Journal of Pharmacognosy and Phytochemistry 2019; 8(2): 1333-1337

E-ISSN: 2278-4136
P-ISSN: 2349-8234
JPP 2019; 8(2): 1333-1337 In vitro and in vivo field efficacy of different
Received: 04-01-2019
Accepted: 06-02-2019 fungicides against Alternaria brassicae (Berk.)
HT Valvi
sacc. causing Alternaria leaf spot of cauliflower
Department of Plant Pathology,
College of Agriculture, Dr.
BSKKV, Dapoli, Maharashtra, HT Valvi, AD Saykar and VR Bangar
India
Abstract
AD Saykar
The Alternaria leaf spot caused by A. brassicae (Berk.) Sacc is one of the destructive disease of
Department of Plant Pathology,
College of Agriculture, Dr.
Cauliflower in Konkan region of Maharashtra. In the present study, bio efficacy of seven fungicides
BSKKV, Dapoli, Maharashtra, tested under in vitro conditions against the pathogen. Among that, Mancozeb 75% WP (0.25 %) was
India completely inhibited the growth of the test fungus. It was followed by Propiconazole 25% EC (0.1%)
which showed 96.29 per cent inhibition of the test fungus and was at par with Mancozeb. No sporulation
VR Bangar was observed in Mancozeb 75% WP, Propiconazole 25% EC and Difenconazole 25% EC. Among the
Department of Plant Pathology, different fungicides tested under field conditions, all the fungicidal treatments were significantly superior
College of Agriculture, Dr. over control after last spray. The treatment Mancozeb 75% WP was the most effective as it recorded
BSKKV, Dapoli, Maharashtra, minimum disease incidence (28 %). It was followed by Azoxystrobin 23% EC which recorded 29 %
India disease incidence.

Keywords: Cauliflower, Alternaria brassicae (Berk.) Sacc. Fungicides, Management, Sporulation and
Mycelial growth

Introduction
Several factors are responsible for low production of cauliflower crop, among which diseases
also play an important role. The important diseases of Cauliflower crop are leaf spot, Downy
mildew, Damping off, Club root, Powdery mildew, White rust, Black rot, Bacterial soft rot and
Cauliflower mosaic. Among these diseases Alternaria leaf spot is a serious disease of
cauliflower. About 20 to 80 per cent loss in yield and 59 per cent loss in seed may occur due to
this disease. The disease appears as minute specks on the leaves, which enlarge over a time
and result in substantial lesions with concentric rings where spores are produced. The disease
starts from lower leaves and slowly progresses towards the upper shoots, leaves, petioles, pods
and heads. Defoliation of the outer leaves may occur on severely infected plants and extensive
trimming may be required to remove infected leaves from the cabbage head at harvest. In
susceptible varieties, apart from yield, significant reduction in quality may occur.
During recent years the fungal diseases were found to occur in severe proportions under
favorable environmental conditions particularly during Rabi season. Alternaria leaf spot
caused by Alternaria brassicae has been reported to inflict heavy yield losses (Pattanamahakul
and Strange, 1999; Azevedo et al., 2000; Peruch et al., 2006; Chauhan et al., 2009; Mishra et
al., 2009; Deep and Sharma, 2012 and Sharma et al., 2013) [14, 1, 15, 2, 11, 4, 17].
Alternaria leaf spot is difficult to control because the fungus penetrates in infected seed, plant
debris in soil and collateral hosts such as other cruciferous crops in the vicinity of the main
crop or weeds. One of the most effective and old method for disease control is the use of
fungicides. There are several fungicides which are being commercially available while several
others are being evaluated in different laboratories. Even though cauliflower is not a regularly
cultivated crop in Konkan region, some progressive vegetable growers are switching to this
crop due to its high marketability. Considering importance of the crop and disease, present
study on Alternaria leaf spot of cauliflower was planned and conducted to evolve in vitro and
in vivo management strategy.

Materials and Methods

In vitro evaluation of fungicides
Correspondence
The laboratory experiments in the present study were conducted in the Department of Plant
VR Bangar
Department of Plant Pathology, Pathology, Dr. DBSKKV, Dapoli during 2015-16. The principle involved in poisoned food
College of Agriculture, Dr. technique is to poison the nutrient medium with a fungi-toxicant and then allowing the test
BSKKV, Dapoli, Maharashtra, fungus to grow on it and finally recording the extent of growth (Nene and Thapliyal, 1997) [13].
India The details of treatments used are mentioned below.
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Potato dextrose agar medium (PDA) was used as basal Transplanting of seedlings
medium and dispensed in 100 ml aliquots in each 250 ml Initially light irrigation was given to the experimental plot in
Erlenmeyer conical flasks, which were sterilized at 1.054 the morning so as to achieve optimum moisture in the soil to
kg/cm2 pressure for 20 minutes. The quantity of every facilitate establishment of seedling during transplanting. On
fungicide for each concentration was calculated for 100 ml the same day evening, 25 days old healthy seedling were
medium separately. The weighed quantity of each fungicide carefully uprooted and immediately transplanted in the field.
was added in lukewarm PDA at 40 ± 2°C, mixed thoroughly Light irrigation was given to the plot immediately after
and poured into sterilized Petri plates and allowed to solidify. transplanting for better crop stand.
The mycelial discs of 5 mm diameter were cut from 7 day old
culture of the test fungus with the help of a sterilized cork Schedule of spraying of Fungicide:
borer. A single disc was transferred aseptically to the centre The crop was observed carefully for initiation of the disease.
of each plate already poured with poisoned medium. The First spray of each fungicide treatment was given immediately
plates with PDA without fungicide but inoculated with fungal after disease incidence was noticed. Two more sprays of
culture, served as control. fungicides were given at an interval of 15 days.

List of fungicides tested under in vitro and in vivo against Method of recording observations:
the pathogen. Five plants per treatment per replication were randomly
selected for recording disease incidence of Alternaria leaf
Name of the fungicide and Conc. (%) spot. Initial observations were recorded before first spray and
T. No. Trade name
formulation tested final observations were recorded 15 days after the last spray
T1 Azoxystrobin 23% EC Amistar 0.1 on the basis of development of spots and lesions by
T2 Carbendazim 50% WP Fungiguard 0.1 considering the whole leaf area as hundred per cent.
T3 Copper hydroxide 77% WP Kocide 101 0.1
T4 Mancozeb 75% WP Dithane M-45 0.25 Observations on disease incidence:
T5 Difenconazole 25% EC Score 0.1
The disease intensity was recorded in 0-5 scale (Conn. et al.
T6 Thiophanate methyl 70% WP Roko 0.1
1990) as described below.
T7 Propiconazole 25% EC Tilt 0.1
T8 Control - -
Score/grade Description
0 No disease
The plates were incubated at room temperature (27 ± 2°C).
1 <5% leaf area affected
Three replications of each treatment were maintained. The 2 6-10% leaf area affected
observation on colony diameter of the fungus were recorded 3 11-25% leaf area affected
when Petri plate in control treatment was fully covered with 4 26-50% leaf area affected
mycelial growth. Per cent inhibition of growth of the test 5 >50% leaf area affected
pathogen was calculated.
Per cent inhibition of growth of the test fungus was calculated Per cent disease intensity (PDI):
by following formula (Horsfall, 1956) [7]. Per cent disease intensity was calculated by the following
formula given by (McKinney, (1923)
Y–Z
X = x 100
Y

Where, X = Per cent inhibition, Y = Growth of fungus in

control (mm), Z = Growth of fungus in treatment (mm)
Per cent disease control (PDC)
Field evaluation of fungicides The per cent disease control was calculated by using the
The field experiment was conducted on variety “Super-fast formula given below:
crop” of cauliflower during Rabi 2015-16 at the Botany farm,
College of Agriculture, Dapoli.

Raising of cauliflower seedlings

The seeds of cauliflower were sown on in portrays containing
cocopeat as growing medium. The trays were staked one Statistical analysis
above the other and the uppermost tray was covered with a The data obtained were statistically analysed by the methods
clean plastic sheet for 4 days. Seeds germinated within 4 days suggested by Gomez and Gomez (1986) [6]. The standard error
and then each tray was arranged separately on raised beds in and critical difference were worked out and the results
shed net. The trays were watered regularly to maintain obtained were compared statistically.
sufficient moisture. The Cauliflower seedlings were ready for
transplanting in 25 days. Results and Discussion
In vitro evaluation of fungicides
Preparation of experimental plot Seven fungicides belonging to different groups were tested for
The experimental plot was prepared with a tractor drawn their efficacy against A. brassicae by employing poisoned
mould board plough by two crisscross ploughings with tractor food technique. The data obtained on the effect of different
operated cultivator. Planking was done to level the field. The fungicides on the vegetative growth and sporulations of A.
experiment was laid in randomized block design. Three brassicae in vitro are presented in Table 1, PLATE-I and
replications were maintained per treatment. depicted in Fig-1.

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Journal of Pharmacognosy and Phytochemistry

The data presented in Table 1 revealed that, among the Azoxystrobin 23% EC (0.1 %) showed 52.96 per cent and
different fungicides tested under in vitro Mancozeb 75% WP 49.25 per cent inhibition of A. brassicae, respectively over
(0.25 %) completely inhibited the growth of the test fungus. It control and were at par with each other. In the treatment of
was followed by Propiconazole 25% EC (0.1%) which Thiophanate methyl 70% WP (0.1%), 38.88 per cent
showed 96.29 per cent inhibition of the test fungus and was at inhibition of the fungus was achieved. Carbendazim 50% WP
par with Mancozeb. The next fungicide in order of merit was (0.1) was appeared as least effective fungicide which showed
Difenconazole 25% EC (0.1 %) which recorded 87.77 per only 27.77 % inhibition of the test pathogen.
cent inhibition. Copper hydroxide 77% WP (0.1 %) and

Table 1: In vitro efficacy of fungicides on growth and sporulation of A. brassicae (Berk.) Sacc.
T. No. Name of the fungicide Conc. (%) Mean colony diameter (mm) Per cent inhibition over control Sporulation
T1 Copper hydroxide 77% WP 0.1 42.33 52.96 +
T2 Azoxystrobin 23% EC 0.1 45.66 49.25 +
T3 Difenconazole 25% EC 0.1 11.00 87.77 -
T4 Propiconazole 25% EC 0.1 3.33 96.29 -
T5 Mancozeb 75% WP 0.25 0.00 100 -
T6 Thiophanate methyl 70% WP 0.1 55 38.88 ++
T7 Carbendazim 50% WP 0.1 86.66 3.70 +++
T8 Control 90.00 - ++++
S.Em.± 0.91
C.D.at 1% 3.85
Sporulation
- = No sporulation, +++ = Good,
+ = Poor, ++++ = Excellent.
++ = Fair,

No sporulation was observed in Mancozeb 75% WP (0.25%), brassicae and A. brassicicola. Hossain and Mian (2004)
Propiconazole 25% EC (0.1%) and Difenconazole 25% EC reported that Mancozeb, Carbendazim and Propiconazole
(0.1%). Poor sporulation was recorded in Copper hydroxide inhibited the mycelial growth of the A. brassicicola infecting
77% WP (0.1%) and Azoxystrobin 23% EC (0.1%). Fair cabbage. Surviliene et al., (2006) reported that Azoxystrobin
sporulation was observed in Thiophanate methyl 70% WP showed significant inhibitory effect on mycelial growth of the
(0.1%). Good sporulation was observed in Carbendazim 50% Alternaria spp. Mishra et al., (2009) [11] reported that
WP (0.1%) as compared to control which showed excellent Propiconazole was the most effective against A. brassicae
sporulation. The result findings are in agreement with those which showed maximum per cent inhibition of mycelial
reported by Kumar et al., (1999) [9] who reported that growth and was followed by Mancozeb and Azoxystrobin.
Carbendazim 0.05 and 0.1 per cent completely inhibited Gaikwad (2013) also reported that Propiconazole,
mycelial growth of A. brassicae. Beatrice et al., (2004) Difenconazole and Mancozeb at different concentrations
showed that difenconazole was most effective against A. completely inhibited the mycelial growth of A. brassicae.

Plate 1: Efficacy of fungicides on growth and sporulation of Alternaria brassicae

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Journal of Pharmacognosy and Phytochemistry

Fig 1: Efficacy of fungicides on growth and sporulation of A. brassicae (Berk.) Sacc.

Management of the disease in field conditions against the disease under field conditions are presented in
The results of the experiment on evaluation of the fungicides Table 2 and depicted in Fig-2.

Table 2: Per cent disease incidence (PDI) and per cent disease control (PDC) against A. brassicae (Berk.) Sacc.
T. No. Treatments Conc. (%) Mean PDI before first spray Mean PDI after last spray PDC
T1 Copper hydroxide 77% WP 0.1 19.60 (26.25)* 36.83 (37.36) 42.45
T2 Azoxystrobin 23% EC 0.1 19.67 (26.32) 29.00 (32.58) 54.69
T3 Difenconazole 25% EC 0.1 20.17 (26.61) 33.50 (35.35) 47.66
T4 Propiconazole 25% EC 0.1 19.83 (26.41) 31.67 (34.24) 50.52
T5 Mancozeb 75% WP 0.25 19.75 (26.38) 28.00 (31.93) 56.25
T6 Thiophanate methyl 70% WP 0.1 19.83 (26.45) 36.00 (36.86) 43.75
T7 Carbendazim 50% WP 0.1 20.00 (26.56) 34.00 (35.67) 46.88
T8 Control 20.15 (26.67) 64.00 (53.13)
S. Em.± 0.54 0.63
C.D.at 5% 1.64 1.91
(*Figures in parentheses indicate arc sin values)

It is apparent from the data presented in Table 2 that, among which recorded 50.52 per cent and 47.66 per cent disease
the different fungicides tested under field conditions all the control, respectively.
fungicidal treatments were significantly superior over control These were followed by treatment T7 (Carbendazim 50% WP)
after last spray. The treatment T5 (Mancozeb 75% WP) was with 46.88 per cent disease control and was at per with T4.
the most effective as it recorded minimum disease incidence Treatment T6 (Thiophanate methyl 70% WP) and T1(Copper
(28 %). It was followed by T2 (Azoxystrobin 23% EC) which hydroxide 77% WP) showed 43.75 and 42.45 per cent disease
recorded 29.00 per cent disease incidence. The treatment T 4 control over control treatment, respectively and were
(Propiconazole 25% EC) was at par with T 3 (Difenconazole statistically at par with each other. Sreedhar et al., (2003) [18],
25% EC). Treatment T7 (Carbendazim 50% WP) showed 34 showed that Azoxystrobin was the most effective fungicides
% disease incidence and it was followed by T 6 (Thiophanate as it reduced disease incidence to the tune of 88 to 93 per
methyl 70% WP) and T1 (Copper hydroxide 77% WP) with cent. Narain et al., (2006) [12] reported that Mancozeb was the
36 and 36.63 per cent disease incidence respectively over most effective fungicide against A. brassicae. Prasad and
control (64 %). Maximum per cent disease control was Lallu (2006) reported that first spray of Carbendazim (0.1%)
achieved in treatment T5 (Mancozeb 75% WP) and T2 + Mancozeb (0.2%) followed by two sprays of Mancozeb
(Azoxystrobin 23% EC) which showed 56.25 per cent and (0.2%) at early date of sowing (October) was the best
54.69 per cent disease control over control, respectively. Both combination in reducing the disease severity on leaves.
the treatments were found statistically at par with each other. Gaikwad (2013) showed that Mancozeb showed the least
The next fungicides in merit of order were treatment T 4 diseases incidence (15.30%) followed by Propiconazole
(Propiconazole 25% EC) & T3 (Difenconazole 25% EC) (18.36%) and Copper oxychloride (20.65%).

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Fig 2: Management of the disease in the field by using different fungicides.

Conclusion 10. Mackinney. A new system of grading plant disease. J

On the basis of the results of present study it can be concluded Agric. Res. 1923; 26:195-218.
that Alternaria leaf spot of cauliflower caused by Alternaria 11. Mishra PK, Rai RK, Saha S, Pandey KK, Singh RP, Rai
brassicae (Berk.) Sacc. is an important disease of cauliflower AB. Effect of fungicides, botanicals and bioagent leaf
in Konkan region. The fungicides viz., Mancozeb (0.25%) and spot of cauliflower caused by Alternaria brassicae
Azoxystrobin (0.1%) are very effective against the pathogen (Berk.) Sacc. Vegetable Science. 2009; 36(3):55-59.
under field conditions. Three sprays of Mancozeb (0.25%) or 12. Narain U, Chand G, Pandey R. Efficacy of fungicides
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