European Scientific Journal October 2014 edition vol.10, No.

30 ISSN: 1857 7881 (Print) e - ISSN 1857- 7431

THE POTENTIAL EFFECT OF TWO

CYANOBACTERIAL SPECIES; ANABAENA
SPHAERICA AND OSCILLATORIA AGARDHII
AGAINST GRAIN STORAGE FUNGI

Wafaa Haggag
Plant Pathology Dept., National Research Center, Dokki, Cairo, Egypt

Azza M. Abd El-Aty

Water Pollution Research Dept., National Research Centre, Cairo, Egypt

Amal A. Mohamed
Plant Biochemistry Dept., National Research Centre, Cairo, Egypt

Abstract
Cyanobacterial secondary metabolites have a diverse antagonistic
activity that lead to disintegration of microbial growth. The present study
was undertaken to explore the inhibitory effect of three different solvents
extracts (methanol, acetone and water) of two cyanobacteria species:
Oscillatoria agardhii and Anabaena sphaerica extracts
against grain
storage fungi. In vitro fungal growth, mycotoxins production of grain
storage fungi (Fusarium equiseti; F. moniliforme; F. solani; F.
proliferatum , F. graminearum; Penicillium digitatum; Aspergillus terreus;
Aspergillus niger; Aspergillus flavus; Aspergillus oryza; Alternaria alternate
and
Alternaria triticina) were significantly(p<0.01) inhibited by
cyanobacterial extracts. HPLC analysis revealed that compelate reduction of
Aflatoxin B1, Zearalenone and Fumonisin B1 with acetone extract of
Oscillatoria agardhii and minimum concentration (0.3g/L) was found with
Ochratoxin A.
Acetone extracts of both
cyanobacteria species have
potential for the suppression of all grain storage fungi and increased seeds
germination of maize, wheat, barley and rice. It was also observed that when
wheat seeds treated with water extract for one hour soaking, significant
(p<0.01) increased in seed germination and reduced the frequency of seeds
storage fungi during 180 days was also observed. These results statements be
a necessary (or) recommended that the both species of Cyanobacterial having
a potential capacity as a source of antifungal activity, which makes them
interesting for screening for natural and highly bioactive secondary
metabolites products and
it may leads to development of new
pharmaceutical agents.

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Keywords: Antifungal activity, Anabaena sphaerica, inhibition zone,

Oscillatoria agardhii, grain storage fungi
Introduction
One of the most important effects of post-harvest decays seed and
feed deterioration by fungi is the induction of mycotoxicoses. This is a
disease of animals and humans following consumption of feeds and foods
invaded by fungi that produce toxic substances called mycotoxins. The
storage fungi, mainly comprising several species of Aspergillus and
Penicillium, do not invade grains to any appreciable degree or extent before
harvest (Mehrotra, 1983), but they can cause severe dis-coloration of seed in
storage resulting in germination failure, dis-coloured or otherwise damaged
embryos or whole seeds, and production of mycotoxins that constitute a
health hazard for man and animals (Dharam Vir, 1974 and Malaker et al.,
2008). Each species or group of species of Aspergillus has its own rather
sharply defined lower limit of moisture content usually between 13 and 18%
for invasion of stored grains. Some common mycotoxicoses caused by
common and widespread fungi such as Aspergillus, Penicillium, Fusarium
and Stachybotrys result in severe illness and death. Aspergillus and
Penicillium produce their toxins mostly in stored seeds, hay or commercially
processed foods and feeds although infection of seeds usually takes place in
the field. Aflatoxin is about the most popular and widespread mycotoxin. Its
name derives from the fact that it was originally found to be produced by
Aspergillus flavus (Agrios, 1978 and Haggag, 2013 a & b), but is now
known to be produced by other species of Aspergillus. Aflatoxin B1 is
produced by Aspergillus terreus, though it may also be produced by
Aspergillus flavus as well as Aspergillus oryzae. It is the most toxic,
carcinogenic and most prevalent of the different aflatoxins (Haggag et al.,
2014 a, b). Algae are one of the chief biological agents that have been
studied for the control of plant pathogenic fungi particularly soil borne
disease. Cyanobacteria are characterized by their capacity to perform
biological nitrogen fixation and oxygenic photosynthesis. As cyanobacteria
are very resistant to extreme environmental conditions, they are assuming
increasing importance in frontier areas of biotechnology. The typical
anabiosis and rapid restoration of activity under favorable conditions are
characteristic of them (Pankratova, 1987). Cyanobacteria have the capability
to produce metabolites both intracellular and extracellular. Secondary
metabolites from cyanobacteria are associated with toxic, hormonal,
antineoplastic and antimicrobial effects (Shweta et al., 2011; Abd El-Aty et
al., 2014). Cyanobacteria from local habitats seem to be a source of
potential new active substances that could contribute to reduction of the

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number of bacteria, fungi, viruses and other microorganisms (Mundt et al.,

2001). Biologically active compounds found in plants appear to be more
adaptable, acceptable and safer than synthetic compounds and display a
wealthy source of potential pathogens control agents.
Moreover,
Cyanobacteria are also reported to produce a wide range of plant growth
regulators such as abscisic acid, ethylene, jasmonic acid, auxin, and
cytokinin-like substances as well as the cytokinin isopentenyl adenine (Kim,
2006). Fungal pathogens cause many diseases in plants leading to destruction
of resources in agriculture. Traditional methods to circumvent fungal
pathogens involve the application of synthetic fungicides and growing
resistant cultivars. The use of crude extract from algae may be an alternative
to conventionally used as antimicrobial to control microbe growth, due to
their secondary metabolites compounds (Abd El-Aty et al., 2014).
Cyanobacteria have drawn much attention as prospective and rich sources of
biologically active constituents and have been identified as one of the most
promising groups of organisms to be able of producing bioactive compounds
(Fish and Codd, 1994 & Schlegel et al., 1999). Therefore, the objective of
the present work was to study the in vitro antifungal activity of three
different extracts (methanol, acetone and water) of Oscillatoria agardhii and
Anabaena sphaerica against grain storage fungi.
MATERIALS AND METHODS
Algae growth and different extracts preparation:
Two algal species were selected to evaluate their antifungal activity
against some species of fungi. These algae belong to cyanobacteria,
Anabaena sphaerica (N2-fixing blue algae) and Oscillatoria agardhii. The
tested algal species were previously isolated from phytoplankton community
structure of River Nile at Ismailia canal, Egypt. BG11 medium was used for
maintenance of Oscillatoria agardhii (Carmichae, 1986) and Nitrogen free
formula for Anabaena sphaerica (Abd El-Aty et al., 2014). The cultured
media were incubated at 30 2 C without aeration and under continuous
illumination of fluorescent lamps with intensity 2500 lux. The cultures were
shaken every day to prevent algal cell clumping and adherence of algal cells
to the containers. Oscillatoria agardhii and Anabaena sphaerica were
characterized for their antifungal activity as follow:
Extracts preparation
The algal pellets (37gm) were extracted using serial Exhaustive
Extraction Method (Das et al., 2010) with methanol, acetone, and water. All
extracts were dried and weighed to estimate the concentration in 1 ml. Dried
extracts were either reconstituted in ethanol or deionized water.

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Isolation and purification of pathogenic fungal strains

Fungi were isolated from stored maize, wheat, maize and rice grains.
The grains were surface-sterilized in NaOCl for 2 min and rinsed in two
changes of sterile distilled water. The grains were plated on sterile potato
dextrose agar (PDA) at the rate of 10 grains per plate. Thirty grains were
plated per crop and incubated at room temperature of 25 1oC. Sub-cultures
were made from emerging colonies and pure cultures obtained for
subsequent studies. After incubation the growth characters as well as
percentage of infection were recorded. In order to isolate these fungi into
pure culture, Potato Dextrose Agar (PDA) was prepared and the fungi were
inoculated onto the sterile PDA and incubated for 7 days at the end of which
the fungi were identified based in their colour, spore morphology and
mycelia growth using the light microscope (Begum et al., 2004). The fungi
identification was done according to the methods of Barnett and Hunter
(1972).
Test fungi
Twelve species of Fusarium equiseti; F. moniliforme; F. solani; F.
proliferatum , F. graminearum; Penicillium digitatum; Aspergillus terreus;
Aspergillus niger; Aspergillus flavus; Aspergillus oryza; Alternaria alternate
and Alternaria triticina isolated from above seeds, selected and used as
test fungi for antifungal activity assay.
Antifungal activity assay of Cyanobacteria extracts
The antifungal activity was tested by using the filter paper disc
diffusion method (Saha et al., 1995) employing 24 hours cultures of the
above mentioned organisms.
The agar media were seeded by fungal suspensions adjusted to 105
CFU/ml, sterile filter paper discs (6 mm diameter) were saturated with 250
g/ml of each crude extract. The Petri dishes were placed for 5 h at 4 C till
the metabolites release into the medium. The plates were incubated at 37 C
for 24 h. The antifungal activity was determined by measuring the diameters
of the clean inhibitory zone around each paper disc.
Effect of algal extracts on dry weight and aflatoxin production of
toxigenic fungi
One ml of conidial suspension of fungi (1xl07 conidia/ml) was
inoculated into 100 ml medium amended with each extracts to obtain the
previous concentration. They were then incubated at room temperature for
10 days. Three replications were used for each treatment (including the
control). Observations were made on the dry weight of mycelia of fungi and
aflatoxin production. The dry weight of mycelia was determined by drying

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the fungal colonies in an oven at 700C for 4 days until a constant weight was
attained. Aflatoxin B1 content was determined using a High Performance
Liquid Chromatography method (Christian, 1990).
The contents of individual fungal culture were transferred into
separate glass bottles and extracted with methanol (85%). Bottles were
capped and shacked on a rotary shaker at 120 rpm for 90 min and the
suspensions were centrifuged at 3500 rpm for 20 min. Supernatants were
transferred to glass bottles and kept at room temperature for analysis.
Aflatoxin levels were measured using high performance liquid
chromatography (HPLC) (model: Perkin Elmer series 200 UV/VIS) with a
C18 column that had an internal diameter of 300 x 3.9 mm. The HPLC
apparatus was equipped with a UV detector, and fluorescence was measured
using 365-nm excitation and 430-nm emission wavelengths. The mobile
phase consisted of methanol: acetic acid: water (20:20:60 v/v/v). The total
run time for the separation was approximately 30 min, and the flow rate was
1 ml/min (Christian, 1990).
Seed treatment with algal extracts: Selected seed samples of grains
were treated following dipping method. The seeds were dipped in each algal
extracts with the same concentration for 1 hour in algae previously prepared
extracts (Acetone, water, and methanol). After 1 hour, extracts were drained
out from the petri dishes. The treated seeds were allowed to be dried up on
filter paper for some time and were tested following the standard blotter
method (ISTA. 1976), to observe the growth of different fungal colonies on
the seeds. For each treatment 100 seeds with three replications were placed
on six petri dishes.
Determination of seed germination (%) of fresh and stored wheat seeds
Blotter method: Three pieces of blotting paper were placed in fold in each
Petri dish 9 cm diameter and incubated at 252C. Three replicates were
prepared. The percent germination of wheat seed was calculated after one
week (De Tempe, 1953). The experimental data were recorded from fresh
seeds as well as those stored, after every three months (0, 90 and 180 days)
of storage at room temperature.
Statistical Analysis The data collected from the experiment were
analyzed for test of significance and compared the treatment means
following Completely Randomized Design (CRD) by using Duncan's
Multiple Range Test (DMRT) at 5% level of probability.
RESULTS AND DISCUSSION
Tested fungi
The study shows that maize, wheat, barley and rice grains were
infested to various degrees with storage fungi. A total of twelve species of

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Fusarium equiseti; F. moniliforme; F. solani; F. proliferatum , F.

graminearum; Penicillium digitatum; Aspergillus terreus; Aspergillus
niger; Aspergillus flavus; Aspergillus oryza; Alternaria alternate and
Alternaria triticina isolated from grain seeds. Maize and wheat were the
most infested of all fungi (Table 1). Mycotoxin assay revealed that several
toxins were produced i.e.
aflatoxin, fumonisin, Ochratoxin A and
zearalenone were identified in this study. Most the fungi produced one toxin
or the other (Table 2).
Table (1). Frequency of grain storage fungi associated with grains
Isolates
Fusarium equiseti
F. moniliforme
F. solani
F. proliferatum
F. graminearum
Penicillium digitatum
Aspergillus terreus
Aspergillus niger
Aspergillus flavus
Aspergillus oryza
Alternaria alternate
Alternaria triticina

Maize
7.2
9.2
8.4
11.4
4.2
6.4
6.7
15.3
12.4
3.2
11.5
-

Percentage %
Wheat
Barley
3.2
2.1
1.4
0.7
3.1
3.2
2.4
2.4
3.3
6.6
8.4
1.3
11.4
2.4
5.4
4.2
6.4
3.2
15.3
2.4
4.6
2.4

Rice
1.3
1.3
12.3
-

Table (2). Mycotoxins produced by fungi isolated from stored grains in liquid media.
Isolates
A. flavus
A. terreus
A. oryzae
Fusarium sp.
A. niger
P. italicum

Amount
Aflatoxin B1
Aflatoxin B1
Aflatoxin B1
Zearalenone
Ochratoxin A
Fumonisin B1

Mycotoxin (g/L)
20.42
11.21
7.54
4.41
18.6
12.71

Antifungal activity
Antifungal potentiality of solvents and aqueous extracts of
cyanobacterial extracts of Oscillatoria agardhii and Anabaena sphaerica
were tested against twelve species of Fusarium equiseti; F. moniliforme;
F. solani; F. proliferatum , F. graminearum; Penicillium digitatum;
Aspergillus terreus; Aspergillus niger; Aspergillus flavus; Aspergillus
oryza; Alternaria alternate and Alternaria triticina in vitro as growth
reduction and mycotoxins production (Table 3, 4 and 5). It was concluded

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Table (3). Antifungal activity of Oscillatoria agardhii and Anabaena sphaerica extracts against fungal grains pathogens

Isolates

Fusarium equiseti
F. moniliforme
F. solani
F. proliferatum
F. graminearum
Penicillium digitatum
Aspergillus terreus
Aspergillus niger
Aspergillus flavus
Aspergillus oryza
Alternaria alternate
Alternaria triticina
LSD

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Acetone
23.3
24.1
20.7
22.5
18.7
11.4
13.5
11.3
10.8
9.8
23.4
22.4
1.6

Diameter of inhibition zone (mm)

Oscillatoria agardhii
Anabaena sphaerica
Methanol
Water
Acetone
Methanol
13.4
9.4
16.2
10.3
16.7
7.9
15.5
12.4
17.9
6.5
16.3
11.4
18.8
7.7
15.2
12.4
15.6
8.1
14.2
11.3
8.8
5.5
9.8
7.6
10.8
7.3
11.3
9.8
8.8
8.1
8.1
16.8
17.6
1.5

6.3
6.6
6.4
9.9
8.6
0.8

9.7
8.7
7.7
10.8
12.5
1.3

6.4
6.6
6.2
8.8
9.8
1.2

Water
7.7
6.6
5.5
5.8
9.1
6.6
6.6
5.5
5.5
6.6
9.5
9.3
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Table (4). Antifungal activity of Oscillatoria agardhii and Anabaena sphaerica extracts against fungal grains pathogens
Isolates

Fusarium equiseti
F. moniliforme
F. solani
F. proliferatum
F. graminearum
Penicillium digitatum
Aspergillus terreus
Aspergillus niger
Aspergillus flavus
Aspergillus oryza
Alternaria alternate
Alternaria triticina

Acetone
0.00
0.02
0.01
0.04
0.02
0.04
0.03
0.04
0.03
0.05
0.04
0.05

Mycelial inhibition (Dry Mycelial Weight method mg/l)

Oscillatoria agardhii
Anabaena sphaerica
Methanol
Water
Control
Acetone
Methanol
Water
0.9
6.3
39.8
09.0
2.1
8.8
0.9
6.3
38.7
09.0
3.3
8.7
1.0
6.4
35.3
1.1
3.3
9.6
2.4
8.3
30.3
1.2
3.4
8.3
2.3
8.4
36.6
09.0
2.4
8.3
2.4
7.3
23.3
09.0
3.2
10.8
2.3
8.2
22.3
08.0
2.3
11.3
2.0
8.2
24.4
09.0
2.2
12.2
2.0
9.6
20.3
09.0
2.5
12.8
1.3
8.8
21.3
07.0
1.5
11.1
2.2
4.8
34.4
09.0
2.4
10.1
1.1
4.3
33.3
09.0
1.4
10.4

Control
39.8
38.7
35.3
30.3
36.6
23.3
22.3
24.4
20.3
21.3
34.4
33.3

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Table (5). Antifungal activity of Oscillatoria agardhii and Anabaena sphaerica extracts against mycotoxins produce
Isolates

A. flavus
A. terreus
A. oryzae
Fusarium sp.
A. niger
P. italicum

435

Mycotoxin

Aflatoxin B1
Aflatoxin B1
Aflatoxin B1
Zearalenone
Ochratoxin A
Fumonisin B1

Mycotoxins production (g/l)

Acetone
0.0
0.0
0.0
0.0
0.3
0.0

Oscillatoria agardhii
Methanol
Water
0.4
3.4
0.5
3.2
0.5
1.3
1.0
1.4
2.3
2.4
2.0
1.4

Control
20.42
11.21
7.54
4.41
18.6
12.7

Acetone
0.5
0.5
0.5
0.8
1.5
0.4

Anabaena sphaerica
Methanol
Water
1.2
6.4
1.5
7.2
0.9
5.3
1.9
4.3
2.7
5.4
2.5
2.4

Control
20.42
11.21
7.54
4.41
18.6
12.71

European Scientific Journal October 2014 edition vol.10, No.30 ISSN: 1857 7881 (Print) e - ISSN 1857- 7431

that the diameter of inhibition zone depends mainly on the type of the algal
species, type of solvent used and the tested fungal species. The experimental
analysis of antifungal effects indicated that all tested fungi showed higher
sensitivity to the acetone extract of Oscillatoria agardhii with the higher
zone with Fusarium spp. and Alternaria spp. as shown in Table (3). Whereas
the methanol extract showed moderate activity against all fungal species.
The same results were also shown in liquid medium, significant
(p<0.01) inhibited of mycelial growth was observed in Oscillatoria agardhii
extract followed by Anabaena sphaerica (Table 4). The highest growth
reduction was recorded with Fusarium spp. and Alternaria spp. Acetone
extract is the best solvent in reducing fungal growth and mycotoxin
production (Table 5). Complete reduction of Aflatoxin B1, Zearalenone and
Fumonisin B1 with acetone extract of Oscillatoria agardhii and minimum
concentration was found with Ochratoxin A. Biological natural products
isolated from cyanobacterial extracts of Oscillatoria agardhii and Anabaena
sphaerica extracts are recommended as potential as antimicrobial (Abd ElAty, et al., 2014 ). Broad ranges for the in vitro anti-fungal activities of
different cyanobacterial extracts of Oscillatoria agardhii and Anabaena
sphaerica extracts. The results obtained recorded the higher antifungal
activity for acetone extract of Oscillatoria agardhii as indicated by inhibition
in dry weight and mycotoxin production. The phenolic compound released
from dried crudes of cyanobacterial extracts may be answerable for their
antifungal properties as confirmed earlier by (Abd El-Aty, et al., 2014) who
found that phenolic compounds responsible for the antibacterial activities of
these extracts. Other scientists have also reported that phenolic compounds
from different plant sources could inhibit various food-borne pathogens
(Plaza et al., 2009; Osman et al., 2011). The lack of activity in other extract
in the disc diffusion assay doesnt mean that the extracts have no active
ingredients. Though the screening by disc diffusion assay has its own advent
as it is simple, less time consuming and requires only a small quantity of
material.
Efficacy of different extract on seed germination and stored wheat seeds
In this experiment different extracts were used in controlling seedborne fungi of four grain seeds. In all grain seeds, the highest germination
was found when the seeds were treated with cyanobacterial of Oscillatoria
agardhii and Anabaena sphaerica extracts and it increased germination
compared with untreated control (Table 6). Among the extracts, best
performance in terms of reducing percent seed-borne infection was obtained
through treating the seeds of acetone extract of Oscillatoria agardhii. This
may be due to the impact of these antifungal compounds on seeds
germination. Reduction in germination of wheat seeds during storage might

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be due to activity of the storage fungi. It was also observed that wheat seeds
of the aqueous extract soaked for one hour duration soaking, significant
(p<0.01)increased in seed germination. It was also observed that when
wheat seeds treated with water extract for one hour soaking,
significant(p<0.01) increased in seed germination and reduced the frequency
of seeds storage fungi during 180 days was also observed (Table 7 & 8).
Among the extracts, best performance in terms of reducing frequency of
seed-borne fungi was obtained through treating the seeds of acetone extract
of Oscillatoria agardhii.
These results are similar to those reported by Caire et al. (1993), Fish
and Cood (1994), Borowitzka (1995) and Rizk, ( 2006) who reported that
the extracts of Nostoc muscorum significantly(p<0.01) inhibited the growth
of Candida albicans , Sclerotium rolfsii Sacc., Rhizoctonia solani Khn. and
Fusarium verticillioides and Sclerotinia sclerotiorum. Cyanobacteria
produce exopolysaccharides that can function as energy sources for fungi
and produce plant growth regulators, which are abscisic acid, ethylene,
jasmonic acid, auxin, and cytokinin-like substances, the cytokinin
isopentenyl adenine. These substances can influence fungal growth (rdg
and Pulz, 1996). Also, Abo-Shady et al. (2007) also reported that
cyanobacteria filtrates strongly inhibited the phytopathogenic fungi isolated
from leaves, stems, and roots of Faba bean. Moreover, mycelial growth of
several phytopathogenic fungi such as Fusarium oxysporum, Penicillium
expansum, Phytophthora cinnamomi, Rhizoctonia solani, Sclerotinia
sclerotiorum, and Verticillium albo-atrum were inhibited by the methanol
extracts of the cyanobacterium Nostoc strain ATCC 53789 (Biondi et al.,
2004).
The field fungi like Alternaria, and Fusarium invade seeds as they
are developing on the plants in the field or after they have matured, but
before they are harvested. These fungi usually do not continue to grow in
grains after harvest, but may remain alive for years in grains stored at low
moisture content and low temperature (Christensen and Kaufmann. 1979).
In general, the damage caused by field fungi is done by the time the grains
are harvested, although invasion of the developing or mature embryos of
cereal seeds by Fusarium may result in development of discoloured embryos
during storage. The storage fungi, mainly comprising several species of
Aspergillus and Penicillium, do not invade grains to any appreciable degree
or extent before harvest, but they can cause severe discolouration of seed in
storage resulting in germination failure, discoloured or otherwise damaged
embryos or whole seeds, and production of mycotoxins that constitute a
health hazard for man and animals.

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Table (6). Effect of Oscillatoria agardhii and Anabaena sphaerica extracts on germination of grain sedes

Seeds
Acetone

Oscillatoria agardhii
Methanol
Water

Control

Acetone

Anabaena sphaerica
Methanol
Water

Control

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Seeds
germin
ation

%
infected
seeds

Maize

96.4

4.3

91.6

8.5

78.4

11.3

56.5

33.2

91.4

8.8

87.6

9.4

77.4

19.5

56.5

33.2

Whet
Barley
Rice
LSD

98.4
98.6
98.4
0.4

3.2
2.4
2.5
0.7

93.3
96.6
95.5
0.6

7.5
6.6
4.4
0.6

79.5
80.1
77.8
0.7

10.3
9.4
8.3
0.6

69.8
74.8
86.5
1.3

24.2
21.4
18.8
2.4

89.7
91.7
89.8
0.5

7.6
6.7
9.6
0.6

89.6
90.6
86.6
0.7

8.4
7.6
8.4
0.8

80.7
83.3
85.4
0.8

18.5
16.4
17.6
0.9

69.8
74.8
86.5
1.3

24.2
21.4
18.8
2.4

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Table (7). Fungi isolated from seeds of Triticum aestivum treated with

Isolates

Fusarium equiseti
F. moniliforme
F. solani
F. proliferatum
F. graminearum
Penicillium digitatum
Aspergillus terreus
Aspergillus niger
Aspergillus flavus
Alternaria alternate
Alternaria triticina
LSD

439

Fresh
seeds
0.2
0.4
1.0
1.2
2.1
2.4
1.4
0.3
0.6
0.2

Acetone
Day
90
0.4
0.4
1.4
1.4
2.5
2.4
1.5
0.4

Day
180
1.3
1.4
2.7
4.1
4.7
5.6
5.7
1.0
1.8
0.6

Oscillatoria agardhii extract by Agar Plate Method during different period of storage

Fungi isolated from seeds

Methanol
Water
Fresh Day
Day
Fresh
Day
seeds
90
180
seeds
90
0.6
1.8
3.8
1.5
3.2
0.8
1.7
2.4
2.8
2.9
0.7
1.4
1.8
2.8
3.6
2.1
3.3
1.4
3.1
4.4
4.3
5.3
3.3
3.7
6.4
4.8
7.7
2.8
3.6
4.4
5.4
6.2
1.9
2.1
3.2
6.8
7.1
1.3
1.4
1.5
4.8
6.7
1.5
2.0
2.8
3.8
0.5
0.7
0.9
0.7
0.9

Day
180
5.5
4.2
11.4
5.6
7.3
9.6
8.4
9.8
8.4
6.2
1.2

Fresh
seeds
3.2
1.4
3.1
2.4
3.3
8.4
11.4
5.4
6.4
2.5
2.4
1.2

Control
Day
90
4.5
8.8
6.9
7.7
11.5
14.6
10.8
12.4
10.8
9.8
7.8
1.4

Day
180
8.7
9.6
8.8
15.4
17.8
24.2
16.7
21.4
18.8
14.6
13.4
1.6

European Scientific Journal October 2014 edition vol.10, No.30 ISSN: 1857 7881 (Print) e - ISSN 1857- 7431

Table (8). Fungi isolated from seeds of Triticum aestivum treated with
Isolates

Anabaena sphaerica extract by Agar Plate Method during different

Fungi isolated from seeds

Methanol
Water

Acetone

Control

Fusarium equiseti
F. moniliforme
F. solani

Fresh
seeds
0.8
0.9
-

Day
90
1.9
1.6
-

Day
180
3.2
2.0
-

Fresh
seeds
1.7
2.2
-

Day
90
3.0
2.7
-

Day
180
4.5
3.2
-

Fresh
seeds
1.9
2.8
-

Day
90
3.8
3.5
-

Day
180
6.9
5.2
-

Fresh
seeds
3.2
1.4
3.1

Day
90
4.5
8.8
6.9

Day
180
8.7
9.6
8.8

F. proliferatum
F. graminearum
Penicillium digitatum
Aspergillus terreus
Aspergillus niger

0.4
1.9
1.7
2.3
2.7

1.0
2.5
2.1
2.7
3.6

1.8
3.7
3.4
5.4
4.9

0.9
2.2
2.3
2.8
2.4

1.8
3.7
4.2
5.4
5.3

7.4
5.3
8.3
8.4
7.1

1.7
2.9
5.3
6.3
7.4

2.9
3.8
7.3
6.7
7.2

12.1
5.9
9.9
9.9
9.4

2.4
3.3
8.4
11.4
5.4

7.7
11.5
14.6
10.8
12.4

15.4
17.8
24.2
16.7
21.4

Aspergillus flavus
Alternaria alternate
Alternaria triticina

1.3
1.1
0.4

2.6
1.9
1.1

3.8
1.9
1.7

2.8
3.8
3.8

5.3
5.7
6.8

8.8
8.1
7.4

8.8
5.3
4.3

8.1
7.7
5.8

11.8
9.1
7.2

6.4
2.5
2.4

10.8
9.8
7.8

18.8
14.6
13.4

LSD

0.5

0.6

0.9

0.7

0.9

1.1

0.9

1.2

1.4

1.2

1.4

1.6

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CONCLUSION
Cyanobacteria are wonderful organisms with diverse range of
potentials. Extraction with different solvents could affect the antifungal
activities of Oscillatoria agardhii and Anabaena sphaerica. The most
efficient solvent for antifungal activity is acetone. The present results lead to
the suggestion that Cyanobacterial algal compounds could be considered as
promising seed treatment against plant pathogenic fungi. This applicable
study characterized as clean, cheep, and fungicide alternative and being
friendly environment.
The acetone extract which exhibits high antifungal activity against seeds
storage fungi should be purified and characterized completely to explore its
potential application as grain preservative.
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