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Evaluation of Microbial Antagonists for the Management of Wilt/Root Rot and

Damping-off Diseases in Chilli (Capsicum annuum)

Article in International Journal of Plant Research · December 2015

DOI: 10.5958/2229-4473.2015.00091.9

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 VEGETOS Vol. 28 (4) : 102-110 (2015)

10.5958/2229-4473.2015.00091.9

Evaluation of Microbial Antagonists for the Management of Wilt/

Root Rot and Damping-off Diseases in Chilli (Capsicum annuum)

Gh. Hassan Dar*, G. Hassan Mir, Humaira Rashid, Waseem Ali Dar and Misba Majeed

Received: 05.08.2015 / Revised: 16.11.2015 / Accepted: 09.12.2015 / Published online: 31.12.2015

This article is published in open access at www.vegetosindia.org

Abstract most important vegetable crop grown in allover

The present study was aimed to assess world consumed both unripe (green) and ripe
wilt and root rot/damping off prevalence in chilli (red) form as an indispensable condiment, be-
(Capsicum annuum) in Kashmir and to examine sides pungency due to the presence of capsaicin
the potential of various bio-agents in manage- and vitamins such as vitamin A and C, the green
ment of these diseases. Survey was carried out in fruit is also known to contain vitamin P (rutin) in
three districts of Kashmir viz., Pulwama, Budgam appreciable proportion (Muthukrishnan et al.
and Srinagar during the year 2013. The damping 2002). India is the second largest exporter of
off and wilt/root rot incidence in chilli growing chilli in international market producing 1.446
areas varied from 7 to 36 and 10 to 54%, respec- million tones of ripe dry chillies with an area of
tively with highest disease incidence in district 0.869 million ha under its cultivation
Pulwama and lowest in district Srinagar. The (Muthukumar et al. 2010, Anonymous 2013). In
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pathogens responsible for wilt/root rot were Jammu and Kashmir State, the chilli is grown on
Rhizotonia solani, Sclerotium rolfsii, Fusarium an area of 3280 ha with annual production of
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solani, F. oxysporium, F. pallidoroseum and Phy- 6480 metric tones (Anonymous, 2010) which is
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tophthora capsici. Pythium sp. was observed to quite low when compared with the other states
be the incitant of damping off. Fifteen isolates of of India. Amongst the major constraints in chilli
Trichoderma species and 2 isolates of Pseudo- production in the state, the losses caused by
monas fluorescens and Bacilus sp. each were various diseases wilt/root rot and damping off,
isolated locally and evaluated against above caused by a complex of pathogens viz., Fusarium
pathogens (except P. capsici) in vitro. Trichoder- sp., Phytophthora capsici, Sclerotium rolfsii, Rhi-
ma viride isolate TK1, T. harzianum isolate TK8, zoctonia solani and Pythium sp., are devastating
Pseudomonas fluorescens isolate Ps1 and Bacil- affecting the crop right from seed sowing to ma-
lus sp. isolate B1 were most effective in inhibit- turity (Najar 2001, Sahi and Khalid 2007, Tayyaba
ing the mycelia growth of pathogens. In vivo Sultana et al. 2014), which has forced many culti-
wilt/ root rot disease management through use vators to abandon its cultivation. The several
of effective bioagents revealed that all the four Fusarium species viz., F. oxysporum and F. solani
biocontrol agents, used individually or in combi- have been reported as major pathogens causing
nation, significantly improved seed germination wilt disease in solanaceous crops like chilli, to-
(62.7-76.3%) as against control (57.7%). The fun- mato, brinjal in Gujrat and Uttar Pradesh
gicide treatment of 2-(methoxycarbamoylamino) (Yelmame et al. 2010, Dwivedi and Enespa 2013),
-benzimidazole (carbendazim 50 WP) had less S. rolfsii wilt reported from Rajasthan chilli grow-
damping-off incidence of 2% as against 6.0 to ing areas (Kalmesh and Gurjar 2001), Phy-
13.7% in bioagent treatments and 18.7% in con- tophthora capsici are associated with pepper
trol. Similarly, wilt/root rot was significantly re- wilting (Sanogo and Carpenter 2006) in South-
duced in bioagent or chemical treatments than western United States.
control. Disease control is achieved through the
Keywords: Disease management, microbial an- use of fungicides which are hazardous and lead
tagonists, wilt complex, damping-off, Capsicum to severe environmental pollution. Management
annuum of diseases through biological components is
one of the effective sustainable approaches. Var-
Introduction ious antagonistics Trichoderma viride and T. har-
Chilli (Capsicum annuum L.) is one of the zianum have proved promising as a biocontrol

Division of Plant Pathology, S.K. University of Agricultural Sciences & Technology of Kashmir, Shalimar,
Srinagar 190 025, Jammu & Kashmir (India)
*Corresponding author E-mail: ghasangar@gmail.com

102
 Evaluation of Microbial Antagonists for the Management of Wilt/Root Rot

agent of various chilli diseases like wilt/root/ hyphae/ mycelial colour and sclerotium, etc.
collar rot and damping off (Hanson and Howell were observed in Petri plates. Spore shape, size
2004, Bajwa et al. 2004, Patel et al. 2014). Varsh- and colour were also noted. The identity of
ney and Chaube (1999) found Pseudomonas flu- pathogens was established by comparison with
orescens as a good biocontrol agent against authentic descriptions (Parmeter and Whitney
several chilli pathogens including Rhizoctonia 1970, Nelson et al. 1981, Booth and Sutton 1984,
solani. The combination, T. harzianum + P. fluo- Krownland and Staglini 1988). The frequency of
rescens, was most effective in reducing disease the occurrence of each isolate was also worked
incidence (66.7% more efficient than the control) out.
(Rini and Sulochana 2006). The P. fluorescens For pathogenicity, apparently healthy
and Trichoderma spp. are biocontrol agents chilli seeds cv. Kashmir Long - 1 were sown in
which possess plant growth promoting ability pot-trays and allowed to germinate under
coupled with antagonistic effect in phytopatho- greenhouse conditions. The healthy germinated
gens (Kumar et al. 2012, Dewangan et al. 2014). seedlings were then transplanted to the earthen
In view of above, the present investigation was pots (1 cm dia.) containing artificially infected
aimed to assess the prevalence of wilt/root rot soil with specific pathogen (sick soil). To prepare
disease of chilli in Kashmir valley and manage a sick soil, spore suspensions of pathogens were
the disease through use of locally isolated bio- prepared a fresh from young axenic cultures and
control agents. inoculum load of 1000-2000 propagules/ml
used to infect soil (Sindhan et al. 1999). In check
Materials and Methods only sterile water was applied. The plants were
Survey for root rot/wilt disease status in chilli regularly monitored for the development of typ-
A pilot survey was conducted during ical disease symptoms and Koch’s postulates
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2013 in randomly selected eight blocks, com- followed. The established pathogens were main-
mercially important chilli growing belts, in three tained on PDA medium at 4°C till further use.
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districts of Kashmir valley viz., Pulwama, Budgam Isolation, characterization and identification
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and Srinagar to assess the incidence of damping of antagonists

-off and wilt/root rot in chilli nurseries and field Rhizospheric soils from major vegetable
crops. A total number of ninety six villages in growing areas dominantly growing Solanaceous
these blocks were surveyed during May (for crops were collected and rhizospheric fungi and
nurseries) and August (for field crop). The plant bacteria isolated using dilution plate technique.
showing damping-off and wilt/root rot symp- The fungal and bacterial pure cultures were
toms in nursery/field were taken to laboratory raised on PDA and King’s B medium, respectively
for pathogen isolation and microscopic studies. (Elad et al. 1981). The antagonists were identi-
The disease incidence in nursery/fields was as- fied on the basis of their morpho-cultural char-
sessed as per the method of Johnston and Both acteristics such as growth rate, colour and colo-
(1983) using the formulae: ny appearance, mycelial and spore characters
No. of infected plants such as shape, size, etc. and compared with au-
Disease incidence (%) = ________________________________________ x 100 thentic descriptions (Rifai 1969, Bissett 1991,
Total number of units assessed
Gams and Bissett 1998, Samuels 2006). For bac-
Isolation, characterization and identification terial identification morphological features such
of pathogens as colony type, bacterial shape, size and growth
Chilli plants exhibiting typical symptoms characteristics of the isolates on King’s B medi-
of green wilting, browning of vascular tissue, um (King et al. 1954) were assessed. Biochemical
rotted roots were collected during survey. The characteristics including catalase test, fluores-
diseased collar and root portions of chilli were cent pigment production and siderophore de-
first surface sterilized, cut into small bit of 5 mm tection were determined as per the standard
size and placed on potato dextrose agar (PDA) methods (Pickett et al. 1991, Goszczynska et al.
medium plates. These plates were incubated at 2000, Aneja 2001).
28±1°C for 7 days and monitored daily for the Both fungal and bacterial isolates were
growth of fungi. The fungi were isolated by sin- screened for their biocontrol potential, if any,
gle spore technique and pathogenicity estab- against chilli root rot/wilt/damping-off patho-
lished by following the Koch’s postulates. Mor- gens. For this, the antagonist and pathogen cul-
phological characteristics of cultures were stud- ture discs (5 mm size) were grown on 90 mm
ied by microscopic observation under com- pertiplates containing PDA and King’s B media
pound and stereoscopic microscopes at different about 50 mm apart (opposite to each other). The
magnifications and growth characteristics like plates were incubated at 28±1°C and regularly
colony colour, texture, growth, pigmentations, monitored. The isolates were cultured with path-

103
 Hassan Dar Gh. et al.

Table 1. Disease incidence of wilt/root rot complex in chilli in vegetable areas districts of Kashmir
Vil- Wilt/Root rot Damping off
Dis- Blocks lages Pathogens observed incidence (%) incidence (%)
tricts sur-
veye Rang Mean Rang Mean
d e e
(No.)
Kakapora 17 Fusarium pallidoroseum, F. oxysporum, F.
solani, Pythium sp. Phytophthora capsici. 14-54 40 9-35 22
Sclerotium rolfsii
Pul- Pulwama 20 Fusarium pallidoroseum, Rhizoctonia
wama solani, F. oxysporum, F. solani, Pythium 15-37 24 11-31 19
sp.,
Phytophthora capsici
Pampore 12 Fusarium pallidoroseum, F. oxysporum,
Rhizoctonia solani, Pythium sp. 11-38 22 5-27 14
Overall mean 49 - 11-54 28.6 5-35 18.3
Kanipora 16 F. oxysporum, F. solani, Pythium sp., 17-48 33 10-33 21
Phytophthora capsici, Sclerotium rolfsii
Bud- Chadoora 11 Fusarium pallidoroseum, F. oxysporum, F. 14-35 22 11-28 16
gam solani, Pythium sp.
Charai-Sharief 8 Fusarium pallidoroseum, F. oxysporum, F. 10-37 26 7-21 10
solani, Rhizoctonia solani,Pythium sp.
Overall mean 35 - 10-48 27.0 7-33 15.6
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Pantha- Chowk 6 Fusarium pallidoroseum, F. oxysporum, F. 18-46 28 10-36 14

Srina- solani, Pythium sp., Phytophthora capsici
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gar Sonawar 6 F. oxysporum, F. solani, Pythium sp., 17-31 20 8-30 12

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Rhizoctonia solani
Overall mean 12 - 17-46 24.0 8-36 13.0

ogens to observe the interaction between them. of 1 cm dia. at the bottom were filled with steri-
The mycelial growth inhibition was estimated as lized amended soils. Inoculum disc (5 mm) of
per the method of Morton and Stroube (1955). each pathogen were slotted with cork borer and
Also, the antagonists/pathogens interac- transferred to sterile soil in plastic containers.
tions were examined under stereo-microscope For incubation, these plastic containers were
for hyper-parasitism, if any. The effective bio- transferred to the plastic trays containing sterile
control agents were mass multiplied on wheat water at the bottom (1 cm height) and covered
bran, boiled for 10 minutes before wheat the with plastic sheet so as to create humidity for 4
rupture of seed, filtered through muslin cloth days. The control without pathogenic inoculums
and then transferred to 250 ml conical flasks for was also maintained.
sterilization in autoclave at 15 psi pressure for In vitro screening of antagonist against path-
20 minutes. After proper sterilization the fungal ogenic fungi
antagonists were inoculated on wheat bran and Fifteen isolates of Trichoderma spp.
incubated at 27+2oC (Simon and Anamika 2011). namely TK1 to TK15 and two isolates each of
The bacterial anatgonists were mass multiplied Pseudomonas fluorescens and Bacillus sp., were
in molasses broth medium (molasses: water, 1:5 isolated from the soils of different vegetable
ratio) and transferred to 250 ml conical flasks for fields during the survey and maintained on
sterilization. After proper sterilization the antag- modified potato dextrose agar (PDA) (for fungi)
onists were inoculated and incubated for 24 and King’s B medium (for bacteria) (Mukherjee
hours (Younis et al. 2010) 1991). The isolates were studied for antagonistic
Preparation of sick soils potential against major fungal pathogens of
The surface soils were collected from chilli individually on media by using dual culture
Shalimar (Kashmir) and thoroughly mixed and technique (Coskuntuna and Özer 2008). Discs (5
sieved. The sorghum flour was added @ 25 g/kg mm) of fifteen isolates of Trichoderma (TK1 to
soil to boost the organic matter content in soil TK15) and two isolates each of P. fluorescens
so that it may support the growth of pathogens. (Ps1 & Ps2) and B. subtilis (B1 & B2) and patho-
The amended soil was sterilized at 15 Psi for 2½ gens were allowed to grow on PDA
hours consequently for two days. The plastic ( Trichoderma sps.), KB (Pseudomonas
containers of 2 kg capacity with drainage holes fluorescens and Bacillus subitlis) in petriplates

104
 Evaluation of Microbial Antagonists for the Management of Wilt/Root Rot

Table 2. Impact of various locally isolated bioagents on mycelial growth inhibition (%) of chilli wilt/root
rot pathogens
Root rot/wilt pathogens
Sclerotium Rhi-
Fungal/bacterial species rolfsii zoctonia Pythium Fusarium palli- Fusarium Fusarium
solani sp. doroseum oxysporum solani

Trichoderma viride TK1 76.4 (+) 73.2 (+) 61.2 (+) 79.1 (+) 74.2 (+) 78.2 (+)
T. harzianum TK2 36.2 (-) 23.2 (-) 12.1 (-) 34.6 (-) 40.1 (-) 37.2 (-)
T. viride TK3 45.7 (-) 54.1 (-) 35.3 (-) 52.6 (+) 62.5 (+) 67.2 (+)
T. viride TK4 42.3 (-) 38.7 (-) 31.9 (-) 54.2 (+) 45.7 (-) 58.2 (+)
T. harzianum TK5 21.3 (-) 15.3 (-) 25.1 (-) 23.8 (-) 31.5 (-) 34.6 (-)
T. viride TK6 33.0 (-) 23.2 (-) 29.3 (-) 12.1 (-) 15.3 (-) 23.2 (-)
T. harzianum TK7 37.2 (-) 33.3 (-) 21.3 (-) 15.3 (-) 12.1 (-) 21.3 (-)
T. harzianum TK8 71.4 (+) 67.5 (+) 62.3 (+) 74.6 (+) 70.1 (+) 75.3 (+)
T. viride TK9 61.0 (+) 65.3 (+) 53.2 (+) 67.3 (+) 45.3 (-) 70.1 (+)
T. viride TK10 57.2 (+) 54.8 (+) 45.8 (-) 61.2 (+) 69.7 (+) 35.8 (-)
T. viride TK11 54.8 (+) 45.1 (-) 39.6 (-) 54.8 (-) 76.4 (+) 54.1 (+)
T. harzianum TK12 65.3 (+) 42.3 (-) 59.8 (+) 53.2 (-) 60.4 (+) 45.7 (-)
T. harzianum TK13 23.2 (-) 38.2 (-) 32.7 (-) 34.6 (-) 33.3 (-) 25.1 (-)
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T. harzianum TK14 31.2 (-) 21.3 (-) 31.5 (-) 36.2 (-) 37.2 (-) 15.2 (-)
T. viride TK15 62.2 (+) 59.8 (+) 54.1 (+) 62.1 (+) 70.1 (+) 56.4 (+)
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65.0 (+)
Pseudomonas fluorescens Ps1 59.8 (+) 61.0 (+) 65.2 (+) 71.4 (+) 69.8 (+)
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Ps. fluorescens Ps2 29.0 (-) 32.0 (-) 21.3 (-) 15.2 (-) 33.3 (-) 42.3 (-)
Bacillus sp.B1 62.3 (+) 54.8 (+) 54.8 (+) 59.8 (+) 68.0 (+) 65.0 (+)
Bacillus sp.B2 23.3 (-) 31.5 (-) 34.6 (-) 12.1 (-) 34.6 (-) (-)
*The (+) indicates the presence of zone of inhibition while (-) indicates the absence of zone of inhibition

and incubated for 7 days at 28±1°C to study ing. The seed sowing was done in the first week
their mycelial growth inhibition. The per cent of April and all package of practices followed,
inhibition of mycelial growth of test fungus and except disease control measures. The damping
bacteria was assessed as per Vincent (1947). The off incidence was estimated 20, 25 and 30 days
disease incidence in nursery/fields was assessed after seed sowing.
as per the method of Johnston and Both (1983). The field experiments were conducted at
Evaluation of biocontrol agents against path- the main campus Shalimar in the year 2013. The
ogens soil at the site was silty-clay loam. The field was
Pot experiments were conducted to eval- well prepared and plots of 2 m x 2 m size inocu-
uate the biocontrol potential of isolated rhizo- lated with most effective fungal and bacterial
spheric fungi and bacteria, singly and in combi- biocontrol agents, individually or in combination
nation. Chilli seeds cv. “Kashmir Long-1”, pro- with fungal antagonist. There were 12 treat-
cured from the Division of Vegetable Sciences, ments including standard checks (carbendazim
SKUAST-Kashmir, were sown in 7.5 litre capacity 50 WP and untreated control). The experiment
containers containing artificially infected soil. was conducted in a randomized block design
Four effective antagonists were selected for the with each treatment replicated three times. The
purpose of evaluation. Also, a fungicide namely chilli cv. ‘Kashmir Long-1’ was transplanted in 1st
carbendazim 50 WP [chemical name: 2- week of June and standard recommended prac-
(methoxycarbamoylamino)- benzimidazole] and tice was followed as per SKUAST-Kashmir, ex-
untreated control treatments were maintained cept plant protection measures. The disease inci-
for comparison. The experiment comprised of 12 dence in nursery/fields was recorded on 30th,
treatments of various locally isolated biocontrol 60th and 90th day after seedling transplantation
agents, arranged in a completely randomized as per the method of Johnston and Both (1983)
design with each treatment replicated four No. of infected plants
times. The observations were recorded for seed Disease incidence (%) = ———————–——— x 100
Total No. of units assessed
germination 12, 15 and 18 days after seed sow-

105
 Hassan Dar Gh. et al.

Table 3. Effect of bioagent inoculation, singly or in combination, on seed germination and damping-off in chilli
Seed germination (%) Damping off (%)
Treatments* 12 DAS 15 DAS 18DAS 20 DAS 25DAS 30 DAS
b b
Trichoderma viride TK1 63.00 69.67 75.67b 2.67 d
4.00 g
6.00e
Trichoderma harzianum TK8 61.33c 64.67c 70.33c 3.67d 5.67f 9.00d
f d
Pseudomonas fluorescens Ps1 56.67 60.33 66.00de 4.00 cd
6.33 ef
9.33d
Bacillus sp. B1 54.67g 58.33g 62.67g 5.33b 8.00d 11.00c
T. viride + T. harzianum 64.33b 69.33b 76.33b 1.33e 2.33h 4.67e
T. viride + P. fluorescens 59.67de 61.67e 65.33ef 5.67b 9.67b 15.00b
cd d
T. viride + Bacillus sp. 60.33 63.33 66.67d 5.00 bc
7.33 de
11.33c
T. harzianum + P. fluorescens 56.33f 58.67g 64.33f 5.67b 9.33bc 12.33c
T. harzianum + Bacillus sp. 60.00cde 63.33d 69.33c 3.67d 5.67f 8.67d
P. fluorescens + Bacillus sp. 58.67e 62.00e 65.67de 6.33ab 8.33cd 13.67bc
a a
Carbendazim 50WP @ 2 g/kg seed 70.33 78.33 85.67a 0.00 f
0.33 i
2.00f
Untreated control 51.67h 54.33h 57.67h 7.33a 13.33a 18.67a
Over all mean 55.26 58.85 63.60 3.98 6.27 9.46
LSD0.05 1.50 1.12 1.17 1.07 1.27 1.41
Values within the same column having a common letter(s) do not differ significantly (P≤ 0.05)
*Soil application of individual bioagents @ 2 g/kg soil [2x108 cfu] or in combination @ 1 g each /kg soil prior to the seed sowing.
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Moreover, the chilli seeds prior to sowing were treated with bioagents @ 4 g/kg seed in individual bioagent treatments and @ 2
g each bioagent/kg seed in dual bioagent treatments; DAS = Days after seed sowing ; **The values superscripted with the same
letter in the same column do not statistically differ from each other as per DMRT.
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All the data was analysis for analysis of North West Pacific to the use of untreated seeds
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variance as per the standard statistical proce- while Najar and Ahmad (2012) attributed it to
dures (Gomez and Gomez 1984). Statistical com- monoculture practice. Frequent irrigation and
puter package MSTAT was used for computing excessive use of nitrogenous fertilizers are re-
the data and mean compared using Duncan’s ported to be amongst the important predispos-
multiple range test. ing factors that give fillip to the perpetuation of
pathogen in Uttarakhand and Jammu & Kashmir
Results and Discussion states (Mamta et al. 2012, Najar and Ahmad
The damping-off disease prevailed in 2012).
the surveyed districts of Kashmir and its inci- The wilt/root rot disease caused wither-
dence varied from 7 to 36% with mean incidence ing and chlorosis, initially in the lower older
of 15.6%, while wilt/root rot incidence varied leaves and then affecting upper young leaves
from 10 to 54% with mean wilt/root rot inci- and other parts. During severe cases the whole
dence of 26.5% (Table 1). The highest damping- plant died within a few days. The symptoms ob-
off incidence was observed in Kakapora block served are in agreement with those of Tariq et
(Pulwama) followed by Kanipora block al. (2012). During the survey of chilli growing
(Budgam). The lowest damping-off incidence areas in nurseries/fields of Kashmir valley, six
was recorded in Charari Sharief block. Wilt/root fungi were found associated with root/rot/wilt
rot incidence was highest in Pulwama block fol- complex namely Rhizoctonia solani, Sclerotium
lowed by Kanipora block and least wilt/root rot rolfsii, Fusarium pallidoroseum, F. oxysporum, F.
incidence was noticed in Sonawar block. District- solani, Pythium sp. and Phytophthora capsici.
wise maximum damping-off and root rot/wilt The isolation frequencies of pathogens revealed
incidence was observed in Pulwama followed by that F. oxysporum, F. pallidoroseum and Pythium
Budgam and Srinagar. The lack of availability of sp. were dominant pathogens with isolation fre-
treated seeds, excessive irrigation, non-judicious quencies of 27, 22 and 18%, respectively. These
use of fertilizers and improper crop manage- were followed by Rhizoctonia solani, F. solani,
ment practices appeared as the main reasons for Sclerotium rolfsii and Phytophthora capsici with
more prevalence of disease in Pulwama. The isolation frequencies of 12, 11, 6 and 4%, respec-
practice of monoculture followed in the vegeta- tively. El-Mohamady et al. (2014) have reported
ble growing areas may also be one of the rea- Fusarium species to be dominant pathogen fol-
sons for high damping-off and root rot/wilt in lowed by Rhizoctonia solani and Sclerotium
Kashmir valley. McGee (1992) has attributed rolfsii responsible for damping off and root rot
higher wilt/root rot prevalence in vegetables in in various solanaceous crops including tomato.

106
 Evaluation of Microbial Antagonists for the Management of Wilt/Root Rot

Table 4. Field evaluation of antagonists against chilli wilt/root rot complex

Wilt/root rot incidence (%) days after seedling transplan-

Treatments* tation
30 60 90
Trichoderma viride TK1 7.33ef 10.00ef 22.00efg
Trichoderma harzianum TK8 3.67g 9.67ef 15.00hi
c bc
Pseudomonas fluorescens Ps1 14.00 20.33 27.67ce
d b
Bacillus sp. B1 10.33 23.33 33.67b
g f
T. viride + T. harzianum 4.33 7.34 11.34i
de de
T. viride + P. fluorescens 8.67 14.00 23.34ef
de d
T. viride + Bacillus sp. 9.67 15.34 22.00efg
fg ef
T. harzianum + P. fluorescens 5.67 10.34 16.34ghi
T. harzianum + Bacillus sp. 8.00def 16.33cd 19.34fgh
b b
P. fluorescens + Bacillus sp. 17.67 24.00 31.87bc
i g
Carbendazim 50 WP @ 2 g/kg seed 0.00 0.00 2.67j
a a
Untreated control 25.67 54.33 79.00a
Overall mean 9.06 16.13 23.06
LSD0.05 2.78 4.79 5.61
Values within the same column having a common letter(s) do not differ significantly (P≤ 0.05)
*Soil application of individual bioagents @ 2 g/kg soil [2x108 cfu] and dual bioagents @ 1 g each/kg soil prior to the seed sowing. Also, the seedlings
prior to transplantation were dipped in bioagents slurry containing 4 g bioagent/L water for individual application and @ 2 g each bioagent / L water
in dual application. **The values superscripted with the same letter in the same column do not statistically differ from each other as per DMRT.
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The observation recorded on the inhibition of acetaldehyde (Bunkar and Kusum, 2001). Other
pathogens in dual culture revealed that the an- reports reveal that Pseudomonas fluorescens,
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tagonists tested inhibited the growth of patho- Bacillus sp. and Trichoderma sp. reportedly
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gens. Trichoderma viride isolate TK1 and T. har- suppress Fusarium species (Akhtar et al. 2010).
zianum isolate TK8 proved most effective All the antagonists tested individually or
against all the pathogens tested, followed by T. in combination showed significant beneficial
viride isolates TK9 and TK12, Pseudomonas impact on seed germination when applied as
fluorescens isolate Ps1 and Bacillus sp isolate B1 both seed and soil treatment (Table 3). The seed
[Table 2]. Trichoderma isolates TK2, TK5, TK6, germination generally increased with time. On
TK7, TK13 and TK14 and bacterial isolates Ps2 18th DAS, the seed germination in individual an-
and B2 showed lesser antagonistic effect on tagonists tested ranged from 62.7 to 75.7%
Pythium sp., S. rolfsii, F. pallidoroseum, F. while in dual antagonist treatments the treat-
oxysporum and R. solani. Trichoderma isolates ment of T. viride + T. harzianum resulted in
TK1, TK8, TK9, TK10, TK11, TK12 and TK15 highest seed germination (76.3%) as compared
showed mycoparasitic activity by growing over to untreated control (57.6%). The use of car-
some of the pathogens viz., R. solani, F. bendazim 50 WP proved most effective with
pallidoroseum, F. oxysporum, F. solani, S. rolfsii 85.7% seed germination which is in agreement
and P. capsici. Our findings are in agreement with Tariq et al. (2012).
with Ngo et al. (2006), Amin et al. (2010) and The damping-off disease incidence was
Jeyaseelan et al. (2012) who reported that the significantly reduced when antagonists or chem-
application of Trichoderma sp. effectively ical was used. The highest disease incidence of
controls a large number of soil born fungi 18.7% was observed in control and least inci-
including S. rolfsii, R. solani, Pythium sp., F. dence of 2% in carbendazim treatment. In bio-
pallidoroseum, F. oxysporum and F. solani in agent treatments the damping-off ranged from
vegetables. Hanson and Howell (2004) and 6 to 13.7% with maximum reduction due to the
Bajwa et al. (2004) have reported that some combined application of T. viride + T. harzianum
Trichoderma species and strains show good (4.67%). Gomathi et al. (2011) have also ob-
mycoparasitic activity so can effectively be used served bioagents to be effective in minimizing
as biocontrol agents against several phyto- the disease incidence in brinjal and other sola-
pathogens under wide range of adverse naceous crops. Khabbaz and Abbasi (2014) have
environmental conditions. Trichoderma sp. reported that indigenous microbes from agricul-
inhibits pathogenic invasion through the tural soils can be developed and formulated as
phenomenon of parasitism, antibiotics, biofungicides for minimizing the early crop loss-
competition, lysis of pathogenic hyphae, es caused by seedling damping-off and root rot
production of organic metabolites like enodin, diseases in vegetable crops.
chitinase and volatile inhibitory substance like Seedling and soil treatment with differ-
107
 Hassan Dar Gh. et al.

ent bio-agents, individually or in combination, plant pathogens in vitro. J Phytol 2: 34-37.

revealed significant decrease in wilt/rot root as
against untreated control on 30, 60 and 90th day Aneja KR (2001). Experiments in microbiology
after seedling transplantation (Table 4). Howev- plant pathology tissue culture and mushroom
er, carbendazim 50 WP treatment proved most production technology (3rd ed.). New Age Inter-
effective in combating the wilt/root rot problem national Publishers, New Delhi, India.
which is in line with the observations of Sitara
and Hasan (2011). Bioagents treatments, individ- Anonymous (2010). Report from Government of
ually or in combination, showed 11.3 to 33.7% Jammu & Kashmir. Directorate of Agriculture,
wilt/root rot incidence as against 79.0% in un- Srinagar, Jammu & Kashmir, India.
treated control on 90th days after seedling trans- Anonymous (2013). Production and area under
plantation. Amongst the bioagents tested the
chillies and peppers dry in India for the year
combined application of T. viride + T. harzianum 2011. Food and Agriculture Organization (FAO),
proved significantly superior over other bio-
Rome, Italy.
agent treatments. The findings are supported by
Mujeebur and Shahana (2002) and Wani (2005) Bajwa R, Mukhtar I and Anjum T (2004). In vitro
who found combined application of biogents biological control of Fusarium solani - Cause of
effective in arresting the root rot/wilt disease in wilt in Dalbergia sissoo Roxb. Mycopath 2: 11-
chilli and other solanaceous crops. Biocontrol 14.
agents exhibit both antagonistic as well as plant
growth promoting activity, therefore could be Bissett J (1991). A revision of the genus Tricho-
more effective in combating plant diseases derma. II. Infrageneric classification. Canadian J
(Akkopru and Demix 2005, Borrero et al. 2006, Bot 69: 2357-2372.
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Shabir-U-Rehman et al. 2013). It has been

opined that antagonistic fungi can induce sys- Booth C and Sutton BC (1984). Fusarium palli-
Members Copy, Not for Commercial Sale

temic resistance in plants thus boost plant de- doroseum the correct name for F. semitectum.
www.IndianJournals.com

fense response to diverse pathogen attacks Trans Br Mycol Soc 83:702-704

(Harman et al. 2004).
Conclusions Borrero C, Ordovas J, Trillas MI and Aviles M
The present study revealed that damping (2006). Tomato Fusarium wilt suppressiveness:
off and wilt/root rot are most predominant dis- The relationship between the organic plant
eases in Kashmir valley. These diseases can be growth media and their microbial communities
effectively managed through use of fungal an- as characterized by biology. Soil Biol Biochem
tagonists viz., Trichoderma harzianum and T. 38: 1631-1637.
viride. The biocontrol agents have good poten-
Bunkar RN and Kusum M (2001). Integration of
tial to reduce the crop losses due to these soil
biocontrol agents and fungicides for suppres-
borne diseases.
sion of dry root rot of Capsicum fructescens. J
Acknowledgements
Mycol Pl Pathol 31: 330-334.
The authors are thankful to the Universi-
ty Grants Commission, New Delhi for providing Coskuntuna A and Özer N (2008). Biological
financial assistance under a Major Research Pro- control of onion basal rot disease using Tricho-
ject. derma harzianum and induction of antifungal
compounds in onion set following seed treat-
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