Notes and brief articles

by A. alternata, A. fiavus, A. niger, A. tamarii, C. PAVLIDES, G ., VARDAVAKIS, E. & LAVRENTIADES, G . (198 5)

cladosporioides, and total Aspergilli.
The writer wishes to express his gratitude to Dr
W. Gams, Centraalbureau voor Schimmelcultures,
Baarn, The Netherlands, for his encouragement
and correction of the manuscript.
REFERENCES
PARK, D. (1973). A modified medium for isolation and
enumeration of cellulose-decomposing fungi. Trans-
actions of the British Mycological Society 60, 148-151.
(unp ublished). Notes on the vegetation and soil profiles
near Polygyros (Chalkidiki, N. Greece).
PUGH, G . J. F . ( 1964). An investigation of soil-borne
cellulose-decomposing fungi in Greece. Annals of
Institute Phytopathology Benaki, N .S. 7, 19-27.
SOIL SURVEY STAFF (1975). Soil Taxonomy . Soil conserva-
tion Service. U.S. Department of Agriculture, U.S.A .
VARDAVAKIS, E. & LAVRENTIADES, G . ( 1982). Cellulolytic
microfungi in some soils of the Anthemous valley in
Northern Greece. Nova Hedungia 36, 715-724.

EFFECT OF PIPTOCEPHALIS SPECIES ON GROWTH AND SPORULATION

OF PILAIRA ANOMALA

BY S. N. WOOD AND R. C. COOKE

Botany Department, University of Sheffield, Sheffield S10 2TN

In agar culture mycoparasitism by either Piptocephalis fimbriata or P. freseniana reduced

growth and sporulation of the coprophile Pilaira anomaia over a temperature range of 15-3 0 "C.
It is, however, argued that in nature the presence of mycoparasites on herbivore faeces
probably has little effect on development of coprophilous Zygomycotina.
A common feature of the fruiting sequence on Cultures of P. anomala, originally isolated from
herbivore dung is the rapid but short-lived sheep droppings, and dual cultures of Piptocephalis-
appearance of mucoralean sporangiophores. Early P . anomala were maintained in standard Petri
cessation offruiting of mucoralean species, usually dishes on 15 ml aliquots of Lab M malt extract-
after only a few days, may be attributed to the peptone agar (M EP ; 3 % malt extract, o'S %
greater ability of coprophilous Ascomycotina and mycological peptone, 1'2 % agar ) at 15 °C. For
Basidiomycotina for resource capture and the experiments, the inoculum consisted of solidified
susceptibility of Zygomycotina to direct physical spore suspensions which were prepared as follows.
antagonism by them (Ikediugwu & Webster, P. anomala sporangiophores were obtained from
1970a, b). However, other factors might also be 14-d cultures by removing them with sterile
involved. In this regard it has been noted that forceps to a 25 ml square-bottomed universal vial
sporulation of mucoralean saprotrophs on dung is containing 10 ml sterile distilled water plus 0'2 ml
frequently accompanied by that of closely-related Tween 80, followed by homogenizing for 1 min to
biotrophic mycoparasites, for example species of break the sporangial walls. The homogenate was
Piptocephalis de Bary, some of which are capable of centrifuged for 60 s at low speed and the resulting
severely affecting development of their mucoralean supernatant, containing only sporangiospores, was
hosts (Harper & Webster, 1965; Curtis et al., 1978; decanted and centrifuged at high speed for 5 min.
Evans, Lewis & Cooke, 1978; Evans & Cooke, The spore pellet was resuspended in sterile distilled
1981). Although the ecological significance of this water and retained. Piptocephalis spores were
kind of mycoparasitism remains undetermined, the removed from 14-d Piptocephalis - P. anomala
close association of mycoparasites and potential cultures using sterile distilled water followed by
hosts in dung may indicate a possible role for the filtration through three layers of sterile muslin to
former in reducing development of the latter. As remove P. anomala sporangia. Spore densities were
part of a study on the autecology of the coprophile not determined. Three vials each containing 15 ml
Pilaira anomala (Cesati) Schroeter, the effect of of molten MEP agar were held a145° in a water bath
Piptocephalis fimbriata Richardson & Leadbeater and to each was then added either 5 ml of P.
(IM I 205706) and P. freseniana de Bary (IM I anomaia sporangiospores or 2'5 ml of this plus
141598) on growth and sporulation of this 2'5 ml of spores of either of the two Piptocephalis
ephemeral species was determined. Both Pipto- species. After thorough mixing the agars were
cephalisspeciesusedhavcbeenrecordedfromdung. poured into Petri dishes and incubated at 15°
Trans. Br. mycol. Soc. 86 (4), (1986) Printed in Great Britain
 Notes and brief articles

Table 1. Sporulation of P. anomala at different temperatures in the presence and absence of Piptocephalis
species, Each figure is the mean with S.E.M. of the number of sporangia along a single colony radius; four radii
scored on each of duplicate colonies
Temperature (0C)

10 is 20 2S 30
P. anomala 164'O±S'S 197'6±6'S 98'4±S'3 63'4±2'8 0
P, anomala-i Pi fimbriata 132'4±6'1 164'9±7'1 44'4±4'S* 28'4±4'4* 0
P, anomala-s-Pi freseniana 177'6±4'7 S9'1±S'3t 24'6±2'1t 9'4±o'8t 0
Significantly different from P, anomala alone at *, S % level; t, 1 % level.

Table 2. Growth rate, mmj 24 h, of Pilaira anomala at different temperatures in the presence and absence of
Piptocephalis species. Each figure is the mean from five replicates with S.E.M.
Temperature (0C)

10 is 20 2S 30
P, anomala 7'29±o'08 10'16±o'18 13'80±O'18 14'43±O'1Q 3'14±o'08
P, anomala-s Pi fimbriata 6'44±O'03 10'S1±O'09 12'S2±O'07* 12'80±O'12t 1'S3±o'06t
P, anomala-v Pi freseniana 7'41±O'08 10'7S±O'14 14'84±o'08* 16'oS±0'12t 2'02±O'OSt
Significantly different from P, anomala alone at *, S% level; t, 1 % level.

overnight. These were then examined micro-
scopically to confirm that germination had taken
place and 6 rom diam agar disks were cut from them
and used to inoculate a series of dishes containing
15 ml MEP agar. For P. anomala and each
Piptocephalis-P, anomala combination five replicate
dishes were incubated at 10, 15, 20, 25 and 30°.
Colony diameters were measured at 24 h intervals
until either the mycelium filled the dish or for a
maximum of 10 d. In either case the final level of
sporulation of P. anomala was assessed on two
plates, taken at random from each treatment, by
counting the number of sporangia present on three
6 mm diam disks taken from across four radii on
each plate. Disks were cut from the colony edge,
from the median point along the radius and from
a position immediately adjacent to the inoculum
disk. Counts for each radius were totalled and the
means for eight radii obtained.
Results are shown in Tables 1 and 2. Of the
temperatures used, 15° and 25° were the most
favourable for sporulation and hyphal extension of
P. anomala respectively. Both Piptocephalis species
sporulated at all temperatures except 30°, and both
severely reduced sporulation of P. anomala at 20°
and 25°, with P.freseniana additionally doing so at
15°. At 20°, 25° and 30° extension rates of P.
fimbrata-P. anomala colonies were significantly less
than those of P. anomala alone as were those of P.
[reseniana-P, anomala colonies at 30°. However, at
20° and 25° P. freseniana-P. anomala colonies
Trans. s« mycol. Soc. 86 (4), (1986) Printed in Great Britain
advanced significantly more rapidly than did those
of the host alone.
These results indicate that, in agar culture, the
presence of mycoparasites can strongly affect
growth and sporulation of P. anomala, although
this may not represent the situation which obtains
in nature. Unpublished experiments carried out in
this laboratory have demonstrated that Piptocephalis
spores survive passage through the rabbit. However,
even those Piptocephalis species which occur
regularly on faeces may not be strictly coprophilous
but are more likely to have arisen via invasion from
underlying soil or litter (Richardson & Leadbeater,
1972). Since their subsequent development would
depend on the presence of an already established
mucorine flora, their effects on potential hosts
might be much less than would occur ifboth groups
of fungi arrived together, the latter situation being
simulated here in co-inoculation of agar plates. In
addition, whilst Piptocephalis species have been
found to severely affect growth of non-coprophilous
hosts at 15°, this does not occur with respect to P.
anomala at and below that temperature (Evans et
al., 1978). Whatever the reasons for this, it may
indicate curtailment of significant mycoparasite
activity at what might be considered as being
ecologically meaningful temperatures.
Leaving aside ecological considerations, the
observation that P. freseniana-P. anomala colonies
extended at a greater rate than host colonies alone
requires further comment, In earlier studies on the
 Notes and brief articles
effects of Piptocephalis species on host growth it was
found that, after a set incubation period and
depending on the particular mycoparasite-host
combination and temperature, dual colonies some-
times achieved a significantly greater diameter than
corresponding host colonies (Curtis et al., 1978).
However, results were based on a single measure-
ment rather than on regular measurements over a
period of time. Where actual rates of growth have
been calculated, it has been found that the greater
diameter achieved by dual colonies can be
attributed, in some instances, to the occurrence of
a reduction in the lag phase for growth in
Piptocephalis-host inocula, subsequent extension
rates being identical in both dual and host colonies
(Yousif, 1983). This effect has not been explained
but here, by contrast, it is extension rates which are
influenced. This is despite the fact that colonies of
the Piptocephalis species used occupy a central
position within the host colony, the concentric
colony margins remaining in excess of Q' 5 em apart
until the host has filled the Petri dish.
We are pleased to acknowledge financial support
of this work by the Science and Engineering
Research Council.
REFERENCES
CURTIS, F. C., EVANS, G. H., LILLIS, V., LEWIS, D. H. &
COOKE, R. C. (1978). Studies on mucoralean myco-
parasites. 1. Some effects of Piptocephalis species on host
growth. New Phytologist 80, 157-165.
EVANS, G. H. & COOKE, R. C.(1981). Piptocephalis-
induced changes in organization of marginal hyphae in
Mycotypha microspora colonies. Transactions of the
British Mycological Society 76, 343-345.
EVANS, G. H., LEWIS, D. H. & COOKE, R. C. (1978).
Studies on mucoralean mycoparasites. II. Persistent
yeast-phase growth of Mycotypha microspora Fenner
when infected by Piptocephalis fimbriate Richardson &
Leadbeater. New Phytologist 81, 62!Hl35.
HARPER, J. E. & WEBSTER, J. (1964). An experimental
analysis of the coprophilous fungus succession. Trans-
actions of the British Mycological Society 47, 511-530.
IKEDlUGWU, F. E. O. & WEBSTER, J. (1970a). Antagonism
between Coprinus heptemerus and other coprophilous
fungi. Transactions of the British Mycological Society 54,
180-204·
IKEDlUGWU, F. E. O. & WEBSTER, J. (1970b). Hyphal
interference in a range of coprophilous fungi. Trans-
actions of the British Mycological Society 54, 205-210.
RICHARDSON,M. J.&LEADBEATER,G.(1972).Piprocephalis
fimbriata sp.nov. and observations on the occurrence of
Piptocephalis and Syncephalis. Transactions ofthe British
Mycological Society 58, 205-215.
YOUSIF, H. A. (1983). Studies on the haustorial myco-
parasite Piptocephalis xenophila Dobbs & English. Ph.D.
Thesis, University of Sheffield.

DUALITY OF ASPERGILLUS CANDIDUS ON MAIZE KERNELS

BY C. A. FRANKLAND
Department of Biological Sciences, University of Natal, Durban, S.A.
AND H. L. LLOYD
Department of Microbiology, University of Durban-Westville, S.A.

Aspergillus candidus isolates have a duality of colony and sporulation morphology. Transition
from the floccose form with penicilliate microconidiophores to the adpressed form with
aspergillate macroconidiophores is influenced by the nature and availability of carbon. Dual
wild types seem to have a competitive (selective) advantage over the separated components
on natural substrates.

Duality is the manifestation of the dissociated and
separate, morphological expressions of two geneti-
cally-dissimilar components of a heterokaryon
(dikaryon). This phenomenon is invariably reported
to be associated with the isolation and culture of
fungi on artificial media (Hansen, 1938; Jinks,
1959a; Smith & Anderson, 1973; Bull & Bushell,
1976; Fincham, Dam & Radford, 1979) since only
when there is clearly defined colony development
can the differences in dual types be observed. The
association of duality with culturing has led to the
phenomenon being viewed as a cultural artefact
with little or no significance in the natural habitat
of fungi. However, its widespread occurrence and
thus its persistance in wild-type anamorphs
suggests that the composite wild type has a selective
(evolutionary) advantage over its components.
Many aspergilli exhibit dual colony forms in

Trans. Br. mycol. Soc. 86 (4), (1986) Printed in Great Britain