International Journal of Botany

and Research (IJBR)

ISSN(P): 2277-4815; ISSN(E): 2319-4456
Vol. 4, Issue 5, Oct 2014, 29-34
TJPRC Pvt. Ltd.

EFFECT OF FORMULATED CULTURE MEDIA ON GROWTH

OF SOME FUNGAL SPECIES
OMODARA TOLANI RACHAEL1 & ADEBOLU, TINUOLA TOKUNBO2
1

Department of Microbiology, Faculty of Science, Ekiti State University, Ado-Ekiti, Ado-Ekiti, Ekiti, Nigeria
2

Department of Microbiology, School of Sciences, Federal University of Technology, Akure, Ondo, Nigeria

ABSTRACT
Agar culture media were formulated using low-cost carbohydrate sources such as cocoyam (CYDA), yam (YDA),
sweet potatoes (SPDA), irish potatoes (IPDA) and cassava (CDA). The formulated media were used to culture the
following fungal species:
Fusarium moniliforme, F. oxysporium, Aspergillus niger, A flavus, Penincilliun notatum, Rhizopus stolonifer,
Mucor mucedo and Aspergillus fumigatus. The cultures were incubated at 2520C and the radial growth was measured at
24hr, 48hr, 72hr and 96hr. Moreover, the colour, texture and fluffiness of the mycelia were also examined. Morphological
examination was also done to study the effect of these formulated media on the hyphae branching, arrangement of spores,
size of sporangia, cornidiophores and septa formation of the test fungi. Growth on commercial potato dextrose agar served
as control. Result showed that growth on Irish potatoes dextrose agar and yam dextrose agar were better than on potato
dextrose agar. Growth on sweet potato dextrose agar and cassava dextrose agar were almost the same; while least growth
was found on cocoyam dextrose agar. Though all the formulated media supported the growth of fungi, the effects on the
morphology of the fungi varied considerably. Therefore, Irish potatoes dextrose agar and yam dextrose agar can be used
for the culturing of these fungi in the absent potato dextrose agar.

KEYWORDS: Fusarium moniliforme, F. oxysporium, Aspergillus niger, A flavus, Penincilliun notatum, Rhizopus
stolonifer, Mucor mucedo, Aspergillus fumigatus

INTRODUCTION
Culture medium can be defined as a substrate which can support the growth of microorganisms outside its normal
hosts (Roger et al., 1977). The first person to formulate culture medium was Anthony Van Leeuwenhoek in 1965
(Carpenter, 1961). Since that time, various kinds of culture media have been continuously being formulated with great
diversity in their composition. To be able to have good growth, proper nutrient has to be supplied. Microorganisms vary in
their nutritional requirements, so a good culture medium should be able to meet the nutritional requirements of different
microorganism. Deliberate cultivation under laboratory conditions is a pre- requisite to study microbes (Carpenter, 1961),
so in order to monitor the growth activities of microorganism, they must be cultured on an appropriate culture medium.
The primary goal of constructing a culture therefore is to provide balance mixture of the required nutrient at
concentration that will permit good growth of the microorganism. However, it ia advisable to make the nutrient as rich as
possible because many nutrient may become growth inhibitory or toxic as the concentration is raised (Alexopolous and
Mims, 1979). The acceptability and suitability of any media formulation is primarily dependent on careful selection

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Omodara Tolani Rachael & Adebolu, Tinuola Tokunbo

(sampling) and specification through tests. Such test includes identification procedure and compatibility with other
components of the culture media. Though a lot of work has been done on formulation of media for growth of
microorganism especially bacteria and fungi in the bid of searching for cheaper sources of material for their growth, more
so that commercially prepared culture medium are very expensive, more is still desirable. Media formulation should be a
continuous work perhaps one may come up with very good ones that will not only support the growth of these organisms
but also improve their yield and quality of useful metabolites e.g. enzymes amino acids, antibiotics and so on produced by
some of them.
This work was done not only to study whether culture media formulated from different tubers will support the
growth of some fungi species commonly encountered in our surrounding but to study the effects of these media on the
morphological characteristics as well.

RESEARCH METHODS
The tubers used for the study were bought at Oja-Oba market, Akure. The tubers were yam (Discorea rotundata),
cocoyam (Xanthosoma sagithefolium), sweet potato (Ipomea batata), Irish potato (Solanium tuberosum) and cassava
(Manihot esculanta).
Preparation of Tubers for Extract
Fresh tubers used for the study i.e yam, cocoyam, sweet potato, Irish potato cassava were peeled and washed
separately with distilled water. This was done for three times. The tubers were then allowed to dry at room temperature for
five minutes and cut into smaller pieces. Two hundred and fifty grams of each of the different types of tubers were
weighed separately into different 1000mls flask and each made up to 1000mls with distilled water and boiled for 30mins.
The tuber extract were filtered using different Whatman No1 filter papers.
Preparation of Tubers Extract Dextrose Agar
The tubers extracts dextrose agar i.e yam dextrose agar (YDA), cocoyam dextrose agar (CYDA), sweet potato
dextrose agar (SWDA), Irish potato dextrose agar(IPDA) and cassava dextrose agar (CDA) were prepared separately by
measuring 500mls of each of the extract into separate conical flasks. Ten grams (10g) of the dextrose was added followed
by 5g of agar powder into each flask. Each mixture was placed on the hot plate for 15minutes so that the agar can dissolve.
Sterilization of the extract was done in autoclave at 121oC for 15 minutes. This was later poured aseptically into separate
sterilized Petri dishes. Inoculation was done in done after the media had set.
Inoculation of the Media
The organisms used for the study were Fusarium moniliforme, Fusarium oxysporium, Aspergillus
niger,Aspergillus flavus,Penicillium notatum Rhizopus stolonifer,Mucor mucedo and Aspergillus fumigatus.
Pure cultures of these organisms were used for the study. The culturing was done at room temperature for
96hrs.The culturing was done using sterilized cork borer to cut the plate containing the pure culture after wish the sterilized
wire loop was used to transfer the cut portion to the center of the prepared media plates. The diameter of the cork borer was
0.4cm. Inoculation was done in triplicates for each of the formulated media. The prepared commercial dextrose agar was
also inoculated in the same manner. The plates were incubated at room temperatures by inverting the Petri dishes.
The diameter of the growth was measured at every 24hrs using vennier calipers for four days.
Impact Factor (JCC): 1.6913

Index Copernicus Value (ICV): 3.0

31

Effect of Formulated Culture Media on Growth of Some Fungal Species

Moreover after 48hrs sterile wire was used to pick part of the mycelia on the culture media and was placed on
a sterile slide moistened with lactophenol blue, the slide was viewed under the microscope at a magnification of 100.
This was done for the growth of the formulated media and the organism on the commercially prepared potato dextrose agar
(PDA).
Analysis of Tubers
The moisture, protein, carbohydrate, fat, crude fibre and ash content of tubers were determined by (AOAC, 2000).
The oven dry method was used was for determining moisture content, for protein content determination, the micro kjedahl
method (2000) was used while the carbohydrate content was determined using the direct acid hydrolysis method. The fat
content was determined using graph metric method, the Weede method was used for determining the crude fibre content
and the ash content was determined using the incineration method over an open flame.
Determination of pH of the Media
The pH determination was done by pH meter immediately after preparation i.e. a digital pH meter (Janway pH
meter p165 model 3510) with a probe was used for the pH measurement of the media.

RESULTS AND DISCUSSIONS

The physico-chemical characteristics of the media prepared are shown in table 1. Sweet potato dextrose agar had
the least pH value of 5.65 while yam dextrose agar and cocoyam dextrose agar had the highest pH value of 5.85.
The commercially prepared potato dextrose agar, however had a greater value of 5.90. All these pH values fall within the
acidic region. The colour of most of the prepared media was light grey except Irish potato dextrose agar (IPDA) that was
pale yellow and cocoyam dextrose agar (CYDA) that was actually grayish in colour. The appearance of most of the
prepared media was transparent except for yam dextrose agar (YDA) and cocoyam dextrose agar (CYDA) that were
slightly turbid.
Table 1: Physico-Chemical Characteristics of the Formulated Media
Medium
YDA
CYDA
SWDA
IPDA
CDA
PDA

pH Value
5.85
5.70
5.65
5.75
5.85
5.90

Medium Colour
Light grey
Grey
Light grey
Pale yellow
Light grey
Yellow

Medium Clarity
Cc
Cc
Cc
Cc
Cc
Cc

Cc

Absence of particles

slightly turbid

++++ =

YDA =

Yam dextrose agar

CYDA =

not turbid
Turbid
Cocoyam dextrose agar

SWD

Sweet potatoes dextrose agar

IPDA

CDA

Cassava dextrose agar

PDA

Turbidity
+
++++
_
_
_

Irish potatoes dextrose agar

Potatoes dextrose agar

The proximate composition of the various carbohydrate tubers used to replace potato dextrose agar is presented in
table 2. The result shows that Irish potato had the highest moisture content of about 80%. This was followed by yam with
moisture content of 75% while the moisture of sweet potato and cocoyam were almost the same i.e. 70% and 70.20%
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Omodara Tolani Rachael & Adebolu, Tinuola Tokunbo

respectively. The moisture content of cassava was the least i.e. 62.80%. In terms of carbohydrate content, the carbohydrate
content of cassava was the highest (34%) followed by Irish potato (29%), sweet potato (27%) and cocoyam (24%) while
yam had the least carbohydrate content of 14.75%. Irish potato had the highest protein content (3.00%) followed by sweet
potato and yam with the same amount of protein content of 2.25%, followed by cocoyam with protein content of 1.70%
while cassava had the least protein content of 0.25%. The highest percentage of fat was recorded in cocoyam (0.40%),
followed by cassava (0.25%) and sweet potato (0.25%) and the least value was recorded in Irish potato (0.15%)
The highest percentage content of ash was recorded in cocoyam (1.7%) followed by yam (1.3%) and sweet potato (1.05%)
while the least value was recorded for Irish potato (0.95%).Table 2: Proximate composition of the tubers
Table 2
Types of
Tubers
Cassava
Cocoyam
Irish potato
Sweet
potato
Yam

Composition of Tubers (%)

0.26
0.40
0.15

Crude
Fibre
1.60
0.90
2.50

1.00
1.30
0.95

2.25

0.20

1.50

1.05

2.25

0.18

1.00

1.70

Moisture Content

Carbohydrate

Protein

Fat

62.80
70.20
80.00

34.00
24.00
29.00

0.25
1.70
3.00

70.00

27.00

75.00

14.50

Ash

As shown in table 3 below, all the media supported the growth of all organisms used namely Fusarium
oxysporium, Aspergillus niger, Aspergillus flavus, Penicillum notatum, Rhizopus stolonifer, Mucor mucedo and Aspergillus
fumigaitus.
The growth performance of some of these fungi species was even better on the formulated media than the
commercially prepared PDA. For instance, Fussarium moniliforme, Fusarium oxysporium, Asprregillus niger, Rhizopus
stolonfer, Mucor mucedo and Aspergillus Fumigatus grew best on Irish potato dextrose agar (IPDA) and yam dextrose agar
(YDA) than on commercially prepared potato dextrose agar (PDA) (p0.05). the reason for this performance may be due to
the high protein content of the tubers. It is however evident from the result that cocoyam is not suitable substitute for
potato in the preparation of a fungus culture medium because it is not transparent hence growth measurements was difficult
to estimate. But for cultural and morphological studies, most of these formulated media were not good enough because
they affected the cultural characteristics and morphological characteristics of these fungi species. The only exceptions were
Irish potato dextrose agar (IPDA) and yam dextrose agar (YDA) that did not affect these characteristics. Therefore in the
absence of commercially formulated potato dextrose agar (PDA), laboratory prepared Irish potato dextrose agar (IPDA)
and yam dextrose agar (YDA) can be used for the cultivation of fungi species. Irish potato dextrose agar (IPDA) and yam
dextrose agar (YDA) were the only formulated media that did not alter the cultural characteristics and only Irish potato
dextrose agar (IPDA) did not alter the morphological characteristics of the fungal species used in this study. Therefore,
they are being recommended for the use in research purposes. Much work is still desired to study the effect of these media
on the physiology of these organisms. Table 3: Radial growth (mm) of test fungi on formulated media

Impact Factor (JCC): 1.6913

Index Copernicus Value (ICV): 3.0

33

Effect of Formulated Culture Media on Growth of Some Fungal Species

Table 3
Test Fungi

PDA

Fusarium
19.50
moniliforme
Fusartium
37.38
oxysporium
Aspergillus
42.98
niger
Aspergillus
37.35
flavus
Penincillum
19.05
notatum
Rhizopus
39.55
stolonifer
Mucor
37.50
mucedo
Aspergillu
36.65
fumigatus
YDA = Yam dextrose agar

YDA

MEDIA
SPDA
CDA

CTDA

IPDA

19.25

18.50

17.25

15.75

21.25

42.48

37.73

40.05

37.53

40.38

43.68

43.15

24.73

24.90

37.03

42.33

34.60

20.23

38.27

16.05

15.53

11.38

13.33

17.18

40.00

32.80

34.25

29.50

42.05

34.13

33.00

32.63

28.70

44.40

44.30

37.73

40.05

37.53

42.88

SWD = Sweet potatoes dextrose agar

CDA = Cassava dextrose agar

45.75

CYDA = Cocoyam dextrose agar

IPDA = Irish potatoes dextrose agar
PDA = Potatoes dextrose agar

REFERENCES
1.

Alexopolus, C. J. & Mims. C. W. (1979). Introductory mycology, (3rd ed.). New York: John Wiley.

2.

A. O. A. C. (1990). Official Method of Analysis, Washington D. C: Association of Official Analytical Chemist

Inc.

3.

Carpenter, P. L. (1961). Microbiology (1st ed.). London: Oxford University Press

4.

Roger, V. S, Edward, A. A. and John, L. I. (1977). General Microbiology, (4th ed.). London: Macmillan Press
limited.

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