Journal of Phytopathology and Pest Management 6(1): 78-98, 2019
pISSN: 2356-8577 eISSN: 2356-6507
Journal homepage: http://ppmj.net/
∗
Corresponding author:
Waleed M. A. Abd-Elmagid,
E-mail: waleedabdel-magid.5419@azhar.edu.eg
78
Copyright © 2019
Impact of applying certain bio-agents and
plant extracts to control root and pod rot
peanut pathogens
Mohamed M. El-Sheikh Aly
1
, Ali H. ElShaer
2
, Ahmed R. Abdallah
3
, Thrwat A. Aldahtory
3
, Waleed M. A. Abd-Elmagid
1*
1
Agricultural Botany Department, Faculty of Agriculture, Al-Azhar University, 71524 Assiut, Egypt
2
Plant Pathology Research Institute, Agricultural Research Center, Giza, Egypt
3
Agricultural Microbiology Department, Faculty of Agriculture, Minia University, Minia, Egypt
Abstract
Keywords: peanut, plant extracts, Fusarium spp., pod rots, root rot.
The current study was performed to evaluate the efficacy of some bacterial and fungal
bio-agents and certain plant extracts i.e. garlic, neem and mint against Fusarium spp.,
the causal agent of peanut root and pod rot diseases. In vitro tests clarified that
Penibacillus polymyxa (BP) and Pseudomonas fluorecensce (PF2) achieved the highest
inhibition percentage of the tested pathogenic fungi followed by Bacillus subtilis (BS1)
and P. fluorecensce (PF1) while, P. fluorecensce (PF3) and Bacillus megaterium (BM2)
came in the last order. Addition of suspensions of Bacillus (B1), P. fluorecensce (P11),
inoculum of T. harzianum (T10) and combination of all bio-control agents (Mixture) to
infected soil significantly increased yield of peanut plants (Giza 6 cv), such as fresh
weight (gm /plant), dry weight (gm /plant), plant height (cm /plant), number of pods /pot
and weight of pods /pot. Regarding plant extracts, garlic extract was the most effective
plant extract in suppressing the mycelial growth of the pathogenic fungi than any other
treatment. Mint extract showed the lowest effect on reduction of linear growth of the
tested pathogens. Concerning root rot disease management, addition of bio-agents in
vivo resulted in distinct disease reduction obtained by P. fluorecensce (PF2) and Mixture
treatments followed by Penibacillus polymyxa (BP) and T. harzianum (T10), respectively.
In the concern of pod rot disease, the mixture and T. harzianum (T10) treatments
recorded the highest disease reduction as compared to control while, P. fluorecensce
(PF2) and Penibacillus polymyxa (BP) came in the last order. Treated seeds of peanut
(Giza 6 cv.) with certain plant extracts (garlic, neem and mint) at concentration of 30%
significantly reduced the severity of root and pod rot diseases compared to control.
Garlic extract gave the highest reduction of root and pod rot severities on peanut plants
followed by neem extract then mint extract, respectively.
El-Sheikh Aly et al., 2019
79
1. Introduction
Arachis hypogaea
L. (Groundnut or
Peanut) is one of the important crops all
over the world. In Egypt it is one of the
major oil seed crops and considered as
one of the most important, export and
edible oil crops as well as in many
countries of the world. In addition to this,
belonging to leguminous plants, peanut
has the ability to fix nitrogen from the
atmosphere biologically into the soil
which enriches the soil and these benefits
the succeeding crop. Peanut seeds are
characterized by their high oil content
(50%), which is utilized in different
industries beside containing 26–28%
protein, 20% carbohydrates and 5% fiber
(Fageria et al., 1997).
The peanut crop is
vulnerable to soil borne diseases since
both roots and pods of the plant grow in
soil. Diseases caused by soil borne fungal
pathogens reduce yields and the quality
of the harvested pods and it affect the
crop plant until harvest. Pathogens attack
all plant parts of groundnut and restrict
plant development throughout the
growing season as well as reducing seed
quality in post-harvest storage
(Mahmoud
et al., 2006).
Root and pod rot diseases of
peanut are serious worldwide diseases
attacking roots and fruits underground
(Hilal et al., 1990).
Fusarium
spp is
known as a pathogen causing different
symptoms of infected roots and pods
(Hussin Zeinab, 2011; Marei, 2000)
.
Fusarium solani
is one of the most
important pathogens causing brown root
rot in peanuts. Under conductive
conditions like drought stress, the losses
could reach 95% of production in some
fields. Fungicide treatments are not
effective enough to control the disease
beside the harmful effect of excessive and
misuse of fungicides which lead to
environmental pollution and causes
damage to the ecosystem. Recently many
researches tried to develop alternative
control strategies in order to reduce
dependency on synthetic fungicides.
Pseudomonas fluorescence
is an effective
candidate for biological control of soil
borne plant pathogens owing to its
versatile nature, rhizosphere competence
and multiple modes of action beside
being endophytic in the plant system
(Diby et al., 2001).
Pseudomonas
spp.
also can induce systemic biochemical and
ultra-structural changes in the roots that
lead to a greater ability of the host plant
to defend itself against root infecting
pathogens (Sarma et al
.,
2000). It was
noticed that in greenhouse experiments
P. flurescens
and
B. subtilis
significantly
reduced the incidence of all types of
peanut pod rot caused by
R. solani,
S.rolfsii, M. phaseolina, Fusarium
spp.
and
Aspergillus
spp. (Mahmoud, 2004).
Trichoderma
spp. are one of the most
important biological control agents and
one of the most frequently isolated soil
fungi present in plant root ecosystems.
They colonize the root and rhizosphere of
plant and suppress plant pathogens by
different mechanisms, such as
competition, mycoparasitism, antibiosis
and induced systemic resistance,
improvement of the plant health by
promoting plant growth, and stimulation
of root growth
(Mohidden et al., 2010).
Trichoderma harzianum
,
T. hamatum
and
B. subtilis
reduced the mycelial growth of
R. solani, F. solani
and
M. phaseolina
.
Trichoderma hamatum
mainly grew over
the mycelium of the tested pathogens (El-
Sayed et al
.,
2009). It is well known that
plants have the ability to synthesize
aromatic secondary metabolites, like
phenols, phenolic acids, quinones,
flavones, flavonoids, flavonols, tannins
and coumarins (Cowan, 1999).
Therefore, many scientists have reported
the use of plant extracts for controlling
certain fungal diseases (Touba et al
.,
2012; Al-Askar & Rashad, 2010). The
objective of this research was the
El-Sheikh Aly et al., 2019
80
development of alternative control
strategies to reduce dependency on
synthetic fungicides. Also to evaluate
Bacillus
spp.,
Psedomonas florescence
and
Trichoderma
species as potential
biocontrol agents to reduce the impact of
the disease under greenhouse conditions.
2. Materials and methods
2.1 Seeds
Seeds of peanut that have been used in
this study were cultivar Giza 6. They
were obtained from Agriculture Research
Center (ARC), Giza, Egypt.
2.2 Pathogenic fungi source
Virulent
Fusarium
isolates used in this
study were previously isolated, identified
and
proved their pathogenic ability to
induce root and pod rot of peanut in
former study (Abdallah et al., 2016).
2.3 Isolation of biocontrol agents
Isolation trials were conducted to isolate
Pseudomonas
and
Bacillus
colonizing
rhizosphere soil of peanut growing in
different location of Minia, Assiut and
Sohag governorates, Egypt. Isolates were
picked up on the suitable media. All
Pseudomonas
and
Bacillus
isolates were
purified by successive streaking on N-
deficient modified king medium and
nutrient agar media,
respectively,
using
the techniques adopted by King et al
.
(1954) for
Pseudomonas
and nutrient
agar medium
(Oxoid Manual 1965) for
Bacillus.
The purified isolates were
maintained on the same media and
confirmed by microscope examination
for Gram stained young cells (18-24
hours old). Then the purified isolates
were stored at 4°C for further studies. As
for
Trichoderma
spp. it was also isolated
from healthy peanut plants rhizosphere
using dilution plates technique according
to Warcup (1955).
Trichoderma
isolates
were identified according to their
morphological characteristics using
identification keys
(Rifai, 1969), also
two identified isolates obtained were
from Assiut University Mycological
Center, Egypt.
2.4 Antagonistic effect of isolated
microorganisms against
Fusarium
isolates
in vitro
2.4.1 Bacterial biocontrol agents
Six bacterial isolates
i.e.
Pseudomonas
(PF1, PF2, PF3),
Bacillus
(BP, BS1,
BM2) were investigated for their effect
against the growth of
Fusarium
isolates
under Lab conditions. Petri dishes
(containing PDA medium) were streaked
with the bacterial growth which obtained
from 24hours old cultures at the
periphery using sterilized needle and left
for 24 hours. One disk of the pathogen
was placed at the center of each plate,
then plates were incubated at 27°C.
Plates without bacterial inoculation were
served as control treatment. Each
treatment contained three replicates.
When growth of the pathogen covered
the plate surface (9.0 cm in diameter) of
control treatment, antagonistic effect was
determined by measuring the free
inhibition zone, then percentage of
mycelial growth inhibition was
calculated according to the formula:
Percentage of mycelial growth inhibition % = [A – B /A] × 100
Where: A = length of hyphal growth of
the control, B = length of hyphal growth
of the treated.
El-Sheikh Aly et al., 2019
81
2.4.2 Fugal biocontrol agents
Petri-dish was divided into equal halves.
The first half was separately inoculated
with standard disc (5 mm) of
Trichoderma
spp.,
previously isolated
from peanut rhizosphere. The second half
was inoculated with an equal disc of each
of
Fusarium spp.
Isolates.
Each
treatment was replicated three times and
inoculated plates with the pathogen
only
were used as control. All Petri-dishes
were incubated at 27 °C and data were
recorded when control treatments cover
the plates. Antagonistic percentage was
calculated according to the following
scale index: 0-4, 0= no antagonism; 1=
slight antagonism; 2= moderate
antagonism; 3= high antagonism and 4=
overgrowth (Hassan, 1992).
2.4.3 Effect of culture filtrate of
antagonistic fungi on growth of
Fusarium
isolates
Trichoderma
harzianum
isolates (No. 2,
8 and 10) were grown on Czapek′s liquid
medium. Each flask containing 100 ml of
the medium was inoculated separately
with 5 mm agar disc obtained from 7
days-old culture
.
Then, the flasks were
incubated at 27°C for 30 days. The
obtained cultures were used to study the
effect of the culture filtrates of
antagonistic fungi on mycelial growth of
F. oxysporum
(I),
F. solani
(V) and
F.
solani
(VII) isolates. The mycelial
growth was eliminated and the filtrate of
each fungus isolate was centrifuged for
60 min at 3000 rpm to separate the fungal
growth (Mohamed et al., 2008). Filtrates
were sterilized by Seitz filter and added
to the medium at the rate of 10, 25 and
50% (v/v), and mixed thoroughly before
solidification at 40-50°C. Petri-dishes
were inoculated with equal discs
Fusarium oxysporum
(I),
Fusarium
solani
(V) and
Fusarium solani
(VII).
Petri dishes received medium only
without culture filtrate were used as
control. Four replicates were used for
each treatment. All treatments were
incubated at 27°C for 7days. Data were
recorded as diameter of linear growth
when the control plates were completely
covered by the fungal mycelium.
Percentage of reduction in growth was
calculated according to the following
formula (Abd El-Khair & Haggag
Wafaa, 2007):
Growth
inhibition
%
=
((Growth in control –
Growth in each treatment) / Growth in control) ×
100
2.5 Effect of aqueous plant extracts on
growth of pathogenic fungi
Aqueous plant extracts were prepared
from cloves of garlic (
Allium sativum
),
leaves of Neem (
Azadirachta indica
) and
leaves of mint (
Mentha
). The plant parts
were washed several times with sterilized
distilled water , cut into small pieces,
then 100 gram of each were macerated in
100 ml distilled water by using mortar,
resulting extract was squeezed twice
through four layers of cheese cloth, then,
centrifuged at (4000 rpm for 9 min ), and
sterilized using Seitz filtrate (Hassan,
2006). Sterilized filtrates of aqueous
plant extracts were kept in dark bottles in
refrigerator until use. Flasks (250 ml)
contained 100 ml of sterilized PDA
medium were melted. After that, plant
extracts were added to the PDA medium
to obtain concentrations 10, 20 and 30%,
mixed and poured in sterilized Petri
dishes (20 ml /plate). Plates were
El-Sheikh Aly et al., 2019
82
inoculated with equal discs (6 mm in
diameter) of
Fusarium
isolates taken
from 4 days old cultures. Four replicates
were used for each treatment. Control
treatment was obtained by culturing the
tested fungi on PDA medium without
addition of aqueous plant extracts. The
inoculated plates were incubated at 27
±2°C until the fungal growth covered the
plate surface of the control treatments.
The percentage of mycelial growth
reduction was calculated using the
following formula:
Percentage of growth reduction
=
[(growth in control –
growth in treatment) /growth in control] × 100
2.6 Greenhouse experiments
2.6.1 Effect of applying biocontrol
agents to control peanut root and pod
rot diseases
Pot experiments were carried out under
greenhouse conditions in Agricultural
Botany Department, Faculty of
Agriculture, Al-Azhar University (Assiut
branch), Assiut, Egypt during 2015 and
2016 growing seasons. It was done to
study the effect of
Penibacillus polymyxa
(BP),
Pseudmonas fluorescens
(PF2) and
Tichoderma harzianum
(T10) on
controlling root and pod rot of peanut.
Each bacterial suspension (1x10
8
cfu/ml)
was prepared by dilution plate assay as
described by Callan et al
.
(1990).
Bacterial cells from agar cultures of each
isolate were inoculated into nutrient broth
(NB) and centrifuged at 3000 rpm for 5
min, the supernatant was discarded and
the precipitate was re-suspended in 100
ml sterilized distilled water. The
suspension was re-centrifuged for 5 min.
and the precipitate was finally suspended
in sterilized distilled water. Bacterial
concentrations were determined
according to their turbidity using
spectrophotometer. Inoculum of
Trichoderma harzianum
was prepared
according to Abdel-Moneem (1996) by
inoculating sterilized conical flasks
(1000 ml) which contain 75 g sorghum
cereal, 25 g clean sand, 2 g sucrose and
200 ml water with equal discs (0.5 cm)
taken from 7 days old cultures grown on
PDA medium at 27°C. The inoculated
flasks were incubated at 27°C for two
weeks. Bacterial isolates and
T.
harzianum
were applied as soil treatment
by adding 100 ml of bacterial
suspensions (10
8
cfu /ml) and
T.
harzianum
at the rate of 2% (w/w) for
each pot. Inocula of the pathogenic
fungal isolates were prepared by growing
them in sterilized conical flasks (1000
ml) containing sand and sorghum
medium, then incubated at 27°C for 21
days. Sterilized pots (30 cm in diameter)
were filled with infested sand clay soil at
the rate 2% (w/w) 7 days before sowing.
Pots were left for one week, watered and
mixed thoroughly to ensure distribution
with the tested fungi. Eight seeds of Giza
6 cv. were sown in each pots and the
biocontrol agents were applied as
mentioned early. Untreated pots were
served as control. Each treatment was
replicated four times. Disease severity of
root rot was recorded after 90 days from
sowing. The arbitrary (0-5) disease index
scale as described by Grunwald et al
.
(2003) and Hussin Zeinab (2011) was
adopted, where: 0= No visible
symptoms, 1= slight hypocotyls lesions,
2= lesions coalescing around epicotyls
and hypocotyls, 3= lesions starting to
spread into the root system with root tips
starting to be infected, 4= epicotyls,
hypocotyls and root system almost
El-Sheikh Aly et al., 2019
83
completely infected and 5= completely
infected root
.
Pod rot severity was
recorded by adopting the scale based on
area of spots covered on pods and percent
disease index (PDI), calculated mean
disease index. Pods were grouped into
six categories described by Wheeler
(1969), Where 0=non disease symptoms
(disease free), 1=spots cover 1-10%, 2=
spots cover 10-30%, 3= spots cover 30-
50%, 4= spots cover 50-70% and 5=
spots cover >70%. The percentage of
disease severity of root rot and the
percentage of pod rot disease were
estimated using the following formula:
Disease severity (%) = ∑ [(n x V)/5 × N)] × 100
Where, n= number of root or pod within
each infection category, V= numerical
values of infection categories, N= total
number of root or pods examined, 5=
constant, the highest numerical value.
At the end of the experiment some
growth parameters such as plant fresh
and dry weights, plant height, and
number of pods and weight of pods were
evaluated.
2.6.2 Effect of aqueous plant extracts
on the incidence of peanut root rot and
pod rot diseases under greenhouse
conditions
This experiment was conducted to study
the effect of aqueous plant extracts of
garlic, neem and mint on the incidence of
root and pod rots caused by
Fusarium
spp. on peanut plants cv. Giza 6, under
greenhouse conditions during 2015 and
2016 growing season. Preparation of
aqueous plant extracts has been done as
mentioned before. Seeds of cv. Giza 6,
were immersed in the highest
concentration (30%) of each extract for
15 minutes. Sterilized pots (30 cm in
diameter) were filled with sterilized sand
clay soil. Infested soil with each tested
pathogen was carried out as previously
mentioned. Four replicates were used for
each treatment. Each replicate contained
four pots (8 seeds/ pot). Untreated seeds
with aqueous plant extracts were used as
control. After 90 days, diseases severity
was recorded as mentioned before. At the
end of the experiment several growth
parameters such as plant fresh and dry
weights, plant height, and number of
pods and weight of pods were estimated.
2.7 Statistical analysis
Comparison of means was performed
using Fisher’s protected least significant
difference (LSD) at p≤0.05 (Gomez &
Gomez, 1984) and the standard error was
calculated using the statistical analysis
software “CoStat 6.4” (CoStat, 2005)
3. Results and Discussion
3.1 Identification of the biocontrol
agents
Bacterial biocontrol agents isolated from
peanut soil rhizosphere were identified
according to their morphological and
biochemical characteristics as: 3 isolates
as
Bacillus
spp., one isolate as
Penibacillus polymyxa
(BP), one isolate
as
Bacillus subtilis
(BS1), one isolate as
Bacillus megaterium
(BM2) and 3
isolates as
Pseudomonas
florescence
(BF1, BF2 and BF3). Fungal biocontrol
agents were also isolated from peanut
soil rhizosphere; eight isolates were
obtained from rhizosphere soil of peanut
plants plus two isolates obtained from
Assiut University Mycological Center
El-Sheikh Aly et al., 2019
84
(AUMC), Egypt. Fungal isolates were
identified as
Trichoderma harzianum
(1,
2, 5, 7
,
8, 9 and 10)
and
Trichoderma
hamatum
(3, 4 and 6) according to their
cultural and microscopical characters as
described by Rifai (1969).
Table 1: Effect of some antagonistic bacteria on the mycelial growth of the tested fungi in vitro.
Mycelial growth inhibition (%)
Code
Bacterial isolates
F. solani (VII)
F. solani (V)
33.6
58.5
BP
7
Bacillus spp.
30
40.7
BS1
7.8
12.3
BM2
18.3
21.2
PF1
Pseudomonas fluorescence
35
41.8
PF2
8.6
14.3
PF3
0
0
Control
2.20
2.01
L.S.D at 5%
3.2 Antagonistic capability of bacterial
bioagents against mycelial growth of
the pathogenic fungi
in vitro
Data in Table (1) and Figure (1) indicated
that the antagonistic isolates of bacteria
were able to inhibit the mycelial growth
of
F. solani
(V),
F. solani
(VII) and
F.
oxysporum
(I) as compared with the
control.
Penibacillus polymyxa
(BP) gave
the greatest reduction of mycelial growth
of
F. solani
(No. V), followed by isolates
Pseudomonas fluorescens
(PF2),
Bacillus
subtilis
(BS1) and
P. fluorescens
(PF1) as
reached 58.5%, 41.8%, 40.7% and 21.2%
respectively.
Bacillus megaterium
(BM2)
showed the lowest mycelial growth
inhibition of
F. solani
(V), followed by
P. fluorescens
(PF3) as recorded 12.3%
and 14.3% respectively.
Pseudomonas
fluorescens
(PF2),
Penibacillus polymyxa
(BP) followed by
B. subtilis
(BS1)
showed the highest degree in suppressing
mycelial growth of
F. solani
(VII), as
reached 35%, 33.6% and 30%
respectively.
P.fluorescens
(PF1) showed
moderate effect of inhibiting the mycelial
growth of
F. solani
(VII) while,
B.
megatirum
(BM2) followed by
P.
fluorescens
(PF3) gave slight effect on
the mycelial growth of the same fungus
as reached 7.8% and 8.6% respectively.
In the same context,
Penibacillus
polymyxa
(BP),
P.fluorescens
(PF1) and
Bacillus subtilis
(BS1) gave the greatest
inhibition of mycelial growth of
F.oxysporium
(I) as recorded 63.8%,
62.2% and 52.3% respectively.
B.megatirum
(BM2) and
P.fluorescens
(PF1) exhibited moderate effect of
inhibition of the same fungus as reached
49.5% and 47.8%, followed by
P.fluorescens
(PF3) as recorded 33%.
These results are similar to those
obtained by El-Mougy
et al.
(2011)
who
examined the influence of the
antagonistic isolates of
B. subtilis
and
P.
fluorescens
and their culture filtrates
against soil-borne root rot pathogens
R.
solani
and
F. solani in vitro
. The tested
antagonists reduced the linear growth of
fungal pathogens. Pandya
et al.
(2009)
evaluated some antagonists (
B. subtilis
,
T. viride
and
T. harzianum
against
mycelial growth of
F. solani in vitro
. All
the antagonists were effective against
F.
solani
and might be very useful as
potential biological control agents.
Also,
El-Sheikh Aly et al., 2019
85
Mahmoud
et al
. (2006) tested seventeen
bacterial isolates
in vitro
for their
antagonistic effect against the pathogens
F. solani
and
M. phaseolina
. The most
effective isolates in reducing the
mycelium growth of pathogenic fungi
were
P. fluorescens
(Pf2) followed by
B.
subtills
(BS1) and
Bacillus
spp. (BP). In
this respect, Abdel-Monaim (2011) tested
certain bio-control agents (
B. subtilis, B.
megaterium, T. viride
and
T. harzianum)
which isolated from chickpea
rhizosphere, against
R. solani, F. solani
and
S. sclerotiorum
.
Bacillus subtilis
,
B.
megaterium
,
T. viride
and
T. harzianum
gave the highest effect against the tested
fungi
in vitro
.
3.3 Effect of
Trichoderma
harzianum
and
T. hamatum
on linear growth of
the pathogenic fungi
3.3.1 Dual culture
Seven isolates of
Trichoderma
harzianum
and three isolates of
T. hamatum
were
tested to study their effect to inhibit
mycelial growth of
F. solani
(V),
F.
solani
(VII) and F.
oxysporum
(I)
in vitro
on Czapek's agar medium. Data in
Figure (2) indicate that all the tested
isolates of both
Trichoderma
species
significantly reduced the mycelial growth
of
F. solani
(V),
F. solani
(VII) and
F.
oxysporum
(I) although exhibited
different degrees of reaction. The
mycelium of
Trichoderma
spp. grew
rapidly over the mycelium of the tested
pathogens and prevented their
development.
Trichoderma harzianum
T2, T8 and T10 were the most effective
in inhibiting the pathogenic fungal
growth, while isolate T9 exhibited the
lowest effect. On the other hand,
T.
hamatum
isolates showed moderate
effects on the pathogenic fungi mycelial
growth.
Trichoderma harzianum
T2, T8
and T10 were selected for further studies.
Such results are in line with those
reported by Ha (2010).
Antagonistic
effect might be due to direct influence of
antagonistic fungi against pathogens
through coiling their hyphae around the
hyphae of the pathogens to prevent their
continued growth (Adekunle et al., 2006)
and/or production of antagonistic
substances which can play an important
role in lyses of cell wall components of
the pathogenic fungi to help the
antagonists to penetrate the host hyphae
and grow on it as a hyper parasite
(Papavizas et al., 1984). This can be
explained in the light of results recorded
by Abd El-Moity (1981),
who stated that
T. harzianum
works through different
mechanisms,
i.e.
production of gliotoxin,
mycoparasitism and growing very fast
and act as barrier between susceptible
plant tissues and virulent pathogens.
Mycoparasitism by
T. harzianum
is a
complex process, involving recognition
of the host, attachment to the mycelium,
coiling round the hyphae, partial
degradation of the cell wall and
penetration of the host mycelium (Seema
and Devaki, 2012).
Scanning electron
microscopic observation of parasitism of
T. harzianum and T. hamatum
on
R.
solani
revealed that the hyphae of
Trichoderma
coil around the host.
T.
harzianum
attached to host mycelium by
forming hooks and
Trichoderma
produces appressoria at the tips of short
branches.
El-Sheikh Aly et al., 2019
86
Figure 1: (A) Effect of Bacillus spp. isolates on mycelial growth of the tested fungi in vitro where
BP=Penibacillus polymexa, BS1=Bacillus substiles and BM2= Bacillus megaterium. (B) Effect of
Pseudomonas fluorescence isolates on mycelial growth of the tested fungi in vitro.
Figure 2: Effect of Trichoderma harzianum (isolates 1, 2, 4, 5, 7, 8, 9 and 10) and T. hamatum (isolates 3, 4 and
6) on inhibiting mycelial growth of the tested pathogenic fungi.
3.3.2 Effect of culture filtrates of
T.
harzianum
isolates on linear growth of
Fusarium
isolates
in
vitro
Isolates of
F. solani
and
F. oxysporum
were used to study their reaction against
toxin produced by
T. harzianum
. Data in
Table (2) that the addition of culture
filtrate of
T. harzianum
isolates T2, T8
and T10 separately to the medium
significantly affected the mycelial growth
of the tested fungi. The reduction of the
mycelial growth of the tested fungi
increased with increasing culture filtrate
concentration. The culture filtrate at the
concentration of 50% gave higher
reduction of the mycelial growth of the
Fusarium
isolates.
Fusarium oxysporum
was significantly reduced for a large
degree with different concentrations of
T
.
harzianum
culture filtrates comparing
with isolates of
F. solani
(V) and
F.
solani
(VII). Generally, the best isolate
of
T. harzianum
which affected the
mycelial growth of the pathogenic fungi
was
T. harzianum
(T10) with all tested
concentrations followed by
T. harzianum
(T2). While, the lowest reduction was
obtained by using
T. harzianum
isolate
(T8). These results are in conformity
El-Sheikh Aly et al., 2019
87
with the results obtained Haran et al
.
(1995).
T. harzianum
is known to
produce relatively high concentrations of
cell-wall degrading enzymes as β-1, 3-
glucanasea and different chitinolytic
enzymes. Several enzymes have been
purified and characterized
in vitro
.
Howell (2003) noted that enzymes such
as chitinases and glucanases produced by
the biocontrol agents are responsible for
suppression of the plant pathogens.
These enzymes function by breaking
down the polysaccharides, chitin, and β-
glucanase that are responsible for the
rigidity of fungal cell walls, thereby
destroying cell wall integrity.
Table 2: Effect of culture filtrates of T. harzianum isolates on mycelial growth of the tested pathogenic fungi.
Isolate No.
Conc.
Reduction of mycelial growth (%)
Mean
F. solani (V)
F. solani (VII)
F.oxysporum (I)
Trichoderma harzianum No. 2
10%
15
17
23
18.3
25%
29
25
40
31.3
50%
33
33
43
36.3
Trichoderma harzianum No. 8
10%
20
17
28
21.6
25%
24
19
31
24.6
50%
34
30
45
36.3
Trichoderma harzianum No. 10
10%
18
19
27
21.3
25%
19
21
32
24
50%
34
34
47
38.3
Control
0
0
0
-
LSD at 5%
Isolate (I)
3.03
3.78
2.19
-
Conc. (C)
1.36
1.84
2.24
-
(I) X (C)
2.73
3.68
4.49
-
3.4 Effect of certain plant extracts on
the mycelial growth of the tested fungi
in vitro
Plant extracts of garlic, neem and mint
were tested to investigate their ability to
inhibit the mycelial growth of
F. solani
(V),
F. solani
(VII) and
F. oxysporum
(I)
in vitro
. Data in Table (3) and Figure (3)
showed that all tested plant extracts
reduced the mycelial growth of
F. solani
(V),
F. solani
(VII) and
F. oxysporum
(I),
with the tested 10, 20 and 30 %
concentrations. The largest percentage of
reduction was obtained with 30%
concentration, while the lower effect of
plant extracts on growth of pathogenic
fungi was obtained with concentration of
10%. Treatment with garlic extract at
concentration of 30% gave the highest
effect on the reduction of the mycelial
growth of
F. solani
(V),
F. solani
(VII)
and
F. oxysporum
(I), isolates which
recorded 90.5, 90.8 and 91.8%
respectively.
Neem extract with the
concentration 30% recorded higher effect
on
F. solani
(V),
F. solani
(VII) and
F.
oxysporum
(I) isolates, where the values
reached it 80.2, 77.8 and 88.9%
respectively. Mint extract at
concentration 30% recorded the lowest
reduction on the mycelial growth of the
pathogenic fungi as values of 60.2, 60.7
and 65.6% respectively. These results are
in line with results of
El-Sharkawy
(2006) and Mukhtar (2007).
The
maximum reduction of the mycelial
growth by plant extracts may be due to
the presence of antifungal compounds in
the extracts (Anusha, 2003).
El-Sheikh Aly et al., 2019
88
Figure 3: Effect of culture filtrates of Trichoderma harzianum isolates on the mycelial growth of a) F.
solani (V), b) F.solani (VII) and c) F.oxysporium (I) in vitro.
Table 3: Effect of different plant extracts on mycelial growth of the tested pathogenic fungi in vitro.
Plant extracts
Conc.
Reduction of pathogenic fungi mycelial growth (%)
Mean
F. solani (V)
F. solani (VII)
F. oxysporum (I)
Garlic
10%
47.3
45.7
50
47.6
20%
56.5
52.8
60
56.4
30%
90.5
90.8
91.8
91
Neem
10%
38
37.5
40.4
38.6
20%
51.5
52
56.2
53.2
30%
80.2
77.8
88.9
82.3
Mint
10%
9.3
8.5
12.5
10.1
20%
19.4
18.5
22.3
20
30%
60.2
60.7
65.6
62.2
Control
0
0
0
-
LSD at 5%
Concentration (A)
3.48
2.87
3.09
-
Treatments (B)
4.05
4.78
4.02
-
Interaction (A × B)
6.96
5.75
6.18
-
Ahmed and Sultan (2000) mentioned that
garlic and onion extracts showed toxic
effects on mycelial growth of the
Rhizoctonia
solani
,
Macrophomina
phaseolina
and
Fusarium
spp.
in vitro
.
Shalin and Dohroo (2003) studied the
activity of 13 plant extracts of broccoli
leaves, eucalyputs, neem, pine, ginger,
turmeric rhizome, chili seeds, tulci seeds,
ocimum sanctum, cotton, sarson caka,
basil and garlic cloves
in
vitro
against
Fusarium
oxysporium
. These extracts
gave different effects on mycelial growth
of
Fusarium
oxysporum
f.sp.
pisi
causing
the wilt of pea plants.
3.5 Estimation of different bio-agents
effect on incidence of root rot disease
of peanut under greenhouse conditions
Data presented in Table (4) showed
that
treated soil with bacterial bio agents
significantly reduced the disease
severity of root rot disease of peanut
compared with the control. The soil
treated with the mixture of bio-bacteria
and
P. fluorescens
(PF2) gave higher
effects in decreasing root rot disease
incidence caused with
F. solani
(V) and
F. solani
(VII) more than the other bio-
agents under greenhouse conditions
during 2015 and 2016 growing seasons.
Penibacillus polymyxa
(BP), followed
by
T. harzianum
(T10) showed
moderate effects in decreasing disease
severity caused with the tested
El-Sheikh Aly et al., 2019
89
pathogenic fungi. Concerning
F.
oxysporum
(I),
Penibacillus polymyxa
(BP) exhibited the highest disease
reduction followed by mixture whereas,
P. fluorescens
(PF2) gave moderate
effect in the term of disease reduction
while, the lowest disease reduction was
obtained by
T. harzianum
(T10). Such
results are in line with those reported by
El-Habbaa et al. (2001; 2002)
who
suggested that addition of
Rhizobium
and
B. subtilis
to soil before sowing
peanut seeds significantly reduced root-
rot disease of peanut caused by
S.
rolfsii
,
R. solani, M. phaseolina
and
F.
solani
. Ibrahim
et al.
(2008) tested the
effects of certain bacterial isolates
against
R. solani, Sclerotium rolfsii
,
F.
solani
and
M. phaseolina
causing root
rot disease of peanut, in greenhouse
experiments. The most effective isolates
in reducing the diseases of peanut were
P. fluorescens
followed by
B. subtills
and
Bacillus
sp.
Kishore and Podile
(2002) also found that different isolates
of
Trichoderma
spp. were identified as
biocontrol agents of groundnut stem rot
and other soil-borne diseases. Vargas et
al.
(2008) mentioned that inoculation
with
Trichoderma
spp. recorded the
lowest incidence of peanut root rot
disease caused by
F. solani
.
3.6 Effect of different bio-agents on
incidence of pod rot disease of peanut
under greenhouse conditions
It was shown from data in Table (5) that
the soil treated with each of tested bio-
agents significantly reduced the disease
severity of pod rot disease of peanut
compared with the control. The infested
soil with mixture of bacteria gave the
highest reduction of pod rot disease of
peanut as compared with the control
and the other bio-agents tested
separately during 2015 and 2016
growing seasons. At the same time,
T.
harzianum
(T10) and
P. fluorescens
(PF2) exhibited moderate effects in
reducing pod rot disease caused with all
tested pathogenic fungi during two
successive seasons.
Penibacillus
polymyxa
(BP) gave the lowest
reduction of pod rot disease severity
during two growing seasons. The
obtained data revealed that
F.
oxysporum
was affected with bioagents
for a large degree compared with
F.
solani
fungus, so the reduction of pod
rot disease was clear during growing
seasons.
Table 4: Effect of different bio-agents on root rot disease of peanut under greenhouse conditions during 2015
and 2016 growing seasons.
Biocontrol agents
Root rot disease severity (%)
F. solani (V)
F. solani (VII)
F. oxysporum (I)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Penibacillus polymyxa (BP)
36
35.1
35.55
35
33.8
34.4
22
20.8
21.4
Pseudomonas fluorescens (PF2)
25
24
24.5
29.2
28
29
30
29.2
29.6
Trichoderma harzianum (T10)
37.5
36.4
36.95
37.5
36.7
37.1
30
38.8
34.4
Mixture
29
27.8
28.4
26
25
25.5
24.4
23
23.7
Control
73
74
73.5
77
78
77.5
65.5
64
64.75
L.S.D at 5%
2.63
2.04
-
2.78
2.52
-
1.78
2.4
-
El-Sheikh Aly et al., 2019
90
Table 5: Effect of different bio-agents on pod rot disease of peanut under greenhouse conditions during 2015
and 2016 growing seasons.
Biocontrol agents
Pod rot disease severity (%)
F. solani (V)
F. solani (VII)
F. oxysporum (I)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Penibacillus polymyxa (BP)
32
31
31.5
32.8
32.1
32.45
27
26.8
26.9
Pseudomonas fluorescens (PF2)
23.5
22.2
22.85
25
23.8
24.4
20
21
20.5
Trichoderma harzianum (T10)
23.5
22
22.75
21
21.8
21.4
20.5
20
20.25
Mixture
16.5
15.3
15.9
17.5
16.8
17.15
16
15
15.5
Control
56
58
57
66
65
65.5
52
54
53
L.S.D at 5%
2.78
2.58
-
1.97
1.89
-
3.26
2.7
-
The results obtained are in agreement
with both Mahmoud (2004)
who found
that
B. subtilis
and
P. fluorescens
significantly reduced incidence of all
types of pod rots caused by
Fusarium
spp. Ahmed (2006) and Ibrahim et al.
(2008) revealed the superiority of
T.
harzianum
followed by commercial
products
Rhizo-N and Plant-guard which
showed high reduction of pod rot disease
of peanut caused by the
R. solani, M.
phaseolina, S. rolfsii
and
F. moniliforme
.
3.7 Effect of bio-agents on growth
parameters of peanut infected with
F.
solani
(V) under greenhouse conditions
Data in Table (6) showed that adding
mixture of biocontrol agents to infested
soil with
F
.
solani
isolate (V) gave the
best result in increasing shoot vigor as
increased both fresh and dry weight (g
/plant) followed by
Trichoderma
harzianum
and
Pseudomonas
fluorescens
. It also gave the best result in
increasing plant height (cm /plant)
followed by
Trichoderma
harzianum
,
Penibacillus
polymyxa
while
Pseudomonas fluorescens
gave the
lowest value. In the same time the
highest number of pods /pot was
achieved by the application of mixture
treatment followed by
Pseudomonas
fluorescens
and
Trichoderma
harzianum
.
Mixture of bioagents achieved the
highest weight of pods /pot followed by
Pseudomonas fluorescens
and
Penibacillus
polymyxa
. Generally,
mixture treatment gave the highest
values of growth parameters of peanut
plants infected with
F
.
solani
(V). Such
results are in line with those reported by
Bolar et al. (2000)
who reported that
peanut plants treated with bioagents as
seed treatment were more health and
produced higher yield compare with
control plants. This might be due to that
bioagent act through different
mechanisms. These mechanisms include
nutrient and growth regulator substances
and some of these antagonists when
sprayed on plant surface, prior real infect
led to stimulate plant resistant and
enforce treated plants to produce some
metabolites which depress pathogens
(Abd-El-Moneim & Maisa, 2011).
Several modes of action of the efficiency
of bioagents on reducing plant diseases
have been described, including
competition for nutrients, antibiosis,
induced resistance, mycoparasitism,
plant growth promotion and rhizosphere
colonization capability.
El-Sheikh Aly et al., 2019
91
Table 6: Effect of bio-agents on growth parameters of peanut infected with Fusarium solani (V) under
greenhouse conditions during 2015 and 2016 growing seasons.
Biocontrol agents
Plants infected with F. solani (V)
Fresh weight (g/plant)
Dry weight (g/plant)
Plant height (cm/plant)
Number of (pods/pot)
Weight of (pods/ pot)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Penibacillus polymyxa
17
19
18
6
6.2
6.1
29
34
31.5
33
31
32
57
55
56
Pseudomonas fluorescens
20.5
22
21.25
6.9
7
6.95
34
38
36
23
26
24.5
45.5
48
46.75
Trichoderma harzianum
16
19
17.5
5.7
6
5.85
33
37
35
23
26
24.5
48
50
49
Mixture
31
33
32
9.5
7.5
8.5
39
42
40.5
35
37
36
65
67
66
Control
9
10
9.5
3
3.3
3.15
23
21
22
10
11
10.5
20.5
22
21.25
L.S.D at 5%
4.49
4.08
-
0.4
0.51
-
4.11
4.58
-
4.72
4.45
-
4.59
5.58
-
3.8 Effect of bio-agents on growth
parameters of peanut infected with
F.
solani
(VII) under greenhouse
conditions
It′s shown from data in Table (7) that
adding bioagents mixture treatment in
infested soil with
F
.
solani
isolate (VII)
gave the highest values of both fresh and
dry weights (g /plant), followed by
Pseudomonas fluorescens
and
Penibacillus polymyxa
. Data also showed
that mixture treatment gave the best
results in plant height (cm/plant)
followed by
Pseudomonas fluorescens
,
Trichoderma harzianum
and
Penibacillus
polymyxa
, respectively. In this respect,
treatment with mixture of bioagents was
the best among all other treatments in
increasing yield represented in both
number and weight of pods followed by
Penibacillus polymyxa
,
Trichoderma
harzianum
and
Pseudomonas fluorescens
respectively. In general, mixture
treatment was the most effective
treatment applied when added in soil
infested with
F. solani
(VII) to enhance
growth parameters of peanut plants as
compared with other treatments and the
control. Zafari et al. (2008) reported that
using beneficial micro-organisms as
biocontrol agents led to enhancement of
plant growth parameters. Such
enhancement may be due to induce plant
resistance
,
produce extracellular enzymes
and antifungal or antibiotics, which
decrease biotic stress on plant, and
produce growth promoter’s substances
(Szczech and Shoda, 2004). In addition,
Egamberdiyeva (2005) hypothesized that
there are several mechanisms by which
rhizosphere bacteria may stimulate plant
growth, such as production of plant
growth substances, nitrogen fixation,
phytohormones, vitamins, solublizing
minerals besides, their role in direct
inhibition of pathogen growth and
suppression of diseases caused by micro-
organisms and increased plant growth
and yield.
Table 7: Effect of bio-agents on growth parameters of peanut infested with F. solani (VII) under greenhouse
conditions during 2015 and 2016 growing seasons.
Biocontrol agents
Plants infected with F. solani (VII)
Fresh weight (g/plant)
Dry weight (g/plant)
Plant height (cm/plant)
Number of (pods/pot)
Weight of (pods/ pot)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Penibacillus polymyxa
18
17
17.5
6.5
6
6.25
28.5
34
31.25
23
27
25
50.5
58
54.25
Pseudomonas fluorescens
21
22
21.5
7
7.3
7.15
29
31
30
28
28
28
63.5
60
61.75
Trichoderma harzianum
20
23
22
7
7.4
7.2
29
34
31.5
26
29
27.5
48.5
46
47.25
Mixture
31.5
30
32.25
8
9
8.5
32
34
32
36
34
35
70
64
67
Control
9
11
10
3.2
3.7
3.45
25
21
23
6
9
7.5
12.5
16
14.25
L.S.D at 5%
3.89
3.67
-
0.68
0.52
-
3.84
4.46
-
3.89
2.72
-
5.22
5.17
-
El-Sheikh Aly et al., 2019
92
3.9 Effect of applying bio-agents on
growth parameters of peanut infested
with
F
.
oxysporum
(I) under
greenhouse conditions
Data in Table (8) exhibited that mixture
treatment gave the best increase fresh and
dry weights (g /plant) followed by
Pseudomonas
fluorescence
,
Trichoderma
harzianum
and
Penibacillus polymyxa
.
Concerning plant height (cm /plant), the
best record was achieved also by
treatment with mixture of bioagents
followed by
Penibacillus polymyxa
,
Pseudomonas
fluorescence
then
Trichoderma
harzianum
. Mixture
treatment gave the highest number of
pods /pot followed by
Trichoderma
harzianum
and
Penibacillus polymyxa
as
compared with control.
Pseudomonas
fluorescence
gave moderate effects. Also
mixture treatment was the best treatment
in increasing weight pods/pot followed
by
Penibacillus polymyxa
,
Trichoderma
harzianum
and
Pseudomonas
fluorescence.
In general treatment plants
with mixture of biocontrol agents gave
the highest values of growth parameters
of peanut plants infected with
F
.
oxysporum
(I). Application of
microorganisms to control diseases,
which is a form of biological control, is
an environment-friendly approach
(Lugtenberg & Kamilova, 2009). In
general, competition for nutrients, niche
exclusion, induced systemic resistance
and antifungal metabolites production are
the chief modes of biocontrol activity in
PGPR
.
Many rhizobacteria have been
reported to produce antifungal
metabolites like, HCN, phenazines,
pyrrolnitrin, 2, diacetylphloroglucinol,
pyoluteorin, viscosinamide and tensin
(Bhattacharyya & Jha, 2012). Interaction
of some rhizobacteria with the plant roots
can result in plant resistance against
some pathogenic bacteria, fungi, and
viruses.
Table 8: Effect of bio-agents on growth parameters of peanut infected with F. oxysporum (I) under greenhouse
conditions during 2015 and 2016 growing seasons.
Biocontrol agents
Plants infected with F. oxysporum (I)
Fresh weight (g/plant)
Dry weight (g/plant)
Plant height (cm/plant)
Number of (pods/pot)
Weight of (pods/ pot)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Penibacillus polymyxa
18
21
19.5
6
7
6.5
31
40
35.5
26
28
27
48
50
49
Pseudomonas fluorescens
23
24.5
23.75
7
7.5
7.25
30
33
31.5
21
21
21
43
48
45.5
Trichoderma harzianum
20
21
20.5
6.8
7.2
7
29
31
30
28
32
30
44
50
47
Mixture
32
35
33.5
9.5
8.5
9
47
40
43.5
38
34
36
66
60
63
Control
10
9
9.5
3.5
3.3
3.4
26
27
26.5
10
10
10
18.5
15
16.75
L.S.D at 5%
5.22
4.62
-
0.51
0.33
-
4.59
4.81
-
4.63
4.08
-
4.99
6.09
-
3.10 Effect of applying plant extracts
on root and pod rot diseases of peanut
in vivo
Peanut seeds cv. Giza 6 were treated with
aqueous solution of three plant extracts,
i.e.
garlic, neem and mint to study their
effects with the highest concentration
30% on root and pod rot diseases
incidence caused with
F. solani
(V),
F.
solani
(VII) and
F. oxysporum
(I)
isolates under greenhouse conditions
during 2015 and 2016 growing seasons.
Data represented in Tables (9 and 10)
revealed that soaking peanut seeds before
sowing in each of tested plant extract
significantly reduced root and pod rots
diseases severity compared with the
control. Garlic extract gave higher
reduction of root and pod rot diseases
El-Sheikh Aly et al., 2019
93
severity followed by neem and mint
extracts. Treated seeds with each plant
extract and sown in infested soil with
F.
oxysporum
(I) showed the highest disease
severity of root rot disease followed by
F.
solani
(VII) then
F. solani
(V). On the
contrary, seeds treated with each plant
extract and sown in infested soil with
F.
solani
(VII) showed the highest disease
severity percentage of pod rot disease
followed by
F. solani
(V), while
F.
oxysporum
(I) gave the lowest disease
severity of pod rot disease. Such results
are in agreement with those reported by
Syed
et al.
(2012). They reported that
plant extracts have important roles in
biologically based management
strategies for controlling plant diseases.
Hassan (2006)
studied the effects of five
plant extracts (camphor, garlic cloves,
basil,
Lantana camara
and neem) against
F. solani
and
F. oxysporum
caused root
rot disease of pea plants. All plant
extracts at any concentration reduced the
mycelial growth of the tested pathogenic
fungi. Under greenhouse conditions,
garlic extract gave the highest effect of
reducing root rot disease on pea plants.
Table 9: Effect of plant extracts on root rot disease of peanut under greenhouse conditions
during 2015 and 2016 growing seasons.
Plant extracts
Root rot disease severity (%)
Mean
F. solani (V)
F. solani (VII)
F. oxysporum (I)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Garlic
37.5
35
36.25
43
41
42
44
43
43.5
40.58
Neem
41
40.5
40.75
44.15
42.5
43.3
45
44
44.5
42.85
Mint
37
40
38.5
41.6
40
40.5
53
52
52.5
43.9
Control
73
75
74
77
79
78
65.5
65
65.25
71.42
L.S.D at 5%
3.76
2.42
-
4.03
2.80
-
4.7
2.94
-
-
Table 10: Effect of plant extracts on pod rot disease of peanut under greenhouse conditions
during 2015 and 2016 growing seasons.
Plant extracts
Pod rot disease severity (%)
Mean
F. solani (V)
F. solani (VII)
F. oxysporum (I)
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Garlic
29
28
28.5
29.5
28.7
29.1
24
23.4
23.7
27.1
Neem
32.5
32.3
32.4
34.5
34.5
34.5
31
32
31.5
32.8
Mint
36
35
35.5
36.5
36.2
36.35
29
29.5
29.25
33.7
Control
56
58
57
66
68
67
52
54
53
59
L.S.D at 5%
2.48
5.15
-
6.05
5.79
-
9.08
5.68
-
-
3.11 Effect of different plant extracts
on growth parameters of peanut plants
This study was carried out using plant
extracts on fresh weight of shoots, dry
weight, plant height, number of pods and
weight of pods. Data in Table (11)
revealed that fresh and dry weights of
plant shoot increased to varying degrees
when sprayed with plant extracts. Garlic
extract gave higher increases of fresh and
dry weights compared with neem and
mint extract
s
as well as control plants,
which infected with the same pathogens.
Also, mint extract gave moderate effect
of fresh and dry weight, while neem
extract came in the last during 2015 and
2016 growing seasons. Concerning the
effect of plant extracts on plant height,
data exhibited that the used concentration
of plant extracts was more effective,
wherever the height of peanut plants
El-Sheikh Aly et al., 2019
94
obviously increased compared with
control plants and infected with the tested
fungi during two growing seasons. The
same trend was obtained by adding garlic
extract followed by neem extract, while
mint extract gave the lowest plant height.
Table 11: Effect of plant extracts on growth parameters of peanut plants under greenhouse conditions during 2015
and 2016 growing seasons.
Plant extracts
Pathogenic fungi
Fresh weight g/plant
Dry weight g/ plant
Plant height cm /plant
Number of pods / pot
Weight of pods/pot
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
2015
2016
Mean
Garlic
F. solani (V)
17
21
19
6
6
6
28
34
31
28
32
30
47
52
49.5
F. solani (VII)
17
23
20
6.3
6.5
6.4
26
30
28
20
25
22.5
41
50
45.5
F. oxysporum (I)
17
23
20
6.3
7
6.65
35
40
37.5
18
22
20
35
43
39
Neem
F. solani (V)
16
14
15
5.8
6
5.9
30
31
30.5
21
23
22
46
46
46
F. solani (VII)
14
15
14.5
4.8
5
4.9
26
27
26.5
18
22
20
43.5
45
44.25
F. oxysporum (I)
19
19
19
6.5
7
6.75
36
37
36.5
18
21
19.5
36
40
38
Mint
F. solani (V)
14
16
15
4.5
5
4.75
29
30
29.5
25
23
24
51
46
48.5
F. solani (VII)
17
19
18
6
6
6
31
31
31
25
22
23.5
46
44
45
F. oxysporum (I)
17
17
17
6
6.2
6.1
36
35
35.5
15
17
16
31
33
32
Control
F. solani (V)
9
9
9
3.1
3
3.05
23
22
22.5
10
11
10.5
20.5
22
21.25
F. solani (VII)
9
8
8.5
3.4
3
3.2
25
22
23.5
6
9
7.5
12.5
18
15.25
F. oxysporum (I)
10
9
9.5
3.5
3
3.25
26
23
24.5
10
9
9.5
18.5
18
18.25
LSD at 5%
Plant extract (A)
1.44
2.66
-
0.28
0.32
-
3.31
2.48
-
2.14
2.83
-
1.90
3.52
-
Pathogen (B)
1.47
1.75
-
0.25
0.23
-
1.79
1.80
-
1.69
2.06
-
2.64
2.92
-
Interaction (A×B)
2.94
3.49
-
0.49
0.46
-
3.57
3.6
-
3.39
4.13
-
5.27
5.84
-
On the other hand, data indicated that
number of pods /pot increased when
peanut plants received plant extracts. In
this respect, garlic extract gave a positive
effect on number of pods, followed by
mint extract, while neem extract
exhibited lower effect in increasing pods/
pot.
So, the plant extracts of garlic
followed by neem when added on peanut
plants positive by affected weight of pods
/plant more than mint extract and control
plant. Generally, plant extracts exhibited
the higher increase of fresh, dry weight of
peanut plants, plant height number of
pods and weight of pods when compared
with growth parameters of control.
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