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ANTIFUNGAL SCREENING OF
SOME COMMON WEED EXTRACTS
AGAINST DOMINANT PLANT
PATHOGENIC FUNGI
A PROJECT REPORT
SUBMITTED
IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE DEGREE OF
MASTER OF SCIENCE
IN
APPLIED MICROBIOLOGY
By
GAURAV KUMAR PAL
Under the supervision of
DR. S.K. SHAHI
Lecturer
Bio-resource Tech Laboratory
DEPARTMENT OF MICROBIOLOGY
CHAUDHARY CHARAN SINGH UNIVERSITY
MEERUT-250004
2012
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Acknowledgements Acknowledgements
With the utmost appreciation, I would like to concede respected supervisor
Dr. S.K. Shahi, Dr. S.K. Shahi, for their guidance, selfless efforts, sparing their precious time for
guidance and supervision and always being ready for pointing out mistakes and
giving suggestions to improve them.
I am also grateful to Head of Department Dr. P.K
necessary facilities in the Department for carrying out the present investigation.
His dedication towards the pursue of knowledge and readiness to instill the same
in his students has always been encouraging.
Dr. P.K. Sharma. Sharma for providing me
Special thanks to the teachers in the Department, namely, Dr. D. C.
Sharma, Dr. D. K. Sharma, Dr. Anjali Malik, Dr. Dilshad Ali, Mr. D. Prakash,
Mr. Ajay Shukla and Mr. Rahul Sharma and for introducing me to the world of
microbes, for helping me to enhance my potentials and develop scientific approach
to delve into the galaxy of microbes.
There are no words to express my appreciation and affection to my friends
Brijesh Kumar, Amit, Ritu etc.for their co-operation and warm company during
my tough time and their support and encouragement.
M.Sc.who always encouraged and showed me the right direction to move me.
I also acknowledge my thanks to my loving friends and all classmates of
brothers who took pain to bring me to this stage and for their enormous help and
healthy suggestions. I am extremely grateful to all my relatives whose constant
inspiration & patience and loving emotions helped me invisibly.
I express my deepest love and affection to my parents my sisters and
( (GAURAV GAURAV KUKUMA MAR R PAL)PAL)
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DEDICATED DEDICATED
TO TO
MYMY
BELOVED BELOVED
PARENTSPARENTS
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चौधरी चरण स िंह विश्िविद्यालय मेरठ
Ch. Charan Singh University, Meerut
P.K. Sharma
M. Sc. (Ag.), M. Phil., Ph.D., N.E.T.
BOYSCAST Fellowship Awardee, Scotland, U.K.
Head
Coordinator (Applied Microbiology)
Coordinator (Bioinformatics)
Coordinator (Medical Microbiology)
Department of Microbiology
C.C.S. University, Meerut
250005 (India)
Tel : +91-121-2600909 (Off.)
Website: www. ccsuniversity.ac.in
E-mail: pks264@rediffmail.com
Date:
TO WHOM IT MAY CONCERN
This is to certify that Mr. Gaurav Kumar Pal has been a student of M.Sc. (Applied
Microbiology) in Department of Microbiology, Ch. Charan Singh University, Meerut,
(UP) during the session 2010-2012. He has carried out his project work on the topic
“Antifungal screening of some common weed extracts against dominant plant
pathogenic fungi’’ under the supervision of Dr. S.K Shahi in the Department. He worked
here from 1st March, 2012 to 15th July, 2012.
I wish him all success in his future endeavours.
(P.K. Sharma)
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CHAUDHARY CHARAN SINGH UNIVERSITY
Department of Microbiology
Bio-Resource Tech Laboratory
MEERUT, 250004- INDIA
: 09319394547 (M), Email: shahi.sk@rediffmail.com,
sushilkshahi@gmail.com
Date:……………….
Certificate
This is to certify that Mr. Gaurav Kumar Pal, M.Sc (Applied Microbiology),
Department of Microbiology, CCS University, Meerut has done more than four months
(w.e.f. 1.3.2012 to 15.7.2012) M.Sc. Project entitled ‘Antifungal screening of some
common weed extracts against dominant plant pathogenic fungi’’ in the Department
of Microbiology, CCS University, Meerut under the supervision of the undersigned in the
Department.
(S.K. Shahi)
Supervisor
Dr. Sushil Kumar Shahi
M.Sc, Ph.D, FIYSA, FPSI, FSPP, FBSI
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CONTENTS
1. INTRODUCTION 1-6
2. REVIEW OF LITERATURE 7-39
2.1. Plant diseases 7
2.1.1. Plant diseases caused by fungi 9
2.2. Wilt Disease 11
2.2.1. Fusarium oxysporum
12
2.2.2. Fusarium moniliforme
13
2.2.3. Fusarium udum
14
2.3. Early Blight Disease 15
2.3.1. Alternaria solani
15
2.4. Plant product as antimicrobial agents 17
2.5. Major groups of antimicrobial compounds from plants 21
2.5.1. Phenolics and polyphenols 22
2.5.2. Quinones 22
2.5.3. Flavones, flavonoids, and flavonols 24
2.5.4. Coumarins 27
2.5.5. Terpenoids and essential oils 29
2.5.6. Alkaloids 31
2.5.7. Lectins and polypeptides 32
2.5.8. Mixtures 33
2.6. Antimicrobial activity of some plants 33
3. MATERIAL AND METHODS 40-51
3.1. Materials 40
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3.1.1. Glassware’s, Plastic wares and Chemicals 40
3.1.2. Media used 42
3.1.2.1Sabouraud dextrose agar media 42
3.1.2.2 Sabouraud dextrose broth media 43
3.1.2.3. Rose bengal chloramphenicol (RBC) agar media 44
3.1.2.4. Potato dextrose agar media 44
3.1.3. Collection of sample for isolation of fungi 45
3.1.4 Plant Materials 45
3.2. Methods 46
3.2.1. Isolation of pre harvesting and seed borne plant
pathogenic fungi 46
3.2.2. Maintenance of fungal cultures 47
3.2.3. Identification of Isolates 47
3.2.3.1. Microscopic and taxonomical study of dominant fungal
pathogens 47
3.2.4. Comparative efficacy of plant extract with chemical fungicide 47
3.2.5. Preparation of plant extracts 48
3.2.6. Antimicrobial screening of plant extracts against isolated
strains by modified poison food method 49
3.2.6.1. Preparation of McFarland standard 49
3.2.6.2. 3.2.6.2. Antimicrobial screening of plant extracts 49
3.2.7. Determination of MIC by microtiter plate assay 50
3.2.7.1. Preparation of resazurin solution 50
3.2.7.2. Preparation of plates 51
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4. RESULTS 52-65
4.1. Collection of infected plant parts and seeds samples and isolation
of the fungal strains 52
4.1.1. Identification of fungi isolated from various infected fruits 53
4.2. Isolation and identification of fungal microorganism 55
4.3. Antibiogram of chemical fungicide (Mancozeb) against pathogenic fungi 56
4.4. Comparison of antifungal activity of plant extracts 57
4.5. Antifungal screening of collected plants extracts against Alternaria strain
GP-6. 58
4.6. Antifungal screening of collected plants extracts against Fusarium strain
GP-2 60
4.7. Antifungal screening of collected plants extracts against F. strain GP-4 62
4.8. MIC of solvent extracts of different plants against Alternaria strain
GP-6
64
4.9. MIC of solvent extracts of different plants against Fusarium
strain GP-2
65
4.10. MIC of solvent extracts of different plants against Fusarium
strain GP-4
67
5. DISCUSSION 69-71
6. CONCLUSION 72-73
7. REFERENCES 74-96
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LIST OF TABLES
Table No. Table title Page No.
Table 2.1 Antifungal activity of various plants against pre harvesting
and seed borne plant pathogenic fungi
34-39
Table .3.1 List of plants collected from the Meerut region 45-46
Table 4.1 Collection of samples and respective code 52
Table 4.2 Identification of isolated fungal strains 53-54
Table 4.3. Identification of isolates fungal microorganism 55-56
Table 4.4 Antibiogram of different mancozabe concentrations against
tetested fungi
56-57
Table 4.5 Antifungal screening of collected plants extracts against
Alternaria strain GP-6
58-59
Table 4.6 Antifungal screening of collected plants extracts against
Fusarium strain GP-2
60-61
Table 4.7 Antifungal screening of collected plants extracts against
Fusarium strain GP-4
62-63
Table 4.8 Minimum inhibitory concentration against Alternaria strain
GP-6
64
Table 4.9 Minimum inhibitory concentration against Fusarium strain
GP-2.
65-66
Table
4.10
Minimum inhibitory concentration against Fusarium strain
GP-4
67
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LIST OF FIGURES
Figure No. Figure title
Figure 3.1 Collected plant from Meerut region
Figure 4.5 Antifungal screening of collected plants extracts against Alternaria
strain GP-6
Figure 4.6 Antifungal screening of collected plants extracts against Fusarium
strain GP-2
Figure 4.7 Antifungal screening of collected plants extracts against Fusarium
strain GP-4.
Figure 4.8
Minimum inhibitory concentrations determination of different plants
extracts.
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Figure 4.5 Antifungal screening of collected plants extracts against
Alternaria strain GP-6.
A.
Showing fungal growth inhibition of Alternaria strain GP-6 by different
extracts of plant Achyranthes aspera.
B.
Showing fungal growth inhibition of Alternaria strain GP-6 by different
extracts of plant Parthenium hysterophorus.
C.
Showing fungal growth inhibition of Alternaria strain GP-6 by different
extracts of plant Cannabis sativa.
D.
Showing fungal growth inhibition of Alternaria strain GP-6 by different
extracts of plant Calotropis gigantean.
E.
Showing fungal growth inhibition of Alternaria strain GP-6 by different
extracts of plant Chenopodium album.
F.
Showing fungal growth inhibition of Alternaria strain GP-6 by different
extracts of plant Canada thistle.
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Figure 4.5 Antifungal screening of collected plants extracts against
Alternaria strain GP-6.
G. Showing fungal growth inhibition of Alternaria strain GP-6 by different extracts
of plant Phalaris minor.
H. Showing fungal growth inhibition of Alternaria strain GP-6 by different extracts
of plant. Cynoden dactylon.
I. Showing fungal growth inhibition of Alternaria strain GP-6 by different extracts
of plant Argemone maxicana.
J. Showing fungal growth inhibition of Alternaria strain GP-6 by different extracts
of plant Ageratum conyzoides.
K. Showing fungal growth inhibition of Alternaria strain GP-6 by different extracts
of plant Lantana camera.
L. Showing control fungal growth of Alternaria strain GP-6.
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Figure 4. 6 Antifungal screening of collected plants extracts against
Fusarium strain GP-2 .
A. Showing fungal growth inhibition of Fusarium strain GP-2 by different
extracts of plant Achyranthes aspera.
B. Showing fungal growth inhibition of Fusarium strain GP-2 by different
extracts of plant Parthenium hysterophorus.
C. Showing fungal growth inhibition of Fusarium strain GP-2 by different
extracts of plant Cannabis sativa.
D. Showing fungal growth inhibition of Fusarium strain GP-2 by different
extracts of plant Calotropis gigantean.
E. Showing fungal growth inhibition of Fusarium strain GP-2 by different
extracts of plant Chenopodium album.
F. Showing fungal growth inhibition of Fusarium strain GP-2 by different
extracts of plant Canada thistle.
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Figure 4.6. Antifungal screening of collected plants extracts against Fusarium
strain GP-2
G. Showing fungal growth inhibition of Fusarium strain GP-2 by different extracts of
plant Phalaris minor.
H. Showing fungal growth inhibition of Fusarium strain GP-2 by different extracts of
plant. Cynoden dactylon.
I. Showing fungal growth inhibition of Fusarium strain GP-2 by different extracts of
plant Argemone maxicana.
J. Showing fungal growth inhibition of Fusarium strain GP-2 by different extracts of
plant Ageratum conyzoides.
K. Showing fungal growth inhibition of Fusarium strain GP-2 by different extracts of
plant Lantana camera.
L. Showing control fungal growth of Fusarium strain GP-2.
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Figure 4. 7. Antifungal screening of collected plants extracts against
Fusarium strain GP-4.
A. Showing fungal growth inhibition of Fusarium strain GP-4 by different extracts of
plant Achyranthes aspera.
B. Showing fungal growth inhibition of Fusarium strain GP-4 by different extracts of
plant Parthenium hysterophorus.
C. Showing fungal growth inhibition of Fusarium strain GP-4 by different extracts of
plant Cannabis sativa.
D. Showing fungal growth inhibition of Fusarium strain GP-4 by different extracts of
plant Calotropis gigantean.
E. Showing fungal growth inhibition of Fusarium strain GP-4 by different extracts of
plant Chenopodium album.
F. Showing fungal growth inhibition of Fusarium strain GP-4 by different extracts of
plant Canada thistle.
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