The Allahabad Farmer Vol. LXX, July - 2014 No.

Effect of bio-agent and chemical against the pigeonpea

wilt pathogen Fusarium udum Butler in vitro
Kunwar Zeeshan Khan and Shashi Tiwarit

Received 30-07-2012
Accepted 20-12-2013

ABSTRACT
Pigeonpea wilt caused by Fusarium udum Butler is the most important disease
and causes upto 100% yield loss in the susceptible cultivars. Effect of two
antagonists, Trichoderma viride and Trichoderma harzianum and one botanical
neem cake powder were compared with fungicide Carbendazim against Fusarium
udum in vitro. Trichoderma viride was best in inhibiting the growth of pathogen
by 72.40 percent and neem cake least affected among the bio-agents with 58.52
percent inhibition. Fungicide Carbendazim, completely inhibit the growth of
pathogen at all concentration (100, 250, 500 ppm).

Key Words: - Pigeonpea, Fusarium udum, Trichoderma sp.

INTRODUCTION
Pigeonpea [Cajanus cajan (L.) Millsp.] is one of the major grain legume crops
of tropics and sub-tropics countries and most extensively grown legume crop in India,
suffers from wilt disease caused by Fusarium udum. The fungus can survive on the
infected plant debris in the soil for about three years and causes serious yield losses,
sometimes even upto 100 per cent in susceptible cultivars Kiprop et al., (2002). The
disease can occurs at any stage of the crop. The total production loss due to this
disease in India alone as estimated to be approximately 97,000 tones per year Saxena
et al., (2010).
Fusarium wilt is characterized by wilting of the affected plants and characteristic
internal browning or blackening of the xylem vessels extending from the root system
to stems Amin et al., (1976). In partially wilted plants purple bands appear. When
the bark of such bands is peeled off, browning of the wood is seen. Wilted plants when
split open, exhibit brown discoloration of the vascular bundles. Effective disease
Student, tAssistant Professor
Department of Plant Protection, Allahabad School of Agriculture, SHIATS, Allahabad 211007 (U.P.)

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Kunwar Zeeshan Khan and Shashi Tiwari

control requires application of combination of broad spectrum fungicide with bioagents. The present paper reports the results of laboratory studies taken under using
dual culture method and poisoned food technique in order to find out an effective
bio-agent and a chemical against Fusarium udum Butler, the pigeonpea wilt
pathogen.

MATERIALS AND METHODS

The pathogen was isolated from wilted pigeonpea plant from research farm of
SHIATS, Allahabad and maintained on Potato dextrose agar medium. The antagonists,
Trichoderma viride and Trichoderma harzianum were isolated from the plant
rhizosphers soil by serial dilution technique on PDA medium. The antagonistic effect
of Trichoderma sp. against the pathogen was tested by the dual culture technique
Dennis and Webster, (1971).
In the dual culture experiment, each treatment was replicated thrice and control
was maintained. A mycelium disc of 5 mm diameter of the pathogen was placed at
end of Petri plate with PDA. An opposite end 5 mm diameter disc of antagonists was
placed. Radial growth of the pathogen was measured and per cent inhibition was
calculated by the following formula:
Growth of the pathogen
Growth of the pathogen
in the control plate
in the presence of antagonist
Inhibition percent =
100
Growth of the pathogen
in the control plate
To evaluate the effect of Carbendazim (at 100, 250, 500ppm) and Neem cake
powder (at 3%, 5%, 7%), concentrations were tasted by poison food technique Nene
and Thapliyal, (1993).
The PDA medium was prepared with double the recommended strength and
sterilized in an autoclave. Carbendazim solution was prepared double the test
concentration. Carbendazim solution was mixed thoroughly with molten PDA medium
in a sterilized Petri plate under aseptic conditions and allowed 5 to 10 minutes to
solidify. The neem cake powder were also mixed into the molten PDA at 3, 5 and
7 percent concentration in sterilised Petriplates under aseptic condition and allowed
to solidify. Appropriate controls were maintained using distilled water. From seven
day old culture plates, 5 mm discs were cut from outer margin of vigorously growing
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fungus with sterilized cork borer and transferred to the centre of the plates containing
fungicidal medium. Three replications were maintained for each treatment. The whole
procedure was carried out under aseptic conditions. The Petri plates were than
incubated at 2520C. The diameter of fungal colony was measured when the growth
of the fungus in controls plates was complete. Finally per cent inhibition was calculated
by the formula.
Diameter of colony
Diameter of colony
in the control plate
in the treated plate
Inhibition per cent =
Diameter of colony
in the control plate

100

RESULTS AND DISCUSSION

The results of dual culture technique revealed the effectiveness of all the
antagonists with varying degrees of inhibition of the growth of the pathogen, (Table
1). Among the antagonists, Trichoderma viride was the best in inhibiting the growth
of the pathogen by 72.40 per cent followed by Trichoderma harzianum 67.90 per cent.
Both the treatments were statistically different from each other. Similar results were
obtained by the Goudar and Kulkarni (1998).

Table 1: Effect of Trichoderma sp. on the radial growth of Fusarium udum in

in vitro (Dual culture technique)
Treatment

Radial growth of
Fusarium udum (mm)*

Percentage inhibition
on control

T0 Control

90

T1 Trichoderma viride

24.83

72.40

T2 Trichoderma harzianum

28.83

67.96

*Mean of three replications

S.Ed ()

0.50

CD at 5%

1.24
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Kunwar Zeeshan Khan and Shashi Tiwari

Fig. 1: Effect of Trichoderma sp. on the radial growth of Fusarium udum in

vitro (Dual culture technique)
Table 2: Effect of bio-agent and carbendazim on the radial growth of Fusarium
udum in vitro (Food poisoned technique)
Treatment

Concentration

Radial growth of
Fusarium udum (mm)*

T0 Control

Percentage inhibition
on control

90

T1 Carbendazim

100 ppm

100

T2

250 ppm

100

T3

500 ppm

100

T4 Neem cake

3%

45.08

49.91

T5

5%

37.33

58.52

T6

7%

38.83

56.85

*Mean of three replications

S.Ed ()

1.10

CD at 5%

2.37
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Fig. 2: Effect of bio-agent and carbendazim on the radial growth of Fusarium

udum in in vitro (Food poisoned technique)

The result (Table. 2) of poisoned food technique revealed that both neem cake
and carbendazim treatments inhibited the radial growth of pathogen significantly as
compared to control at different concentrations. However carbendazim inhibited the
growth of fungus at all concentrations followed by neem cake powder at 5%
concentration (58.52%).
Similar result was reported by Raju et al., (2008) in the respect to inhibition
of the mycelia growth of Fusarium udum. Therefore it was concluded that the
antagonist, Trichoderma viride and systematic fungicide, carbendazim can be
successfully utilized for the control of pigeonpea wilt pathogen.

ACKNOWLEDGEMENT
Authors are thankful to Dr. Shafaat Ahmad, Professor and Head, Department of
Plant Pathology, Sam Higginbottom Institute of Agriculture, Technology & Sciences,
Allahabad 211007 (U.P.) INDIA for his kind guidance, motivation and unconditional
support for this work.
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Kunwar Zeeshan Khan and Shashi Tiwari

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