[ 45 ]
Trans. Br. mycol. Soc. 79 ( I ) 45-64 (1982) Printed in Great Brit ain
TAXONOMIC STUDIES ON AQUATIC HYPHOMYCETES IV .
PURE CULTURE AND TYPIFICATION OF VARIOUS SPECIES
By E. DESCALS
Department of B iological Sciences, Uni versity of Exeter
Criteria based on ontogeny are given for clear distinctions between Culicidospora aquatica and
C. gravida. Po/yc/adium equiseti, Tricladium castaneicola and T. patulum, and the phialidic
states of T . splendens and T . terrestre, believed to be sperrnatial, are described from pure
culture. A lectotype is proposed for T . angulatum,
The classification of the' Ingoldian hyphomycetes'
becomes more difficult as recently described
species show strong similarities with terrestrial and
dernatiaceous hyphornycetes and coelomycetes .
Conidia of various hyphomycetc species resemble
each other so closely that more detailed pure
culture observations under imp roved conditions
for sporulation are now needed (Descals, Nawawi
& Webster , 1976). This paper discus ses various
species of "aquatic hyphomycetes ' which were
isolated during the course of recent taxonomic
studies (D escals & Webster , 1982). Some of
them had pre viousl y been described on ly from
nature, and were poorly defined. Their growth in
pure culture has enabled us to obser ve other
characters which are of taxonomic importance, and
to increase the sparse type material available. The
techn iques used were those detailed in De scals,
Webster & Dyko (1977), although emphasis has
been placed on allowing the pure cultures to
sporulate above the water level, in a saturated
atm osphere, for rather long per iods.
CULICIDOSPORA AQUATICA Petersen, Bull. Torrey
Bot. Club 87: 342-347 (1960). (F igs 1, 12, 13)·
Pure culture: from single conidium on MA 2 %
under standard conditions. Colony dark brown,
growth 6 cm diam /j months, aer ial mycelium
lanose, brownish-grey, with few pale grey patches,
diffusible pigment reddish-brown . Sp orulation
sparse on colony slivers in sterile distilled water in
a Petri dish at 15°C for 2 days, but abundant after
7 days at 11-14°. Conidiophores either on aerial
mycelium or immersed hyphae lateral, mononerna-
tous , in dense aggregations, simple, del icate,
cylindrical,cells few, septa indistinct . Conidiogcnous
cells apical, single, integrated, detachment scar
flat, proliferations (or 3, percurrent, cylindrical
20-30 x 3'7-4"4 JIm . Conidia solitary, aerogenous,
staurosporous, septa often indistinct , primary body
clavate , curving through 45° at the insertion of the
lateral branches; 47-57 x 7'5-10 Jim , apical cell
AND J. WEBSTER
hemispherical, cells 4-5, detachment scar broad
and flat ; branches: one apical, two lateral and one
caudal, straight, acicular but with protoplasm, cells
1-(3), apical and lateral branches budding out
before release , base abruptly constricted,
47-100-(175) x 0'5 mm , lateral branches at same
level, arising dors ally, usually on the sub-apical
cell, synchronous and soon after the apical branch,
diverging perpendicularly, caudal branch broad-
based, 75-(160) x 2'5- 3'7 JIm . Released at a septum
and by irregular rupture of the outer wall, which
remains as an infundibular membranous structure
with torn edges , in progressive series of up to 3 or
4 enclo sing the subsequen t percurrent
proliferations .
Phial idic state: produced sparsely under the same
conditions . Conidiophores on mycelial hyphae,
mononematous , branched, turning dark, septa
thick, 100-250 I'm long , branches few, lateral,
aris ing singly, in loose terminal clusters, antrorse at
a narrow angle and ascending. Conidiogenous cells
phialidic, single, apical, straight or ascending ,
ellipsoid, often irregularly-shaped, dark, collarette
attenuate, marg in slightly constricted,
15-17 x 3-4 I'm. Conidia bacilliform, unicellular,
turning dark , 2-3 x f : 2-1 .5 psn, aggregating in
large gloeoid mass es at the collarette, germination
not seen.
Specimen examined : From foam, Becka Falls, Becka
Brook, near Manaton, Devon, England, 21 Feb. 1974,
Descals, Herb. Exet er 3519.
CULICIDOSPORA GRAVIDA Petersen, Mycologia 55 :
24-25 (1963). (F igs 2, 14, 15).
Pure cultur e : from single conidium on MA 2 %
under standard conditions. Colony black, margin
wh ite, narrow, strongly lobed, sectoring abun-
dantly, growth slow, aerial mycelium greyish black ,
felty. Sporulation abundant (but with conidial
branches incompletely developed) on the dry
colon y in sealed Petri di shes , more typical on
colon y slivers submerged in sterile distilled water
 Aquatic hyphomycetes. IV

Fig. 1. Culicidospora aquatica. (Pure culture Herb. Exeter 3519. Phase-contrast, cotton-blue lacto-phenol. )
A-M, percurrent proliferations and trumpet-shaped remains . D, first conidium released from uppermost
septum, followed by a short proliferation (e) and the initiation of a conidiogenous cell (c) (also seen in E,
F, H and I). G, developing conidium not yet separated from its outer wall. B, development of fourth conidium.
N, metasclerotia supporting the conidiophores. Os-P, phialidic state: phialidcs and phialospores in gloeoid
mass on the collarette. (Dotted lines indicate plasmolysed cytoplasmic membrane.)
 E. D escals and J. W ebster
at 15-1 8° for 8 da ys, appearing on cu t sur faces.
Conidiophore: on mycelial hyphae or on torulose
cells, apical or lateral, macronematou s, mono nern -
atous, in den se aggregation s, simple or sparsely
branched, slightl y fu scous, sti pe broad ening
slightly near the apex. Conidiogenous cells int e-
gr ated, apical , clavate , 5-10 x 3"5-6 utn, detach-
ment scar ap ical, broad, prolifer at ions percurrent,
up to 4 aft er 1Z days incubation. Conidi a solitary ,
aer ogenous , staurosporous, fu scou s, pr otopla sm
granular, primary bod y clavate, 36-48 x <}-11 pm,
cu rved 45° at the lateral branch inser tio ns, ap ical
cell hem ispherical, cells 4-5, detachment scar
broad , convex, branche s st raight, acicular (or
br oader with obtuse ap ex), protoplasmic ,
20-27 x 2-2'5 p.m, apical branch bu dd ing out, base
abruptly con stricted, cells 1-2, late ral branches
(0}-2-(4 ), budding out, synchronous, dorsal and at
the same level, usually from the subap ical cell,
slightly retrorse or perpendicular, cells 1-2, caudal
branch growing through the detachment scar, base
broad , unicellular, 6-12 x 3-4'2 utt» .
Specimen examined: from foam below waterfall, in
stream near Bettws-y-Coed, Snowdonia, N . Wales, 17
May 1976, D escals, Herb. Exeter 3486.
D yko (1977) claimed that C. aqual ica and C.
gravida are conspecific. Marvano va ( 1970) had
pre viously exam ined the typ es in an att em pt to
comp ar e their method of proliferation . The
holot ype of C. aquatica has ver y young sp or ulating
structu res. In C. gravi da , however, several ' ann el-
lide-like percurrent pr olife rations ar e present.
These were also detected on her isolate when
sporulating in a hanging dr op . In our isolates of
both species, pr oliferation is percurrcn t, but the
d ifferen ces in the method of release are so
fundamental that we ha ve no doubt that we are
dealing with two d istinct species.
The two fungi ar e regularly detected in foam
samp les in Great Britain . When grown in pure
culture, they form dark colonies, with characteristic
dark cells on which the conidiopho res are frequently
found. Culicidospora aquatica can produce a reddish
diffu sibl e pigment , Sporulation occurs in dense
aggregations often on cut surfaces of agar. The
con idiophores are sparsely branched, and conidio-
gen ou s cells are integrated and apic al, with several
percurrent extensions. Their coni dial morphology,
with ap ical, late ral an d cauda l br an ches, is un ique
among th e hyphomycete s, th ough a similar form
can be found in th e Melancon iales iPestaloz eina
Sacc.), Culicidospora aquatica on ly sporulates
abundantly when submerg ed, while C. grav ida will
form larg e ma sses of conidia in con de nsation water
on th e surface of the colon ies in sea led Petri dishe s.
T he con idial branche s, th ough, develop po or ly
47
under th ese condi tio ns . Even when pr oduced
underwater , the arms frequently remain sho rter
than in nature, whi le C. aquatica form s typ ical
conidia. In th e latter species, these are delicate,
colourless and hyalin e. The fungus has a micro-
coni dial state which is dernatiaceou s and rather
thick-walled . We have not seen th e equivalent stat e
in C. gra vida. It s con idia are dens ely granular an d
fuscous, sh orter and fatter. The main difference lies
in the method of release. The processes of in itiatio n
itself is basically th e same in both cases, bur in C.
gravida release is simply by separation at a septu m,
while in C. aquatica th ere is, in addi tion, a ripping
apart of an outer wall on the main axis, wh ich leaves
beh ind a char acte rist ic trumpet-l ike, membran ous
structure. The tearing is irregular, and is best seen
with phase contrast, As a result of percurrent
proliferations , up to four of these casings can appear
arising from cons ecutive detachment scars. Such a
method of release has never been seen in C. gra vida .
This type of ontogeny is not represented in any
of the sectio ns of H ughes (1953) or Tubaki (1958).
Apparently a sim ilar pr ocess is seen in Chaetendo-
ph ragmia, Deighton iella, Conioscypha, Stigmina,
Endophragmiopsis, Endophragmia and Cacumi-
sporiu m, and mu st not be confused with a separating
cell, where the tearing or dissolution of the lateral
wall ne ver occu rs in the con idium proper. D yko
(1977) exam ined he r isolate of C . aquat ica and also
found th ese membranous empty walls. In her
material, th ey app eared singly, with up to one
percurrent prolifer ation. This may be just a matt er
of length of incubation peri od s un derwater.
There is a need to designate a n eot ype for
Culicidosporaaquatica, preferably sho wing pr olifer-
ation and the microconidial state. D irect observa -
tion of development, with either N omarski
interference or pha se cont rast microscop y, shou ld
help confirm the above interpretation of release. It
is pr obable that th e periclinal sheathing of the
lateral wall on the ma in bod y occu rs gradually and
that the breakage of the resultant outer wall tak es
place due to turgor pressure of the main body as
well as by th e early projection of the cauda l
extension, always pre sent, and seen on one occasion
while the conidium was still within the split outer
wall (see Fig . 1 D ).
POLYCLADIUM EQUISETI Ingold, Trans. Br. M yc ol,
S oc. 4Z : 112-114 (1959). (F igs 3, 20, 23).
Pu re culture from single con idium on MA 2%,
at standard condition s. Colony dark brown , margin
diffu se, growth mode rate , aerial mycel ium low and
dark. S porulation abundant on 3-momh-old
colony slivers in steri le d istilled water aerated for
10 days . Conidiophores mononematou s, up to
600 x 5 pm . C onidiogenous cells intercalary, often
 Aquatic hyphomycetes. IV

Fig . 2. Culicidosporagravida. (Pure culture Herb. Exeter 3486.) A-N, ontogeny, detachment and percurrent
extensions. I-M, conidiophores and supporting structures (hyphae and rnetasclerotia). N, conidium soon after
release. O-X, detached conidia. Q, R, X, typical conidia. (K and L drawn to scale B; remainder to scale A.)
 E. Descals and J. Webster 49

A: lO0l'm
B: 200 I'm

Fig. 3. Polycladium equiseti. (Pure culture, Herb. Exeter 3408.) A-F, ontogeny. Unlabelled figures: habit
drawings of detached conidia . A to scale A; B-F to scale B; unlabelled drawings to scale C. Arrows indicate
conidiophore apex or lateral detachment scars.
5° Aquatic hyphomycetes. IV
consecutive, integrated, detachment scars single
and directly below septa. Conidia pleurogenous,
progressive, staurosporous, spanning up to
800 !tm, main body cylindrical with broad base and
obtuse apex, 300-500 x 5-6 lim, antrorse at an
acuteangle, curved at branch insertions, detachment
scar broad, septa indistinct in water, branches in 1-2
orders, lateral, single, in various planes, diverging
at broad angles, 50-300 x 3'5-5 !tm, primary bran-
ches (1)-4-(5), straight, or curved at secondary
branch insertions, secondary branches 0-4, straight,
released at a basal septum, dispersed singly or in
tangled aggregations underwater.
Specimen examined : From foamin BleaTarn, Cumbria,
12 May 1974, Webster & Descals,Herb. Exeter 3408.
Several conidia of this species were isolated from
the type locality, from both foam and decaying
Equisetum internodes. This is the first description
from pure culture, in which it sporulates well upon
submersion. The discovery of a species of Scorpio-
sporium (Herb. Exeter 3522) (Descals & Webster,
1982) prompted the isolation of this species, as
they have similar conidia. It has also enabled
comparison with S . rangiferinum (Descals &
Webster, 1982). Polycladium equiseti seems to
be rather localized and restricted to moorland
habitats. Frequent surveys on Dartmoor and
Exmoor have failed to detect it. It is unlikely that
its substrate is limited to Equisetum. Such high
specificity has not been reported for other sapro-
phytic Ingoldian fungi.
Our description, from authenticated material,
matches that of Ingold (1959). This author claims
that sporulation is exclusively pleurogenous. Lateral
conidia are evident in our material, i.e, lateral
detachment scars can be seen along the conidio-
phore. But it is not clear whether the conidiophore
apex may eventually produce a conidium, or
whether it remains vegetative. There are no signs
of percurrent proliferations or of apical scars , which
would prove aerogenous sporulation, but it is
difficult to discern between the conidia and the
conidiophore while these are developing, as the
whole arborescent structure is made up of branches
of similar width. Aerogenous conidia can only be
detected by the presence of apical scars on a
completely spent conidiophore.
TRICLADIUM SPLENDENS Ingold, Trans. Br. Mycol.
Soc. 25: 385-389 (1942). (Figs 4,22 ).
Pure culture from single conidium on MA 2 %
under standard conditions. Colony black, margin
white , growth moderate, aerial mycelium cottony,
dark. Sporulation aerial or aquatic, abundant on
colony slivers partly submerged in sterile tap water
in a Petri dish at 15-18° for 3 days . Conidiophores
mononematous, sparsely branched. Conidiogenous
cells monoblastic, proliferations percurrent, Conidia
typical.
Phialidic state: colourless, very sparse on same
material after 7 days. Sporophores on mycelial
hyphae or macroconidia, mononematous, aquatic,
prostrate, branched in an apical head, main axis
100 x 2'5 pm, delicate, multicellular, branches sev-
eral, single, penicillate or verticillate, in 1-2 orders,
antrorse, 9-15 x 2'5 !tm, cells few. Sporogenous cells
phialidic, numerous, apical , single or more typically
penicillate, straight or curved inwards, ellipsoid,
cellarette (if present) straight and cupulate,
8-12 x i : 5-2 '5 I'm. Phialospores unicellular, first
spore irregularly ovate to ellipsoid, 3'5 x 1'7 !tm,
subsequent spores globular, 1'2-2 lim, released at
the base of the cellarette, gloeosporous, germination
not seen .
Specimens examined: From decaying Pinus twig in R.
North Teign at Batworthy, near Chagford, Devon,
through deciduous woods and moorland, Aug, 1974,
Descals, Herb. Exeter 3529; on dicotyledonous leaf, R.
Teign, near Dunsford, Devon, through deciduous wood,
17 Jan. 1974, Descals, Herb. Exeter 3515, 3516.
This well-known species is common in Great
Britain and has also been reported from Eastern
Europe, U.S.S.R., N. America, Australia and
South Africa. A record from Hawaii (Anastasiou,
1964), if correct, is interesting in being the only one
from the tropics. Tubaki (1960), in Japan,
collected, from foam, conidia with a primary body
45 I'm long and branches 25!tm long. Tricladium
splendens usually has much larger conidia, and
Tubaki's fungus may have been Ingoldia biappen-
diculata Arnold or Tricladium attenuarum Iqbal.
Pure culture characters are as reported by Ingold
(1942). Percurrent proliferations were seen by
Greathead (1961) in hanging-drop preparations.
We have confirmed their presence in our isolates
after long incubation in standing water.
We include the first description of its phialidic
state. It is present only sparsely on agar and not
easily detectable, but has been seen in several
isolates. The teleomorph has been recently discov-
ered on Fagus cupules (Abdullah, Descals &
Webster, 1981), and belongs to Hymenoscyphus
S. F. Gray (an inoperculate discomycete). The
material on which this association is based has been
proposed as lectotype for the anamorph.
TRICLADIUMANGULATUM Ingold, Trans . Br. Mycol.
Soc. 25: 389-393 (1942).
Pure culture from single conidium on MA 2 %
under standard conditions. Colony orange-brown,
growth slow, aerial mycelium lanose, white.
Sporulation aquatic, abundant on colony slivers
partly submerged in sterile distilled water in a Petri
 E. Descals and J. Webster 51

E
::t
o E
V) ::t
:ll g

c. 200 I'm

Fig. 4. Tricladium splendens. (Pure culture, Herb. Exeter 3529.) A-K, Tricladium state. B, C, F and G,
conidophore branching. C, D and E, percurrent conidiophore extensions (cg.) H and I, conidial lateral
branches and main axis, probably acting as separate propagules, J and K, polar germination. K, secondary
conidia. L, phialidic state: sporophores and two types of phialospores. M, phialidic state on germinating
macroconidium. N, dark, thick-walled propagules of unknown nature. (A-F and L to scale B; G-J and M
to scale A; K to scale C.)
52 Aquatic hyphomycetes. IV

Fig. 5. Tricladium angulatum, (Pure culture, Herb. Exeter 3398.) A-Q, ontogeny and proliferation. K, unusual
conidiogenous cell. A, B, H, L, N, 0, P, Q, various degrees of sympodial proliferation. R-Y, detached
conidia. Y, unusual third lateral. R, T, V and X, polar germination. Z, habit drawing with sporulation on
hyphal cord. (2 to scale B; remainder to scale A.)
 E. Descals and J. Webster
dish at 15-18° for a few days. Conidiophores often
on mycelial cords, mononematous, in loose aggreg -
ations, prostrate, simple, apex eventually swollen,
100 x 2'5-5 {lm, multicellular. Conidiogenous cells
apical, single, integrated, detachment scars distinct,
proliferations (0) to few, sympodial, very short and
clustered. Conidia aerogenous, solitary or com-
monly fasciculate, in close succession, septa often
indistinct, staurosporous, main body narrowly
fusoid, curved at branch insertions, 48--63 x 2' 5-
3'5 {lm, cells 3-4, detachment scar truncate,
branches lateral, 2 ( - 3), appearing before release,
progressive, single, in different planes, perpendicu-
lar, straight (to slightly curved), conical, base broad,
first lateral 32-45 x 2'5-3'5 {lm, second lateral
18-33 x 1'2-1'5 {lm, cells 1-2, typically with 1
septum near the base, released at a basal septum,
germinating readily on isolation medium.
Specimens examined: From foam in Kirby Beck at
Kirby-Malham, Yorkshire, Mar. 1974, Descals, Herb.
Exeter 3398,authenticated lectotype, 1M1 254851; foam
in Crook Beck, East Slope, Pen-y-Ghent, Yorkshire, 30
Mar. 1974,Descals Herb. Exeter 3395.
There is no type designated for this species. Our
isolate agrees with Ingold's (1942) except in colony
pigmentation, which in our case is orange-brown
rather than white. But pigmentation often only
develops after a certain age. Our material has been
authenticated and is herein proposed as lectotype.
Tricladium splendens Ingold (1942), the type
species, resembles T. angulatum in conidial branch
arrangement and sequence. But proliferation is
percurrent, while in T. angulatum it is sympodial.
The two fungi do not appear to be closely related
but, as remarked by Dyko (1977), its redisposition
in Scorpiosporium Iqbal (1974) is not yet justified.
Price (1964) described a South Australian isolate
and labelled it T . gracile Ingold prox., although it
shows the platform-like swelling caused by pro-
liferations (Fig. 9). Marvanova (pers. comm.) also
reports an isolate with sporulation intermediate
between these two.
Tricladium patulum Marvanova & Marvan (1963)
(see below) bears very short sympodulae, somewhat
similar to those in T. angulatum, but the conidio-
phores are penicillate, much more like A . tetra-
cladia Ingold (1942). The conidiogcnous cells
in T . varium Jones & Stewart (1972) proliferate by
short sympodulae but appear on much shorter
extensions. These could be interpreted as lateral on
micronematous conidiophores.
TRICLADIUM CASTANEICOLA Sutton, Trans. Br.
Mycol. Soc. 64: 422-424 (1975).
Pure culture from single conidium on MA 2 'Yo
under standard conditions. Colony dark brown,
53
margin white, growth 8'5 em diarn/z; months, aerial
mycelium grey, abundant, cottony. Sporulation
abundant on 4-month-old colony slivers partly
submerged in sterile distilled water in a Petri dish
at 15-18° for 7 days. Conidia produced above the
water level in dense, gloeoid aggregations, on newly
growing mycelium. Conidiophores on mycelial
hyphae or coils, micronernatous or macronematous,
mononematous, apical or lateral, simple (to sparsely
branched), 12-48 X 2'5-3 '5 pm, stalk sinuous (to
straight), base broadened, cells 1 to several,
branches diverging variously, cells 1 (to few) .
Conidiogenous cellssingle, apical, straight or curved,
ampulliform or integrated, detachmentscar distinct,
proliferations (0) to several, sympodial, rachis-like.
Conidia solitary, aerogenous, staurosporous, main
body straight (or ascending), cylindrical to obclav-
ate, straight or slightly bent at branch insertions,
100-126 x 3'5-4'5 pm, multicellular, detachment
scar truncate, branches lateral, budding out in 1(-2 )
orders, progressive, near the centre of the parent
structure, in different planes, diverging at broad
angles, straight, obclavatc, base smoothly constric-
ted, 40-65 x 2'5-3 '5/'ID, primary branches
(1)-2- 3- (4), secondary branches 0(-1-2), on the
lowest primary branch, released at a basal septum,
germinating readily on isolation medium. Secondary
conidia on detached primary conidia, near the
detachment scar, resembling primary conidia,
release not seen.
Specimen examined : From sterilized Fagus leaf under-
water in stagnant pond for 4 weeks, Stoke Woods,
University of Exeter, 5 June 1976, Fisher, Herb. Exeter
3542, authenticated.
This is a first description from pure culture.
Sutton (1975) described this species on Castanea
cupules from a sandy heath and from a ditch. It
forms lateral, erect, sparsely branched conidio-
phores, with conidiogenous cells mostly integrated,
terminal or lateral, polyblastic, (with up to 3
detachment scars) and with sympodial prolifera-
tions. Conidia are composed of a straight, septate,
main body with a truncate base and an acute apex
bearing 0-2 straight laterals with smoothly con-
stricted bases.
Our authenticated isolate originated from a moist
habitat, but sporulated in pure culture only above
water level. The description differs from Sutton's
only in the higher degree of conidial branching,
with a secondary branch sometimes appearing on
a lower primary. The primary body can also be
slightly curved at branch insertions. Our description
is based on more abundant material from pure
culture which, as in other species, tends to display
greater morphological variation.
Tricladium castaneicola resembles T. splendens
54 Aquatic hyphomycetes, IV

Fig. 6. Tricladium castaneicola. (Pure culture, Herb. Exeter 3542.) A-V, ontogeny and proliferation. X, hyphal
coil with conidiophores. Y, habit drawing of conidiophore. (A-X drawn to scale A; Y to scale B.)
 E. Descals and J. Webster 55

.. ~
10011ln

Fig. 7. Tricladium castaneicola (same material). A, secondary conidium arising near the detachment scar of
the primary conidium. B, C, E , F, G, J and K, conidia with unusual number and arrangement of laterals.
D, H, I, Land M , typical conidia with either 2 or 3 primary laterals.
 Aquatic hyphomycetes. IV

Fig. 8. Tricladium patulum. (Pure culture, Czechoslovak Collection of Micro-organisms F 06576.) A,

ontogeny and proliferation. B-E, typical conidia. F, conidium with three primary laterals. G, conidium with
one secondary lateral. H, spores of unknown origin.
 E. Descals and J. Webster 57

Fig. 9. Tricladium patulum. (Pure culture, Herb. Exeter 3544.) A, habit drawing of typical conidiophore head.
~H, details of conidium ontogeny and proliferation. E: sympodial branching of conidiophore. I-M, detached
conidia. I and M, typical conidia. J, K and L, unusual conidia with third primary lateral and/or secondary
lateral. N : chains of metasclerotia in mycelium. (A and J-M to scale B; remaining to scale A.)
 Aquatic hyphomycetes. IV

Fig. 10. Tricladium terrestre. (Herb. Exeter 3245.) A-], ontogeny and proliferation.
 E. Descals and J. Webster 59

Fig. 11. Tricladium terrestre (same material). A-F, detached conidia. F, sketches showing primary body
Carrow), detachment scar at its base Ce). G, phialidic state: Phialides and phialospores. C, D and E, most
common conidial morphology. CA-E drawn to scale A; F to scale B; G to scale C.)
60 Aquatic hyphomycetes. IV
(the type species) only in conidial branching
pattern (see above). However, it is similar to T.
terrestre Park in the type of rachis-like proliferation
and in the highly variable branching habit of its
conidia (see below).
TRICLADIUM PATULUM Marvanova & Marvan, Acta
Musei Silesiae Ser.A 12: 111-113 (1963). (Figs
8,9,21).
Pure culture from single conidium on MA 2 % at
standard conditions. Colony creamy brown, growth
6 em diamj i month, surface radially convoluted,
withscattered, immersed, dark hyphal aggregations,
metasclerotia colourless, catenate, aerial mycelium
white and mealy. Sporulation very sparse on agar in
a sealed Petri dish, but abundant on 7-month-old
colony slivers partly submerged in sterile distilled
water in a Petri dish at 15-18° after 3 days, on or
underneath the meniscus. Aerial conidia in gloeoid
masses. Conidiophore: on mycelial hyphae, apical,
macronematous, mononematous, branched,
5So x 3'5-5 flm, spanning 150 flm, stalk prostrate,
delicate, cylindrical, base broad (to swollen),
multicellular, branches penicillate, antrorse at a
narrow angle, straight or slightly curved, cylindrical
to clavate, base slightly constricted, unicellular.
Conidiogenous cells apical, single, integrated,
2S-27 x 3-3 'S um, polyblastic, detachment scars
truncate, apical, 1-4, proliferations few, sympodial.
Conidia (1)-3-(4), aerogenous, fasciculate, in close
succession, staurosporous, septa indistinct, main
body slightly geniculate at branch insertions,
cylindrical, apex conical, base slightly constricted,
110-140 x 3-4 flm, primary branches lateral, 2(-3),
budding out in succession before release, antrorse,
in different planes, cylindrical to obclavate, slightly
pendulous, apex conical, base constricted and
curved, 7~S x 4-S usn, first branch longer, multi-
cellular, secondary lateral infrequent, on the
lowermost primary lateral, diverging at a broad
angle, released at a basal septum, dispersed singly
or in clusters, germinating readily on isolation
medium.
Specimens examined: From foam in stream near
Bettws-y-Coed, Snowdonia National Park, N. Wales,
through mixedwood, 24 Oct. 1976 , Descals,Herb. Exeter
3544; from foam in R. Aber, near car park, N. Wales,
through a deciduouswood, 23 Oct. 1976, Descals, Herb.
Exeter 3501.
This fungus was described from nature on a
Fagus leaf. In our experience, this substrate
consistently yields very few conidia of Ingoldian
hyphomycetes, though aero-aquatic hyphomycetes
sporulate abundantly on it. Whether the former
cannot colonize it efficiently or, alternatively,
cannot sporulate on it, is not known. Marvanova
(pers. comm.) later isolated the fungus, but there
is, as yet, no published descripton from pure
culture. Furthermore, the iconotype was not
clearly printed, although the original drawings
were kindly supplied by the author.
Conidia of this fungus superficially resemble
those originally illustrated for T. eccentricum
Petersen (1962), but at higher power they are quite
distinct. They are not commonly detected in foam,
although Ingold (1975 ) reported them from stream
foam in the Scottish Highlands.
Our isolate closely matches Marvanova's des-
cription and iconotype, as well as isolate F 06576
from her collection (Fig. 8), except for a higher
degree of conidiophore branching. In our material,
profusely branching heads appear after several
days incubation.
The classification of this fungus in Tricladium is
mainly based on similarity of conidial characters
and will need revision. Articulospora tetracladia
Ingold, the type species, strongly resembles this
fungus in its penicillate conidiophores and in
conidial morphology, especially when the five-
armed conidium is considered. In T. attenuatum,
the conidial branches also arise by budding
successively at different levels. Tricladium patulum
is also similar to T . angulatum (see drawings).
TRICLADIUM TERRESTRE Park, Trans. Br . Mycol.
Soc. 63: 179-183 (1974). (F igs 10, 11, 18,19).
Pure culture from single conidium on MA 2 % at
standard conditions . Colony dark brown, with
concentric rings , growth 8'5 cm/e weeks, aerial
mycelium pale grey, low . Sporulation very abun-
dant on ro-monrh-old colony slivers partly sub-
merged in sterile distilled water in a Petri dish at
15-18° for 7 days, at water level. Conidiophores on
mycelial hyphae, gregarious, macronematous,
mononematous, prostrate or ascending,
100-15° x 2-5 usn, (simple) to branched, (0) to
several branches, arising unilaterally and randomly

Figs 12-19. Light-micrographs. Fig. 12. Culicidospora aquatica. (Pure culture, Herb. Exeter 3519.)
Conidial development. Fig. 13. C. aquatica (samematerial). Infundibuliform sheath enclosing percurrent
proliferation. (Same scale as Fig. 12.) Fig. 14. C. gravida. (Pure culture, Herb. Exeter 3486.) Detached
conidium. Fig. 15. C. gravida (samematerial). Percurrent proliferation, without enclosing sheath. (Same
scaleas Fig. 14.) Fig. 16. Tricladium angulatum . (Pure culture, Herb. Exeter 3398.) Proliferationon short,
platform-like sympodula. Fig. 17. T. angulatum (same material). Detached conidium. (Same scale as
Fig. 16) . Fig. 18. T. terrestre, (Pure culture, 1M! 216345 and Herb. Exeter 3245.) Ontogeny. Fig. 19.
T. terrestre (samematerial.)Detached conidium, with probably secondaryconidiumdevelopingon the main
body. (Same scaleas Fig. 18).
 E. Descals and J. Webster 61

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/ I1 00
62 Aquatic hyphomycetes . IV

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N
 E . Descals and J. Web ster
in 1-2 orders, straight or curved, cells (1) to few.
Con idiogenous cells apical, single, often sinuous,
integrated, 15 x 3-4 p.m, monopolyblastic, prolifer-
ation sympodial, detachment scars truncate . Conidia
aer ogenous, solitary or with two appearing in close
succession, staurosporous, main bod y cylindrical,
geniculate at branch insertions, multicellular,
12er220 x 4-6 utn, detachment scar truncate, flat,
laterals budding out in succession, in 1-2( - 3)
orders, single (or seldom in pairs ), d orsal , in
different planes, straight or slightly geniculate at
secondary branch insertions , slightly obclavate,
base smoothly constricted, 65-200 x 3-4 utn, multi-
cellular, septa indistinct, primary laterals (1)-3-(4),
secondary laterals er(1-3 ), tertiary lateral rarely
pre sent, released at a basal septum, dispersed on
water, germination on isolation medium or on
water. Secondary conidia occasional, single, sessile,
ar ising near the detachment scar, simil ar to primary
conidia.
Phialidic Slate: colourless, on the same material.
Sporophores aquatic, mononernatous, macronern-
atous, prostrate, simple or with a sparsely branched
head , stalk multicellular, often strongl y curved
apically, branches few, in t-2 orders, short, single ,
antrorse at narrow angle s, cells 1 to few, septa
indistinct. Sporogenous cells phialidic, few, apical
and single or penicillate, or lateral and single ,
ampulliform, ~12 x 2'5-3 utn, collarette (when
pre sent) cylindrical to slightly cup-shaped. Phialo-
spores tear-shaped, 2 p.m diam, detachment scar
truncate, gloeosporous, germination not seen.
Specimen examined: From foam in R. Dee below the
waterfall at Linn of Dee, Aberdeenshire, Scotland, Sept.
1975, Descals, IMI 216345 and Herb. Exeter 3245,
authenticated.
This is a first record of the ph ialidic state of T.
terreste , a fungus which is not easy to identify. The
irregularly branched conidiophores and smooth
basal constrictions of the con idial branches are
qu ite similar to those of Tricladium castaneicola
Sutton, but the conidia are much larger and more
complex. These two species may eventually be
segregated into a common genus along with
Ramulispora spp . with branched conidia . The
holotype of Varicosporium giganteum Crane
(1968), which superficially resembles our fungus,
has been examined. The swollen conidial branch
apices , illustrated in the original description, are
only present on some con idia , obviously as a result
of incipient germination, but, nevertheless, other
differences are obvious : the conidial branches are
much more symmetrical and delicate and br oad
based. The other species in Varicosporium show no
resemblance to T. ter rest re: V. delicatum Iqbal
(1971), V. aquaticum Vischnevskaja (1955) (with
didymospores rather than staurospores, according
to N ilsson, 1964), and V . helicosporum Nawawi and
V. macrosporum Nawawi (1974b) will eventually
have to be segregated too. Varicosporium splend ens
Nawawi (1974 a) will be transferred to Dendrosporo-
myces (N awawai & Webster, 1982) Varicosporium
trimo sum Wolfe (1976) and the Varicosporium
state of Hymenoscyphus v aricosporioides Tubaki
(1966) need further work, as their ontogeny is not
clear .
We are grateful to Dr L. Marvanova (Czechos-
lovak Culture Collection of Micro-organisms) for
providing herbarium specimens, Professor C. T .
Ingold for examining our slides, Dr D. Park and Dr
B. C. Sutton who examined and authenticated
herbarium slides, Dr P . J. Fisher who supplied us
with a collection, and Professor A. Nawawi who
deposited valuable herbarium specimens at Exeter
Herb. The late Dr P. H . B. Talbot kindly supplied
us with pertinent literature.
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