D M Pawar

K U Solanky

R B Rajput

S R Suradkar

V P Prajapati

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Save Nature to Survive
8(2): 627-629, 2013 (Supplement on Medicinal plants)
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627
EFFECT OF FUNGAL AND BACTERIAL BIOAGENTS AGAINST
ALTERNARIA ALTERNATA (FR.) KEISSLER IN VITRO CONDITION
R. B. RAJPUT*, K. U. SOLANKY, V. P. PRAJAPATI, D. M. PAWAR AND S. R. SURADKAR
Department of Plant Pathology, N. M. College of Agriculture,
Navsari Agricultural University, Navsari - 396 450, Gujarat, INDIA
e-mail: vir_311985@yahoo.com
INTRODUCTION
Brinjal or egg plant (Solanum melongena L.) belongs to the
kingdom: Plantae, division: Magnoliophyta, class:
Magnoliopsida, order: Solanales, family: Solanaceae. Egg plant
is the second major vegetable crop next to potato in India. It is
considered as a “King of vegetables”.Total production of brinjal
is about 32 million tonnes in the world wherein India is world’s
second largest producer after China. It contains about 1.4 g
proteins, 4.0g carbohydrates, 0.3g fat, 18 mg calcium, 2.0mg
potassium and 0.9mg iron per 100 g of edible portion. It also
provides vitamins like A, B and C. (Choudhary and Gaur,
2009). In India, the crop is grown about 5, 66, 000 hectares
and produces 95, 96, 000 metric tons of fruits (Anon, 2008).
West Bengal, Orissa, Bihar, Karnataka Maharashtra and Gujarat
are the major brinjal growing states of the country. Brinjal
crop is attacked by number of fungal, bacterial, viral and
phytoplasmal diseases, during various growth stages which
reduce its yield and quality of fruits. Diseases like Alternaria
Leaf spot (Alternaria alternata (Fr.) Keissler), Damping off
(Pythium aphanidermatum (Eds.) fitz.; Phytophthora spp.;
Rhizoctonia spp.), Phomopsis blight (Phomopsis vexans Sacc.
and Syd.) Harter., Cercospora Leaf spot (Cercospora solani-
melongenae Chupp., C.solani.), Verticillium wilt (Verticillium
dahliae Kleb), fusarium wilt (Fusarium solani (Mart.) App and
Wollenw, Bacterial wilt (Ralstonia solanacearum Smith), Little
leaf (Phytoplasma.), Mosaic virus, Root-knot nematodes
(Meloidogyne javanica (Treub) Chitwood).Among all the fun-
gal diseases, Alternaria leaf spot, Alternaria leaf blight and
fruit rot diseases are of regular occurrence in moderate to
severe proportion in India and causes extensive damage to
the quality of fruits (Pandey and Vishwakarma 1999). Infected
seeds caused reduction in seed germination and yield loss up
to 30-50 Per cent. (Thippeswamy et al., 2005). Alternaria leaf
spot caused by Alternaria alternata, Alternaria solani and Al-
ternaria melongenae, heavily damage the plants. Cool and
humid weather, coupled with cloudiness which favors the
occurrence and spread of the disease. When humid condi-
tions prevailing at ground level, lower leaves are first attacked
and infection spreads to the upper leaves and fruits. The dis-
ease causes characteristics leaf spot with concentric rings.
The spots are mostly irregular and collapse to cover a large
leaf area. Considering the seriousness of the problem, the
present investigation was carried out. The hazardous effects
of chemicals used in plant disease management have diverted
plant pathologists to find out the alternative techniques of
plant disease control which may cause little or no adverse
effect on environment. Notable success of disease manage-
ment through the use of antagonistic bioagents in the labora-
tory, glass house and field has been achieved during past
several years. On the basis of this information, there is possi-
bility of development of biological control for plant diseases.
Now a day, the commercial formulation of some of the
biocontrol agents has already become available in the mar-
ket. In the present study, attempts have been made to identify
antagonistic bioagents against Alternaria alternata in vitro
condition.
MATERIALS AND METHODS
Eight known fungal and bacterial bioagents (antagonists) viz.,
ABSTRACT
Investigation on leaf spot disease (Alternaria alternata (FR.) Keissler) of Brinjal (Solanum melongena L.) under
south Gujarat condition was carried out to find out suitable management strategies. Due to hazardous effect of
chemical fungicides, search for safer alternative to control the pathogen is better choice. This led to trials on the
use of bioagents to control the pathogen. The eight known bioagents were evaluated by dual culture, pathogen at
periphery and pathogen at the centre technique to monitor antagonistic effect. The results revealed that out of all
the eight bioagents used, three bioagents viz., Trichoderma viride (IARI isolate) (74.77%, 69.04% and 79.45%),
Trichoderma viride (Navsari isolate) (74.14%, 66.08%, and 76.99%) maximum growth inhibition in dual
culture, pathogen at periphery and pathogen at the centre methods respectively), T. harzianum (Junagadh isolate)
(71.25%, 59.96% and 74.78%) maximum growth inhibition in dual culture, pathogen at periphery and pathogen
at the centre methods respectively, showed strong antagonistic effect to inhibit the mycelia growth of the
pathogen significantly.
KEYWORDS
Biological control
Brinjal
Alternaria alternata
Fungal and Bacterial
bioagents
Received on :
14.01.2013
Accepted on :
27.04.2013
*Corresponding
author
628
Trichoderma viride Pers Ex. Grey. (IARI isolate), Trichoderma
viride (Navsari isolate), T. harzianum Rifai (Junagadh isolate),
Bacillus subtilis Ell. (Navsari isolate), Pseudomonas fluorescens
Migula. (Navsari isolate), Aspergillus niger Link. (IARI isolate),
Gliocladium virens Miller. (Junagadh isolate) and Chaetomium
globosum Kunze. (Navsari isolate) were tested in vitro against
Alternaria alternata. The culture discs measuring 5mm of test
organism and pathogen were cut aseptically from the colony
of pure culture grown on PDA medium and kept at different
positions according to different techniques employed in the
present investigation. In dual culture technique (Dennis and
Webster, 1971), culture discs of test organisms and the
pathogen were placed opposite to each other at 4cm apart in
the Petri plate containing 20mL PDA aseptically and real
antagonistic properties of the test bioagents were exhibited .In
Pathogen at the periphery technique (Asalmol and Awasthi,
1990), the culture disc of the pathogen placed aseptically
4cm away radially at four corners keeping one disc of test
organism at centre in the plate containing 20mL PDA
aseptically. In Pathogen at the centre the culture disc of the
pathogen was placed in the center and four similar discs of
the test organisms were placed 4cm away from the pathogen
at the periphery in the Petri plate containing 20mL PDA
aseptically. The culture discs of the pathogens were kept at
respective places of pathogen in each technique without
bioagent served as control. All the treatments were incubated
at room temperature (27 ± 2ºC) and after 6 days the radial
growth of the test organism and pathogen was measured. CRD
design with three repetitions of each treatment was employed
in the present experiment. The per cent growth inhibition (PGI)
was calculated by using formula given as below:
PGI =
Where,
PGI = Per cent growth inhibition
DC = Average diameter of mycelial colony of control set (mm)
DT = Average diameter of mycelial colony of treated set (mm)
RESULTS
All the antagonists under test were significantly superior over
control in all the techniques against A. alternata however in
Dual culture technique, Out of eight antagonists tested, T.
viride (IARI isolate) (74.77%) and T. viride (Navsari isolate)
(74.14%) showed maximum growth inhibition of the pathogen
and appeared to be the most superior over all the antagonists
tested. Next best in order of merit was T. harzianum (Junagadh
isolate) (71.25%) followed by B. subtillis (Navsari isolate)
(69.73%), G. virens (Junagadh isolate) (67.23%), A. niger (IARI
isolate) (64.55%) and rest of the antagonists showed
comparatively least growth inhibition (Table 1). In Pathogen
at the periphery techniques, T. viride (IARI isolate) gave
maximum growth inhibition (69.04%) and appeared to be the
most superior antagonists against A. alternata over all the
antagonists tested. Next best in order of merit was T. viride
(Navsari isolate) (66.08%) which was statistically at par with T.
harzianum (Junagadh isolate) (59.96%) and Bacillus subtilis
(Navsari isolate) (58.32%) followed by G. virens (Junagadh
isolate) (57.38%), A. niger (IARI isolate) (55.55%) while rest of
the antagonists showed comparatively least growth inhibition
(Table 2). In Pathogen at the center technique, maximum
inhibition was found in T. viride (IARI isolate) (79.45 %). Which
was statistically at par with T. viride (Navsari isolate) (76.99 %)
next best in order of merit was T. harzianum (Junagadh isolate
(74.78%) and B. subtillis (Navsari isolate) (73.14%). The rest
of the antagonists showed comparatively least growth
inhibition (Table 3).
DISCUSSION
It appears from the results that all the antagonists tested by
Table 1: Antagonistic effect of different microorganisms against
Alternaria alternata under dual culture method
Sr. Test organism Average colony Per cent
No. diameter of growth
pathogen (mm) inhibition
1 Trichoderma viride 13.50 74.77
(IARI isolate )
2 Trichoderma viride 13.83 74.14
(Navsari  isolate )
3 Trichoderma harzianum 15.38 71.25
(Junagadh isolate)
4 Bacillus subtilis 16.19 69.73
(Navsari isolate)
5 Gliocladium virens 17.53 67.23
(Junagadh isolate)
6 Aspergillus niger 18.96 64.55
(IARI isolate )
7 Pseudomonas fluorescens 23.16 56.70
(Navsari isolate)
8 Chaetomium globosum 25.16 52.96
 (Navsari isolate)
9 Control 53.50
S.Em ± 0.56
C.D. at 5% 1.68
C.V. % 4.49
Table 2: Antagonistic effect of different microorganisms against
Alternaria alternata under pathogen at periphery method
Sr. Test organism Average colony Per cent
No. diameter of growth
pathogen (mm) inhibition
1 Trichoderma viride 16.36 69.04
(IARI isolate)
2 Trichoderma viride 17.93 66.08
(Navsari  isolate)
3 Trichoderma harzianum 21.16 59.96
(Junagadh isolate)
4 Bacillus subtilis 22.03 58.32
(Navsari isolate)
5 Gliocladium virens 22.53 57.38
(Junagadh isolate)
6 Aspergillus niger 23.50 55.55
(IARI isolate)
7 Pseudomonas fluorescens 26.16 50.50
(Navsari isolate)
8 Chaetomium globosum 27.46 48.05
(Navsari isolate)
9 Control 52.86
S.Em ± 0.61
C.D. at 5% 1.83
C.V. % 4.19
 100 (DC-DT)
 DC
R. B. RAJPUT et al.,
629
EFFECT OF FUNGAL AND BACTERIAL BIOAGENTS
three different methods were effective against A.alternata and
useful as potential biological control agents. Among them, T.
viride (IARI isolate), T. viride (Navsari isolate) and harzianum
(Junagadh isolate) proved to be effective antagonist against A.
alternata. This may be due to undeniably its mode of action
like competition, antibiosis and mycoparasitim and it possess
some important secondary metabolites and antibiotics like
viridin, harzianiol and so many. These are in harmony with
earlier workers viz., Ghosh et al. (2002) and Akbari and
Parakhia (2007), who observed strong antagonism of T. viride
on A. alternata. Patel (1991) and Sempere et al. (2007)
observed strong antagonism on A. alternata with T. harzianum
and T viride. Among the bacterial antagonist Akbari and
Table 3: Antagonistic effect of different microorganisms against
Alternaria alternata under pathogen at centre method
Sr.No. Test organism Average colony Per cent
(Antagonists) diameter of growth
pathogen (mm) inhibition
1 Trichoderma viride 10.86 79.45
(IARI isolate )
2 Trichoderma viride 12.16 76.99
(Navsari  isolate)
3 Trichoderma harzianum 13.33 74.78
(Junagadh isolate)
4 Bacillus subtilis 14.20 73.14
(Navsari isolate)
5 Pseudomonas fluorescens 14.36 72.82
(Navsari isolate)
6 Aspergillus niger 16.36 69.04
(IARI isolate)
7 Gliocladium virens 18.23 65.51
(Junagadh isolate)
8 Chaetomium globosum 21.56 59.21
(Navsari isolate)
9 Control 52.86 —
S.Em ± 0.57
C.D. at 5% 1.70
C.V. % 5.15
Parakhia (2007) observed good antagonism of Bacillus subtilis
with A. alternata. The present findings in complete agreement
with the findings of above workers. Hence it can be
recommended after rigorous testing in the pot and field
condition against the pathogen for management of Brinjal leaf
spot disease.
REFERENCES
Akbari, L. F. and Parakhia, A. M. 2007. Management of Alternaria
alternata causing blight of sesame with fungicides. J. Mycol. Pl. Pathol.
37(3): 426-430.
Anonymous 2008. Indian Horticultural Data Base 2008. pp. 5-157.
Asalmol, M. N. and Awasthi, J. 1990. Role of temperature and pH in
antagonism of Aspergillus niger and Trichoderma viride against
Fusarium solani Proc. All India Phytopathol. Soc., (West Zone).
M.P.A.U., Pune. pp. 11-13.
Choudhry, B. and Gaur, K. 2009. The development and regulations
of Bt Brinjal in India. Isaaa Brief 38. pp: 1-2.
Dennis, C. and Webster, J. 1971. Antagonistic properties of species
groups of Trichoderma III hyphal interaction. Trans. Br. Mycol. Soc.
57: 363-369.
Ghosh, C., Pawar, N. B., Kshirsagar, C. R. and Jadhav, A. C. 2002.
Studies on management of leaf spot caused by Alternaria alternata on
gerbera J. Maharashtra Agric. Univ. 27(2): 165-167.
Pandey, K. K. and Vishwakarma, S. N. 1999. Morphological and
symptomatological variations in Alternaria alternata causing leaf blight
in brinjal. J. Mycol. Pl. Pathol. 29(3): 350-354.
Patel, M. J. 1991. Studies on leaf blight of onion caused by Alternaria
alternata (Fr.) Keissler. M.Sc. (Agri.) thesis submitted to Gujarat
Agricultural University, Sardar Krushinagar.
Sempere, F. and Santamarina, M. P. 2007. In vitro biocontrol analysis
of Alternaria alternata (Fr.) Keissler under different environment
conditions. Mycopathologia. 163(3): 183-190.
Thippeswamy, B., Krishnappa, M., Chakravarthy, C. N. 2005.
Location and Transmission of Phomopsis vexans and Alternaria solani
in Brinjal, Indian phytopath. 58(4): 410-413.
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Effect of fungal and bacterial bioagents against Alternariaalternata (fr.) Keissler in vitro condition.

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Effect of fungal and bacterial bioagents against Alternariaalternata (fr.) Keissler in vitro condition.

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