The Effect of Trichoderma viride

and Pseudomonas fluorescence

on Tobacco (Nicotiana tabacum)
Wilt Disease.

By
K.N.A Jayawardana
 INTRODUCTION
Tobacco (Nicotiana tabacum.L) is an
important cash crop.
Tobacco wilting is a prominent
disease.
Wilting may occur at any stage of
the crop.
Causal agents of wilt may be
bacteria, fungi and rarely viruses.
• Fungus: Fusarium oxysporum
• Bacterium: Pseudomonas
Symptoms of wilt:
Management of the disease:

• Field sanitation throughout the

cropping season.
• Crop rotation with cereals.
• Avoid susceptible crops.
• Chemical control
» Fungicides (Carbendazim)
Problems with chemical
control:

• Environmental pollution.
• Human health hazards.
• Resistance build up against
fungicides.
• Undesirable consequences on non
target organisms.
• High cost.
Advantages of biological
control:

• Durable.
• Effective.
• Risk free.
• Less environmental pollution.
• Cost effective.
Potentials of Trichoderma and
Pseudomonas as biocontrol agents

• High reproductive capacity.

• Ability to survive under unfavorable
conditions.
• Efficiency in the utilization of
nutrients.
• Capacity to modify the rizosphere.
• Ability to promote plant growth
Mechanisms of biological
control:
• Indirect mechanisms:
 Competing for nutrients and space.
 Modify the environmental conditions.
 Promote plant growth and plant
defense mechanism.
 Antibiosis.
• Direct mechanisms:
 Mycoparasitism.
 Production of sideropores.
 OBJECTIVES
• Specific objective:-
• To investigate the biocontrol
potential of Trichoderma viride and
Pseudomonas fluorescence against
controlling tobacco wilt under field
conditions.
• General objective:-
• To introduce an effective and
environmentally friendly biocontrol
mechanism to overcome tobacco wilt
disease with the reduction in cost of
cultivation.
 METHODOLOGY
• Duration: April to September 2006
during yala season

• Location: Mahaweli system

“C”, Dehiattakandiya
(Sandunpura), wilt disease
prevailed fields in
previous five cropping
seasons.
• Experimental design -factorial split plot
design with six treatments and three
replicates.
• Factors -Biocontrol agent
» Trichoderma viride
» Pseudomonas fluoresence
Ploughing depth
» Deep ploughing (8”-10”)
» Shallow ploughing (6”)
Type of organic fertilizer
» biocontrol agent + Cow dung
» biocontrol agent + Compost
» biocontrol agent alone
• First treatment application
-As talc based powder mixed
with cow dung, compost or without
any fertilizer with the field planting.
• Treatments in first application:-
T1. Trichoderma + Cow dung
T2. Trichoderma + Compost
T3. Trichoderma alone
T4. Pseudomonas + Cow dung

T5. Pseudomonas + Compost

• Preparation of mixtures in first treatment
application

 100 -200 kg of farmyard manure / compost was

mixed with 5 kg of Trichoderma viride /
Pseudomonas fluorescence.
 Mixed thoroughly and add water to attain a
moisture level of 40%.
 Kept the mixture in a shade for 48 hrs for the
product to be incubate.
 Bagged the mixture just prior to the application.
 Applied the mixtures in the planting holes.
 This amount of mixture was enough for applying
over 1 ha of land.
 This application was done only once.
Preparation of mixtures in first
treatment application:
200 kg of either
cow dung or compost
+
5 kg of Trichoderma viride /
Pseudomonas fluorescence

Add water, 40%.

Kept the mixture in a shade for 48 hrs

Applied to the planting holes (enough

-1 ha)
 Field layout
Tv + Tv + Tv + Pf + Pf+ Pf +
CO CD CO CO CD CO
Tv Tv + Tv Pf Pf + Pf Tv -
Trichoderma
CD CD
Pf -
Tv + Tv Tv + Pf + Pf Pf + Pseudomonas
CD CO CD CO
Pf + Pf+ Pf + Tv + Tv + Tv +
CO CD CO CO CD CO
CD - Cow
Pf Pf + Pf Tv Tv + Tv Dung
CD CD CO - Compost
Pf + Pf Pf + Tv + Tv Tv +
CD CO CD CO

Deep Shallow
• Field was prepared in to two
ploughing depths; Deep (8”-10”) and
shallow (6”) using a disc plough
• The field was separated into four
blocks that received each biocontrol
agent under both ploughing levels.
• Nine plots were raised in each block.
• In each block only one biocontrol
agent was applied as different
treatments.
• The biocontrol agents were applied
to the plots according to the three
methods mentioned
• They were applied to the planting
holes.
• Tobacco seedlings of variety K326
were planted one week after the
treatment application.
• Seventy plants were raised in each
plot.
• Second treatment application
As a soil drench application to the
base of the plant with the disease
incidence.

Applied at the rate of 30 ml of the

liquid pure solution (2×106cfu/ml) per
• Method of inoculation of the
disease was the natural infection.
• Data were recorded as number of
wilted plants per each plot.
• Data were recorded after the
disease incidence in weekly for
four weeks.
• Data were analyzed by Minitab
software analyzing package.
RESULTS AND DISCUSSION
• The first symptoms of the wilt
disease were observed at three
weeks after field planting.
The effect of biocontrol agent:
• No significant difference was observed in
number of wilted plants under the
application of Trichoderma viride and
Pseudomonas fluorescence
35
Second treatment
Mean of the percentage wilted

30
T1
25
T2
20 T3
15 First treatment T4
T5
10
T6
5

0
1 2 3 4 5 6 7 8 9 10
Time (weeks)
The effect of ploughing depth
Lesser number of wilted plants were
observed in deep ploughed fields over
shallow ploughed fields
50
Mean of the percentage wilted

45
40
35
30 Deep ploughing
25
20 Shallow ploughing
15
10
5
0
1 2 3 4
Weeks after disease incidence
The effect of method of
application:
• The application of both biocontrol
agents with compost showed a lesser
number of wilted plants.

Method of Ploughing
application level
Deep shallow
Cow dung 16.19 24.17
Compost 5.71 19.11
Without fertilizer 8.45 22.02
Practical difficulties:

• Contamination of biocontrol agents

» Separate by big bunds
» Separate irrigation

• Weeds
» Weeding once a fortnight

• Maintenance of soil moisture

» Irrigate once in 3-4 days / moisture 50-
60%
 CONCLUTIONS
• Disease can be manage by deep
ploughing
• Disease can be manage with a
very low incidence level by
applying Trichoderma viride or
Pseudomonas fluoresence with
compost
• Repeated application of the
biocontrol agents once a fortnight
will effectively suppressed the
Recommendations:

• Compost application followed by

soil drench application of
Trichoderma viride or
Pseudomonas fluoresence with the
field planting under deep
ploughing is recommended
 ACKNOWLEDGEMENTS
• The director, Ceylon Tobacco Company,
Kandy
• Mr. K. H. M. I Karunarathna, Computer
service unit, Makandura
• Mrs. Auchithya Disanayaka, Coconut
Research institute, Lunuwila
• Tobacco farmers in village Sandunpura