Vol.

58, Pan 1 1972

Trans. Br. mycol. Soc. 58 (I), 1-4 (1972)
Printed in Great Britain

CULTURE STUDIES
ON HYPOCREA AND TRICHODERMA
V. HYPOCREA PSYCHROPHILA SP.NOV.
By E. MULLER, B. AEBI,
Institut fur Spezielle Botanik, Eidgenossischen Technische Hochschule,
Zurich
AND J. WEBSTER
Department ofBiological Sciences, The Unioersity, Exeter
(With I Text-figure)

A new species of Hypocrea growing in the Swiss Alps on Rhododendronferrugineum

and Vaccinium myrtillus is described. The fungus is unusual in its low-temperature
optimum for growth and sporulation. Single ascospore cultures yielded conidia
belonging to the form-genus Gliocladium.

In September 1968 and again in September 1970, we have collected in

the subalpine region on rotting twigs of Rhododendronferrugineum L. a species
of Hypocrea with golden yellow stromata. Cultures from both collections
are identical in morphology and behaviour. They are psychrophilic and
produce a characteristic conidial state of the Gliocladium type. In the
herbarium of the Swiss Federal Institute of Technology (ZT) we have
found more collections of the same species. This fungus is considered to
be a new species, for which we propose the name H. psychrophila.

Hypocrea psychrophila sp.nov.

Status perfectus. Stromata peritheciorum hemisphaerica vel lobata, rufa, ad 4 mm
diam, ostiolis obscuriore proiecturis. Cortex stromae ex cellulis 4-5 pm diam, crasse
tunicatis compositus. Medulla pseudoparenchymatica ad basim cellulis ad 28 pm longis
et 5-7 pm crassis composita. Perithecia numerosa, ampullacea, ad 200 pm altitudine,
150 pm diam, saepe dense aggregata et lateralibus applanata. Asci numerosi, cylin-
dracei, 180-200 x 5 pm, octospori, paraphyses non ades. Ascosporae hyalinae, uniseriatae,
bicellulatae, normaliter divisiae interiore ascorum. Cellulae ascosporarum numerosis
spines parvulis ornatae. Cellulae superiorae subglobosae vel truncatae - ovoideae, 4'5-
5 x 4-4'5 pm; cellulae inferiorae plus minusve cylindraceae, 6-7 x 3'5-4 pm.
Status imperfectus. Conidiophora in cultura pura prodita, ramosa, saepe aggregata,
genus Penicillium simila; phialides solitariae vel in penicillus aggregatae, in parte
superiore conidiophororum praeditae, conicae vel curvatae, 3-1 I x 3-3'5 pm. Conidia
non catenulata, oblonga, cylindracea vel ovoidea, saepe ad basim truncata, glabra,
hyalina, 3'5-6 (8'5) x 2.8-3'5(3.8) pm. Hab. in ramis emortuis Rhododendri ferrugineae,
Helvetia, Valesia, in silvis nomine 'Aletschwald' prope Brigua, 12 Sept. 1968, leg. B. Aebi
et E. Muller (TYPE K).

Perithecial stromata up to 4 mm diam, golden yellow in colour, hemi-

spherical or lobed, smooth, ostioles somewhat darker, slightly raised,
Vol. 57, Part 3 was issued 24 December 1971
2 Transactions British Mycological Society
shining. The stromata arise as pustules which burst through the bark of
the host, Rhododendron ferrugineum. Between the bark and the wood, the
mycelium forms a conspicuous subiculum of closely packed hyaline cells,
5-9.um diam, with very thick walls (up to z usx: thick). In the xylem of
the host the mycelium is thin-walled and 3-5 .urn diam. The cortex of the
perithecial stroma is ofsmall cells, 4-5 .urn diam, closely packed and thick-
walled. Medulla pseudoparenchymatous, at the base composed of densely
packed thick-walled cells resembling those of the subiculum, composed
of segments up to 28.um long x 5-7 pm, with fatty globules. The lower
part of the stroma may also include fragments of the host cortex. The
medulla of the body of the stroma is made up of thinner-walled, wider
cells radiating outwards from the point of origin of the stroma, measuring
up to 50 pm long and up to 15 pm wide. Perithecia numerous, tightly
packed, so that the perithecial walls may be flattened, about 200 pm
high and about 150 pm diam. Asci numerous, cylindrical, 180-200 x 5 pm.
Paraphyses not seen. Ascospores hyaline, uniseriate, bicellular, often
dividing into part-spores within the ascus. Each part-spore bears
numerous fine spines on its surface. The upper part-spores are subglobose,
to truncate-ovoid, and measure 4'5-5 x 4-4'5 pm. The lower part-spores
are rather longer and narrower, cylindrical or tapering, and measure
6-7'5 x 3'5-4 pm.
CULTURES

Colonies slow-growing at room temperature, growing best at tempera-

tures near 15°C, with only slight growth above 20°. The appearance of
the colony is watery, with the mycelium mostly confined to the sub-
stratum, and only the conidiophores projecting. With age the medium
may be tinged yellow. Mycelium within the substratum only infrequently
branched, anastomosing, septate, with numerous small granules visible,
about 5 .urn diam. Chlamydospores not seen.
Aerial mycelium sparse, consisting of long, stolon-like hyphae about
5 pm diam, branching at their tips to form clusters of phialides. More
compact white clumps of conidiophores, about 1-2 mm diam, may also
be formed. The diffuse conidiophores bear lateral branches singly and
the laterals may either bear phialides directly, or produce further
branches on which the phialides are borne. The ultimate branches may
be closely crowded and each ends in a candelabrum-like cluster of one to
about seven phialides. The whole arrangement is somewhat reminiscent
of a Penicillium. Phialides skittle-shaped, and the outer members of a
cluster are often incurved so that the spore balls from a cluster of phialides
may cohere to form a single mass. Phialides measure 3-11 x 3-3'5 pm.
Conidia are variable in shape, oblong-eylindrical to ovoid, often with
a truncate base, smooth, usually biguttulate, white in mass, 3'5-6(8'5) x
2.8-3'5(3.8) pm.
In 6-week-old cultures grown in the light at 10-15 °C yellow perithecial
stromata develop.
Hypocrea psychrophila. E. Muller, B. Aebi and J. Webster 3

!!' II
A
Imm

:::::;,: ,)
~:; ~: . ~ ~

~ ' ,"
E
..
, ~ ~

A'-.
'.
B
L-...J
20 /lm

D lOpm

Fig.!. Hypocreapsychrophila (M.R.S.D. 3031). A, V.S. Stroma, B, ascospores. C, portion

of outer cortex. D, portion of subiculum composed of thick-walled cells; the stippled
cells are part of the host periderm. E and F, Conidiophores. G, Phialides and conidia.
C, D to same scale j B, G to same scale.

1-:2
4 Transactions British Mycological Society

TEMPERATURE RELATIONS

Growth is possible between 0° and 24°. The optimum temperature is

about 15°. Grown in lower temperatures (up to 12°), Hypocrea psychrophila
forms a colourless mycelium. Between 15° and 21° there is a red pig-
mentation of the medium, and at 24° growth is very weak, so that again
no pigmentation may be observed. Conidia are produced between 3°
and 21°.
The type specimen (Mycological Herbarium of Sheffield University 303 1)
was collected on Rhododendronferrugineum, Kanton Wallis, bei Brig, alter
Belalpweg, by E. Muller and B. Aebi, 12 Sept. 1968. It is now deposited
in Herb. K.
Additional collections: on Rhododendron ferrugineum, Kanton Wallis,
Aletschreservat near Brig, Switzerland, 8 Sept. 1970; on R. ferrugineum,
Kanton Graubunden, Arosa, near station of suspension railway to Weiss-
horn, Switzerland, 30 Aug. 1959, leg. E. Rahm; on Vaccinium myrtillus
Kanton Graubunden, Arosa, 'Schwarzsee ', Switzerland, 20 Oct. 1960,
leg. E. Rahm. These specimens are in the Herb. ZT.
The morphology of the conidial apparatus is similar to Hypocrea gelatinosa
(Tode ex Fr.) Fr. (Webster, 1964) and therefore morphologically inter-
mediate between the form genera Trichoderma and Gliocladium. The
phialides are similar to those typical for Trichoderma (Rifai, 1969), but
their arrangement is more penicillioid as in Gliocladium. The species of
Trichoderma known at present produce no pigment or are green-pigmented.
Red pigments are unknown in Trichoderma; they may be produced in
Gliocladium but are not typical for the conidial stage of Hypocrea gelatinosa,
which in any case has green conidia.
Hypocrea psychrophila therefore is a species easily distinguishable because
of its golden-yellow perithecial stromata, its psychrophilic growth and the
red pigmented Gliocladium-like conidial state with whitish conidia. The
thick-walled cells at the base of the perithecial stroma are also unusual in
Hypocrea and may possibly be related to its alpine habitat.

REFERENCES
RIFAI, M. (1969). A revision of the genus Trichoderma. Mycological Papers no. I 16, pp. I-56.
WEBSTER, J. (1964). Culture studies on Hypocrea and Trichoderma. I. Comparison of the
perfect and imperfect states of H. gelatinosa, H. TUfa and Hypocrea sp. I. Transactions of
the British Mycological Society 47, 75-96.

(Acceptedfor publication 1 February 1971)