec-
imens that gave
T. polysporum
anamorphs in culture
could not be distinguished from the isotype speci-
men of
H. pachybasioides.
Most of the species in the polysporum clade had a
more or less strong tendency to form discrete, white

320 M
YCOLOGIA
pustules on CMD or SNA. On PDA, conidial produc-
tion tended to be continuous and not in pustules.
The pustules of
H. pachybasioides
/
T. polysporum
were
dense, and spiraled hairs were produced abundantly
from each pustule. Within the polysporum clade the
pustules of
H. pilulifera
/
T. piluliferum,
G.J.S. 91-60
and G.J.S. 99-222 were constructed more loosely and
conidiophores with more or less long, fertile axes
were conspicuous. This was especially true of G.J.S.
99-222, in which long, narrow, plumose conidio-
phores with short lateral branches were conspicuous
(F
IGS
. 18, 75–79)
In the minutisporum clade there was a less well-
developed tendency to form robust pustules. The co-
nidiophores, while densely aggregated, instead
formed in regions of continuous sporulation. In
T.
minutisporum
discrete conidiophores were not con-
spicuous when viewed with the stereo microscope
(F
IGS
. 19–21) and with the compound microscope
fertile branches were relatively short and had short
internodes between fertile branches (F
IGS
. 95, 96).
In G.J.S. 99-198 conidial pustules were cottony and
individual conidiophores were seen easily with the
stereo microscope (F
IG
. 23). Conidiophores in this
isolate comprised a long fertile axis that produced
short, fertile lateral branches along its length (F
IGS
.
107–112) or they produced one or a few phialides at
the tip but then were sterile for a long distance (F
IG
.
106). In this isolate (typical of
Trichoderma
) branches
more distant from the tip tended to be longer and
to rebranch more, ultimately producing a complex,
pyramidal fertile system. The phialides produced in
less crowded parts of the conidiophore usually were
longer than were those produced lower on the co-
nidiophore where phialides were more crowded.
All conidia were smooth. Conidia of all members
of the polysporum clade were hyaline, white in mass
except for conidia of the ex-type isolate of
T. croceum,
which were yellow. Conidia of all members of the
minutisporum clade were green.
Conidia of most species were ellipsoidal with a
length/width ratio of ca. 1.5. Conidia of
H. piluli-
fera
/
T. piluliferum
were subglobose, L/W
5
ca. 1.1.
Conidia of most species had a mean of size of ca. 3.0
3
2.1
m
m. Of the
H. pachybasioides
/
T. polysporum
subclades, only G.J.S. 99-207 (New Zealand) had
overall larger conidia (mean
5
3.5
3
2.3
m
m) than
other members of the clade. Conidia of
H. pilulifera
/
T. piluliferum
were wider than those of most species
studied, but as was already mentioned conidia of that
species were subglobose. In the minutisporum clade,
conidia of G.J.S. 99-198 (New Zealand) were slightly
longer (95% CI
5
3.4–3.6
m
m) than conidia of other
members of the clade (95% CI
5
3.2–3.3), all of
which originated in temperate eastern North Amer-
ica and Europe.
Conidiophores of
H. pachybasioides
(F
IGS
. 24–27,
46, 47) and G.J.S. 91-60/G.J.S. 99-188 (F
IGS
. 65, 66)
comprised a spiralled, thin-walled-septate, sterile hair
with subacute to rounded tips, from the base of
which arose fertile branches. The hairs of
H. pachy-
basioides
tended to be rugose. All other cultures pro-
duced conidiophores that had a discernible main
axis that bore phialides at the tip and produced fer-
tile lateral branches along the length. The branches
tended to increase in length with distance from the
tip and often were paired.
Phialides of most species were short and wide, with
a mean length of 4.9–5.8
m
m and L/W ratio 1.7–1.8.
Phialides of G.J.S. 91-60 and G.J.S. 99-222 were some-
what longer and narrower, L/W
5
2.0 and 2.3, re-
spectively.
Chlamydospores were produced within 10 d on
CMD by some collections of all clades. They were
terminal or intercalary and subglobose.
Within the polysporum clade four ascospore iso-
lates (G.J.S. 86-540, G.J.S. 90-126, G.J.S. 93-38, G.J.S.
99-244) formed a well-supported sister clade of the
clade that included the ex-type isolates of
T. polyspo-
rum
and
T. croceum.
These ascospore collections had
slightly but significantly smaller ascospores than is
typical of
T. polysporum
and slightly but significantly
slower growth (after 72 h on PDA maximum
5
20
mm and 25–30 mm, respectively). These ascospore
isolates originated from widely separated geographic
stations (United States, New Zealand). However,
there was continuous variation in growth rate in this
clade with each subclade having a slightly different
rate of growth. The optimum temperature for all iso-
lates except G.J.S. 99-207 was 25 C; the optimum for
G.J.S. 99-207 was 20 C.
The ex-type isolate of
T. croceum
produced yellow
conidia (F
IGS
. 15, 16) but it was almost identical to
several strains in RAPD and ITS 1
1
2 sequences, all
of which had hyaline conidia.
The isolate G.J.S. 99-222, which was alone in a basal
position in the polysporum clade, grew more slowly
than any of the isolates that were studied, reaching a
maximum radius of approximately 15 mm after 72 h
on PDA at 20 and 25 C.
Within the minutisporum clade most isolates ge-
netically were identical or very close to the ex-type
isolate of
T. minutisporum
(DAOM 167069) and there
was very little divergence in their respective growth
rates, the optimum temperature being 25 C on PDA.
The only other subclade that included more than
one culture was that including G.J.S. 90-82, and all
five isolates in that clade had the same growth char-
acteristics. G.J.S. 90-112 and ATCC 28012 had slightly

321L
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:
H
YPOCREA
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RICHODERMA SPECIES WITH PACHYBASIUM
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different growth rates from the rest, the former being
slower and the latter being faster. Greater differences
were seen at 30 C, where G.J.S. 90-198 was slowest
(10 mm),
T. minutisporum
ex-type group, G.J.S. 90-
112 the next slowest (ca. 15 mm), and all the rest
reached a radius of 27–30 mm on PDA after 72 h.
DISCUSSION
This research included members of the morpholog-
ically defined
Trichoderma
sect.
Pachybasium
(Sacc.)
Bissett (Bissett 1991b). This section is now known to
be paraphyletic (Kindermann et al 1998, Kullnig-Gra-
dinger et al 2002). Our work demonstrates that the
white-conidial species of
Trichoderma
/
Hypocrea
and
the green-spored species without apical elongations
form a strongly supported clade (B5) in pachybasium
B in which at least six different species can be distin-
guished. Although evidence for this clade confirms
earlier investigations by Kindermann et al (1998) and
Kullnig-Gradinger et al (2002), support for its fine
structure largely came from the analysis of single-
copy loci (viz.
tef1, ech42
). It is interesting to note
that sequence analysis of the internally transcribed
spacer regions 1 and 2 of the rDNA cluster, which
have been shown to result in a reliable phylogeny in
earlier studies (Kuhls et al 1996; Lieckfeldt et al 1998,
1999), failed to resolve the phylogeny of subclade B5.
A closer investigation of the ambiguous nucleotide
area revealed that it was due mainly to the occur-
rence of an extended AT-rich stretch within a GC-
rich sequence in the 5
9
end of ITS1. This does not
occur in other species of
Trichoderma
and not in phy-
logenetically close species in genera such as
Hypo-
myces, Nectria
or
Gibberella
(C.P. Kubicek, unpubl)
and thus is specific for subclade B5. The origin of
this sequence probably is due to an initial T
→
A ex-
change. Such an exchange can be found in some
members of pachybasium B, such as
T. oblongisporum
Bissett,
T. tomentosum
Bissett or
H. semiorbis
Berk.,
which is an invariable hallmark sequence (TCAT) in
species belonging to sect.
Trichoderma.
In a phylo-
genetic tree based on ITS1 sequence analysis only,
subclade B5 therefore occurs as a sister clade of sect.
Trichoderma
(Kindermann et al 1998). Species of po-
lysporum subclade B5 bear a duplication of this re-
gion, with C
→
T exchanges in some isolates. On the
other hand, a variable (n
5
2–4) stretch of As adja-
cent to the G-rich stretch occurs in the minutisporum
clade, which can be explained by preferential inser-
tion of A in depurinated places (Dianov et al 1992).
All in all, this indicates that this region is a major hot
spot for mutations in the B5 subclade of
Trichoder-
ma
/
Hypocrea
species and in fact there is little nucle-
otide variation in the remaining sequence of ITS1
and 2. The reason for this is unknown. However,
Moore et al (1991) demonstrated that various fami-
lies of AT-rich triplet repeats form secondary struc-
tures that escape DNA repair, which, because of the
proliferated secondary structure of the internally
transcribed spacer regions of rDNA, might be an ex-
planation.
Recombination might be a further reason for the
variability of this region. A much higher variability
was observed in the polysporum clade than in the
minutisporum clade, which agrees with the results
from the partition homogeneity test. These data re-
veal that there is considerable genetic exchange with-
in
H. pachybasioides
but not among
H. pachybasioides,
H. pilulifera, H. parapilulifera
or
H. stellata,
thus jus-
tifying our recognition of the latter three as species
distinct from
H. pachybasioides.
Recombination within the minutisporum clade ap-
pears to be rare, and this cluster is characterized by
a significant clonal element despite the inclusion of
a significant number of ascospore isolates of
T. min-
utisporum
in the analysis. One possible explanation
would be that
T. minutisporum
is homothallic. Few
species of
Hypocrea
undergo sexual reproduction in
vitro. Of those, bipolar heterothallism has been dem-
onstrated only in
H. jecorina
Berk. & Broome/
T. ree-
sei
E.G. Simmons (Lieckfeldt et al 2000b). The more
usual situation, first exemplified (Mathieson 1952,
Perkins 1987) by
Hypocrea spinulosa
Fuckel (
5
Chromocrea spinulosa
[Fuckel] Petch), has been de-
scribed as ‘‘mating-type shifting’’ wherein half the
spores in an ascus are self fertile and the other half
are self sterile but cross compatible with the self-fer-
tile cultures. This system is strongly suggested for
H.
citrina
(Pers. Fr.) Fr. (
5
H. pulvinata
Fuckel) (Can-
ham 1969) and
H. poronioidea
A. Mo¨ller (Samuels
and Lodge 1996). However, we do not know of any
strictly homothallic species of
Hypocrea.
Hyaline conidia are uncommon in
Trichoderma.
Species related to
H. poronioidea
(Samuels and
Lodge 1996),
H. pulvinata
Fuckel (Rifai and Webster
1966) and
H. sulphurea
(Schw.) Fr. (Samuels unpub-
lished) produce acremonium-like anamorphs that
have hyaline conidia held in drops of clear liquid.
These are never formed en mass and never appear
white; they are not morphologically pachybasium-
like. Bissett (1991a) proposed
Trichoderma
sect.
Hy-
pocreanum
Bissett to accommodate them. The
H. ci-
trina
group of species falls within a large clade that
is sister of Pachybasium B5, where they form a sub-
clade (termed ‘‘Pachybasium B1’’ by Kullnig-Gradin-
ger et al 2002) that is sister of several species that
have a typical pachybasium-like morphology.
Hypo-
crea poronioidea
produces a green-conidial
Trichoder-
ma
in addition to the hyaline-conidial, acremonium-

322 M
YCOLOGIA
like synanamorph. The phylogenetic position of
H.
poronioidea
is not known. Samuels and Lodge (1996)
suggested that these acremonium-like anamorphs
could be spermatial because species of the
H. citrina
group and
H. poronioidea
form stromata in culture
and incipient stromata of
H. poronioidea
are associ-
ated with such conidia in culture.
The species of
Trichoderma
that have white conidia
all have a pachybasium-like morphology. Green co-
nidial pigment, coupled with the pachybasium-like
morphology, seems to have been lost only once in
Hypocrea
because the species that we have included
in this study appear to share a common ancestor. We
know of only one species with ‘‘white’’ conidia that
was not included in this study, viz.
H. placentula
Grove (Spooner and Williams 1990). On the basis of
the morphology of its teleomorph and anamorph, we
predict it will be closely related to
H. pachybasioides
and
H. pilulifera.
The ‘‘bright greenish-yellow’’ or ‘‘rosy-buff’’ coni-
diogenous pustules and somewhat narrower conidia
of
T. croceum
Bissett distinguished it from
T. polyspo-
rum
(Bissett 1991b). Unfortunately, only a single iso-
late (DAOM 167068) was cited in the original de-
scription of
T. croceum.
Widden (1979) isolated
Trichoderma
species from soil particles. He identified
a number of morphological species, including LP63.
Although each morphospecies comprised more than
one isolate, LP63 is given in the DAOM catalogue of
cultures as the only available isolate of
T. croceum.
The morphospecies LP63 was isolated rarely from
soil particles in a deciduous forest of southern On-
tario, although the protologue of the new species
T.
croceum
indicates that it was isolated from soil under
Pinus,
according to Widden (1979). Thus the phe-
notypic variability of this unusual yellow-conidial
form is not known and its provenance is suspect.
Nonetheless, conidia of the ex-type culture of
T. cro-
ceum,
in addition to being yellow, are slightly but sig-
nificantly longer and wider than those of the several
collections that are morphologically
T. polysporum.
However, comparison of all isolates identical to
DAOM 167068 in ITS 1
1
ITS 2 sequences and
.
80% similar in RAPD profiles (T
ABLE
II) revealed
that the conidia of the ex-type culture of
T. croceum
fell within the range of all of the isolates. We con-
clude that
T. croceum
is synonymous with
T. polyspo-
rum,
the older name. The constancy of the yellow
conidia suggests that the ex-type strain may represent
a clonally isolated population of
T. polysporum.
Syn-
onymy of
T. polysporum
and
T. croceum
also has been
suggested by Lee and Hseu (2002) on the basis of
UP-PCR and ITS1 sequence analysis.
Yellow conidia occur in at least two additional and
unrelated species of
Trichoderma,
viz.
T. crassum
Bis-
sett and
T. fasciculatum
Bissett. Chaverri et al (2003)
found that
Trichoderma crassum
is closely related to
T. virens
( J. Miller et al) Arx, which has green conid-
ia, and that
T. fasciculatum
is a synonym of the green-
conidial
Hypocrea strictipilosa
Chaverri & Samuels/
T.
strictipile
Bissett. Conidia of
T. stromaticum
Samuels
et al remain white or yellow for a long time and often
do not become green (Samuels et al 2000). In a ge-
nus wanting in useful phenotypic characters, the tax-
onomically seductive trait of yellow conidia unfortu-
nately may be consistent only at the strain level.
Several collections of
Hypocrea
produced
T. polys-
porum
and one of them (G.J.S. 99-90, from Australia)
was identical to a strain of
T. polysporum
(CBS 820.68,
from Germany) considered by Rifai (1969) to be typ-
ical of the species. These
Hypocrea
collections are in-
distinguishable from
H. pachybasioides
Doi (1969),
and we concur with Komatsu (1976) and Gams and
Bissett (1998) that the teleomorph of
T. polysporum
is
H. pachybasioides.
Rifai (1969) reported that
T. polysporum
is com-
mon and cosmopolitan. Some authors have reported
it to be more frequent or even restricted to cooler
climates and soil around conifers (Danielson et al
1973, So¨derstro¨m and Ba˚a˚th 1978, Widden 1979,
Widden and Abitol 1980, Smith 1995). However, the
collections of teleomorphs were found on a diversity
of dicotyledonous trees, never on conifer wood. Our
own collections included isolates from temperate
Northern America, Europe, Japan and Australia, and
we are not aware of any correctly identified isolates
from subtropical or tropical areas. In this regard it is
intriguing that our own Northern Hemisphere col-
lections were made over several years during which
H. pachybasioides
was found infrequently, while the
Australasian collections reported here all were found
during a single two-week collecting trip in 1999. This
observation suggests a greater diversity of the species
in Australasia. A similar Australasian richness was
found for the
H. schweinitzii
(Fr.) Sacc. complex, with
anamorphs in
Trichoderma
sect.
Longibrachiatum
(Kuhls et al 1997). These observations suggest that
the New Zealand, Australia and the Pacific area could
be a center of genetic diversification of
Hypocrea.
As emphasized above, most of the genetic diversity
within
H. pachybasioides
/
T. polysporum
probably is
due to recombination. However, for four isolates
(G.J.S. 86-540, G.J.S. 93-38 [both New York], G.J.S.
90-126 [North Carolina], and G.J.S. 99-244 [New Zea-
land]) some phenotypic differences were seen, be-
cause their ascospores are significantly smaller than
in those of other collections and they grow more
slowly on PDA than the other isolates in the clade.
They also form a well-supported clade that is sister of
the rest of the isolates that are morphologically
H.

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RICHODERMA SPECIES WITH PACHYBASIUM
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pachybasioides
/
T. polysporum.
In the absence of other
phenotypic differences, we do not propose a separate
taxon for them, yet consider them to be clonal iso-
lates in the process of speciation.
Trichoderma minutisporum
previously has not been
linked to a teleomorph. The isolate CBS 901.72 was
nearly identical to the type of
T. minutisporum
(DAOM 167069) in forming a clade with 96% boot-
strap support (F
IG
. 2); additional ascospore isolates
from North America were identical phenotypically
but genetically slightly different from the ex-type iso-
late and from each other (F
IG
. 2). In characters of
the teleomorph, this species scarcely can be distin-
guished from many other
Hypocrea
species; it is its
anamorph that distinguishes it, and no described
Hy-
pocrea
has been shown to have an anamorph with the
morphology of
T. minutisporum.
The teleomorph is
proposed here as the new species
H. minutispora.
Contrary to
H. pachybasioides
/
T. polysporum,
which is
widely distributed in north temperate regions, being
known from Asia, North America, Europe and Aus-
tralasia,
H. minutispora
/
T. minutisporum
is apparent-
ly a species of temperate North America and north-
ern Europe.
The single isolate G.J.S. 99-198 from New Zealand
is genetically distinct within the minutisporum clade.
Its anamorph is similar to
T. minutisporum
but it can
be distinguished from
H. minutispora
/
T. minutispo-
rum
in having larger conidia and in the structure of
its conidial pustules. It is described below as the new
species
H. lacuwombatensis.
The isolates G.J.S. 91-60 and G.J.S. 99-188, and
G.J.S. 99-222 are genetically distinct within the polys-
porum clade. The first two of these apparently are
related closely to
H. pilulifera
/
T. piluliferum
(F
IG
.2)
but differ in having ellipsoidal conidia. It is proposed
here as the new species
H. parapilulifera.
The isolate
G.J.S. 99-222 is distinguished by pustules that com-
prise long fertile branches from which short lateral
branches arise; many of these branches extend be-
yond the surface of the pustule giving the pustule a
stellate aspect (F
IG
s. 17, 18). It is proposed here as
H. stellata.
We do not propose species names for the
Trichoderma
anamorphs of these
Hypocrea
species be-
cause they are known only from one or two collec-
tions and have not been encountered in nature as
their anamorphs.
DESCRIPTIONS OF THE SPECIES
1. Hypocrea pachybasioides Doi, Bull. Natn. Sci.
Mus. Tokyo 12:685. 1969.
F
IGS
. 4–6, 13–16, 24–27, 33–51
Anamorph:
Trichoderma polysporum
(Link : Fr.) Rifai,
Mycol. Pap. 116:18. 1969. F
IGS
. 13–16, 24–27, 45–
51
[
Sporotrichum polysporum
Link, Mag. Ges. Naturf.
Freunde Berl. 7:34. 1815: Fries, Syst. Mycol. 3:424.
1832.
(see Rifai [1969] for extensive anamorph synony-
my)
Stromata solitary, sometimes gregarious but rarely
cespitose, (0.3–)0.5–2.0(–6.0)
3
(0.2–)0.5–1.5(–4.5)
mm, shape of stromata variable, mostly rounded to
elongate, or sometimes irregular in outline, particu-
larly when aggregated or cespitose, pulvinate; usually
with margins of the stromata fully attached to the
substratum, but sometimes slightly constricted at the
base; color variable among different collections and
different stages of the same collection, mostly brown
to reddish-brown, some collections yellow or light
brown particularly when young, collections from Ja-
pan purple-brown to black-brown; color of most stro-
mata nearly uniform, but margins of young stromata
often white. All tissues KOH-. Stromatal surface vari-
ously wrinkled or creased, plane but infrequently
slightly tuberculate from perithecial apices. Ostiolar
openings mostly visible as purple-brown to blackish-
brown flat or slightly raised dots (F
IGS
. 4–6). Cells of
the stroma surface in face view elongate, angular or
irregular in outline, (2–)4–8(–18)
3
2–5(–8.5)
m
m,
usually reddish-brown, rarely gray to whitish-gray, cell
walls 0.5–1.0
m
m thick (F
IG
. 35). Stroma surface re-
gion (10–)20 –40(–50)
m
m thick, cells elongate, an-
gular or compressed, occasionally rounded, (2–)3–
5(–10)
3
(1–)2–4(–7)
m
m, cell walls 0.5–1.0
m
m
thick (n
5
10), brown, KOH- (F
IGS
. 36, 37). Hairs
arising from the stroma surface scattered and incon-
spicuous, hyphal, (4–)5–7(–9)
m
m long, (2.0–)2.5–
3.5(–6.0)
m
m wide at the base, hyaline to light brown
(F
IG
. 38). Cells immediately below the stroma surface
hyphal, thin-walled, hyaline. Tissue below the peri-
thecia tending to comprise compact
textura intricata
or intertwined hyphae, less frequently cells pseudo-
parenchymatous, (3–)6–10(–22)
3
(2–)4–5(–8)
m
m,
thin-walled, hyaline, KOH- (F
IGS
. 39, 40). Perithecia
immersed in the stroma, densely disposed, mostly
globose to subglobose, laterally compressed and pyr-
iform to clavate when greatly compacted, (75–)150–
270(–346)
m
m high, (53–)100–150(–232)
m
m wide,
ostiolar canal (18–)50–80(–116)
m
m long, cells of the
perithecial wall brown or reddish-brown to hyaline;
ostiolar region not sharply delimited from the sur-
rounding tissue of the stromal surface (F
IGS
. 33, 34).
Asci cylindrical, (48–)75–100(–133)
3
(3.5–)4.5–
6.0(–8.5)
m
m, tip thickened and with a pore (F
IGS
.
41–43). Part-ascospores hyaline, uniseriate, finely spi-
nulose, dimorphic. Distal part-ascospores globose to
subglobose or conical, (2.2–)3.2–4.5(–6.0)
3
(2.0–)

324 M
YCOLOGIA
F
IGS
. 4–12.
Hypocrea
stromata (herbarium specimens, stereo microscope). 4–6.
H. pachybasioides.
Stromata diverse in
appearance, either solitary or aggregated, from vinaceous to light brown or buff; note ostiolar openings usually visible as
dark or viscid dots against the lighter ground of the stroma. F
IGS
.45G.J.S. 90-116, 5 5from TMI 8217, 6 5TMI 8215.
F
IG
.7.
H. parapilulifera
(G.J.S. 91-60). F
IG
.8.
H. pilulifera
(SHD 2965). F
IG
.9.
H. stellata
(G.J.S. 99-222). F
IGS
. 10, 11.
H.
minutispora
(10 5G.J.S. 90-115, 11 5G.J.S. 90-82). F
IG
. 12.
Hypocrea lacuwombatensis
(G.J.S. 99-198). Scale bars: F
IGS
. 4–7,
9–12 51 mm, 8 50.5 mm.

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F
IGS
. 13–23. Tufts and pustules of
Trichoderma
anamorphs of
Hypocrea
species, all from CMD at 20–21 C within 10 d.
F
IGS
. 13–16.
H. pachybasioides
/
T. polysporum
(13 5G.J.S. 99-207, 14 5G.J.S. 99-221, 15, 16 5DAOM 167068, the ex-type
culture of
T. croceum
). F
IG
. 17.
Hypocrea parapilulifera,
G.J.S. 91-60. F
IG
.18
H. stellata,
G.J.S. 99-222. F
IGS
. 19–21.
H. minu-
tispora
/
T. minutisporum
(19 5DAOM 212372, 20 5DAOM 191102, 21 5DAOM 178046). F
IGS
. 22, 23.
H. lacuwombatensis,
G.J.S. 99-198. All stereo. Scale bars: F
IGS
. 13, 16, 20, 23 50.1 mm, 18, 19, 21 50.2 mm, 14, 15, 17, 22 51 mm.

326 M
YCOLOGIA
F
IGS
. 24–32. Line drawings of conidiophores and conidia of
Trichoderma
anamorphs of
Hypocrea
species. 24–27.
H.
pachybasioides
/
T. polysporum.
Note cork-screw-shaped, sterile, often spinulose elongations of conidiophores (24 5G.J.S. 90-
63, 25 5G.J.S. 86-540, 26, 27 5G.J.S. 88-44). F
IG
. 28.
Hypocrea parapilulifera,
G.J.S. 91-60. F
IGS
. 29–32.
H. minutispora
/
T.
minutisporum
(29, 32 5G.J.S. 90-80, 30 5CBS 901.72, 31 5G.J.S. 90-81). Scale bars 510 mm.

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F
IGS
. 33–44.
Hypocrea pachybasioides
/
T. polysporum.
33, 34. Median longitudinal sections through perithecia within stro-
mata (33 5TMI 8217, 34 5TMI 8221). 35. Angular cells at the surface of the stroma seen in face view, G.J.S. 99-155. 36,
37. The surface region of the stroma formed of angular cells; intertwined hyphae form immediately below the surface region
(36 5G.J.S. 90-116, 37 5G.J.S. 99-223). 38. The surface of the stroma showing one of the scattered hairs arising, G.J.S. 99-
207. 39, 40. Cells of the interior of the stroma below perithecia, typically compacted and with a strong tendency to be hyphal
(F
IG
. 39, G.J.S. 99-181); less frequently the cells are pseudoparenchymatous (F
IG
. 40, G.J.S. 89-135). 41–43. Asci with asco-
spores. Thickening of the ascal apex can be seen in F
IGS
. 42 (G.J.S. 90-116) and 43 (G.J.S. 88-44, stained with 1% [aq.]
phloxine); a pore can be seen in F
IG
. 43. 44. Discharged part-ascospores, G.J.S. 88-59. The distal parts are ellipsoidal to
subglobose and the proximal parts are wedge-shaped. All D
IC
. Scale bars: F
IGS
. 35, 43, 44 510 mm, 36–42 520 mm, 33, 34
5100 mm.
3.0–4.0(–5.7)
m
m. Proximal part-ascospores mostly
subglobse to oblong, sometimes wedge-shaped or at-
tenuated toward the base, tending to be more oblong
toward the base of the ascus, (2.5–)3.7–5.2(–7.2)
3
(1.7–)2.5–3.5(–4.5)
m
m(F
IG
. 44).
Cultures and anamorph.
Optimum temperature for
growth on PDA 25 C, no growth occurring at 35 C.
Colony radius on PDA at 25 C after 3 d in darkness
(14–)18–34(–46) mm,
,
15 mm at 30 C. Conidia typ-
ically not forming on PDA in darkness within 6 d, or
rarely after 4 d at 20 C; yellow pigment sometimes
forming in colony reverse after 4 d in darkness at 20
and 25 C. No odor detected on PDA or CMD. Colony
radius on CMD at 25 C in light after 6 d 65 mm,
within 10 d small white hyphal tufts scattered
throughout the colony and conidia forming on the
tufts after 13 d. Conidial pustules white to cream-col-
ored, solitary or rarely aggregated, pulvinate, (0.2–)
0.8–1.5(–13.5) mm diam, dense, appearing velvety
owing to numerous, projecting, sterile extensions of
conidiophores (hairs; F
IGS
. 13,16, 45–47), produc-
tion of pustules increasing to 30 d. Hairs flexuous,
sinuous to corkscrew-shaped, typically attenuated to
a narrow tip, septate, thin-walled, roughened by nu-
merous small warts near the apices, unbranched for
a great distance, sometimes producing a single phial-
ide at the tip (F
IGS
. 24–27, 46, 47). Fertile branches
of conidophores arising from the base of the hairs at

328 M
YCOLOGIA
F
IGS
. 45–51.
Hypocrea pachybasioides
/
Trichoderma polysporum,
anamorph. 45. Part of a conidial pustule showing the cork-
screw-like extensions of conidiophores projecting beyond the pustule, ATCC 19650. 46, 47. Extensions of conidiophores;
typically ornamented and sterile but often smooth and sometimes producing a phialide at the tip as in F
IG
. 47 (46 5G.J.S.
90-28, 47 5G.J.S. 99-219). 48. Phialides, Tr 54. 49. Conidia, Tr 80. 50. Phialides clustered on short, broad branches at the
base of sterile extensions of conidiophores, DAOM 167068. 51. Chlamydospores, Tr 54. F
IGS
. 52–55.
Hypocrea pilulifera
/

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LIKE CONIDIOPHORES
←
Trichoderma piluliferum,
conidiophores and conidia. 52–54. Conidiophores. Note convergently arranged phialides and sterile
conidiophore bases. (52 5SHD 2974 dry culture, 53, 54 5CBS 814.68). 55. Conidia, CBS 814.68. F
IGS
. 46–50, 52–55 5
DIC; 51 5BF. Scale bars: F
IG
.4550.5 mm; 46–48, 52–54 520 mm; 49, 50, 55 510 mm; 51 550 mm.
approximately 90
8
to the hairs, or conidiophores
lacking sterile extensions. 1
8
conidiophore branches
mostly short, comprising one or few cells, and in-
creasing in length with distance from the tip, pro-
ducing phialides directly and also rebranching to
produce 2
8
branches. 2
8
branches typically unicellular
and producing 1–4 phialides from the tip. Phialides
lageniform and more or less constricted to form a
neck, (3.0–)3.7–6.7(–13.5)
m
m long, (1.7–)2.5–3.5
(–5.0)
m
m at the widest point, L/W
5
(0.9–)1.5–
2.2(–5.0), (1.0–)1.7–2.7(– 4.0)
m
m wide at the base.
Cell supporting the phialides (1.7–)2.5–4.5(–9.7)
m
m
wide (F
IGS
. 48, 50). Conidia ellipsoidal, occasionally
oblong or subglobose, (1.2–)2.5–3.5(–5.5)
3
(1.0–)
1.5–2.5(–3.0)
m
m, L/W
5
(0.5–)1.0–2.0(–2.5), hya-
line, smooth (F
IGS
. 26, 49). Chlamydospores terminal
or intercalary within hyphae, globose or subglobose,
(3.0–)5.5–9.0(–23.5)
3
(2.8–)5.5–8.0(–16.0)
m
m,
smooth or somewhat spinulose (F
IG
. 51).
Known distribution.
Australia, Canada, Germany,
Italy, Japan, Korea, New Zealand, Switzerland, United
Kingdom (England), United States.
HOLOTY PE. JAPAN. Otsuno, Kochi City, on bark,
3 May 1966,
Y. Doi TNS.D-77
(TNS-F-190528, holo-
type not available, ISOTY PE: NY!).
Additional
Hypocrea
specimens examined.
AUSTRALIA.
NEW SOUTH WALES: Blue Mountains, Morton National
Park, vicinity of Bundnadoon, Fairy Bower Track, on bark,
19 Aug 1999,
G.J.S. et al 8740
(BPI 74667, culture G.J.S. 99-
155); Fairy Bower Track, on bark, 19 Aug 1999,
G.J.S. 8741
(BPI 746680, culture G.J.S. 99-159). VICTORIA: vic. Hea-
lesville, Toolangi State Forest, Myers Creek Road, Wirra-
walla Rainforest Walk and Myrtle Walking track in Myrtle
Gully, altitude. 575 m, on bark of recently dead tree, 23 Aug
1999,
G.J.S. 8599
(BPI 746812,culture G.J.S. 99-219); Otway
Ranges, Melba Gully State Park, Madsen’s Track along Jo-
hanna River, altitude 350 m, on bark, 27 Aug 1999,
G.J.S.
8636
(BPI 746848; culture G.J.S. 99-221 5BPI 112261);
Yarra Ranges National Park, at intersection of road to Don-
na Buang and Acheron Way, Donna Buang Gallery along
stream, altitude ca. 600 m, on bark of
Nothofagus
sp.,
G.J.S.
8607
(BPI 746821, culture G.J.S. 99-223). CANADA. QUE-
BEC: Gatineau State Park, on decorticated wood, possibly
on an ascomycete, 20 June 1987,
R.P. Korf
(NY). ITALY.
VITERBO PROV.: Bomarzo, along river Vezza, on wood of
Quercus
sp., Nov 2002,
W. Gams
(CBS 111723, specimen
and culture). JAPAN. HIKUI PREF.: Konjyo-cho, Nanjyo-
gun, 18 Jul 1968,
M. Komatsu
(TMI 8218). HYOGO-PREF.:
Fukuzaki-cho, kanzaki-gun, 23 Feb 1965,
M. Komatsu
(TMI
8215). HYOYO-PREF.: Nahokaichi, Kasumi-cho, Kinosaki-
gun, 13 May 1968, Akiyama (TMI 8258). TOTTORI PREF.:
4 Jul 1968,
M. Komatsu
(TMI 8220, 8221); Bogaki, Tottori-
shi, 13 Apr 1964,
M. Komatsu
(TMI 8084); Tottori-shi, 4 Jul
1968 (TMI 8217). NEW ZEALAND. BULLER: Lewis Pass,
St. James Walkway, altitude ca. 900 m, 428249S 1728249E, on
bark of
Nothofagus menziesii,
9 Sep 1999,
G.J.S. & S. Dodd
8736
(BPI 842229; culture G.J.S. 99-90 5CBS 112257).
WESTLAND: vicinity of Franz Josef, trail to Lake Wombat,
438259S 1708219Eto438219S 170899E, mixed podocarp for-
est, altitude 200–275 m, on bark, 3 Sep 1999,
G.J.S. & S.
Dodd 8684
(BPI 746623; culture G.J.S. 99-207 5CBS
112256); WESTLAND: vicinity of Hokitika, Lake Kaniere,
Track along S side of lake between Sunny Bight and Hiiker
Creek, mixed podocarp with
Dacrydium cupressinum
(Rimu),
Prumnopitys taxifolia
(Matai), and
Metrosidros um-
bellata
(Southern Rata), altitude 0 m, 438S 171879E, on bark
of small branches of recently dead tree,
G.J.S. & S. Dodd
8672
(BPI 746612, culture G.J.S. 99-224). SWITZERLAND.
KT. GRAUBU
¨NDEN: vicinity of Davos, Dischmatal, altitude
1500–1700 m, on decorticated wood of
Picea
sp., 4 Sep
1990,
G.J.S.
(BPI 1107148; culture G.J.S. 90-28 5BBA
70310 5CBS 112262). UNITED STATES. MARYLAND:
Garrett County, 5 miles North of Barton, Little Savage River
ravine, on bark, 23 Sep 1989,
G.J.S. et al
(NY; culture G.J.S.
89-121 5CBS 112258). NEW YORK: Dutchess County, east
side of Pawling, Pawling Nature Reserve, Nature Conser-
vancy, on bark, 8 Oct 1990,
G.J.S. & C.T. Rogerson
(BPI
1107154; culture G.J.S. 90-63 5CBS 112260); Hamilton
County, Long Point, Racquette Lake, on decorticated wood
of
Fagus,
6 Sep 1986,
J.H. Haines
(NY; culture G.J.S. 86-540
5BBA 70311 5CBS 112267); Schuyler County, Van Etten,
Arnot Forest, Cornell University, Bonfield road, vicinity of
Bonfield Creek, on decorticated wood, 9 Oct 1993,
K.F. Ro-
drigues
(BPI 802508 culture G.J.S. 93-38). NORTH CARO-
LINA: Clay County, Standing Indian Campground, off U.S.
64, on Aphyllophorales on
Betula,
15 Oct 1990,
Y. Doi, A.Y.
Rossman & G.J.S.
(BPI 1107182; culture G.J.S. 90-116 5
CBS 112259); Jackson County, Ellicott Rock Trail, on re-
cently dead tree, 1989,
G.J.S. et al
(NY, culture G.J.S. 89-
135); Macon County, Blue Valley, off Clear Creek Road,
along Overflow Creek, 358009N, 838159W, on fungi on bark,
16 Oct 1990,
Y. Doi, A.Y. Rossman & G.J.S.
(BPI 1107168,
culture G.J.S. 90-126); Swain County, Great Smoky Moun-
tains, National Park, 5 Mile North of Deep Creek Camp
Ground, Indian Creek Trail, on bark, 27 Sep 1988,
K.F. Ro-
drigues, C.T. Rogerson, G.J.S., E. Parmasto & R.H. Petersen
(NY, G.J.S. 88-44); Swain County, Great Smoky Mountains
National Park, 3.6 mile N of Deep Creek Camp ground,
Indian Creek Trail, on decorticated wood, 27 Sep 1988,
K.F.
Rodrigues et al
(NY, culture G.J.S. 88-59).
Commentary. Hypocrea pachybasioides
is possibly the
easiest species of the genus to identify when its ana-
morph,
T. polysporum,
is known. Despite the absence

330 M
YCOLOGIA
of strongly diagnostic characters in the teleomorph,
the anamorph of this common species is almost in-
variant. Several isolates of
T. polysporum
are listed in
the culture catalogue of the Centraalbureau voor
Schimmelcultures, including isolates from Germany,
Korea, the Netherlands, and U.K.; we have not stud-
ied those isolates but do not doubt their identity.
The shape, degree of aggregation and color of
stromata of
H. pachybasioides
are variable. Typically
stromata are rounded to elongate; when aggregated
they are irregular in outline. Most of the collections
from Japan were cespitose in lines, and stromata of
one Japanese collection were somewhat effused over
the substratum. The stromata of most collections are
reddish-brown (i.e., G.J.S. 99-90, 99-116, 99-155, 99-
207, 99-219, 99-220, 99-221, 99-222, 99-223, TMI
8215, TMI 8218); immature stromata tend to be yel-
low to yellowish-brown (i.e., G.J.S. 88-44, 88-59, 89-
135, 90-63, 99-159, TMI 8258) and old stromata are
dark brown to rusty (i.e., G.J.S. 99-224, TMI 8217,
8220, 8221). The specimen TMI 8084, from Japan,
was unusual in having pale vinaceous stromata; as-
cospores of this collection were smaller than is typical
of the species.
Trichoderma polysporum
frequently is cited in the
biological control and ecology literature (Domsch et
al 1980). It is one of the slowest-growing species in
the genus. Its phenotypic similarity to the distantly
related, common soil fungus
Tolypocladium inflatum
W. Gams (
5
To. niveum
[O. Rostrup] Bissett, nom.
rejic.) has led to its misidentification (Samuels 1996).
The species has been identified as the producer of
the immunosuppressant cyclosporin A, but it is now
known that cyclosporin A is produced by
To. inflatum
(Horsburgh et al 1980, Thali 1995). Iida et al (1999)
isolated a peptidic immunosuppressant from
T. polys-
porum.
Komatsu (1976) found that
H. pachybasioi-
des
/
T. polysporum
is common on and inside of bed
logs used in cultivation of shiitake mushrooms in Ja-
pan and was strongly antagonistic to the mycelium,
severely diminishing mycelial growth and mushroom
production. Nelson (1982) found
T. polysporum
to be
a normal member of the soil biota of Douglas fir
(
Pseudotsuga menziesii
) and he and his collaborators
isolated it from roots and stumps of Douglas fir that
were infected with
Phellinus weirii
(Nelson et al 1987,
1995; Goldfarb et al 1989).
Trichoderma polysporum
has shown some promise in biological control of
Pyth-
ium aphanidermatum
( Jackisch-Matsuura and Menez-
es 1999) and
Phytophthora cinnamomi
(Kelley 1977).
Barbosa et al (2001) found that
T. polysporum
was
highly antagonistic to
Cladosporium cladosporioides
on
passion fruit. A combination of arbuscular mycorrhi-
zal fungi,
Rhizobium
and
T. polysporum
slowed fusar-
ium-induced seed rot and damping-off in
Dalbergia
sissoo
while promoting tree growth (Singh et al
2002).
2. Hypocrea
pilulifera
J. Webster & Rifai, Trans. Br.
Mycol. Soc. 51:511. 1968. F
IGS
. 8, 52–55
Anamorph. Trichoderma piluliferum
J. Webster & Rifai,
Mycol. Pap. 116:16. 1969. F
IGS
. 52–55
The type specimen of
H. pilulifera
(SHD-M 2965,
K!) comprises 4 pieces of
Juncus
culms, each 1.0–1.5
cm long. Only one stroma remains. It is discoidal,
broadly attached and not constricted at the base, an-
chored to the substratum by a dense halo of white
hyphae. The surface is pale orange and the ostiolar
openings are slightly papillate and darker orange
(F
IG
. 8). Because only one stroma remains, a micro-
scopic preparation was not made. The stroma is
KOH-. Part-ascospores were described and illustrated
by Webster and Rifai (1968) as being dimorphic, with
the distal part subglobose and the proximal part el-
lipsoidal to wedge-shaped, hyaline and spinulose, 4–
6
3
3.0–4.5
m
m.
No perithecia remain on a paratype cited by Web-
ster and Rifai (SHD-2638, K[m] 64378!). The ex-type
culture of
T. piluliferum
was derived from ascospores
of this specimen. This specimen comprises one piece
of rotten, decorticated wood (
Betula
) that has a few
clumps of white conidia. There are also four dry cul-
tures (malt-extract agar) with 3–4 mm diam, white
conidial pustules. No phialides remain in these dry
cultures. Conidia (based on dry conidia from SHD
2974 and living culture CBS 814.68) are globose to
subglobose, (2.5–)2.7–3.2(–3.5)
3
(2.5–)2.5–3.0
(–3.2)
m
m, L/W
5
(0.9–)1.0–1.2(–1.3), hyaline and
smooth. The ex-type culture of
T. piluliferum
has
been preserved as CBS 814.68. The following descrip-
tion is based on that culture.
Cultures and anamorph.
Optimum temperature for
growth on PDA 20–25 C, no growth occurring at 30
C. Colony radius on PDA at 20–25 C after 3 d in
darkness 21–23 mm and not reaching 60 mm after 6
d. Colony radius on CMD at 20 and 25 C in light
after 5 d ca. 65 mm, sterile. Conidia not forming on
PDA in darkness within6datanytemperature but
forming on PDA incubated at 20 C under light within
2 wk and on SNA within 17 d; conidia slower to form
at 25 C. Pale yellow pigment forming in PDA after
96 h. No odor detected on any medium. Conidial
production sparse, in minute, white, cottony tufts.
Conidiophores branching more or less symmetrically
near the tip and with a short stipe; branches arising
at 90
8
, progressively longer with distance from the tip;
1
8
branches often rebranching at right angles. Phial-
ides arising at the tips of the 1
8
and 2
8
branches in a
convergent whorl of 2– 4, lageniform, often constrict-

331L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
ed sharply below the tip, (4.5–)5.0–6.7(–8.0)
m
m,
width of the phialides at the widest point (2.2–)2.7–
3.7(–4.2), L/W
5
(1.3–)1.5–2.3(–3.2), base of the
phialides (1.5–)2.0–2.7(–3.5)
m
m. Cell supporting
the phialides (2.5–)3.0–4.0(–5.5)
m
m wide (F
IGS
. 52–
54). Conidia globose to subglobose, base often trun-
cate, 2.5–3.5
m
m diam, L/W ratio 0.9–1.3, hyaline,
smooth (F
IG
. 55).
Known distribution.
Germany, Denmark, U.K.
HOLOTY PE. UNITED KINGDOM. DERBYSHIRE:
Glossop, Chunal Moore, on dead culms of
Juncus ef-
fusus,
11 Jul 1965,
J. Webster
(SHD-M 2965, K[m]
64379!).
Additional specimens examined
(none with perithecia, an-
amorph only). UNITED KINGDOM. DERBYSHIRE: Chat-
sworth Park, on
Fomes annosus
(5
Heterobasidion annosum
),
13 Oct 1963,
J. Webster & M.A. Rifai
(SHD-M 3024, K[m]
64380); Kinder, Edale Croos, on
Juncus effusus,
17 Aug
1969,
J. Webster
(K[m] 644040). YORKSHIRE: near Clith-
eroe, Lancashire., Dunsop Bridge, on dead wood of
Betula,
23 Sep 1962,
J. Webster
(SHD-M 2638, K[m] 64378; culture
CBS 814.68; SHD-M 2639, K[m] 64381).
Commentary. Hypocrea pilulifera
/
T. piluliferum
is
easily recognized in the genus when the anamorph
is known because it is the only known species that
has globose, hyaline conidia. It is unfortunate that
the ex-type culture of
T. piluliferum
(on wood) was
not derived from the type specimen of
H. pilulifera
(on
Juncus
). From the original description of
H. pi-
lulifera
(Webster and Rifai 1968) it is not clear wheth-
er the described anamorph originated from the type
collection or the paratype, which is the source of the
type of
T. piluliferum.
In our experience it would be
unusual for a species of
Hypocrea
to occur on both
Juncus
and rotting wood, although the anamorph
drawn by Webster and Rifai (1968: F
IG
. 1c, d) does
have subglobose conidia. Nonetheless,
T. piluliferum
is a distinctive species that thus far is restricted to
northern Europe. The culture catalogue of Centraal-
bureau voor Schimmelcultures (http://www.cbs.
knaw.nl) lists cultures isolated from forest soil in Den-
mark and Germany and roots of
Pseudotsuga menziesii
in the Netherlands, and the original authors report-
ed collections of
H. pilulifera
from basidiomata of
Fomes annosus
(
5
Heterobasidion annosum
), culms of
Juncus effusus
and wood of
Betula
and
Fagus.
Hypocrea placentula
W.B. Grove originally was de-
scribed from herbaceous tissue, including culms of
Juncus effusus
in the U.K. This species and its ana-
morph were described by Spooner and Williams
(1990). The anamorph of
H. placentula
differs from
T. piluliferum
in having ellipsoidal to oblong conidia.
Conidia of the new species
H. parapilulifera
and
H.
stellata,
both of which occur on wood, are also white
and ellipsoidal.
In being restricted to northern Europe, H.
piluli-
fera
/
T. piluliferum
is similar to the unrelated
H. au-
reoviridis
/
T. aureoviride
(Lieckfeldt et al 2001).
3. Hypocrea parapilulifera B.S. Lu, Druzhinina & Sa-
muels, sp. nov. F
IGS
. 7, 17, 28, 56–68
Stromata solitaria vel gregaria, pulvinata, rotunda vel ob-
longa, 1.0–2.5(–3.5) 3(0.7–)1.5–2.5(– 4.0) mm, ostiolo
conspicuo. Asci cylindrici, (72–)75–95(–100) 3(3.7–)4.0–
5.5(–6.7) mm, apice incrassato, poro apicali praediti. Ascos-
porae hyalinae, bicellulares, ad septum disarticulatae; parte
distali globosa vel subglobosa, (2.8–)3.5–4.5(–5.0) 3(2.7–)
3.2–4.0(– 4.7) mm, parte proximali ellipsoidea vel oblonga,
(3.5–)4.0–5.2(–6.0) 3(2.5–)2.7–3.5(– 4.0) mm, hyalinae,
verrucosae. Anamorphe
Trichoderma.
Phialides (3.5–)4.5–
7.5(–11.0) 32.4–3.5(–4.0) mm. Cellulae subterminales con-
idiophori (2.5–)3.0–5.0(–10.5) mm. Conidia hyalina, aggre-
gata alba, ellipsoidea vel oblonga, 2.5–3.7(–4.5) 31.7–2.0
(–2.5) mm, L/W 5(1.2–)1.5–1.9(–2.3), laevia.
Holotype.
BPI 112832 (ex-type culture G.J.S. 91-60
5
CBS 112771).
Etymology.
referring to a similarity to
H. pilulifera.
Anamorph. Trichoderma
sp. F
IGS
. 17, 28, 62–68
Stromata solitary or aggregated, rounded or elon-
gate, occasionally irregular in outline, 1.0 –2.5(–3.5)
3
(0.7–)1.5–2.5(–4.0) mm, margin slightly free from
substratum, yellowish-brown to brown with white-yel-
low margin, surface smooth, perithecial apices barely
protruding. All tissues KOH-. Ostiolar openings visi-
ble as slightly raised, purple-brown spots (F
IG
. 7).
Cells of the stroma surface in face view round, elon-
gate or angular, (2.5–)4.0–7.5(–16.5)
3
(1.5–)2.5–
3.5(–5.5)
m
m, reddish-brown, cell-walls 0.5–1.0
m
m
thick (F
IG
. 58). Stroma surface (22–)25–45(–60)
m
m
thick, cells round, elongate, or occasionally angular,
(1.5–)2.5–3.5(–5.5)
3
(1.5–)2.0–3.0(–5.0)
m
m, cell
walls 0.5–1.0
m
m thick, brown (F
IG
. 57). Hair-like ex-
tensions of cells at the stroma surface inconspicuous,
scattered, 3.5–4.5
m
m long, 2.5–3.5
m
m wide at base,
hyaline to light brown. Cells immediately below stro-
ma surface, hyphal, thin-walled. Tissue below peri-
thecia tending to be of a compact
textura intricata
or
occasionally pseudoparenchymatous, (2.4–)4.5–
10.0(–18.0)
3
(2.5–)3.5–6.5(–7.5)
m
m, hyaline (F
IG
.
59). Perithecia immersed in stroma, densely dis-
posed, globose to subglobose, (154–)180–240(–260)
m
m high, (70–)90–125(–155)
m
m wide, ostiolar canal
(58–)65–90(–103)
m
m long, wall light brown to yel-
lowish-brown; ostiolar region not sharply delimited
from the surrounding stroma tissue (F
IG
. 57). Asci
cylindrical, (72–)75–95(–100)
3
(3.7–)4.0–5.5(–6.7)
m
m, tip thickened and with a pore (F
IGS
. 60, 61).
Part-ascospores uniseriate, hyaline, finely spinulose,
dimorphic; distal part-ascospores globose to subglo-

332 M
YCOLOGIA
F
IGS
. 56–68.
Hypocrea parapilulifera
and its
Trichoderma
anamorph. 56. Median longitudinal section of a perithecium
immersed in a stroma. 57. Ostiolar region of a perithecium and the stroma surface. 58. Cells of the stroma surface in face
view. 59. Hyphal cells of the interior of the stroma below a perithecium; perithecial base seen on the right. 60, 61. Asci and
part-ascospores. 62. Pustules. Note sterile extensions of conidiophores projecting beyond the pustule. 63–68. Sterile extensions
of conidiophores, phialides and conidia. F
IGS
. 56–62 from G.J.S. 91-60, 63–68 from G.J.S. 99-188. F
IGS
. 56–61, 64, 65 5DIC;
62 5stereo; 63, 66, 67 5FL; 68 5PC. Scale bars: F
IG
.565100 mm, 57–60, 65, 67, 68 520 mm; 61, 63, 64 510 mm; 62
51 mm, 66 540 mm.

333L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
bose, (2.8–)3.5–4.5(–5.0)
3
(2.7–)3.2–4.0(– 4.7)
m
m,
proximal part-ascospores ellipsoidal to wedge-
shaped, (3.5–)4.0–5.2(–6.0)
3
(2.5–)2.7–3.5(–4.0)
m
m(F
IG
. 61).
Cultures and anamorph.
Optimum temperature for
growth on PDA 20–25 C, no growth occurring at 30
C. Colony radius on PDA after 3 d in darkness 20–
25 mm. Conidia typically forming within 2–4 wk on
CMD and SNA under light in white pustules. Pustules
scattered around the periphery of the colony, 0.5–1
mm diam, hemispherical,
6
cottony, with conspicu-
ous sterile hairs (F
IGS
. 17, 62). Sterile extensions of
conidiophores (hairs) sinuous, septate, thin-walled,
typically attenuated to a narrow tip, with conspicuous
granular accretions over the terminal 80–100
m
m,
branching ca. 100
m
m below the tip. Fertile branches
of conidiphores arising from the base of the hairs at
approximately 90
8
to the hairs, or conidiophores
lacking sterile extensions (F
IG
. 28). 1
8
conidiophore
branches comprising one or a few cells, progressively
longer with increasing distance from the tip of the
conidiophore, producing phialides directly or pro-
ducing 2
8
branches. 2
8
branches unicellular and pro-
ducing 1–3 phialides along the length and at the tip.
Phialides short and broad, (3.5–)4.5–7.5(–11.0)
m
m,
width of the phialides at the widest point 2.4–3.5
(–4.0)
m
m, L/W of phialides
5
1.0–4.1, base of the
phialides 1.4–3.0(–3.5)
m
m(F
IGS
. 28, 63–68). Cells
supporting phialides (2.5–)3.0–5.0(–10.5)
m
m. Co-
nidia ellipsoidal to oblong, 2.5–3.7(–4.5)
3
1.7–2.0
(–2.5)
m
m, L/W
5
(1.2–)1.5–1.9(–2.3), hyaline,
smooth (F
IGS
. 28, 64). Chlamydospores globose to
subglobose, terminal or intercalar y in hyphae,
(4.0–)6.0 –8.5(–10.5)
3
(3.7–)5.5–8.0(–9.5)
m
m, hy-
aline.
Known distribution.
Australia, United States.
HOLOTY PE. UNITED STATES. VIRGINIA: Giles
County, Mountain Lake Biological Station, Little
Spruce Bog, 37
8
22
9
N, 80
8
31
9
W, altitide 1170 m, on
decorticated wood, 17 Sep 1991,
G.J.S. et al
(BPI
112832, culture G.J.S. 91-60).
Additional specimen examined.
AUSTRALIA. VICTORIA:
between Yarram and Traralgon, Tarra Valley, Tarra-Bulga
National Park, Tarra Rainforest Walk, altitude 250 m, on
bark of
Nothofagus
sp., 22 Aug 1999,
G.J.S. 8556
(BPI
746770; culture G.J.S. 99-188 5CBS 112264).
Commentary.
Under our conditions
Hypocrea par-
apilulifera
produces conidia very slowly and in small
numbers. Although the Australian collection
G.J.S.
8556
is old and in poor condition, there is no doubt
about its identity with the North American collection.
4. Hypocrea stellata B.S. Lu, Druzhinina & Samuels,
sp. nov. F
IGS
. 9, 18, 69–81
Stromata solitaria, pulvinata, rotundata, oblonga vel ir-
regularia, 1.6–12.8 31.3–2.4 mm, ostiolo conspicuo. Asci
cylindrici, (72–)80–90(–100) 34.5–6.0(–7.0) mmmm, apice
incrassato, poro apicali praediti. Ascosporae bicellulares, hy-
alinae, spinulosae, ad septum disarticulatae; parte distali
globosa vel subglobosa, 3.2–4.0(– 4.5) 33.0–3.7(–4.0) mm,
parte proximali oblonga vel cuneata, (3.0–)3.5–4.5(–6.7) 3
(2.2–)2.5–3.2 mm. Anamorphe
Trichoderma.
Phialides cylin-
dricae vel ampulliformes, (4.5–)5.5–8.0(–9.5) 3(2.0–)2.7–
3.5(–3.7) mm, L/W 5(1.5–)1.8–2.8(–3.1). Cellulae subter-
minales conidiophori (2.7–)3.5–5.0(–6.2) mm. Conidia hy-
alina, late ellipsoidea vel ovoidea, (2.0–)2.5–3.5(– 4.0) 3
1.7–2.2(–2.7) mm, L/W 5(1.0–)1.2–1.6(–1.7), laevia.
Holotype. Samuels & S. Dodd 8671
(PDD, ISO-
TY PE: BPI 746610; ex-type culture G.J.S. 99-122
5
CBS 112265).
Etymology. stellata
referring to the long, projecting
conidiophores.
Anamorph.
Trichoderma
sp. F
IGS
. 18, 75–81
Stromata solitary, 1.6–12.8
3
1.3–2.4 mm, rounded
or elongate or somewhat irregular, pulvinate, slightly
constricted at the margin, brown, surface plane, peri-
thecial apices barely visible. All tissues KOH
2
. Osti-
olar openings appearing as dark brown or black dots
(F
IG
. 9). Cells of the stroma surface in face view
rounded, elongate or angular, (2.5–)3.0 –4.5(–6.2)
3
(1.7–)2.0–3.5(– 4.5)
m
m, grayish-brown to brown,
cell-walls 0.5–1.0
m
m thick (F
IG
. 71). Stroma surface
20–40
m
m thick, cells elongate or occasionally an-
gular, (1.5–)2.5–3.0(–3.5)
3
1.2–2.5
m
m, cell-walls
0.5–1
m
m thick (n
5
10), brown (F
IG
. 70). Cells im-
mediately below the stroma surface hyphal, thin-
walled, hyaline. Tissue below the perithecia tending
to be pseudoparenchymatous, (2.0–)2.5–3.5(–4.0)
3
1.7–2.6
m
m, or hyphal, cells thin-walled, hyaline (F
IG
.
72). Perithecia immersed in the stroma, densely dis-
posed, globose, subglobose or oblong, 275–365
m
m
high, 105–290
m
m diam, ostiolar canal, 67–104
m
m
long, cells of the perithecial wall light brown; ostiolar
region not sharply delimited from the surrounding
tissue of the stromal surface (F
IG
. 69). Asci cylindri-
cal, (72–)80–90(–100)
3
4.5–6.0(–7.0)
m
m, tip thick-
ened and with a pore (F
IG
. 73). Part-ascospores un-
iseriate, hyaline, dimorphic, spinulose. Distal part-as-
cospores globose to subglobose, 3.2–4.0(–4.5)
3
3.0–
3.7(–4.0)
m
m. Proximal part-ascospores ellipsoidal to
wedge shaped, (3.0–)3.5–4.5(–6.7)
3
(2.2–)2.5–3.2
m
m(F
IG
. 74).
Cultures and anamorph.
Optimum temperature for
growth on PDA 20–25 C, no growth occurring at 30
C. Colony radius on PDA at 25 C after 3 d in darkness
ca. 15 mm. Conidia not forming on PDA in darkness
within 6 d; no pigment or odor noted. Conidia form-
ing profusely on CMD after 1 wk at 20 C under light.
Pustules formed around the margin of the colony,

334 M
YCOLOGIA
F
IGS
. 69–81.
Hypocrea stellata
and its
Trichoderma
anamorph. 69. Submedian longitudinal section of a perithecium im-
mersed in a stroma. The stroma is old and the tissues are disintegrating. 70. Ostiolar region of a perithecium and the stroma
surface region. 71. Cells of the stroma surface in face view near an ostiolar opening. 72. Cells of the interior of a stroma
below a perithecium. The stroma is old and the tissues are disintegrating but the hyphal structure can be seen. 73. Ascus.
74. Part-ascospores. The distal part-ascospores are globose to subglobose and the proximal part-ascospores are wedge-shaped
to oblong. 75–79. Conidiophores and phialides. F
IG
. 75 showing long, completely fertile conidiophores as seen in the stereo
microscope. 80. Conidia. 81. Chlamydospore (arrow). All from G.J.S. 99-222. F
IGS
. 69–74, 80, 81 5DIC; 75 5stereo; 76–79
5FL. Scale bars: F
IGS
. 69, 75, 77 5100 mm; 70, 72, 76, 78, 81 550 mm; 71, 73, 79 520 mm; 74, 80 510 mm.

335L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
white, ca. 1 mm diam, cottony, with fertile rachides
or plumes evident giving the pustule a somewhat stel-
late appearance (F
IGS
. 18, 75). Conidiophores com-
prising a very long and completely fertile axis with
no obvious stipe; phialides arising directly from the
axis near the tip and from 1
8
lateral branches of in-
creasing length with distance from the tip. 1
8
branch-
es arising at 90
8
with respect to the main axis, pro-
ducing phialides directly along the length or in a
penicillus or verticil at the tip and/or producing 2
8
branches at 90
8
.2
8
branches producing phialides di-
rectly laterally and in a terminal penicillus or verticil;
several 1
8
or 2
8
branches often arising at a single
point resulting in a dense fascicle of fertile branches.
Phialides nearly cylindrical to ampulliform and slight-
ly below the middle, (4.5–)5.5–8.0(–9.5)
m
m long,
(2.0–)2.7–3.5(–3.7)
m
m at the widest point, L/W
5
(1.5–)1.8–2.8(–3.1), base of the phialides (1.7–)2.0–
2.5(–2.7)
m
m. Cell supporting the phialides (2.7–)
3.5–5.0(–6.2)
m
m(F
IGS
. 77–79). Conidia broadly el-
lipsoidal to ovoidal, (2.0–)2.5–3.5(– 4.0)
3
1.7–2.2
(–2.7)
m
m, L/W
5
(1.0–)1.2–1.6(–1.7), hyaline,
smooth (F
IG
. 80). Chlamydospores terminal or inter-
calary within hyphae, globose or subglobose, (5.5–)
6.5–8.5(–10.0)
3
(5.0–)5.5–8.0(–8.5)
m
m, smooth or
slightly rough (F
IG
. 81).
Known distribution.
New Zealand.
HOLOTY PE. NEW ZEALAND. WESTLAND: vicin-
ity Hokitika, Lake Kaniere, track along S side of lake
between Sunny Bight and Hooker Creek, mixed po-
docarp with
Dacrydium cupressinum
(Rimu),
Prum-
nopitys taxifolia
(Matai), and
Metrosidros umbellata
(Southern Rata), altitude 0 m, 43
8
S, 171
8
7
9
E, on
bark, 2 Sep 1999,
G.J.S. & S. Dodd 8671
(PDD, ISO-
TY PE BPI 746610; culture G.J.S. 99-122, CBS).
5. Hypocrea minutispora B.S. Lu, Fallah et Samuels,
sp. nov. F
IGS
. 10, 11, 19–21, 82–97
Stromata gregaria vel caespitosa, rarius solitaria, pulvina-
ta, ochracea, bubalina vel vinoso-bubalina, (0.5–)0.7–2.0
(–4.5) 30.5–1.5(–3.0) mm. Ostiolo conspicuo. Ascosporae
hyalinae, bicellulares, spinulosae, ad septum disarticulatae;
parte distali globosa vel subglobosa, (3.2–)3.7–4.7(–6.5) 3
(2.7–)3.5–4.5(–5.5) mm; parte proximali ellipsoidea vel cu-
neiformi, (3.5–)4.0–5.5(–6.7) 3(2.2–)2.7–4.0(–5.5) mm.
Anamorphe
Trichoderma minutisporum
Bissett. Phialides
ampulliformes, (3.0–)4.5–6.7(–11.2) mm3(2.2–)3.0–3.7
(–4.7) mm, L/W 5(1.0 –)1.3–2.2(–3.6); cellulae subtermin-
ales conidiophori (1.7–)3.0–4.5(–8.5) mm. Conidia viridia,
ellipsoidea, (1.7–)2.5–4.0(–9.7) 3(1.5–) 2.0–3.0(–6.5) mm,
L/W 5(0.9–)1.1–1.5(–1.9).
Holotype.
BPI 1109373 (ex-type culture G.J.S. 90 -
81
5
ATCC MYA-2951)
Etymology. minutispora
connects to the anamorph,
T. minutisporum.
Anamorph. Trichoderma minutisporum
Bissett, Can J
Bot 69:2396. 1991. F
IGS
. 89–97
Stromata mostly gregarious to cespitose, less fre-
quently solitary, (0.5–)0.7–2.0(–4.5)
3
0.5–1.5(–3.0)
mm, stromata of variable shape, mostly irregular
when aggregated and the margins of individual stro-
mata overlapping, occasionally rounded or elongate
especially when solitary; margins typically fully at-
tached to the substratum, sometimes constricted at
the base (e.g. CBS 901.72); uniformly light brown to
yellowish-brown, young stromata with white margins.
Stroma surface smooth, wrinkled or creased but oc-
casionally slightly tuberculate from perithecial apices.
All tissues except perithecia wall KOH
2
. Ostiolar
openings visible as slightly raised, purple-brown spots
(F
IGS
. 10, 11, 82). Cells of the stroma surface in face
view angular, elongate or irregular in outline, (2–)4–
7(–22)
3
(1.5–)2.0–3.5(–6.5)
m
m, reddish-brown,
cell-walls 0.5–1.0
m
m thick. Surface region of stroma
(13–)15–20(–22)
m
m thick, cells angular to elongate,
rarely circular, light brown to brown, (1.5–)3.0–8.0
(–13.0)
3
(1.2–)2.5–3.5(–5.0)
m
m, cell-walls 0.5–1.0
m
m thick (F
IGS
. 84–85). Cells immediately below the
stroma surface, hyphal, thin-walled, hyaline. Tissue
below the perithecia compact and hyphal, cells thin-
walled, hyaline (F
IG
. 86). Perithecia immersed in the
stroma, densely disposed, mostly oblong to obpyri-
form, (175–)150–280(–333)
m
m high, (68–)100–
200(–222)
m
m wide, ostiolar canal (52–)60 –80(–114)
m
m long (F
IG
. 83); wall of perithecia hyaline to light
brown, KOH
1
red. Asci cylindrical, (66–)75–100
3
(3.5–)4.5–6.5(–7.5)
m
m, tip thickened and with a
pore (F
IGS
. 87, 88). Part-ascospores uniseriate, hya-
line, finely spinulose, dimorphic to monomorphic;
distal part-ascospores globose to subglobose, (3.2–)
3.7–4.7(–6.5)
3
(2.7–)3.5–4.5(–5.5)
m
m; proximal
part-ascospores subglobose, oblong, ellipsoidal or
wedge-shaped, (3.5–)4.0–5.5(–6.7)
3
(2.2–)2.7–4.0
(–5.5)
m
m(F
IG
. 88).
Cultures and anamorph.
Optimum temperature for
growth on PDA 25 C. Colony radius on PDA at 25 C
after 3 d in darkness (28–)33–50(–55) mm, no
growth occurring at 35 C. Conidia forming on PDA
in darkness as early as 3 d at 25 and 30 C, but in
several cultures green conidia not evident before 7 d
and some cultures remaining sterile; conidia typically
forming in dense discrete or confluent, flat pustules
(F
IGS
. 19–21). Dull yellow or brown-yellow pigment
forming in reverse of several cultures on PDA; pig-
mentation also typically associated with sterile cul-
ture. Conidia typically forming in green pustules on
CMD at 20–25 C in light after 6 d around the margin
of the colony or in one or two concentric rings; less
frequently conidia forming in aerial mycelium with-
out pustules. Pustules 0.5–1.0 mm diam, dense and

336 M
YCOLOGIA
F
IGS
. 82–88. Hypocrea
minutispora
/
Trichoderma minutisporum,
teleomorph. 82. Stroma. Note dark ostiolar openings. 83.
Median longitudinal section through a mature perithecium immersed in a stroma. 84, 85. Surface region of the stroma. F
IG
.
85 is near the ostiolar canal, on the left. Note the hyphae below the surface region. 86. Hyphal cells of the interior of the
stroma below a perithecium. 87, 88. Asci and part-ascospores. Note the thickened apex with an obscure pore in F
IG
. 88. F
IGS
.
82, 85–87 from G.J.S. 90-82; 83, 88 from G.J.S. 99–81; 84 from G.J.S. 99-80. F
IG
.825stereo, 83–86 DIC. Scale bars: F
IG
.82
51 mm; 83 5100 mm; 84–87 520 mm; 88 510 mm.
with the stereo microscope conidiophores appearing
to be columnar, fertile to the tip and individual heads
of conidia visible; occasionally conidiophores with a
few phialides formed at the tip of a straight, sterile
stipe visible in pustules. No odor detected on PDA or
CMD. Conidiophores highly uniformly branched,
branches frequently paired or in threes, arising at or
near 90
8
with respect to the main axis, longer and
more profusely branched with distance from the tip.
1
8
branches producing phialides directly and forming
2
8
branches. 2
8
branches tending to arise in pairs at
90
8
, typically terminating in a whorl of 2–3 phialides
(F
IGS
. 89–96). Phialides arising singly from main axis
and branches or held at or near 90
8
in whorls, tend-
ing to be conspicuously swollen below the sharply
constricted tip, ampulliform, (3.0–)4.5–6.7(–11.2)
m
m long, (2.2–)3.0–3.7(– 4.7)
m
m at the widest point,
L/W
5
(1.0–)1.3–2.2(–3.6), (1.2–)1.7–2.5(–4.0)
m
m
wide at the base. Cells supporting phialides (1.7–)
3.0–4.5(–8.5)
m
m(F
IGS
. 29–32, 90, 95, 96). Conidia
ellipsoidal, (1.7–)2.5–4.0(–9.7)
3
(1.5–) 2.0–3.0
(–6.5)
m
m, L/W
5
(0.9–)1.1–1.5(–1.9), mostly green
to light-green, smooth (F
IGS
. 31, 97). Chlamydo-
spores typically not formed but occasionally abun-
dant, terminal and intercalary within hyphae, round-
ed or somewhat elongate, (4–)5–11(–22)
3
(3–)5–
10(–17)
m
m, smooth or slightly rough.
Known distribution.
Canada, Germany, United
States.
HOLOTY PE. UNITED STATES. NORTH CARO-

337L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
F
IGS
. 89–97.
Hypocrea minutispora
/
Trichoderma minutisporum,
anamorph. 89–96. Conidiophores and phialides. 97. Co-
nidia. F
IGS
. 89 from G.J.S. 90-112; 90, 97 from G.J.S. 99-115; 91 from DAOM 175931; 92 from ATCC 64262; 93 from G.J.S.
99-244; 94 from DAOM 212372; 95 from DAOM 179041; 96 from DAOM 178046. F
IG
. 89, 92–94 5PC; 90, 91, 95–97 5DIC.
Scale bars: F
IGS
. 89–96 520 mm, 97 510 mm.

338 M
YCOLOGIA
LINA: Macon County, Ammons Branch Camp-
ground, off Bull Pen Road, on bark, 14 Oct 1990,
Y.
Doi, A.Y. Rossman & G.J.S.
(BPI 1109373, culture
G.J.S. 90-81, ATCC MYA-2951).
Additional
Hypocrea
specimens examined.
GERMANY.
Teutoburger Wald, Neuenheerse, on decaying wood, 25 Sep
1972,
W. Gams
(CBS 901.72, culture and specimen 5BBA
70329). UNITED STATES. ALABAMA, Franklin County,
Bear Creek Education Center, on decorticated wood, 26
Sep 1992,
G.J.S., C.T. Rogerson & S.M. Huhndorf
(BPI
802851, culture G.J.S. 92-107). KENTUCKY: Daniel Boone
National Forest, Laurel River Lake Recreation Area, Cane
Creek Wildlife Refuge, altitude 350 m, on
Stereum
and de-
corticated wood and bark, 27 Sep 1995,
G.J.S.
(BPI 737757,
culture G.J.S. 95-181). NORTH CAROLINA: Clay County,
Standing Indian Campground, off U.S. 64, on bark of
Acer,
15 Oct 1990,
Y. Doi, A.Y. Rossman & G.J.S.
(BPI 1107140;
culture G.J.S. 90-112 5BBA 70323 5CBS 112254); Stand-
ing Indian Campground, off U.S. 64, on decorticated wood,
15 Oct 1990,
Y. Doi, A.Y. Rossman & G.J.S.
(BPI 1107177;
culture G.J.S. 90-119 5BBA 70301); Standing Indian Camp-
ground, off U.S. 64, on decorticated wood, 15 Oct 1990,
Y.
Doi, A.Y. Rossman & G.J.S.
(BPI 1109381; culture G.J.S. 90-
89 5BBA 70321); Standing Indian Campground, off U.S.
64, on bark of
Quercus,
15 Oct 1990,
Y. Doi, A.Y. Rossman
& G.J.S.
(BPI 1107178, culture G.J.S. 90-115); Macon Coun-
ty, Ellicott Rock Trail, off Bull Pen Road, on decorticated
wood, 14 Oct 1990,
Y. Doi, A.Y. Rossman & G.J.S.
(BPI
1109372, culture G.J.S. 90-80); Macon County, Ammons
Branch Campground, off Bull Pen Road, 358019N, 838089W,
altitude 3000 ft., on decorticated wood, 14 Oct 1990,
Y. Doi,
A.Y. Rossman & G.J.S.
(BPI 1109374; culture G.J.S. 90-82
5CBS 112255); Macon county, Blue Valley, off Clear Creek
Road, along Overflow Creek, 358009N, 838159W, on decor-
ticated wood, 16 Oct 1990,
Y. Doi, A.Y. Rossman & G.J.S.
(BPI 1107190, culture G.J.S. 90-132).
Commentary.
The regularly branched conidio-
phore with short and broad phialides of
T. minutis-
porum
suggests the very common species
T. harzian-
um.
The two species can be distinguished easily by
their conidial morphology, globose to subglobose in
T. harzianum
and ellipsoidal in
T. minutisporum
and
by the presence of more or less compact pustules in
T. minutisporum.
There are small and taxonomically insignificant
differences among the
H. minutispora
collections in
characters of the teleomorph and anamorph. Part as-
cospores of G.J.S. 90-115 are somewhat longer and
wider than is typical. Coloration of stromata ranges
from yellowish-brown (G.J.S. 90-112, 90 -115) to
brown to reddish-brown (G.J.S. 90-82). Ostiolar
openings are conspicuous in most collections but are
invisible in 90-82, which is overmature. Conidium siz-
es of different collections overlap, however conidia
of G.J.S. 90-82 are somewhat shorter and narrower
than is typical.
Trichoderma minutisporum
was among several
Trichoderma
species isolated in Mexico from soil of
mango orchards affected by mango malformation
caused by
Fusarium oxysporum
and
F. mangiferae.
It
limited in vitro growth of the pathogens (Michel-Ac-
eves et al 2001).
6. Hypocrea lacuwombatensis B.S. Lu, Druzhinina &
Samuels, sp. nov. F
IGS
. 12, 22, 23, 98–113
Stromata solitaria, rarius gregaria, pulvinata, rotunda vel
oblonga, rarius irregularia, 2.2–4.7 31.5–4.0 mm, ostiolo
conspicuo. Asci cylindrici, 90–111 36.5–6.8 mm, apice in-
crassato, poro apicali praediti. Ascosporae hyalinae, spinu-
losae, bicellulares, ad septum disarticulatae; parte distali
globosa, 3.0–)4.0 –4.5(–5.0) 3(3.0 –)3.7– 4.7(–5.0) mm, par-
te proximali ellipsoidea vel cuneata, (4.0–)4.5–5.5 3
(3.0–)3.5– 4.7 mm. Anamorphe
Trichoderma.
Phialides am-
pulliformes vel pyriformes, (3.5–)4.0–6.0(–7.0) 3(2.2–)
2.5–3.0(–3.5) mm, L/W 5(1.3–)1.5–3.2(–2.7). Cellulae sub-
terminales conidiophori (1.7–)2.5–4.2(–5.7) mm. Conidia
viridia, ellipsoidea vel ovoidea, (3.0–)3.2–3.7(– 4.0) 32.2–
2.7(–3.0) mm, L/W 5(1.2–)1.3–1.5(–1.6).
Holotype. Samuels & S. Dodd 8682
(PDD, ISO-
TY PE: BPI 746621; ex-type culture G.J.S. 99-198
5
CBS 122668)
Etymology. lacuwombatensis
refers to the collection
site, Lake Wombat, near Franz Josef Glacier, West-
land, New Zealand.
Anamorph. Trichoderma
sp. F
IGS
. 22, 23, 106–113
Stromata solitary, rarely gregarious, mostly round-
ed or elongate, occasionally irregular in outline, 2.2–
4.7
3
1.5–4.0 mm, slightly constricted at the base,
dark brown, somewhat reddish, stroma surface plane,
perithecial protuberances barely visible. All tissues
KOH-. Ostiolar openings visible as small black dots,
sometimes inconspicuous (F
IG
. 12). Cells of the stro-
ma surface in face view elongate or rounded, rarely
angular or irregular in outline, (1.5–)2.5–4.0(–6.0)
3
(1.2–)1.5–3.5(–5.0)
m
m, reddish-brown, cell-walls
0.5–1.0
m
m thick (F
IG
. 99). Surface region of the stro-
ma 20–25
m
m thick, cells elongate, angular or com-
pressed, brown, (2.0–)2.5–3.5(–5.0)
3
(1.2–)1.5–
2.5(–2.7)
m
m, cell walls 0.5–1.0
m
m thick (F
IG
. 100).
Hair-like extensions of cells at the stroma surface,
scattered, 7.5
m
m long, 3.5
m
m wide at the base, hy-
aline to light brown (F
IG
. 101). Cells immediately be-
low stromal surface hyphal, thin-walled, hyaline. Tis-
sue below the perithecia hyphal, cells (2.5–)3.0 –4.5
(–5.5)
m
m wide, thin-walled, hyaline (F
IG
. 102). Peri-
thecia immersed in the stroma, densely disposed, glo-
bose to subglobose, 255–282
m
m high, 168–183
m
m
wide, ostiolar canal 85–112
m
m long (F
IG
. 98). Asci
cylindrical, 90–111
3
6.5–6.8
m
m, tip thickened and
with a pore (F
IGS
. 103, 104). Part ascospores uniser-
iate, hyaline, finely spinulose. Distal part-ascospores

339L
UETAL
:
H
YPOCREA
/
T
RICHODERMA SPECIES WITH PACHYBASIUM
-
LIKE CONIDIOPHORES
F
IGS
. 98–113.
Hypocrea lacuwombatensis
and its
Trichoderma
anamorph. 98–105. Teleomorph. 98. Median longitudinal
section through a perithecium embedded in a stroma. Vertical section of perithecia. 99. Cells of the surface of the stroma
in face view. 100. Ostiolar region of a perithecium and stroma surface. 101. Stroma surface region. Note hyphal cells below
the surface and one of the inconspicuous hairs that arise from cells at the stroma surface (arrow). 102. Hyphal cells of the
internal tissue of the stroma below a perithecium. 103. Asci. 104. Ascal apex. 105. Discharged part-ascospores. 106–112.
Conidiophores and phialides. 113. Conidia. All from G.J.S. 99-198. F
IGS
. 98–105, 113 5DIC; 106–112 5FL. Scale bars: F
IG
.
98 5100 mm; 100–103, 107–112 520 mm; 99, 104, 105, 113 510 mm; 106 550 mm.

340 M
YCOLOGIA
globose, (3.0–)4.0 –4.5(–5.0)
3
(3.0–)3.7–4.7(–5.0)
m
m. Proximal part-ascospores ellipsoidal to wedge-
shaped, (4.0–)4.5–5.5
3
(3.0–)3.5–4.7
m
m(F
IG
.
105).
Cultures and anamorph.
Optimum temperature for
growth on PDA 25 C, no growth occurring at 35 C.
Colony radius on PDA at 25 C after 3 d in darkness
ca. 35 mm,
,
10 mm at 30 C. Conidia forming in a
cottony, central ring on PDA in darkness within 3 d
at 20 and 25 C in darkness. Conidia forming on CMD
within6dat20and25Cinlight in a continuous,
cottony ring around the margin. No odor or pigmen-
tation on PDA or CMD (F
IGS
. 22, 23). Conidiophores
comprising a long axis with phialides at the tip and
sterile below or with fertile branches arising along
the entire length; 1
8
branches arising at 90
8
with re-
spect to the main axis, progressively longer and more
profusely branched with distance from the tip, pro-
ducing phialides directly or producing unicellular 2
8
branches; 2
8
branches terminating in 2–4 phialides
(F
IGS
. 106–112). Phialides convergent in heads, la-
geniform to ampulliform, (3.5–)4.0–6.0(–7.0)
m
m
long, (2.2–)2.5–3.0(–3.5)
m
m at the widest point,
L/W
5
(1.3–)1.5–3.2(–2.7), base of the phialides
(1.2–)1.5–2.2(–2.7)
m
m wide. Cells supporting the
phialides (1.7–)2.5–4.2(–5.7)
m
m(F
IGS
. 111–112).
Conidia ellipsoidal to ovoidal, (3.0–)3.2–3.7(–4.0)
3
(2.2–2.7(–3.0)
m
m, L/W
5
(1.2–)1.3–1.5(–1.6), light-
green, smooth (F
IG
. 113). Chlamydospores scattered,
not abundant, terminal and intercalary in hyphae,
globose or subglobose, often in pairs, (5–)6–8(–10)
3
(4.5–)5.5–7.5(–8.0)
m
m, wall smooth or somewhat
roughened.
Known distribution.
New Zealand.
HOLOTY PE. NEW ZEALAND. WESTLAND: vicin-
ity Franz Josef, trail to Lake Wombat, 43
8
25
9
S
170
8
21
9
Eto43
8
21
9
S 170
8
10
9
E, mixed podocarp for-
est, altitude 200–275 m, on bark of recently dead
tree, 3 Sep 1999,
G.J.S. & S. Dodd 8682
(PDD, ISO-
TY PE: BPI 746621; culture G.J.S. 99-198
5
CBS
122668).
Commentary.
There is strong phenotypic similarity
between
H. lacuwombatensis
and
H. minutispora
de-
spite their genetic differences. Stromata of
H. lacu-
wombatensis
are solitary, dark brown, somewhat red-
dish, but those in
H. minutispora
/
T. minutisporum
are mostly aggregated, light brown to yellowish-
brown. The conidiophores of
H. lacuwombatensis
tend to be longer than those of
T. minutisporum,
and
in the former species conidia tend to form in loose
lawns rather than in dense, flat pustules. Conidia of
H. lacuwombatensis
are slightly larger and wider than
in
T. minutisporum,
3–4
3
2.2–2.9
m
m, L/W
5
1.2–
1.6, versus 2.3–3.1
3
1.7–2.2
m
m, L/W
5
1.2–1.5.
Finally,
H. lacuwombatensis
is known only from New
Zealand while
H. minutispora
/
T. minutisporum
ap-
pears to be a species of north temperate regions.
KEY TO
HYPOCREA
/
TRICHODERMA
SPECIES HAVING
WHITE CONIDIA (IN MASS)
All the species discussed in this paper can be found in an
interactive key to
Trichoderma
at http//nt.ars-grin.gov
1. Sterile hairs present
.........................
2
1. Sterile hairs absent
..........................
3
2. Pustules loosely cottony and individual conidio-
phores visible. Hairs sinuous or straight but not
conspicuously cork-screw-like
...
3.
H. parapilulifera
2. Pustules dense and individual conidiophores not
visible. Hairs conspicuous, cork-screw-like
....
.............
1.
H. pachybasioides
/
T. polysporum
3. Conidia globose, 2.5–3.5 32.3–3.2 mm
........
....................
2.
H. pilulifera
/
T. piluliferum
3. Conidia ellipsoidal to oblong, narrower
..........
4
4. On wood. Conidia 2.5–3.5 31.7–2.5 mm
....
...............................
4.
H. stellata
4. On herbaceous stems. Conidia 2.5–3.0 32.0–2.5
mm
...
H. placentula
(Spooner and Williams 1990)
ACKNOWLEDGMENTS
We are grateful to Ms Carolyn Babcock, and Drs. John Bis-
sett (DAOM), Walter Gams (CBS) and Mitsuo Komatsu
(TMI) for providing us with cultures and/or specimens
from their respective collections. Walter Gams corrected
the Latin descriptions. We appreciate the comments of Dr.
A.Y. Rossman. Bingsheng Lu would like to thank Shanxi
Research Foundation for Returned Overseas Chinese Schol-
ars for its support (2001039). We thank Drs. Wen-ying
Zhuang and Sarah Dodd for their helpful discussion and
suggestions. Ms Luttori Ashley provided able technical as-
sistance. C.P.K. acknowledges support by the Austrian Sci-
ence Foundation (P-12748 MOB). The research was also
supported by the U.S. National Science Foundation PEET
grant to the Department of Plant Pathology, Pennsylvania
State University (Monographic Studies of Hypocrealean
Fungi:
Hypocrea
and
Hypomyces
DEB-9712308).
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