Efficacy Of Some Plant Products On Some Rice Seedlings Disease,
Muhammad et al.,
CONTROL OF SOME RICE AND MAIZE SEEDLINGS DISEASE CAUSING AGENTS (ALTERNARIA
ALTERNATA AND ASPERGILLUS PARASITICUS) IN THE LABORATORY AND FIELD USING LEAF
EXTRACTS AND POWDER OF SOME SAVANNAH PLANTS
Muhammad, A1, Modibbo, U.D.2*, Abdullahi G2 and Dangora, I.I3.
1
2
Department of Pure and industrial chemistry, Bayero University, Kano
Department of Agricultural Education, Federal College of Education (Tech) Gombe
3
Department of Biological Sciences, Federal University Dutse.
*e-mail address: [email protected]
ABSTRACTS
Study on the control of Rice seedlings disease causing Agents (Alternaria alternata and
Aspergillus parasiticus) was carried out at the School of Vocational Education
Laboratory of F.C.E (T), Gombe. Disease rice seedlings were collected from Dadin-kowa
irrigation Scheme. Complete Randomised design (CRD) was used for Laboratory work
while Complete Randomized Block Design (CRBD) was used for field work; each
treatment was replicated four times. The data collected were analyzed using analysis of
variance (ANOVA) and the means that were significant were separated using Duncan
multiple range test (DMRT). In this work, the two fungal species and isolated from the
rice seedlings. Pathogenicity test carried out showed that the isolated organisms were
pathogenic to Rice. A hundred percent stock solution was obtained by soaking
separately 1g of leaf powder of the plant used in the experiment in 10ml sterilized
water for 48hours at room temperature.Treatment of the isolated organisms using the
plant materials both in laboratory and field conditions show that control was achieved
with increasing percentage concentration of the plant materials used in the experiment
from 25%, 50% and 100% respectively. However different plant materials show different
percentage inhibition of mycelial growth at different concentration whereby in all the
experiments at 100% concentration of plant material in the medium Jatropha curcas
gives 98% and 96% percentage inhibition followed by Ficus platyphylla (93% and 94%),
followed by Parkia biglobosa(83% and 85%) and lastly by Balanites eagyptiaca (0%).
Key words: Plant materials, Disease causing Agents
INTRODUCTION
Fungi belonging to several genera including
Aspergillus,Fusarium, Penicillium as well as Alternaria
often contaminate rice and other agricultural
products.Seedling diseases that cause stand
reductions and growth abnormalities can affect rice
(Oryza sativa L.) in fields,seedbeds grown for
transplanting, and seed boxes used in mechanical
transplanting operations (Rush, 1992).So far, efforts
to reduce crop damage by fungi and other microbial
flora involve a combination of crop management
practices and the use of synthetic fungicides.In
recent years, resistant cultivars have become
available (Keller et al., 1997) but their use has lagged
st
behind the use of fungicides, especially in Nigeria
and other developing countries. However, the poor
biodegradability of fungicides and their effects on
non-target organisms render them hazardous.
Consequently, there is a need to find
alternative and or additional means of controlling
fungal damage of our crops. Plant derived products
have become increasingly attractive in this regard
since they represent affordable and safe as well as
effective means of combating some microbial
problems including crop damage in fields and
nursery (Mahmoud, 1994; Basilico,1999;Pinto et al.,
2001).
Proceedings of the 1 International conference on Drylands
115
Efficacy Of Some Plant Products On Some Rice Seedlings Disease,
In this study we have evaluated the efficacy of water
extract of leaf powder of four plants (Balanites
eagyptiaca, Ficus platyphylla, Jatropha curcas and
Parkia biglobosa) against 2 plant pathogenic fungi
which are Altenaria altanata and Aspergillus
parasiticus. Demonstration of the efficacy of plant
materials against phyto fungal crop diseases could
constitute a first step towards maintaining crop
quality. More so, such demonstration could
significantly contribute towards effective strategies
for combating other related microbial crop diseases.
MATERIALS AND METHODS
Soil preparation and Sterilization
Top soil which is sandy loamy was used in this study,
the soil were collected from pilot farms of Dadin
kowa irrigation scheme and it was sterilized using
soil solarisation method for 3 weeks and maintained
for an hour
Incidence of disease
Direct methods measure disease as incidence or
severity.
Disease incidence (I) = Number of infected plant
units per plot * 100
Total number of units assessed
Descriptive scale used for estimating disease
intensity
The grades developed by British Mycological Society
(1947) are as :0
=
Not seen in field
0.1
=
A few spots (1 or 2 spots) in 10.8m
radius
1%
=
up to 10 spots per plant, or general
light spotting
5%
=
about 50 spot per plant, or up to
one leaflet in ten attacked
25 %
=
Nearly every plant with lesions,
plant still remaining normal form
Field may smell of the disease but
look green although every plant is infected
50 %
=
Every plant affected and about half
of the leaf area destroyed by the pathogen.
Field look green flecked with brown
75 %
=
About three quarters of the leaf
area destroyed by the pathogen.
Field looks neither predominantly
brown nor green
95 %
=
Only a few leaves left green, but
stems green
st
Muhammad et al.,
100 % =
All leaves dead, stem dead or dying
Sample preparation
Leaves of the plants (Balanites eagyptiaca, Ficus
platyphylla, Jatropha curcas and Pakia biglobosa)
used in this study were collected from different
locations in Gombe metropolis and its suburb, and
washed thoroughly under running tap water for 25
minutes and separately shade dried for 2 weeks. The
dried leaves were ground separately to fine powder
using mortar and pestle and stored in an air tight
container. A hundred percent stock solution was
obtained by soaking separately 1g of leaf powder of
the plants use in the experiment in 10ml sterilized
water for 48hrs at room temperature.The resultant
was filtered through Whatman filter paper. The
extract was sterilized using filter sterilization
method to avoid contamination. Different
concentration of 25 %, 50 % and 100 % were prepared
by adding sterilized water and different
concentration of plant extract was incorporated to
potato dextrose medium agar for inoculation of the
isolated fungi in sterilized plates.
Amendment Application rate
The leaf powder of the plant used in this study were
separately mixed with 5kg of solarized soil at the
rate of 25 g, 50 g, 75 g and 100 g and the mixtures
were transferred into 30 cm diameter perforated
plastic containers
Pathogenicity test
Twelve perforated plastic containers of dimension
12inches by 14inches were divided into 4 sets of 3
each. In the first set, the pots were filled with
sterilized soil and 20 seeds of rice surfaced sterilized
with 0.1% mercuric chloride (HgCl2) were sown in
each pot. This set serves as control. The sterilization
of the soil was carried out using soil solarization
method. The second set was infested by adding half
Petri plate of a six days old culture of the isolated
fungi at a uniform depth of soil in 2 layers i.e. 3.5 cm
and 7.5 cm distance. The culture was allowed to
grow for 7 days for maximum infestation; sterilized
seeds were sown in it. In the third set, the sterilized
soil and infested seeds were used. In the fourth set
infested soil and infested seeds were used. The
experiment was conducted during the month of
February in the open field and data on percentage of
plant infection and disease severity was recorded
after 35 days of sowing (Table 1).
Proceedings of the 1 International conference on Drylands
116
Efficacy Of Some Plant Products On Some Rice Seedlings Disease,
Isolation of pathogen from diseased plant tissues of
rice seedlings:
Isolation of pathogen:
Pieces of diseased tissues were washed and placedin
a beaker. Squares of tissues approximately 0.5cm
across were cut from the advancing margins of
lesions and placed in calcium hypochlorite solution
for 3 to 5minutes. Small pieces of tissues was
transferred on to hardened agar medium using
sterilized forceps. Agar medium with diseased tissue
were surfaced sterilized to avoid contamination
(Ahmad et al, 2011).
Recording growth of mycelia
Radial growth of the fungi were carried out in solid
medium, inoculum discs of 5 mm diameters from 6
days old fungal culture was cut from the margin and
placed aseptically at the center of the sterilized plate
containing the medium incorporated with 1ml of the
extract of plant material and incubated at 25±10C for
6 days each treatment was replicated 4 times. Radial
growth of isolated fungi was measured. Percentage
inhibition of mycelial growth by plant extracts was
calculated using the formula:
Percentage inhibition of mycelia growth= ((Dc - Dt)/
Dc) * 100
Where, Dc = Diametre of control
Dt = Diametre of test pathogen
The data obtained were analyzed using Duncan
Multiple Range Test
Table 1: Pathogenicity Test showing Different Methods of Inoculation
Treatments
Diseased Seedlings
Healthy Seedlings
Sterilized
soil
+
0
20
Sterilized Seeds
Infested
Soil
+
5
15
Sterilized Seeds
Infested seeds +
14
6
Sterilized Soil
Infested
Soil
+
20
0
Infested Seeds
RESULT AND DISCUSSION
This study revealed that the percentage
inhibition of the test organisms under different
concentration of the different plant extract show
that there is significant reduction in the radial
growth of Alternaria altenata whereby Jatropha
curcas gives the highest percentage inhibition of 61
%, 80 % and 98 % at the concentrations of 25 %, 50 %
and 100 % stock solution respectively. Ficus
platyphylla showed an inhibition of 55 %, 69 % and 93
% at the concentrations of 25 %,50 % and 100 % stock
solution, while Parkia biglobosa exhibited 45 %, 69 %
and 83 % at the same concentrations while Balanites
egyptiaca gave smaller percent inhibition of 10 %, 0 %
and 0 % at 25 % 50 % and 100 % stock solution which is
approximately zero percent.
However in the case of Aspergillus
parasiticus, Jatropha curcas demonstrate the highest
inhibition of 60 %, 80 % and 96 % at the
concentrations of 25 %, 50 % and 100 % stock solution,
followed by Ficus platyphylla with 55 % 78 % and 93 %
percentage inhibition at 25 %, 50 % and 100 % stock
solution concentrations. Parkia biglobosa gave 45 %,
69 % and 83 % percentage inhibition at the same
st
Muhammad et al.,
Percentage disease
0
25
70
100
concentration as above. Balanites eagyptiaca gives
0% percentage inhibitions at all concentration of
stock solution of its leaf extract (Table 2 and 3).
The in vitro analysis of the leaf powder of
the test plants on the test pathogens reveals that
100 g amendment of the leaf powder of the test
plant gives the better result of controlling the
disease pathogen from 10 % incidence at 25 g to as
low as 2 % incidence at 100 g in the case of Alternaria
altenata but in the case of Aspergillus parasiticus the
incidence percentage decrease from 21.70 % at 25 g
amendment rate to 5 % at 100 g amendment rate for
the leaf powder of Jatropha curcas. The leaf powder
of Ficus platyphylla gives 6.5 % incidence at 25 g
amendment rate and 3.5 % incidence at100 g
amendment rate for Alternaria altenata while for
Aspergillus parasiticus the decrease in incidence is
from 6.7 % at 25 g amendment to 3.2 % at 100 g
amendment rate. However, for Parkia biglobosa leaf
powder the decrease in the incidence percentageis
from 11.3 % at 25 g amendment rate to 3 % at 100 g
amendment rate for Alternaria altenata while for
Aspergillus parasiticus the decrease in the incidence
is from 9.7% at 25 g amendment rate to 5 % at 100 g
Proceedings of the 1 International conference on Drylands
117
Efficacy Of Some Plant Products On Some Rice Seedlings Disease,
amendment rate and for Balanites eagyptiaca there
is no any decrease in terms of incidence for both the
test organisms (Alternaria altenata and Aspergillus
Muhammad et al.,
parasiticus) from 25 g amendment rate to 100 g
amendment rate of which the value range is 20 % to
28 % in all the amendment rates (Table 4,5,6 and 7)
Table 2: Efficacy of leaf extract of different plants on percentage inhibition of Alternaria altanatain in- vivo
studies
Treatments
25%
50%
100%
Control
18a(0%)
18a(0%)
18a(0%)
Jatropha curcas
7a (61%)
2.5b (86%)
a
Ficus platyphylla
8 (55%)
3.18b(69%)
a
Parkia biglobosa
9.8 (45%)
5.5b(69%)
a
Balanites eagyptiaca
17.9 (0%)
17.8a (0%)
*Means followed by the same letter within a row do not differ significantly at p > 0.05
0.2b (98%)
12b(93%)
3b(83%)
17.7a(0%)
Table 3: Efficacy of leaf extracts of different plants on percentage inhibition of Aspergillus parasiticus
Treatments
25%
50%
100%
Control
23a(0%)
23a(0%)
23a(0%)
Jatropha curcas
9a(60%)
4.5b (80%)
0.9b(96%)
a
b
Ficus platyphylla
10 (55%)
5.5 (78%)
2b(93%)
a
b
Parkia biglobosa
13 (45%)
7.5 (69%)
4.2b(83%)
a
a
Balanites eagyptiaca
29.9 (0%)
22.6 (0%)
22.4a(0%)
*Means followed by the same letter within a row do not differ significantly at p > 0.05
Table 4: Efficacy of leaf powder of Jatropha curcas at different amendment rate in vitro
Plant leaf powder used
Amendment rate (gm)
Incidence %
Incidence %
Alternaria altenata
Aspergillus parasiticus
Control
30.67
34.89
Jatropha curcas
25
10.00
21.70
50
3.89
15.00
75
2.50
8.00
100
2.00
5.00
Table 5: Efficacy of leaf powder of Ficus platyphylla at different amendment rate in vitro
Plant leaf powder used
Control
Ficus platyphylla
Amendment rate (gm)
25
50
75
100
Incidence %
Alternaria altenata
36.87
6.50
3.89
5.50
3.50
Incidence %
Aspergillus parasiticus
37.89
6.70
15.00
5.00
3.20
Table 6: Efficacy of leaf powder of Parkia biglobosa at different amendment rate in vitro
Plant leaf powder used
Control
Parkia biglobosa
st
Amendment rate (gm)
25
50
75
100
Proceedings of the 1 International conference on Drylands
Incidence %
Alternaria altenata
25.87
11.50
6.50
4.50
3.00
Incidence %
Aspergillus parasiticus
24.47
9.70
5.00
4.79
5.00
118
Efficacy Of Some Plant Products On Some Rice Seedlings Disease,
Plant leaf powder used
Muhammad et al.,
Incidence %
Incidence %
Alternaria altenata
Aspergillus parasiticus
Control
25.87
24.47
Parkia biglobosa
25
11.50
9.70
50
6.50
5.00
75
4.50
4.79
100
3.00
5.00
Table 7: Efficacy of leaf powder of Balanites eagyptiaca at different amendment rates in vitro studies
Plant leaf powder used
Control
Balanites eagyptiaca
Amendment rate (gm)
Amendment rate (gm)
Incidence %
Alternaria altenata
36.75
20.00
25.00
27.50
24.00
25
50
75
100
DISCUSSIONS
From the above result we can infer that in
both the two test organisms Jatropha curcas gives
highest percentage inhibition in all the stock solution
of its leaf extracts with good percentage inhibition
this was corroborated by findings of Siva et al.
(2008) who reported that concentration of jatropha
curcas at 40 % recorded 100 % inhibition of growth of
Alternaria altanata, Aspergillus flavus and other phyto
fungal parasites. Also Rajesh et al. (2012) reported
that leaf extract of Jatropha curcas at 20 %
concentration inhibit sporulation of six plant
pathogenic fungi (Alternaria alternata, Aspergillus
flavus, Aspergillus niger, Fusarium oxysporium,
Rhizoctonia solani and Trichoderm aviride). This is
because
the
plant
materials
used
has
phytochemicals that inhibit the normal metabolic
activities of the fungi.
However, under field trials it can be equally
observed that Jatropha curcas give the highest
percentage inhibition on the test organisms at 100g
amendment rate, followed by Ficus platyphylla and
then Parkia biglobosa and finally Balanites eagyptiaca.
This indicted Jatropha curcas to be the most
effective among the test plants in controlling plant
fungal pathogens, this is corroborated by the
findings of Roshan et al, 2014 on the effect of leaves
of Jatropha curcas as soil amendments on the
growth of Alternaria solani on tomatoes, where by
“3 % concentration was found to be superior in
controlling of disease as well as favored the growth
st
Incidence %
Aspergillus parasiticus
35.20
30.00
27.00
28.00
23.50
of tomato plant over other treatments including the
control.”
CONCLUSION
In conclusion findings of this study reveal
that the leaf powder of the test plants on the test
organisms have strong antifungal compounds and
their addition to the soil control the pathogens
(fungal) that causes rice diseases in field and result
in disease free growth of rice. However, this finding
is germane from the point of view of controlling rice
diseases since the use of synthetic fungicides by
subsistence farmers is plaque with several
limitations. This findings will enrich farmers know
how on how to utilized plants products in controlling
some plant fungal diseases, it will equally help in
designing a model for controlling phytofungal
pathogens base on their phytochemical composition
and to a lesser extent developed a broad spectrum
antifungal formulation for all farming system with
many combination of 2 or more plants materials.
Base on the finding of this research followings are
some of the recommendations projected:
Fungal diseases of plants particularly rice
can be controlled using plant products which are
cost effective. Controlling of plants diseases using
chemicals as well as plants products should be
ecofriendly and the plant based products particularly
Jatropha curcas can be adopted as amendment or
pre sowing treatment of the soil for effective
controlling of fungal pathogens that are ruderals.
Proceedings of the 1 International conference on Drylands
119
Efficacy Of Some Plant Products On Some Rice Seedlings Disease,
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