WELCOME

WELCOME

Suraiya Yasmin
Student
ID: 080802
IN VITRO EVALUATION OF Trichoderma harzianum Rifai.
AND BOTANICAL EXTRACTS AGAINST SOME SOIL AND
SEED BORNE FUNGI
INTRODUCTION
 Trichoderma as bio-control agent are opportunistic, avirulent plant
symbionts

 Plant extracts of many higher plants have been reported to exhibit

antibacterial, antifungal and insecticidal properties

 Biologically based and environmentally safe alternatives are being

investigated for possible use as components in integrated
management programs
OBJECTIVE
To investigate the antagonistic potentiality of T.
harzianum and the effectiveness of selected botanical
extracts against F. oxysporum, B. sorokiniana and S.
rolfsii
MATERIALS AND METHODS
Experiment No.1

Collection of Isolate
Preparation of Potato Dextrose Agar (PDA)
Medium
Multiplication of T. harzianum, B. sorokiniana, F.
oxysporum and S. rolfsii
 In vitro antagonistic effect of T. harzianum on mycelial
growth of B. sorokiniana , F. oxysporum, S. rolfsii
 Dual culture method (Dennis and Webster,1971)
 A triangular shaped colony of each pathogenic fungies
was observed
 Area of the colony was calculated by ½×base×height
 The area of the colony of control plates also measured by
π×(radius)²
 Inhibition percentages of the pathogenic fungies were
calculated by following the formula suggested by Sunder et
al., (1995).
X-Y
% Inhibition = ---------------------- X 100
X
Where X = Mycelial growth of pathogen alone (control)
Y = Mycelial growth of pathogen along with
antagonist
Experimental Design and Data Analysis
 Six replication
 Completely randomized design (CRD)
 MSTAT-C
 Least Significant Difference (LSD)
Experiment No.2
 Preparation of Potato Dextrose Agar (PDA) Medium

 Multiplication of B. sorokiniana, F. oxysporum and S. rolfsii

 Preparation of Botanical Extracts

By following poison food technique (Grover and Moor, 1962)

 Neem Extract
 Garlic Extract
 Ginger Extract
 Botanical Extracts and Their Concentration
Table l. List of plant materials that were used in the experiment

Common Botanical Name Plant Concentration

Name Parts (%)

Used

Neem Azadirachta indica Leaves 5, 10 & 15

Garlic Allium sativum Bulb 5, 10 & 15

In vitro effect of botanical extracts on mycelial growth of B.

sorokiniana, F. oxysporum, S. rolfsii

The inhibition percentages of the tested fungi were calculated

by following the same formula as in Eexperiment No.1

Experimental Design and Data Analysis

 Five replication

 Completely randomized design (CRD)

 MSTAT-C

 Duncan’s Multiple Range Test (DMRT)

RESULTS AND DISCUSSION
Table 2. Inhibition of the growth of tested fungi by T. harzianum
on PDA

T. harzianum

S. rolfsii 76.76

F. oxysporum 88.69

B. sorokiniana 89.20

LSD value - 5.490

 48 hour

Control Dual plate culture

240 hour

Control Dual plate culture

Plate 1. Antagonistic in vitro effect of T. harzianum on mycelial growth of
F. oxysporum
 72 hour

Control Dual plate culture

168 hour

Control Dual plate culture

Plate 2. Antagonistic in vitro effect of T. harzianum on mycelial growth of B.

sorokiniana
 24 hour

Control Dual plate culture

72 hour

Control Dual plate culture

Plate 3. Antagonistic in vitro effect of T. harzianum on mycelial growth of

S. rolfsii
Table 3. Effect of different botanical extracts at different concentration on mycelial
growth inhibition of B. sorokiniana

Botanical Concentration(%) Inhibition percentage

Extracts

Control 0

Ginger 5 10.72ef
10 24.12d
15 38.35ab

Neem 5 24.33d
10 33.40bc
15 40.62a

Garlic 5 4.74f
10 15.88e
15 31.14c

CV(%) 12.62%
LSD value 6.643
Level of significance 0.01
 45 Ginger Linear (Ginger) Neem Linear (Neem) Garlic
Linear (Garlic)

40 f(x) = 1.63 x + 16.49

R² = 1
f(x) = 2.76 x − 3.23
R² = 1
35

30 f(x) = 2.64 x − 9.15

R² = 0.99
Percent Inhibition

25

20

15

10

0
4 6 8 10 12 14 16

Different Concentration

Fig 1. Functional relationship between concentration and mycelial growth inhibition

percentage of botanical extracts of B. sorokiniana
 Control

Ginger 15% Neem 15% Garlic 15%

Plate 4. Effect of different botanical extracts at 15% concentration on mycelial

growth of B. sorokiniana
Table 4. Effect of different botanical extracts at different concentration on
mycelial growth inhibition of S. rolfsii

Botanical extracts Concentration(%) Inhibition Percentage

Control 0

Ginger 5 14.24f
10 17.05e
15 25.76d

Neem 5 4.17g
10 12.28f
15 26.14d

Garlic 5 32.35c
10 53.94b
15 72.20a

CV(%) 3.35%
LSD value 2.039
Level of significance 0.01
 80
Ginger Linear (Ginger) Neem

70 f(x) = 3.99 x + 12.98

R² = 1

60
Percent Inhibition

50

40

30

2.2 x x−+7.77
f(x) = 1.15 7.5
20 R² = 0.98
0.92

10

0
4 6 8 10 12 14 16

Different Concentration
Fig 2. Functional relationship between concentration and mycelial growth inhibition
percentage of botanical extracts of S. rolfsii
 Control

Garlic 15% Ginger 15% Neem 15%

Plate 5. Effect of different botanical extracts at 15% concentration on mycelial

growth of S. rolfsii
Table 5. Effect of different botanical extracts at different concentration on
mycelial growth inhibition of F. oxysporum

Botanical extracts Concentration(%) Inhibition percentage

Control 0

Ginger 5 9.49f
10 31.28c
15 55.80a

Neem 5 17.5e
10 29.62cd
15 56.41a

Garlic 5 5.59g
10 27.82d
15 48.72b

CV(%) 4.28%
LSD value 2.852
Level of significance 0.01
 60 Ginger Linear (Ginger) Neem Linear (Neem)

f(x) = 4.63 x − 14.12

f(x)
R² ==13.85 x − 3.8
R² = 0.95
50
f(x) = 4.31 x − 15.75
R² = 1

40
Percent Inhibition

30

20

10

0
4 6 8 10 12 14 16

Different Concentration

Fig 3. Functional relationship between concentration and mycelial growth inhibition

Percentage of botanical extracts of F. oxysporum
 Control

Ginger 15% Neem15% Garlic 15%

Plate 6. Effect of different botanical extracts at 15% concentration on mycelial

growth of F. oxysporum
 80

72.2
70

60
55.8 56.41

50 48.72

40.62 B. sorokiniana
40 38.35
S. rolfsii
Percent Inhibition

F. oxysporum
31.14
30
25.76 26.14

20

10

0
Ginger Neem Garlic

Fig 4. Functional relationship between 15% concentration and mycelial growth

percentage of botanical extracts of F. oxysporum, B.sorokiniana, S. rolfsii
CONCLUSION
Based on the evaluation of T. harzianum and different
botanical extracts, it can be concluded that-
 T. harzianum was found to be effective against B.
sorokiniana, F. oxysporum, S. rolfsii and that was more than
75%
 Mycelial growth inhibition percentages increased with the
increasing in extracts concentration in all cases. At 15%
concentration garlic maximum inhibited S. rolfsii 72.20%,
neem and ginger maximum inhibited in F. oxysporum
56.41% and 55.80% respectively
 Further investigation may be conducted with more
treatments to confirm this result and generation of
more information.
THANKS
TO
ALL