Mycologia, 98(5), 2006, pp. 678–681.
# 2006 by The Mycological Society of America, Lawrence, KS 66044-8897
Utilization of various carbon sources for the growth of waterborne conidial fungi
S.C. Sati
S. Bisht1
Department of Botany, Kumaun University, Nainital –
263002, India
Abstract: Four isolates of waterborne conidial fungi
(Tetracheatum elegans, Tetracladium marchalianum,
Pestalotiopsis submersus and Flagellospora penicil-
lioides) were investigated for their carbon require-
ment, using eight different carbon sources (viz.
glucose, fructose, sucrose, xylose, starch, cellulose,
dextrin and lactose). All fungi tested grew sparsely on
the basal medium lacking in carbon, which was the
control. However these fungi were found to vary in
their ability to use the supplied sources of carbon.
Glucose and sucrose were found to be suitable
sources of carbon for all four fungal isolates, whereas
fructose proved good for T. marchalianum and P.
submersus. Starch and xylose also supported growth of
T. marchalianum, P. submersus and F. penicillioides.
Cellulose, a polysaccharide, was a poor source of
carbon for the growth of these isolates. Four g/L of
glucose was recorded as the most useful concentra-
tion that gives the maximum dry weight of selected
fungi (262 mg and 400 mg for T. elegans and P.
submersus respectively after 15 d).
Key words: aquatic hyphomycetes, carbon utiliza-
tion, conidial fungi, fungal growth
INTRODUCTION
Waterborne conidial fungi, previously known as
aquatic hyphomycetes (Ingold 1942), represent the
major microbial elements of decaying leaves in
running freshwater. These fungi are characterized
by their magnificent conidial types (Ingold 1975).
Submerged plant litter serves as substrates for the
nutrition of these fungi. They use a wide range of
simple and polymer carbohydrates from abscised
leaves or litter.
In studies on the nutrition of aquatic hyphomycetes
(Ranzoni 1951) reported that thiamine and a carbo-
hydrate source are required for their growth. Aquatic
hyphomycetes can satisfactorily use a wide range of
carbohydrates, which includes starch, cellulose, cello-
biose, sucrose, mannose, xylose, maltose, glucose and
galactose. Thornton (1963) analyzed leaf extract
Accepted for publication 25 Jun 2006.
1
Corresponding author. E-mail: saraswatibisht@yahoo.com
678
carbohydrates, and these leaf extract carbohydrates
were tested for utilization by aquatic hyphomycetes in
vitro. Jones and Stewart (1972) reported that some
aquatic hyphomycetes are able to use cellobiose and
starch and can degrade cellulose effectively.
The occurrence and distribution of these fungi
have attracted attention worldwide (Webster and
Descals 1981, Marvanova 1997), but there is a paucity
of knowledge on their physiology (Sridhar et al 1992,
Sati and Pant 2000). Limited studies on carbon
nutrition of aquatic hyphomycetes are available
(Jones and Stewart 1972; Gulis and Suberkropp
2003, 2004). Carbon nutrition of aquatic fungi,
especially on waterborne conidial fungi, is still an
area that requires investigation.
Thus investigation of the nutritional requirements
of some commonly occurring waterborne conidial
fungi, isolated from the streams of the temperate
zone in Kumaun Himalaya, India, (1600–2000 m asl,
29u59–31u259N, 77u489–81u69E), was carried out to
determine the use of various carbohydrates for the
growth of four selected waterborne conidial fungi.
MATERIALS AND METHODS
Four fungal species of waterborne conidial fungi (viz.
Flagellospora penicillioides Ingold, Pestalotiopsis submersus
Sati and Tiwari, Tetracheatum elegans Ingold and Tetra-
cladium marchalianum de Wildeman) were selected for the
present investigation. Monohyphal cultures of these isolates
were obtained by single spore isolation. The cultures were
maintained at 20 6 2 C on 2% malt extract agar slants with
initial pH 6.5 and in Petri dishes containing malt extract
agar.
The basal medium consisted of KH2PO4, 1g; MgSO4 ?
7H2O, 0.2 g; FeCl3 ? 6H2O, 0.02 g; Difco yeast extract,
1.0 g/L distilled water. To find the optimal concentration
of carbon sources for the best growth of waterborne
conidial fungi, preliminary experiments were conducted
by growing T. elegans and P. submersus in different
concentrations of glucose, which is a good source of
carbon. The range of concentration was taken arbitrarily,
and 4 g/L of glucose in the basal medium was found to be
the desirable concentration that gives maximum dry weight
of waterborne conidial fungi (TABLE I). Eight carbon-
containing compounds (glucose, fructose, sucrose, xylose,
starch, cellulose, dextrin and lactose) were taken separately
in sufficient quantity to create a concentration equivalent to
4.0 g of carbon/L of medium. Three replicates were used in
each experiment for each fungal isolate. Uniform circular
agar blocks (5 mm diam) containing mycelial mat were cut
from a 15 d old culture plate and transferred into sterilized
conical flasks (100 mL cap.) containing 25 mL of basal
 SATI AND BISHT: CARBON SOURCE UTILIZATION 679

TABLE I. Average dry weight yields (6SEM) of two selected marchalianum and P. submersus. Xylose also sup-
fungal isolates in different concentrations of glucose as ported good growth of all fungi except T. elegans.
source of carbon Starch was found to be a good source of carbon for T.
Growth of fungal isolate (after 15 d) elegans and P. submersus in comparison with T.
(Average dry weight in mg.) 6SEM* marchalianum and F. penicillioides. Cellulose was
observed to be a poor source of carbon for all studied
Concentration (g/L) T. elegans P. submersus
fungi (FIG. 1). Dextrin also was found to be a good
2 151.0 6 0.4 202.0 6 0.3 carbon source for the growth of these fungi. Lactose
4 262.0 6 0.8 400.0 6 1.0 supported moderate growth of P. submersus and F.
6 200.0 6 0.5 314.0 6 0.6 penicillioides but was a poor carbon source for T.
8 178.0 6 0.7 209.0 6 0.5 marchalianum and T. elegans (FIG. 1).
* SEM 5 Standard error of mean.
DISCUSSION

medium. The inoculated flasks were incubated at 20 6 2 C
in the dark and casually shaken for aeration. After 15 d of
incubation the net hyphal growth of the fungus in terms of
mycelial dry weight in the basal medium was determined.
Adhered agar medium from the mycelial mat was removed
by straining through a filter paper (Whatman No. 1). The
mycelial mat was rinsed with distilled water 3–4 times to
remove traces of basal medium and it was placed in a drying
oven at 80 C for 4 h. The fungal biomass was weighed with
a digital electronic balance.
RESULTS
The eight carbon sources (glucose, fructose, sucrose,
xylose, starch, cellulose, dextrin and lactose) were
screened for the growth of four selected waterborne
conidial fungi (viz., Flagellospora penicillioides, Pesta-
lotiopsis submersus, Tetracheatun elegans and Tetra-
cladium marchalianum). The growth results of these
fungal isolates are provided (TABLE II). Basal medium
without carbon source (the control) supported little
growth. Four g/L of glucose was the most useful
concentration to enhance fungal growth (262 mg and
400 mg for T. elegans and P. submerses respectively)
(TABLE I). Glucose and sucrose were found to be the
most suitable carbon sources for all inoculated fungi,
whereas fructose was found to be equally good for T.
Waterborne conidial fungi occur abundantly on
abscised, mature and submerged leaf litter in running
freshwater (Sati and Tiwari 1997, Sati et al 2002).
These fungi have nutritional advantage over some of
their close competitors (viz. the zoosporic fungi,
bacteria and zooplanktons) because decaying leaves
and litters contain specific nutrition required by the
conidial fungi (Thornton 1963).
In the present study out of eight carbon sources
glucose supported maximum growth of P. submersus
and F. penicillioides. The remaining fungal isolates
also showed comparatively higher growth. These
observations conform with studies of (Ranzoni
1951) on glucose as a good source of carbon for the
growth of Anguillospora longissima and A. gigantea.
Thornton (1963) similarly found glucose as a good
source of carbon for the growth of Articulospora
tetracladia, Flagellospora penicillioides, Tricladium an-
gulatum, T. splendens, Tetracladium setigerum, Var-
icosporium elodeae and V. aurantiaca.
Starch, fructose and dextrin are good sources of
carbon for the growth of P. submersus and T.
marchalianum (FIG. 1). Sucrose supports maximum
growth of T. elegans as well as of T. marchaliunum and
moderate growth of P. submersus and F. penicillioides.
These results support the findings of (Ranzoni 1951).

TABLE II. Average dry weight yields (6SEM) of water borne conidial fungi after 15 d using different sources of carbon

Growth of fungal isolate (after 15 d) (Average dry weight in mg.) 6SEM*

Carbon Sources Tetracheatum elegans Tetracladium marchalianum Pestalotiopsis submersus Flagellospora penicillioides

Glucose 284.0 6 7.21 100.0 6 2.89 438.0 6 20.13 449.0 6 21.52

Fructose 47.3 6 2.19 139.3 6 9.68 430.0 6 11.55 187.0 6 19.31
Sucrose 90.0 6 0.58 107.0 6 3.51 394.03 6 12.44 366.0 6 52.16
Xylose 38.3 6 5.78 98.0 6 1.15 347.0 6 52.00 334.7 6 12.72
Starch 57.3 6 2.40 89.0 6 0.58 396.3 6 39.01 129.0 6 2.31
Cellulose 6.00 6 0.58 12.3 6 2.03 59.7 6 10.74 39.0 6 1.73
Dextrin 45.3 6 3.53 97.3 6 13.28 422.7 6 58.72 356.7 6 12.67
Lactose 29.7 6 2.96 10.3 6 1.86 361.0 6 29.14 245.3 6 24.88

* SEM 5 Standard error of mean.

680
FIG. 1. Dry weight yields of the four fungal isolates in different carbon sources after 15 d incubation at 20 C.
Thornton (1963) also recorded that fructose, xylose
and starch are good sources of carbon for some
species of aquatic hyphomycetes. He concluded that
starch could be an alternative source of carbon for
these fungi. Mer (1982), while studying the carbon
requirements of zoosporic fungi (watermolds), found
that dextrin is a good source for the growth of
Leptolegnia caudata and Saprolegnia subterranea.
Lactose was found to promote moderate growth of
P. submersus and F. penicillioides; however it resulted
in poor growth for the remaining two fungal species
(FIG. 1).
It is interesting to note that cellulose is a poor
source of carbon for the growth of these hyphomy-
cetous fungi (FIG. 1). This supports (Ranzoni 1951)
who also recorded the absence of growth of A.
gigantea and A. longissima in cellulose as a source of
carbon in medium. These fungi probably lack the
MYCOLOGIA
enzyme necessary to degrade cellulose. The fast
colonization of these fungi on leaf litter might be
explained on the basis of the availability of nutrients
and polysaccharides preferred by them.
ANOVA calculated for the observed data showed
significant variations in the dry weight produc-
tion of different fungal species grown with diff-
erent carbon sources (p , 0.01). This indicates
that different fungal species have different priorities
for their carbon source and one carbon source that is
less preferred by one fungal species could be pre-
ferred by another fungal species. This might be due
to the fact that simple carbon compounds are
assimilated directly while complex ones (i.e. poly-
saccharides) must be converted into simpler forms
before their use. Glucose (a monosaccharide) and
sucrose (a disaccharide) are well used by the
waterborne fungi.
 SATI AND BISHT: CARBON
ACKNOWLEDGMENTS
The authors thank Prof. S.P. Singh, head, Department of
Botany, Kumaun University, Nainital, for providing labora-
tory facilities and Dr M. Belwal for his help in conducting
the experiments. We also thank Prof. S. Chandra (Botany)
and Dr Madhu Joshi (English) for their careful reading of
the manuscript and improving it.
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