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Archives of Applied Science Research, 2011, 3 (3):389-393

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ISSN 0975-508X
CODEN (USA) AASRC9

Preliminary screening of alternative culture media for the growth of

some selected fungi
S. Tharmila, E. C. Jeyaseelan and A. C. Thavaranjit
Department of Botany, Faculty of Science, University of Jaffna, Jaffna, Sri Lanka
______________________________________________________________________________
ABSTRACT
Based on the market value and scarcity of culture media, screening of alternative media for the
growth of fungi is found to be an important task for microbiologist in developing countries. In
this study an attempt was made to test the possibility of using some cheaply available local
materials (Sago, raw dried Palmyrah tuber flour, tubers of sweet potato and cassava) as
alternative media for fungal growth. The sago and Palmyrah tuber flour were able to solidify
without adding agar at the concentration of 10g / 100ml and 16g / 100ml respectively, within 25
minutes. Culture discs of Mucor sp., Penicillium sp., Fusarium sp. and Trichoderma sp. were
transferred on the centre of the test media and the control media, potato dextrose agar (PDA).
The diameters of fungal colony were measured at different time points. The results were
subjected to analysis of variance (ANOVA) (P<0.05) followed by Tukey test. This study
demonstrated that the sago and Palmyrah tuber media were suitable for the growth of all the
tested fungi and the growth of Penicillium sp. on Palmyrah tuber medium was significantly
higher when compared to PDA and sago medium. The results for Trichoderma sp. on sago
medium and Mucor sp. on sago and Palmyrah tuber media were comparatively better than PDA
after 24 hours incubation. However, with the prolonged incubation period the growth rate
increases quickly in PDA than in tested media. Present study clearly shows the possibility of
using cheap locally available sago and Palmyrah tuber media as an alternative media for PDA
for the fungal growth. Further study is needed to elucidate the active principles for the self
solidification and fungal growth of these two media.
Key words: culture media, fungi, PDA, sago media and Palmyrah tuber media.
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INTRODUCTION
The growth of microorganisms in an artificial medium is influenced by several physical and
chemical factors. A nutrient material prepared for the growth of microorganisms in a laboratory
is called culture media and the nutrient composition of a culture medium plays a major role in
microbial growth [1]. For microbiological studies, commercially available ready made culture
media are used in research laboratories. These culture media are standardized and they are in

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different types, some media are used for general purpose and some others are used for specific
growth of certain microorganisms [2].
Potato Dextrose Agar (PDA) is commonly used for the isolation and growth of wide range of
fungi in laboratories and its compositions are well defined [3]. However, in developing countries
microbiological researches are hindered by high cost and scarcity of culture media [4].
Therefore, in recent years some researchers concentrated to screen alternative culture media from
locally available cheap materials [4, 5, 6]. Even though they successfully cultivated different
types of fungi on the alternative media, in all the cases they had to add agar as a solidifying
agent. The cost of agar gives additional burden. Therefore, present study was conducted to test
the ability of four locally available substances to serve as nutritive and self solidifying source for
the growth of different types of fungi.
MATERIALS AND METHODS
Preparation of test powder
500g of Sago, raw dried Palmyrah (Borassus flabellifer Linn.)tuber flour, tubers of sweet potato
(Ipomoea batatas) and cassava (Manihot esculenta) were obtained from local shop. The sago
was ground into fine powder using electric grinder. The raw dried Palmyrah tuber flour was
dispensed in water for 10- 15 minutes and the supernatant was decanted, this was repeated five
times in order to remove an antimicrobial substance present in the flour [7]. Then it was filtered
with muslin cloth, dried and sieved through fine mesh. Sweet potato and cassava tubers were
scraped and dried in shade and then ground in to fine powder.
Test fungi
In this study, four different fungi namely; Mucor sp., Penicillium sp., Fusarium sp. and
Trichoderma sp. were tested. These fungal cultures were obtained from the culture collection of
Department of Botany, University of Jaffna, Jaffna, Sri Lanka. Prior to the study above fungi
were sub-cultured on fresh PDA medium.
Test for the solidification
Media were prepared by adding different weights of sago, Palmyrah tuber, cassava and sweet
potato powders (2g, 4g, 6g, 8g, 10g, 12g, 14g, 16g and 18g) in 100ml solution (50ml distilled
water + 50ml king coconut water) separately and then they were mixed well by stirring and
heating in hotplate stirrer till the solution became homogenous. The media were transferred to
250ml capacity conical flasks and plugged with absorbent cotton and sterilized in an autoclave at
121 OC for 15 minutes under the pressure of 15 lbs/inch2. The conical flasks were taken out and
20ml of sterilized cooled molten media were poured into sterile Petri dishes separately. Media
solidification time was noted for each concentration of each test powder. The procedure was
repeated thrice. PDA medium was prepared following the manufacturers instruction and the
time required for the solidification of PDA also noted. The media composition which showed
equal mean solidification time as the solidification time required for PDA was selected and used
for the fungal growth studies.
Comparison of fungal growth on test media and PDA
Solid media of sago and Palmyrah tuber were prepared with 10g / 100ml and 16g / 100ml
concentration respectively. The test fungi in actively growing pure cultures were taken and 8mm
diameter discs were cut with sterile cork borer and transferred on the centre of sago and
Palmyrah tuber media separately. Control was maintained by transferring fungal discs on PDA
medium. The plates were incubated at 28 OC in dark and the radial growth of mycelia from disc
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onto the medium was measured at 24 hours interval. The study was repeated three times and the
mean radial growth was determined.
Statistical data analysis
The results obtained on Sago, Palmyrah tuber and PDA media were subjected to examine by
analysis of variance (ANOVA) (P<0.05) followed by Tukey test ( = 0.05) by using a software,
SPSS 14.0 for Windows version.
RESULTS
The test for self solidification of the tested powders revealed that the media prepared with
cassava and sweet potato powders did not have enough solidification that favor for the fungal
growth. But, the media prepared using Sago and Palmyrah tuber showed better solidification and
the lowest concentration required for the solidification of sago and Palmyrah tuber were
6g/100ml and 12g/100ml concentration respectively. Furthermore, in these two powders with the
increases in the concentration the time required for the solidification was decreased (Table 1).
Table 1: Mean solidification time for different concentration of sago and Palmyrah tuber media
Weight of test powder(g/100ml)

Mean solidification time

Sago medium
Palmyrah tuber medium
2
Not solidified until 45 min Not solidified until 45 min
4
Not solidified until 45 min Not solidified until 45 min
6
38 min
Not solidified until 45 min
8
30 min
Not solidified until 45 min
10
25 min
Not solidified until 45 min
12
20 min
35 min
14
15 min
29 min
16
12 min
25 min
18
10 min
21 min
25 min
Mean solidification time required for the PDA

The study for the possibility of fungal growth on sago and Palmyrah tuber media showed that all
the four tested fungi were able to grow on both of these media and in all the cases the mean
radial growth increased with time. However, the rate of increases varied with the type fungi. The
test fungi Trichoderma sp. grew well on sago medium compare to Palmyrah tuber and PDA
media after 24 hours incubation, but later the rate of growth increased greatly in PDA and
Palmyrah tuber media. After 72 hours the Trichoderma sp. showed higher mean growth on PDA
and Palmyrah tuber media (Table 2).
Table 2: Mean radial growth of Trichoderma sp. on test and control media.
Mean radial growth (mm)*
24hours
48hours
72hours
96hours
Sago
33.3 1.5a 37.0 1.0b 44.0 1.0c 47.0 1.0c
Palmyrah tuber 14.8 0.8c 32.2 1.0c 59.8 1.3b 78.5 0.5b
PDA
30.5 0.5b 53.0 1.0a 74.8 1.0a 82.0 1.0a
*Values are diameter of fungal colonies (Mean SD), Values with different superscripts in the same column differ
significantly (P < 0.05).
Media

The Mucor sp. showed better growth on sago and Palmyrah tuber media compared to PDA after
24 hours incubation. However, with the increasing incubation period the growth rate highly

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Arch. Appl. Sci. Res., 2011, 3 (3):389-393
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accelerated on PDA and sago rather than Palmyrah tuber (Table 3). The test fungus Fusarium sp.
showed better growth on PDA and followed by Palmyrah tuber and sago (Table 4).
Table 3: Mean radial growth of Mucor sp. on test and control media.
Mean radial growth (mm)*
24hours
48hours
72hours
96hours
Sago
22.5 0.5a 32.0 1.0b 51.3 1.5b 68.3 0.6b
Palmyrah tuber 20.3 0.6b 24.5 0.5c 30.3 0.6c 37.7 0.6c
PDA
15.5 0.5c 35.3 0.6a 65.5 0.9a 80.0 0.0a
*Values are diameter of fungal colonies (Mean SD), Values with different superscripts in the same column differ
significantly (P < 0.05).
Media

Table 4: Mean radial growth of Fusarium sp. on test and control media.
Mean radial growth (mm)*
24hours
48hours
72hours
96hours
Sago
14.8 0.3b 18.0 1.0c 22.7 0.6c
Palmyrah tuber 10.2 0.3b 12.7 0.6c 22.0 1.0b 30.5 0.5b
PDA
11.3 0.3a 20.0 0.0a 25.5 0.5a 32.8 0.8a
*Values are diameter of fungal colonies (Mean SD), Values with different superscripts in the same column differ
significantly (P < 0.05). (-) No growth
Media

Most interesting result was obtained for Penicillium sp. on Palmyrah tuber medium, where it
grew well compare to PDA and sago medium. After 96 hours incubation the mean radial growth
was 61.5 0.5mm, 19.3 0.6mm and 21.7 0.6mm in Palmyrah tuber, sago and PDA media
respectively (Table 5).
Table 5: Mean radial growth of Penicillium sp. on test and control media
Mean radial growth (mm)*
24hours
48hours
72hours
96hours
Sago
13.5 0.5b 17.0 0.0c 19.3 0.6c
Palmyrah tuber 16.3 0.6 34.7 0.6a 47.7 0.6a 61.5 0.5a
PDA
13.8 0.3b 19.5 0.5b 21.7 0.6b
*Values are diameter of fungal colonies (Mean SD), Values with different superscripts in the same column differ
significantly (P < 0.05). (-) No growth
Media

DISCUSSION
There is an increasing difficulty for microbiology researchers for their routine works as well as
for university and especially school students for their microbiological studies to buy imported
commercially available culture media in developing countries like Sri Lanka.
The results of the present study revealed that the sago and Palmyrah tuber media can be used for
the in vitro cultivation of different types of fungi and for the preparation of these two media do
not depend on additional solidifying agent.
The growth of mycelia, spore formation and colony morphology of the test fungi in sago and
Palmyrah tuber media were also found to be similar as in PDA. For instance, the Trichoderma
sp. always produces alternative rings of spores and mycelia in PDA, the same feature was also
observed in these two media.

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The cost for commercially available 500g PDA (Oxoid) is about 137 US$ (LKR 15000) in local
chemical shops. Palmyrah is one of the natural resources in Sri Lanka. It is widely distributed
throughout the tropics. In Sri Lanka it exists in the dry zone, mainly in the North and East
Provinces. Raw dried Palmyrah tuber flour is obtained by grinding the raw dried tuber [7]. The
annual potential of tuber flour from Palmyrah of Northern region is around 3000 metric tons [8]
and the cost for 500g flour is about 0.7 US$ (LKR 80). This illustrates the abundance of this
source in this region. Sago is imported from other countries to Sri Lanka and 500g of this powder
cost about 1 US$ (LKR 120). Therefore, it is clear that the sago and Palmyrah tuber flour media
are cheaper raw materials compared to the cost of commercially available PDA.
Some previous studies also proved the possibility of alternative local materials to replace PDA.
In a former study different fungi were successfully cultured in alternative media using different
cereals to replace potato in PDA [4]. In another study with powdery substances in pod of Parkia
biglandulosa revealed the possibility of using this powder instead of PDA for the fungal growth
[5]. The study carried out with watery endosperm of Cocos nucifera showed that the watery
endosperm was found to be effective for the growth of Colletotrichum gloeosporioides [6].
However, in all above studies they used agar as a solidifying agent. 500g of agar (Oxoid) cost
about 60 US$ (LKR 6500), it causes further expenses. But Sago and Palmyrah tuber flour media
have self solidifying substances and they support the growth of fungi without any alterations in
the solidity of the media.
CONCLUSION
The present preliminary study for the screening of alternative local and cheaper sources for the
fungal culture media shows that among the four tested sources Sago and Palmyrah tuber flour
can be used instead of PDA with very low expenses. Therefore, we recommend this media for
the fungal growth in order to carryout microbiological works. Further studies regarding the
chemical composition of this media and cultivation of other types of fungi are needed to enrich
the value of this finding.
REFERENCES
[1] Tortora GJ, Funke BR, Case CL. Microbiology an introduction, 5th ed., The
Benjamin/Cummings, New York, 1995; pp. 147-150.
[2] Atlas RM. Microorganisms in our world, 1st ed., Von Hoffmann, New York, 1995; pp. 8485.
[3] Sharma G, Pandey RR. Journal of Yeast and Fungal Research, 2010, 1, 8, 157-164.
[4] Adesemoye AO, Adedire CO. World Journal of Microbiology and Biotechnology, 2005, 21,
329-336.
[5] Rajasab AH. Current Science, 2007, 92, 25, 1359-1360.
[6] Marikar FMMT. Micologia Aplicada International, 2009, 21, 2, 63-66.
[7] Balasubramanium K, Jansz ER, Aryasen DD. Palmyrah, International program in chemical
science (IPICS), Uppsala University, Sweden, 1999; pp. 1-5.
[8] Jansz ER, Karunatillaka N, Theivendirarajah K. Extraction and exploitation of palmyrah
tuber starch for agro industry, Proceedings and abstracts of seminar on the new development in
the exploitation in Sri Lanka, Palmyrah Development Board, Sri Lanka, 1992.

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