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IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS)
e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 6 Ver. II (Nov - Dec. 2015), PP 33-41
www.iosrjournals.org
DOI: 10.9790/3008-10623341 www.iosrjournals.org 33 | Page
Efficacy of Some Botanicals in the Control of Fungi Causing post
harvest rot of Yam in Katube market, Obudu, Nigeria.
1
Liamngee kator 2
Akomaye Maria Umgbeb 3
Okoro James Kalu
Department of Biological Sciences, Benue State University Makurdi, Benue State, Nigeria.
Department of Biology, Federal College Of Education, Obudu, Cross River State, Nigeria.
College of Agronomy, Federal University of Agriculture, Benue State, Nigeria.
Abstract: Efficacy of some botanicals in the control of fungi causing post harvest rot of yam in Atube market,
Obudu was conducted. The fungal pathogens isolated from the yam samples were Aspergillus niger,
Botryodiplodia theobromae, Aspergillus flavus, and Aspergillus fumigatus. Water and ethanol extracts of the
plant species at 10g/ml, 20g/ml, 30g/ml, 40g/ml and 50g/ml were tested against the fungal isolates in vitro. The
inhibitory effects of water extracts of Daniellia oliveri, Moringa oleifera and Parkia biglobosa at 50g/ml on
all test organisms ranged from 11.00 to 35.00 and at 10g/ml it ranged from 25.00 to 80.00. The inhibitory effect
of ethanol extract of Daniellia oliveri, Parkia biglobosa and Moringa oleifera at 50g/ml on test organisms
ranged from 6.50 to 34.50 and at 10g/ml it ranged from 8.00 to 77.50. There was no significant difference in the
grand mean inhibitory effect of both water and ethanol extracts on Aspergillus niger and Aspergillus flavus at P
= 0.05. There was significant difference in the grand mean inhibitory effect of water and ethanol extracts on
Aspergillus fumigatus and Botryodiplodia theobromae. The extracts of Parkia biglobosa, Moringa oleifera and
Daniellia oliveri are useful antifungal agents, inhibiting fungal growth at all concentrations.
Keywords; Efficacy, Botanicals, fungi, Atube market, rot, Post harvest.
I. Introduction
Yam (Dioscorea spp) is among the most important staple food in the world especially some parts of the
tropics and subtropics [1]. The most cultivated varieties in Nigeria are the D. rotundata (white yam), D.
cayenesis (yellow yam) and D. alata (water yam) [2]. Nutritionally, yams are mainly carbohydrate foods. It is
one of the most important dietary sources of energy produced in the tropics [3].
White yam (Dioscorea rotundata) is the most important and cultivated variety found in Africa. In
2005, [4] reported that Nigeria produces about 66.6% (26.6 million metric tonnes) of the total world’s yam
production every year. White yam (Dioscorea rotundata) is much preferred to other yam varieties and it
constitutes about 80% of the total yam produced in Nigeria [5].Though this figure is high, demand for yam has
always been more than the supply.
As important as yam is, its production is constrained by many problems ranging from high cost of
production, attack by fungi, nematodes and pests, singly or in combination. These constraints are responsible for
field suppression and tuber quality deterioration in storage [2]. The magnitude of these problems has made
people express fears that yam production in Nigeria may decline substantially in the near future [3].
Rot is major factor limiting the post – harvest life of yams and losses can be very high. It is estimated
that an average of 25- 50% of yam tubers produced in Nigeria are lost to pests and diseases [4]. The disease
causing agents reduce the quantity and quality of yam and make them unappealing to the consumer. Losses due
to post- harvest rot significantly affect farmers’ and traders’ income, food security and seed yams stored for
planting. In Nigeria, over 60% of white yam get rotten when stored for planting [6].
Most rot of yam tubers are caused by pathogenic fungi such as Aspergillus flavus, Aspergillus niger,
Aspergillus fumigatus, Botryodiplodia theobromae, Fusarium oxysporum, Fusarium solani, Penicillium
chrysogenum, Rhizoctonia spp, Penicillium oxacilum, Trichoderma viride and Rhizopus nodosus [7]; [8].
It is therefore important to know the fungal organisms that are responsible for post-harvest rot of white
yam in Atube market, obudu and seek their effective control measures to extend the life span of yam in storage.
II. Materials and Methods
2.1 Sample Collection
About 20 tubers of white yam (Dioscorea rotundata) with symptoms of rot were obtained from Atube
market, Obudu Local Government Area of Cross River State. The yam tubers were packaged in polyethylene
bags and taken to botany laboratory of the Benue State University, Makurdi, where they were assessed for
microbial (fungal) presence.
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 34 | Page
2.2 Media Preparation
The medium used was Potato Dextrose Agar (PDA) prepared according to the manufacturers
instruction. About 39.6g of powdered PDA medium was dissolved in1 litre of sterile distilled water and
sterilized by autoclaving at 1210
C for 15 minutes.
2.3 Isolation of fungal pathogens from rotten yam tubers
Small sizes were cut from yam tubers infected with rot. They were first surface sterilized by dipping
completely in a concentration of 5% sodium hypochloride solution for 2 mins. They were then removed and
rinsed in several changes of sterile distilled water. The yam pieces were then placed on sterile paper towels to
dry for 5 mins. The pieces of yam sections were placed on solidified Potato Dextrose Agar medium. Three
replicates were made for each of the 20 samples of yam tubers. The inoculated plates were incubated at 25-280
C
and observations were made daily for possible microbial growth. After 5 - 7 days of growth, Sub-culturing was
done to obtain pure cultures of the isolates.
2.4 Identification of fungal isolates
The identification of the isolates was done by examining the isolates macroscopically and
microscopically. The colony characteristics such as colony appearance, change in medium colour and growth
rate were observed. Shapes of the conidia and conidiophores were taken note of. These structural features were
matched with standards in [9] and [10].
2.5 Pathogenicity Test
To confirm the pathogenicity of isolates from white yam, pure cultures of the isolates were used to
inoculate three white yams per pathogen with 5mm diameter mycelia plugs of a 4 day old culture. On
appearance of symptoms, the tissues at the margins of the healthy and diseased parts were surfaced sterilized,
excised and plated onto Potato Dextrose Agar (PDA) and incubated at 25-28o
C for 5 - 7 days. At the end of this
period, morphological characteristics and growth patterns observed in each case were compared with the ones of
the original isolates. One tuber of white yam was used for each fungal isolate respectively, replicated three times
and arranged in complete randomized design. Controls were yam tubers inoculated with sterile PDA only.
Following appearance of symptoms, 8 - 12 days post inoculation, rot severity index was assessed on a scale of 0
- 5, where 0 - no disease manifestation, 1 - 1-20% rot, 2 - 21-40% rot, 3 - 41-60% rot, 4 - 61-80% rot, 5 - 81-
100% rot.
2.6 Collection of Plant Material
Fresh leaves of three selected plant species were collected in polyethylene bags and transported to the
botany laboratory of the Benue State University Makurdi for preparation of extracts. They are Parkia biglobosa
(locust bean tree), Moringa oleifera (Drumstick) and Daniellia oliveri (African Copaiba Balsam).
2.7 Preparation of Extracts of Plant Parts
Water and ethanol extracts of each selected plant species was prepared. Each of the leaves were washed
under running tap water and soaked in a 1% solution of sodium hypochloride for 2 minutes. They were rinsed
severally with sterile distilled water and air dried at room temperature for 30 mins. 50g each of the fresh leaves
of the selected trees were weighed for both water and ethanol extractions respectively. The leaves were ground
one after the other using a mortar and pestle. The mortar and pestle were washed after each type of leave was
pounded to avoid mixture of their different components.
The macerates were transferred into beakers and each was soaked in100ml of sterile distilled water and 90%
ethanol respectively for water and ethanol extractions.
2.8 Concentrations of Crude Extracts
Serial dilutions of the crude extracts of the leaves of each plant was prepared to give different
concentrations of the extracts. Extract concentrations of 10g/ml, 20g/ml, 30g/ml, 40g/ml and 50g/ml were thus
obtained for both water and ethanol extractions.
2.9 Antifungal Susceptibility Test in vitro
One ml of the extract concentration for both ethanol and water was dispensed in Petri dishes and 15-
20mls of molten PDA was added. The Petri plates were swirled gently on the work bench to ensure even
dispersion of the extracts. The Agar-extract mixture was allowed to solidify and used for the inhibition of
mycelia growth for the tested fungi. 4mm diameter of mycelia disc obtained from the colony edge of 5day old
culture of each of the test fungi was inoculated centrally onto the medium. Three replications were used for each
fungal isolate. Controls were Petri plates containing PDA with no botanical extract and inoculated with the test
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 35 | Page
fungi. The plates were arranged on laboratory desk following complete randomized design. The Petri plates
were then incubated at 25 - 28o
C for 5-7 days during which measurement of growth was done using a metre rule
at an interval of 24 hours. Inhibition of fungal growth was calculated using the formula R1 - R2 x 100
R1
where R1 = radial growth of the pathogen in the control plates
R2 = growth of the pathogen with treatment
2.10 Data Analysis
Data obtained from the study was analyzed using Analysis of Variance (ANOVA) and the Fishers
Least Significant Difference was used to separate the means at 5% level of significance.
III. Results
3.1 Fungal pathogens isolated from yam samples
A total of four fungi namely Botryodiplodia theobromae, Aspergillus fumigatus, Aspergillus flavus and
Aspergillus niger were isolated from the yam samples as shown in Table 1. Identification of the isolates was
done using recommendations given by [9] and other electronic documentations on the genera isolated.
Table 1: Characterization of Fungal isolates from Yam samples on PDA.
Cultural/microscopic
characteristics
Appearance on PDA Photomicrograph Probable
organism
Colony colour is dark
brown to black. Conidia
are black and densely
packed. Conidiophore
length is 855 – 9 57(µm)
and colour was hyaline to
yellowish or slightly
brown.
Aspergillus niger
Colony colour ranged
from grayish to dark blue
in colour. The conidia
colour was dark green to
blue. Conidiophore
length was 265–365µm
and it was uncoloured to
grayish brown
Aspergillus
fumigatus
Colony colour was parrot
green to deep green.
Conidia were light green
in colour.
Conidiophores length
ranged from 550 – 678
µm .
Aspergillus flavus
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 36 | Page
Colony colour ranged
from white to black.
Mycelium is septate and
the conidia are 2 celled.
Botryodiplodia
theobromae
3.2 Pathogenicity test.
The Pathogenicity test showed that the fungal isolates induced rot in healthy yam tubers after 14 days
of inoculation. The rot severity ranged from 3 – 5 which indicated 41 – 100% of tuber rot as shown in Table 2.
Analysis of Variance revealed a significant difference in the disease causing potential of the fungal isolates on
healthy yam tubers with respect to their controls as shown in Table 3.
Table 2: Disease causing potential of fungal isolates on healthy yam tubers
Fungal isolates/Replicates C R1 R2 R3
Aspergillus niger 0 5 5 4
Aspergillus fumigates 0 4 4 4
Aspergillus flavus 0 4 3 3
Botryodiplodia theobromae 0 5 5 5
Key Severity Scale
C = Control 0 - No infection
R1 = Replicate 1 1 - 1 – 20% of rot
R2 = Replicate 2 2 - 21 – 40% of rot
R3 = Replicate 3 3 - 41 – 60% of rot
4 - 61 – 80% of rot
5 - 81 – 100% of rot
Table 3: Analysis of Variance in the disease causing potential of Fungal Isolates on healthy Yam tubers
Fungal Isolates A. niger A. flavus A. fumigatus B. theobromae
Pathogenicity 5.00a
4.00a
4.00a
5.00a
Control 0.00b
0.00b
0.00b
0.00b
Footnote: Means not tagged with same alphabets are significant at P= 0.05
3.3 Mean inhibitory effect of water extract of leaves of Moringa oleifera on test organisms.
The highest significant inhibitory effect was observed on both Botryodiplodia theobromae and
Aspergillus fumigatus at 50g/ml (11.00) while the least significant inhibitory effect was observed at 10g/ml
(77.50) and (41.00) respectively. On A. flavus, highest inhibitory effect was observed at 50g/ml (13.50) and the
lowest inhibitory effect was observed at 10g/ml (35.00).On A. niger, the highest inhibitory effect was observed
at 50g/ml (36.00) and the lowest inhibitory effect was observed at 10g/ml (60.50) as shown in Table 4.
Table 4: Mean inhibitory effect of Water extract of Moringa oleifera on test organisms.
Conc g/ml A. flavus A. niger B. theobromae A. fumigatus
10 35.00 60.50 77.50a
41.00a
20 29.00 41.00 56.50b
29.50b
30 24.00 59.00 49.50b
21.00b
40 14.50 52.50 44.50b
26.00b
50 13.50 36.00 11.00c
11.00b
Footnote: Means not tagged with same alphabets are significant otherwise they are the same at P=0.05
NS - No significant difference
LSD(0.05) 0.93 0.92 0.91 0.00
LSD(0.05) NS NS 13.57 8.92
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 37 | Page
3.4 Mean inhibitory effect of ethanol extracts of Moringa oleifera on test fungi
The highest significant inhibitory effect was observed on Aspergillus niger at 50g/ml and 20g/ml
(22.50) while the least significant inhibitory effect was observed 10g/ml (53.50). On Botryodiplodia
theobromae, highest significant inhibitory effect was observed at 50g/ml (17.00) and the lowest significant
inhibitory effect was observed at10g/ml (60.00). On A. fumigatus, highest inhibitory effect was observed at
50g/ml (16.00) and lowest inhibitory effect was observed at 10g/ml (60.00). On A. flavus, highest inhibitory
effect was observed at 50g/ml (7.00) while the lowest inhibitory effect was observed at 10g/ml (29.00) as shown
in Table 5.
Table 5: Mean inhibitory effect of Ethanol extract of Moringa oleifera on test organisms.
Conc g/ml A. flavus A. niger B. theobromae A. fumigatus
10 29.00 53.50a
60.00a
60.00
20 19.00 22.50b
35.50b
39.00
30 21.50 23.50b
25.50c
30.00
40 26.50 25.00b
22.50c
45.00
50 7.00 22.50b
17.00c
16.00
Footnote: Means not tagged with same alphabets are significant otherwise, they are the same at P=0.05
NS - No significant difference
3.5 Mean inhibitory effect of water extracts of Daniellia oliveri on test fungi
The highest inhibitory effect was observed at 50g/ml (20.00) on Aspergillus flavus and the least
inhibitory effect at 10g/ml (80.00) was on Aspergillus niger. Significant inhibitory effect was observed on
Aspergillus flavus and Botryodiplodia theobromae. On Aspergillus flavus, all means were significantly different
in their zones of inhibition except at 20g/ml (41.50) and 40g/ml (46.00) while all inhibitory effects on
Botryodiplodia theobromae were insignificant except at 30g/ml (80.00) as shown in Table 6.
Table 6: Mean inhibitory effect of Water extract of Daniellia oliveri on test organisms.
Conc g/ml A. flavus A .niger B. theobromae A. fumigatus
10 69.50a
80.00 50.00a
77.50
20 41.50b
60.50 42.50a
75.00
30 56.30c
45.00 80.00b
65.00
40 46.00b
42.50 39.00ad
34.00
50 20.00d
35.00 29.50cd
31.00
Footnote: Means not tagged with same alphabets are significant otherwise, they are the same at P=0.05
NS - No significant difference
3.6 Mean inhibitory effect of ethanol extracts of Daniellia oliveri on test fungi
There was insignificant inhibitory effect at all concentrations on Aspergillus flavus, Aspergillus niger
and Botryodiplodia theobromae while A. fumigatus had highest significant inhibitory effect at 50g/ml (5.00) and
least significant inhibitory effect at 10g/ml (30.00). On A. flavus highest inhibitory effect was observed at
50g/ml (12.50) and least inhibitory effect was observed at 40g/ml (16.50). On A. niger, highest inhibitory effect
was observed at 50g/ml (9.50) while least inhibitory effect was observed at 40g/ml (24.00). On B. theobromae,
highest inhibitory effect was observed at 50g/ml (6.50) and least inhibitory effect was observed at 40g/ml and
10g/ml (8.00) respectively as shown in Table 7.
Table 7: Mean inhibitory effect of Ethanol extract of Daniellia oliveri on test organisms.
Conc g/ml A. flavus A. niger B. theobromae A. fumigatus
10 36.50 28.50 8.00 30.00a
20 22.50 41.00 15.50 14.00b
30 28.50 47.00 17.50 9.50bd
40 16.50 24.00 8.00 15.00b
50 12.50 9.50 6.50 5.00cd
LSD(0.05) NS 11.54 8.89 NS
LSD(0.05) 11.54 NS 12.98 NS
LSD(0.05) NS NS NS 7.17
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 38 | Page
Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05
NS - No significant difference
3.7 Mean inhibitory effect of water extract of Parkia biglobosa on test fungi
There was insignificant inhibitory effect on all the fungal isolates at all concentrations except on
Aspergillus flavus which had significant highest inhibitory effect at 50g/ml (16.50cm) and least significant effect
at 10g/ml (63.50cm). On A. niger, highest inhibitory effect was observed at 50g/ml (26.50) and lowest
inhibitory effect was observed at 10g/ml (57.50). On A.fumigatus, highest inhibitory effect was observed at
50g/ml (18.00) and the lowest was at 10g/ml (25.50). On B. theobromae, highest inhibitory effect was observed
at 50g/ml (22.50) and the lowest was at 20g/ml (58.00) as shown in Table 8.
Table 8: Mean inhibitory effect of Water extract of Parkia biglobosa on test organisms.
Conc g/ml A. flavus A. niger B. theobromae A. fumigatus
10 63.50a
57.50 57.50 25.50
20 43.50b
56.00 58.00 39.00
30 47.00b
43.50 40.80 35.00
40 35.50b
47.50 48.00 39.00
50 16.50c
26.50 22.50 18.00
Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05
NS - No significant difference
3.8 Mean inhibitory effect of ethanol extracts of Parkia biglobosa on fungal Isolates
There was insignificant inhibitory effect of Parkia biglobosa on the test organisms at all
concentrations. Highest inhibitory effect was observed on A. flavus at 40g/ml and 50g/ml (12.50). Lowest
inhibitory effect was observed on A.niger at 10g/ml (77.50) followed by A. fumigatus (75.00), B. theobromae
(69.00) and A. flavus (41.50) respectively. A. niger and B. theobromae shared highest inhibitory effect at 50g/ml
(34.50) and A. fumigatus had highest inhibitory effect at 50g/ml (27.00) as shown in Table 9.
Table 9: Mean inhibitory effect of Ethanol Extract of Parkia biglobosa on test organisms.
Conc g/ml A .flavus A. niger B. theobromae A. fumigatus
10 41.50 77.50 69.00 75.00
20 43.00 60.00 53.50 63.00
30 33.50 48.00 38.50 52.00
40 12.50 40.00 39.00 35.00
50 12.50 34.50 34.50 27.00
Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05
NS - No significant difference
3.9 Grand mean inhibitory effect of water extracts of the three plant species on the test organisms
There was insignificant inhibitory effect of all the water extracts of the three plants species on
Aspergillus flavus, Aspergillus niger and Botryodiplodia theobromae except on Aspergillus fumigatus where the
effects of M. oleifera and P. biglobosa were significantly different from that of D. oliveri as shown in Table 10.
Table 10: Grand Mean inhibitory effect of Water extracts of all the three plant species on test organisms.
Plant Extract A. flavus A. niger B. theobromae A.fumigatus
M.oleifera 23.20 49.80 47.80 25.70a
D.oliveri 46.66 52.60 48.20 56.50b
P.biglobosa 41.20 46.20 45.36 31.30a
Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05
NS - No significant difference
LSD(0.05) 11.13 NS NS NS
LSD(0.05) NS NS NS NS
LSD(0.05) NS NS NS 21.21
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 39 | Page
3.10 Grand mean inhibitory effect of ethanol extracts of the three plant species on test fungi
There was significant inhibitory effect of all the ethanol extracts of the three plant species on
Botryodiplodia theobromae and Aspergillus fumigatus. Extracts of M. oleifera and P. biglobosa were
significantly different from that of D. oliveri in their effects on B. thoebromae and A. fumigatus as shown in
Table 11.
Table 11: Grand Mean inhibitory effect of Ethanol extracts of all the three plant species on test
organisms.
Plant Extract A. flavus A. niger B. theobromae A. fumigatus
M.oleifera 20.60 29.40 32.10a
38.00a
D.oliveri 23.30 30.00 11.10b
14.70b
P.biglobosa 28.60 52.00 46.90a
50.40a
Footnote: Means not tagged with same letters are significant; otherwise they are the same at P=0.05
NS - No significant difference
3.11 Comparative grand mean inhibitory effect between water and ethanol extracts of the three plant
species on test fungi
There was significant inhibitory effect between water and ethanol extracts of Daniellia oliveri on
Aspergillus niger (21.40) while water and ethanol extracts of Moringa oleifera and Parkia biglobosa had
insignificant inhibitory effect on Aspergillus niger.
There was significant inhibitory effect between water and ethanol extracts of Moringa oleifera on
Aspergillus flavus (19.92) while water and ethanol extracts of Daniellia oliveri and Parkia biglobosa had
insignificant inhibitory effect on Aspergillus flavus. Water and ethanol extracts of Daniellia oliveri showed
significant inhibitory effect on Botryodiplodia theobromae (20.54) while water and ethanol extracts of Moringa
oleifera and Parkia biglobosa had insignificant inhibitory effect on Botryodiplodia theobromae. Water and
ethanol extracts of Daniellia oliveri showed significant inhibitory effect on Aspergillus fumigatus (25.13) while
water and ethanol extracts of Moringa oleifera and Parkia biglobosa had insignificant inhibitory effect on
Aspergillus fumigatus as shown in table 12.
Table 12: Comparative grand Mean inhibitory effect of water and ethanol extracts of the three plant
species on test fungi
Solvents A niger A flavus B theobromae A fumigatus
MO DO PB MO DO PB MO DO PB MO DO PB
Water extract 23.20 46.66a 41.20 49.80a
52.60 46.20 47.80 48.20a
45.36 25.70 56.50a
31.30
Ethanol extract 20.60 23.30b
28.60 29.40b
30.00 52.00 32.10 11.10b
46.90 38.00 14.70b
50.50
LSD (0.05) NS 21.40 NS 17.92 NS NS NS 20.54 NS NS 25.13 NS
Footnote: Means not tagged with same alphabets are significant otherwise they are the same at P=0.05
Key
NS - No significant difference
MO - Moringa oleifera
DO - Daniellia oliveri
PB - Parkia biglobosa
IV. Discussion
The Fungal organisms associated with rot of white yam (Dioscorea rotundata) in this present study
were Aspergillus niger, Apergillus flavus, Aspergillus fumigatus, and Botryodiplodia theobromae. These
organisms have been associated with post harvest rot as reported by [11] , [12], [1].
The pathogenicity test showed that the fungal isolates inoculated into healthy yam tubers caused tuber
rot. This was due to the ability of the fungal pathogens to utilize the nutrients of the yam as a substrate for
growth and development. This result is similar to the report on fungi associated with Nigerian yams by [13],
[11] and [12].
In another set of experiments, the water and ethanol extracts of the three plant species; Parkia
biglobosa, Daniellia oliveri and Moringa oleifera showed antifungal activities against the test fungi at all
concentrations in vitro. This implies that locally occurring plants commonly used in folklore human medicine
also posses anti microbial effects on post harvest fungi. This conform to the results obtained from the studies of
LSD(0.05) NS NS 18.12 21.75
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 40 | Page
[14] and [15] on the bioassay of some plant extracts on some fungal pathogens. This also agrees with the reports
of [16], [17] and [18] on the inhibitory action of plant products employed on the mycelia growth and spore
germination of other pathogenic fungi.
It was observed that susceptibility of the test fungi increased with increasing concentration of the plant
extracts for all the fungal isolates. This may have resulted from variation in the principal active ingredients
present in the extracts. The effectiveness of a plant extract depends on the nature and amount of active
ingredient found in it.
This agrees with the findings of [19] who reported significant inhibitory property of Neem (A. indica)
extract that contains azadrachtin which is known to be fungitoxic against most fungal pathogens just as [20]
found the extract of Ocimum gratissimum to reduce the growth of Rhizopus spp. Also, [21] and [22] reported
that the phytochemical screening of the plant species used in this study showed the presence of alkaloids,
saponins, tannins, flavonoids, glycosides and phenol compounds.
An increase in the concentration of the plant extracts correspondingly decreased the growth of the test
fungi. This is because an increase in the concentration of the extract implied an increase in the active ingredient
of the solu tion which acts on the fungus thereby affecting its physiological processes and consequently
lowering the growth of the fungus.
The ethanol extract of D. oliveri recorded the highest grand inhibitory effect of (11.10) on
Botryodiplodia theobromae and the least was water extract of D. oliveri (56.50) on A. fumigatus. The ethanol
extract of Moringa oleifera recorded grand mean inhibitory effect of (20.60) on Aspergillus niger, (29.40) on
Aspergillus flavus, (32.10) on Botryodiplodia theobromae, (38.00) on Aspergillus fumigatus compared to water
extract which had a mean inhibitory effect of (23.20) on Aspergillus niger, (49.80) on Aspergillus flavus, (47.80)
on Botryodiplodia theobromae and (25.70) on Aspergillus fumigatus.
This is in agreement with [23] who stated that plant extracts in organic solvents provided more
consistent antimicrobial activity compared to water extracts.
The most sensitive test organism in this study was Botryodiplodia theobromae which had an inhibitory
diameter of (11.10) on ethanol extract of Daniellia oliveri. The least sensitive test organism was Aspergillus
fumigatus with an inhibitory diameter of (56.50) on water extract of D. oliveri. The differences recorded in the
fungitoxic activity of the extracts may also be attributed to the solubility of the active ingredients in water and
ethanol or the presence or absence of inhibitors for the fungitoxic principle. Also the effectiveness of the ethanol
extracts than water extracts observed in this study is probably due to its ability to extract more bioactive
compounds from the plant material which are effective on the test organisms.
The active ingredient present in an extract is influenced by many factors which include the age of the
plant, extracting solvent, method of extraction and time of harvesting the plant material.
V. Conclusion
This study has confirmed and established that the extracts of Parkia biglobosa, Daniellia oliveri and
Moringa oleifera are useful antifungal agents, inhibiting fungal growth at all concentrations. This agrees with
earlier reports of [24] on the inhibition of growth and sporulation of fungal pathogens on Ipomea batatas and
Dioscorea species. and [25] on the use of Xylopia aethiopica and Zingiber officinale to control yam tuber rot
caused by F. oxysporum, Aspergillus niger and Aspergillus flavus.
References
[1] RN Okigbo and UO Ogbonnaya, Antifungal effect of two tropical plant leaf extract Ocimum gratissimum and Aframomum
melegueta on post harvest yam (Dioscorea spp) rot, Afr. J. Biotech. 5 (9), 2006, 727-731.
[2] N Amusa, yam diseases and their management in Nigeria, African Journal of Biotechnology 2 (12), 1999, 497-502.
[3] N Ekefan, Diseases of yam tubers, International Journal of Tropical Plant Diseases, 3, 1999, 23 -27.
[4] FAO, Food and Agricultural Organisation, FAO, Rome, 2005, pp23.
[5] JP Tetteh and C. Saakwa, Prospects and constraints of yam production in Ghana (Tyum press, 1994).
[6] SO Adesiyan and RA. Odihirin, Histopathology studies of the yam tuber infected with Scutellonema bradys, International
Biodegradation Bulletin11, 1975, 48-55.
[7] RN Okigbo and FEO. Ikediugwu, Evaluation of water losses in different regions of yam tubers in storage, Nig. J. Exp. Appl
Bio, 3, 2002, 20- 21.
[8] KA Aidoo, identification of yam tuber rot fungi from storage systems at the Kumasi central market, Nig. Journal of botany, 22,
2007, 18-19.
[9] GL Barnett and AL Barry, Illustrated Genera of Imperfect fungi, 3rd edition (Burgess publishing company, 1972) 5-10.
[10] RH Booth, Post-harvest deterioration of tropical root crops, losses and their control, Trop. Sci. 16 (2), 1971, 49- 63.
[11] SK Ogundana, SHZ Naqvi and JA Ekundayo, Fungi associated with soft rot of yams (Dioscorea spp) in storage in Nigeria,
Trans. Br. Mycol. Soc. 54, 1970, 445-451.
[12] RN Okigbo, Variation in photochemical properties of selected fungicidal aqueous extract of some plant leaves in Kogi state,
Nigeria, American Eurossian journal of Sustainable Agriculture 3 (3), 2000,407- 409.
[13] MO Adeniji, Fungi associated with storage decay of yam in Nigeria, Phytopathology 60, 1972, 590-592.
[14] DB Olufolaji and OO Adeyeye, Evaluation of some plant extracts in the control of Curvuluria leaf spot of maize in the
screen house, Journal of sustainable agriculture and Environment 2 (1), 2002 ,78-75.
Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu.
DOI: 10.9790/3008-10623341 www.iosrjournals.org 41 | Page
[15] DB Olufolaji, and BA Ojo, Evaluation of Neem plant extracts in the control of incidence and severity of Antrachnose disease
of Soybean caused by Colletotrichum truncatum, Niger delta Biologia 5 (1), 2005, 24-31.
[16] SA Bankole, and A Adebanjo, Inhibition of growth of some plant pathogenic fungi using some Nigerian plants, Int. J. Trop.
Plant Dis. 13, 1995, 91-95.
[17] JO Uzuegbu, and OK Okoro, Protection of mechanically injured yellow yam (Dioscorea cayenensis) tubers from fungal rot sing
pastes of some local herbs, J. sustainable Agric Environ. 1 (2), 1999,262-266.
[18] R Tiwari, Fungitoxity of volatile constituents of some higher plants against predominant storage fungi, Indian Phytopathol 50,
1997, 548-551.
[19] AD Akpa, B Musa and AT Paswull, Effect of water extract of neem (Azadrachta indica) leaves as a systemic nermaticide,
Nigerian journal of plant protection 5, 1991, 70-74.
[20] RT Amuchi, Fungitoxic effect of extracts of some African plants, Annual Applied Biotechnology 155, 1999, 451-452.
[21] O Emmanuel, A Olayinka, AM Aiyegoro, and IO Anthony,Preliminary phytochemical screening and anti bacteria properties of
crude stem bark extracts and fractions of Parkia biglobosa, Phytochemistry 18, 2013, 8485-8499.
[22] GB Adebayo, SO Oguntoye and EU Agbowo, The phytochemical analysis and anti- bacteria screening of extracts of Daniellia
oliveri stem bark, Science research 4, 2010, 23-26.
[23] J Parekh, R Nair and S Chanda, Preliminary screening of some folklore medicinal plants from western India for potential
antimicrobial activity, Indian journal of traditional medicine 14, 2005, 34-44.
[24] SE Udo, BE Madunagu and CD Isenin, Inhibition of growth and sporulation of fungal pathogens on potato and yam by garlic
extract. Nigerian journal of Botany 4, 2001, 35-39.
[25] RN Okigbo and IA Nmeka, Control of yam tuber rot with leaf extracts of Xylopia authopica and Zingiber officinale, Afr. J.
Biotechnol., 4(8), 2005, 804-807.

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Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu

  • 1. IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 6 Ver. II (Nov - Dec. 2015), PP 33-41 www.iosrjournals.org DOI: 10.9790/3008-10623341 www.iosrjournals.org 33 | Page Efficacy of Some Botanicals in the Control of Fungi Causing post harvest rot of Yam in Katube market, Obudu, Nigeria. 1 Liamngee kator 2 Akomaye Maria Umgbeb 3 Okoro James Kalu Department of Biological Sciences, Benue State University Makurdi, Benue State, Nigeria. Department of Biology, Federal College Of Education, Obudu, Cross River State, Nigeria. College of Agronomy, Federal University of Agriculture, Benue State, Nigeria. Abstract: Efficacy of some botanicals in the control of fungi causing post harvest rot of yam in Atube market, Obudu was conducted. The fungal pathogens isolated from the yam samples were Aspergillus niger, Botryodiplodia theobromae, Aspergillus flavus, and Aspergillus fumigatus. Water and ethanol extracts of the plant species at 10g/ml, 20g/ml, 30g/ml, 40g/ml and 50g/ml were tested against the fungal isolates in vitro. The inhibitory effects of water extracts of Daniellia oliveri, Moringa oleifera and Parkia biglobosa at 50g/ml on all test organisms ranged from 11.00 to 35.00 and at 10g/ml it ranged from 25.00 to 80.00. The inhibitory effect of ethanol extract of Daniellia oliveri, Parkia biglobosa and Moringa oleifera at 50g/ml on test organisms ranged from 6.50 to 34.50 and at 10g/ml it ranged from 8.00 to 77.50. There was no significant difference in the grand mean inhibitory effect of both water and ethanol extracts on Aspergillus niger and Aspergillus flavus at P = 0.05. There was significant difference in the grand mean inhibitory effect of water and ethanol extracts on Aspergillus fumigatus and Botryodiplodia theobromae. The extracts of Parkia biglobosa, Moringa oleifera and Daniellia oliveri are useful antifungal agents, inhibiting fungal growth at all concentrations. Keywords; Efficacy, Botanicals, fungi, Atube market, rot, Post harvest. I. Introduction Yam (Dioscorea spp) is among the most important staple food in the world especially some parts of the tropics and subtropics [1]. The most cultivated varieties in Nigeria are the D. rotundata (white yam), D. cayenesis (yellow yam) and D. alata (water yam) [2]. Nutritionally, yams are mainly carbohydrate foods. It is one of the most important dietary sources of energy produced in the tropics [3]. White yam (Dioscorea rotundata) is the most important and cultivated variety found in Africa. In 2005, [4] reported that Nigeria produces about 66.6% (26.6 million metric tonnes) of the total world’s yam production every year. White yam (Dioscorea rotundata) is much preferred to other yam varieties and it constitutes about 80% of the total yam produced in Nigeria [5].Though this figure is high, demand for yam has always been more than the supply. As important as yam is, its production is constrained by many problems ranging from high cost of production, attack by fungi, nematodes and pests, singly or in combination. These constraints are responsible for field suppression and tuber quality deterioration in storage [2]. The magnitude of these problems has made people express fears that yam production in Nigeria may decline substantially in the near future [3]. Rot is major factor limiting the post – harvest life of yams and losses can be very high. It is estimated that an average of 25- 50% of yam tubers produced in Nigeria are lost to pests and diseases [4]. The disease causing agents reduce the quantity and quality of yam and make them unappealing to the consumer. Losses due to post- harvest rot significantly affect farmers’ and traders’ income, food security and seed yams stored for planting. In Nigeria, over 60% of white yam get rotten when stored for planting [6]. Most rot of yam tubers are caused by pathogenic fungi such as Aspergillus flavus, Aspergillus niger, Aspergillus fumigatus, Botryodiplodia theobromae, Fusarium oxysporum, Fusarium solani, Penicillium chrysogenum, Rhizoctonia spp, Penicillium oxacilum, Trichoderma viride and Rhizopus nodosus [7]; [8]. It is therefore important to know the fungal organisms that are responsible for post-harvest rot of white yam in Atube market, obudu and seek their effective control measures to extend the life span of yam in storage. II. Materials and Methods 2.1 Sample Collection About 20 tubers of white yam (Dioscorea rotundata) with symptoms of rot were obtained from Atube market, Obudu Local Government Area of Cross River State. The yam tubers were packaged in polyethylene bags and taken to botany laboratory of the Benue State University, Makurdi, where they were assessed for microbial (fungal) presence.
  • 2. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 34 | Page 2.2 Media Preparation The medium used was Potato Dextrose Agar (PDA) prepared according to the manufacturers instruction. About 39.6g of powdered PDA medium was dissolved in1 litre of sterile distilled water and sterilized by autoclaving at 1210 C for 15 minutes. 2.3 Isolation of fungal pathogens from rotten yam tubers Small sizes were cut from yam tubers infected with rot. They were first surface sterilized by dipping completely in a concentration of 5% sodium hypochloride solution for 2 mins. They were then removed and rinsed in several changes of sterile distilled water. The yam pieces were then placed on sterile paper towels to dry for 5 mins. The pieces of yam sections were placed on solidified Potato Dextrose Agar medium. Three replicates were made for each of the 20 samples of yam tubers. The inoculated plates were incubated at 25-280 C and observations were made daily for possible microbial growth. After 5 - 7 days of growth, Sub-culturing was done to obtain pure cultures of the isolates. 2.4 Identification of fungal isolates The identification of the isolates was done by examining the isolates macroscopically and microscopically. The colony characteristics such as colony appearance, change in medium colour and growth rate were observed. Shapes of the conidia and conidiophores were taken note of. These structural features were matched with standards in [9] and [10]. 2.5 Pathogenicity Test To confirm the pathogenicity of isolates from white yam, pure cultures of the isolates were used to inoculate three white yams per pathogen with 5mm diameter mycelia plugs of a 4 day old culture. On appearance of symptoms, the tissues at the margins of the healthy and diseased parts were surfaced sterilized, excised and plated onto Potato Dextrose Agar (PDA) and incubated at 25-28o C for 5 - 7 days. At the end of this period, morphological characteristics and growth patterns observed in each case were compared with the ones of the original isolates. One tuber of white yam was used for each fungal isolate respectively, replicated three times and arranged in complete randomized design. Controls were yam tubers inoculated with sterile PDA only. Following appearance of symptoms, 8 - 12 days post inoculation, rot severity index was assessed on a scale of 0 - 5, where 0 - no disease manifestation, 1 - 1-20% rot, 2 - 21-40% rot, 3 - 41-60% rot, 4 - 61-80% rot, 5 - 81- 100% rot. 2.6 Collection of Plant Material Fresh leaves of three selected plant species were collected in polyethylene bags and transported to the botany laboratory of the Benue State University Makurdi for preparation of extracts. They are Parkia biglobosa (locust bean tree), Moringa oleifera (Drumstick) and Daniellia oliveri (African Copaiba Balsam). 2.7 Preparation of Extracts of Plant Parts Water and ethanol extracts of each selected plant species was prepared. Each of the leaves were washed under running tap water and soaked in a 1% solution of sodium hypochloride for 2 minutes. They were rinsed severally with sterile distilled water and air dried at room temperature for 30 mins. 50g each of the fresh leaves of the selected trees were weighed for both water and ethanol extractions respectively. The leaves were ground one after the other using a mortar and pestle. The mortar and pestle were washed after each type of leave was pounded to avoid mixture of their different components. The macerates were transferred into beakers and each was soaked in100ml of sterile distilled water and 90% ethanol respectively for water and ethanol extractions. 2.8 Concentrations of Crude Extracts Serial dilutions of the crude extracts of the leaves of each plant was prepared to give different concentrations of the extracts. Extract concentrations of 10g/ml, 20g/ml, 30g/ml, 40g/ml and 50g/ml were thus obtained for both water and ethanol extractions. 2.9 Antifungal Susceptibility Test in vitro One ml of the extract concentration for both ethanol and water was dispensed in Petri dishes and 15- 20mls of molten PDA was added. The Petri plates were swirled gently on the work bench to ensure even dispersion of the extracts. The Agar-extract mixture was allowed to solidify and used for the inhibition of mycelia growth for the tested fungi. 4mm diameter of mycelia disc obtained from the colony edge of 5day old culture of each of the test fungi was inoculated centrally onto the medium. Three replications were used for each fungal isolate. Controls were Petri plates containing PDA with no botanical extract and inoculated with the test
  • 3. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 35 | Page fungi. The plates were arranged on laboratory desk following complete randomized design. The Petri plates were then incubated at 25 - 28o C for 5-7 days during which measurement of growth was done using a metre rule at an interval of 24 hours. Inhibition of fungal growth was calculated using the formula R1 - R2 x 100 R1 where R1 = radial growth of the pathogen in the control plates R2 = growth of the pathogen with treatment 2.10 Data Analysis Data obtained from the study was analyzed using Analysis of Variance (ANOVA) and the Fishers Least Significant Difference was used to separate the means at 5% level of significance. III. Results 3.1 Fungal pathogens isolated from yam samples A total of four fungi namely Botryodiplodia theobromae, Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger were isolated from the yam samples as shown in Table 1. Identification of the isolates was done using recommendations given by [9] and other electronic documentations on the genera isolated. Table 1: Characterization of Fungal isolates from Yam samples on PDA. Cultural/microscopic characteristics Appearance on PDA Photomicrograph Probable organism Colony colour is dark brown to black. Conidia are black and densely packed. Conidiophore length is 855 – 9 57(µm) and colour was hyaline to yellowish or slightly brown. Aspergillus niger Colony colour ranged from grayish to dark blue in colour. The conidia colour was dark green to blue. Conidiophore length was 265–365µm and it was uncoloured to grayish brown Aspergillus fumigatus Colony colour was parrot green to deep green. Conidia were light green in colour. Conidiophores length ranged from 550 – 678 µm . Aspergillus flavus
  • 4. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 36 | Page Colony colour ranged from white to black. Mycelium is septate and the conidia are 2 celled. Botryodiplodia theobromae 3.2 Pathogenicity test. The Pathogenicity test showed that the fungal isolates induced rot in healthy yam tubers after 14 days of inoculation. The rot severity ranged from 3 – 5 which indicated 41 – 100% of tuber rot as shown in Table 2. Analysis of Variance revealed a significant difference in the disease causing potential of the fungal isolates on healthy yam tubers with respect to their controls as shown in Table 3. Table 2: Disease causing potential of fungal isolates on healthy yam tubers Fungal isolates/Replicates C R1 R2 R3 Aspergillus niger 0 5 5 4 Aspergillus fumigates 0 4 4 4 Aspergillus flavus 0 4 3 3 Botryodiplodia theobromae 0 5 5 5 Key Severity Scale C = Control 0 - No infection R1 = Replicate 1 1 - 1 – 20% of rot R2 = Replicate 2 2 - 21 – 40% of rot R3 = Replicate 3 3 - 41 – 60% of rot 4 - 61 – 80% of rot 5 - 81 – 100% of rot Table 3: Analysis of Variance in the disease causing potential of Fungal Isolates on healthy Yam tubers Fungal Isolates A. niger A. flavus A. fumigatus B. theobromae Pathogenicity 5.00a 4.00a 4.00a 5.00a Control 0.00b 0.00b 0.00b 0.00b Footnote: Means not tagged with same alphabets are significant at P= 0.05 3.3 Mean inhibitory effect of water extract of leaves of Moringa oleifera on test organisms. The highest significant inhibitory effect was observed on both Botryodiplodia theobromae and Aspergillus fumigatus at 50g/ml (11.00) while the least significant inhibitory effect was observed at 10g/ml (77.50) and (41.00) respectively. On A. flavus, highest inhibitory effect was observed at 50g/ml (13.50) and the lowest inhibitory effect was observed at 10g/ml (35.00).On A. niger, the highest inhibitory effect was observed at 50g/ml (36.00) and the lowest inhibitory effect was observed at 10g/ml (60.50) as shown in Table 4. Table 4: Mean inhibitory effect of Water extract of Moringa oleifera on test organisms. Conc g/ml A. flavus A. niger B. theobromae A. fumigatus 10 35.00 60.50 77.50a 41.00a 20 29.00 41.00 56.50b 29.50b 30 24.00 59.00 49.50b 21.00b 40 14.50 52.50 44.50b 26.00b 50 13.50 36.00 11.00c 11.00b Footnote: Means not tagged with same alphabets are significant otherwise they are the same at P=0.05 NS - No significant difference LSD(0.05) 0.93 0.92 0.91 0.00 LSD(0.05) NS NS 13.57 8.92
  • 5. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 37 | Page 3.4 Mean inhibitory effect of ethanol extracts of Moringa oleifera on test fungi The highest significant inhibitory effect was observed on Aspergillus niger at 50g/ml and 20g/ml (22.50) while the least significant inhibitory effect was observed 10g/ml (53.50). On Botryodiplodia theobromae, highest significant inhibitory effect was observed at 50g/ml (17.00) and the lowest significant inhibitory effect was observed at10g/ml (60.00). On A. fumigatus, highest inhibitory effect was observed at 50g/ml (16.00) and lowest inhibitory effect was observed at 10g/ml (60.00). On A. flavus, highest inhibitory effect was observed at 50g/ml (7.00) while the lowest inhibitory effect was observed at 10g/ml (29.00) as shown in Table 5. Table 5: Mean inhibitory effect of Ethanol extract of Moringa oleifera on test organisms. Conc g/ml A. flavus A. niger B. theobromae A. fumigatus 10 29.00 53.50a 60.00a 60.00 20 19.00 22.50b 35.50b 39.00 30 21.50 23.50b 25.50c 30.00 40 26.50 25.00b 22.50c 45.00 50 7.00 22.50b 17.00c 16.00 Footnote: Means not tagged with same alphabets are significant otherwise, they are the same at P=0.05 NS - No significant difference 3.5 Mean inhibitory effect of water extracts of Daniellia oliveri on test fungi The highest inhibitory effect was observed at 50g/ml (20.00) on Aspergillus flavus and the least inhibitory effect at 10g/ml (80.00) was on Aspergillus niger. Significant inhibitory effect was observed on Aspergillus flavus and Botryodiplodia theobromae. On Aspergillus flavus, all means were significantly different in their zones of inhibition except at 20g/ml (41.50) and 40g/ml (46.00) while all inhibitory effects on Botryodiplodia theobromae were insignificant except at 30g/ml (80.00) as shown in Table 6. Table 6: Mean inhibitory effect of Water extract of Daniellia oliveri on test organisms. Conc g/ml A. flavus A .niger B. theobromae A. fumigatus 10 69.50a 80.00 50.00a 77.50 20 41.50b 60.50 42.50a 75.00 30 56.30c 45.00 80.00b 65.00 40 46.00b 42.50 39.00ad 34.00 50 20.00d 35.00 29.50cd 31.00 Footnote: Means not tagged with same alphabets are significant otherwise, they are the same at P=0.05 NS - No significant difference 3.6 Mean inhibitory effect of ethanol extracts of Daniellia oliveri on test fungi There was insignificant inhibitory effect at all concentrations on Aspergillus flavus, Aspergillus niger and Botryodiplodia theobromae while A. fumigatus had highest significant inhibitory effect at 50g/ml (5.00) and least significant inhibitory effect at 10g/ml (30.00). On A. flavus highest inhibitory effect was observed at 50g/ml (12.50) and least inhibitory effect was observed at 40g/ml (16.50). On A. niger, highest inhibitory effect was observed at 50g/ml (9.50) while least inhibitory effect was observed at 40g/ml (24.00). On B. theobromae, highest inhibitory effect was observed at 50g/ml (6.50) and least inhibitory effect was observed at 40g/ml and 10g/ml (8.00) respectively as shown in Table 7. Table 7: Mean inhibitory effect of Ethanol extract of Daniellia oliveri on test organisms. Conc g/ml A. flavus A. niger B. theobromae A. fumigatus 10 36.50 28.50 8.00 30.00a 20 22.50 41.00 15.50 14.00b 30 28.50 47.00 17.50 9.50bd 40 16.50 24.00 8.00 15.00b 50 12.50 9.50 6.50 5.00cd LSD(0.05) NS 11.54 8.89 NS LSD(0.05) 11.54 NS 12.98 NS LSD(0.05) NS NS NS 7.17
  • 6. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 38 | Page Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05 NS - No significant difference 3.7 Mean inhibitory effect of water extract of Parkia biglobosa on test fungi There was insignificant inhibitory effect on all the fungal isolates at all concentrations except on Aspergillus flavus which had significant highest inhibitory effect at 50g/ml (16.50cm) and least significant effect at 10g/ml (63.50cm). On A. niger, highest inhibitory effect was observed at 50g/ml (26.50) and lowest inhibitory effect was observed at 10g/ml (57.50). On A.fumigatus, highest inhibitory effect was observed at 50g/ml (18.00) and the lowest was at 10g/ml (25.50). On B. theobromae, highest inhibitory effect was observed at 50g/ml (22.50) and the lowest was at 20g/ml (58.00) as shown in Table 8. Table 8: Mean inhibitory effect of Water extract of Parkia biglobosa on test organisms. Conc g/ml A. flavus A. niger B. theobromae A. fumigatus 10 63.50a 57.50 57.50 25.50 20 43.50b 56.00 58.00 39.00 30 47.00b 43.50 40.80 35.00 40 35.50b 47.50 48.00 39.00 50 16.50c 26.50 22.50 18.00 Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05 NS - No significant difference 3.8 Mean inhibitory effect of ethanol extracts of Parkia biglobosa on fungal Isolates There was insignificant inhibitory effect of Parkia biglobosa on the test organisms at all concentrations. Highest inhibitory effect was observed on A. flavus at 40g/ml and 50g/ml (12.50). Lowest inhibitory effect was observed on A.niger at 10g/ml (77.50) followed by A. fumigatus (75.00), B. theobromae (69.00) and A. flavus (41.50) respectively. A. niger and B. theobromae shared highest inhibitory effect at 50g/ml (34.50) and A. fumigatus had highest inhibitory effect at 50g/ml (27.00) as shown in Table 9. Table 9: Mean inhibitory effect of Ethanol Extract of Parkia biglobosa on test organisms. Conc g/ml A .flavus A. niger B. theobromae A. fumigatus 10 41.50 77.50 69.00 75.00 20 43.00 60.00 53.50 63.00 30 33.50 48.00 38.50 52.00 40 12.50 40.00 39.00 35.00 50 12.50 34.50 34.50 27.00 Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05 NS - No significant difference 3.9 Grand mean inhibitory effect of water extracts of the three plant species on the test organisms There was insignificant inhibitory effect of all the water extracts of the three plants species on Aspergillus flavus, Aspergillus niger and Botryodiplodia theobromae except on Aspergillus fumigatus where the effects of M. oleifera and P. biglobosa were significantly different from that of D. oliveri as shown in Table 10. Table 10: Grand Mean inhibitory effect of Water extracts of all the three plant species on test organisms. Plant Extract A. flavus A. niger B. theobromae A.fumigatus M.oleifera 23.20 49.80 47.80 25.70a D.oliveri 46.66 52.60 48.20 56.50b P.biglobosa 41.20 46.20 45.36 31.30a Footnote: Means not tagged with same alphabets are significant; otherwise they are the same at P=0.05 NS - No significant difference LSD(0.05) 11.13 NS NS NS LSD(0.05) NS NS NS NS LSD(0.05) NS NS NS 21.21
  • 7. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 39 | Page 3.10 Grand mean inhibitory effect of ethanol extracts of the three plant species on test fungi There was significant inhibitory effect of all the ethanol extracts of the three plant species on Botryodiplodia theobromae and Aspergillus fumigatus. Extracts of M. oleifera and P. biglobosa were significantly different from that of D. oliveri in their effects on B. thoebromae and A. fumigatus as shown in Table 11. Table 11: Grand Mean inhibitory effect of Ethanol extracts of all the three plant species on test organisms. Plant Extract A. flavus A. niger B. theobromae A. fumigatus M.oleifera 20.60 29.40 32.10a 38.00a D.oliveri 23.30 30.00 11.10b 14.70b P.biglobosa 28.60 52.00 46.90a 50.40a Footnote: Means not tagged with same letters are significant; otherwise they are the same at P=0.05 NS - No significant difference 3.11 Comparative grand mean inhibitory effect between water and ethanol extracts of the three plant species on test fungi There was significant inhibitory effect between water and ethanol extracts of Daniellia oliveri on Aspergillus niger (21.40) while water and ethanol extracts of Moringa oleifera and Parkia biglobosa had insignificant inhibitory effect on Aspergillus niger. There was significant inhibitory effect between water and ethanol extracts of Moringa oleifera on Aspergillus flavus (19.92) while water and ethanol extracts of Daniellia oliveri and Parkia biglobosa had insignificant inhibitory effect on Aspergillus flavus. Water and ethanol extracts of Daniellia oliveri showed significant inhibitory effect on Botryodiplodia theobromae (20.54) while water and ethanol extracts of Moringa oleifera and Parkia biglobosa had insignificant inhibitory effect on Botryodiplodia theobromae. Water and ethanol extracts of Daniellia oliveri showed significant inhibitory effect on Aspergillus fumigatus (25.13) while water and ethanol extracts of Moringa oleifera and Parkia biglobosa had insignificant inhibitory effect on Aspergillus fumigatus as shown in table 12. Table 12: Comparative grand Mean inhibitory effect of water and ethanol extracts of the three plant species on test fungi Solvents A niger A flavus B theobromae A fumigatus MO DO PB MO DO PB MO DO PB MO DO PB Water extract 23.20 46.66a 41.20 49.80a 52.60 46.20 47.80 48.20a 45.36 25.70 56.50a 31.30 Ethanol extract 20.60 23.30b 28.60 29.40b 30.00 52.00 32.10 11.10b 46.90 38.00 14.70b 50.50 LSD (0.05) NS 21.40 NS 17.92 NS NS NS 20.54 NS NS 25.13 NS Footnote: Means not tagged with same alphabets are significant otherwise they are the same at P=0.05 Key NS - No significant difference MO - Moringa oleifera DO - Daniellia oliveri PB - Parkia biglobosa IV. Discussion The Fungal organisms associated with rot of white yam (Dioscorea rotundata) in this present study were Aspergillus niger, Apergillus flavus, Aspergillus fumigatus, and Botryodiplodia theobromae. These organisms have been associated with post harvest rot as reported by [11] , [12], [1]. The pathogenicity test showed that the fungal isolates inoculated into healthy yam tubers caused tuber rot. This was due to the ability of the fungal pathogens to utilize the nutrients of the yam as a substrate for growth and development. This result is similar to the report on fungi associated with Nigerian yams by [13], [11] and [12]. In another set of experiments, the water and ethanol extracts of the three plant species; Parkia biglobosa, Daniellia oliveri and Moringa oleifera showed antifungal activities against the test fungi at all concentrations in vitro. This implies that locally occurring plants commonly used in folklore human medicine also posses anti microbial effects on post harvest fungi. This conform to the results obtained from the studies of LSD(0.05) NS NS 18.12 21.75
  • 8. Efficacy of Some Botanicals In the Control of Fungi Causing post harvest rot of Yam in Atube market, Obudu. DOI: 10.9790/3008-10623341 www.iosrjournals.org 40 | Page [14] and [15] on the bioassay of some plant extracts on some fungal pathogens. This also agrees with the reports of [16], [17] and [18] on the inhibitory action of plant products employed on the mycelia growth and spore germination of other pathogenic fungi. It was observed that susceptibility of the test fungi increased with increasing concentration of the plant extracts for all the fungal isolates. This may have resulted from variation in the principal active ingredients present in the extracts. The effectiveness of a plant extract depends on the nature and amount of active ingredient found in it. This agrees with the findings of [19] who reported significant inhibitory property of Neem (A. indica) extract that contains azadrachtin which is known to be fungitoxic against most fungal pathogens just as [20] found the extract of Ocimum gratissimum to reduce the growth of Rhizopus spp. Also, [21] and [22] reported that the phytochemical screening of the plant species used in this study showed the presence of alkaloids, saponins, tannins, flavonoids, glycosides and phenol compounds. An increase in the concentration of the plant extracts correspondingly decreased the growth of the test fungi. This is because an increase in the concentration of the extract implied an increase in the active ingredient of the solu tion which acts on the fungus thereby affecting its physiological processes and consequently lowering the growth of the fungus. The ethanol extract of D. oliveri recorded the highest grand inhibitory effect of (11.10) on Botryodiplodia theobromae and the least was water extract of D. oliveri (56.50) on A. fumigatus. The ethanol extract of Moringa oleifera recorded grand mean inhibitory effect of (20.60) on Aspergillus niger, (29.40) on Aspergillus flavus, (32.10) on Botryodiplodia theobromae, (38.00) on Aspergillus fumigatus compared to water extract which had a mean inhibitory effect of (23.20) on Aspergillus niger, (49.80) on Aspergillus flavus, (47.80) on Botryodiplodia theobromae and (25.70) on Aspergillus fumigatus. This is in agreement with [23] who stated that plant extracts in organic solvents provided more consistent antimicrobial activity compared to water extracts. The most sensitive test organism in this study was Botryodiplodia theobromae which had an inhibitory diameter of (11.10) on ethanol extract of Daniellia oliveri. The least sensitive test organism was Aspergillus fumigatus with an inhibitory diameter of (56.50) on water extract of D. oliveri. The differences recorded in the fungitoxic activity of the extracts may also be attributed to the solubility of the active ingredients in water and ethanol or the presence or absence of inhibitors for the fungitoxic principle. Also the effectiveness of the ethanol extracts than water extracts observed in this study is probably due to its ability to extract more bioactive compounds from the plant material which are effective on the test organisms. The active ingredient present in an extract is influenced by many factors which include the age of the plant, extracting solvent, method of extraction and time of harvesting the plant material. V. Conclusion This study has confirmed and established that the extracts of Parkia biglobosa, Daniellia oliveri and Moringa oleifera are useful antifungal agents, inhibiting fungal growth at all concentrations. This agrees with earlier reports of [24] on the inhibition of growth and sporulation of fungal pathogens on Ipomea batatas and Dioscorea species. and [25] on the use of Xylopia aethiopica and Zingiber officinale to control yam tuber rot caused by F. oxysporum, Aspergillus niger and Aspergillus flavus. References [1] RN Okigbo and UO Ogbonnaya, Antifungal effect of two tropical plant leaf extract Ocimum gratissimum and Aframomum melegueta on post harvest yam (Dioscorea spp) rot, Afr. J. Biotech. 5 (9), 2006, 727-731. [2] N Amusa, yam diseases and their management in Nigeria, African Journal of Biotechnology 2 (12), 1999, 497-502. [3] N Ekefan, Diseases of yam tubers, International Journal of Tropical Plant Diseases, 3, 1999, 23 -27. [4] FAO, Food and Agricultural Organisation, FAO, Rome, 2005, pp23. [5] JP Tetteh and C. Saakwa, Prospects and constraints of yam production in Ghana (Tyum press, 1994). [6] SO Adesiyan and RA. 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