[Khóa luận tốt nghiệp] Isolation, identification and study of biological characteristics of the pathogenic fungi curvularia lunata causing leaf spot to bananas
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CONTENTS
COMMITMENT ...................................................................................................... i
ACKNOWLEDGEMENTS ..................................................................................... ii
CONTENTS ........................................................................................................... iii
LIST OF TABLES ................................................................................................. vi
LIST OF FIGURES ............................................................................................... vii
LIST OF ABBREVIATIONS ............................................................................... viii
ABSTRACT ........................................................................................................... ix
I. INTRODUCTION .............................................................................................. 1
II. LITERATURE REVIEW ................................................................................. 4
2.1. Overview of Bananas ................................................................................ 4
2.1.1. Bananas in food demanding ....................................................................... 4
2.1.2. Bananas as economic importance ............................................................... 4
2.1.3. Diseases causing the banana production loss. .............................................. 5
2.1.3.1. Diseases from bacteria .............................................................................. 5
2.1.3.2. Diseases from virus ................................................................................... 2
2.1.3.3. Diseases from fungi ................................................................................... 3
2.2. Overview of Curvularia lunata .................................................................. 6
2.2.1. Overview of Curvularia species ................................................................. 6
2.2.2. Overview of Curvularia lunata .................................................................. 8
2.2.2.1. Morphology of Curvularia lunata .............................................................. 8
2.2.2.2. Various hosts of Curvularia lunata ............................................................ 9
2.2.2.3. Infection mechanism of Curvularia lunata ................................................ 12
III. MATERIALS AND METHODS ................................................................... 14
3.1. Time and location for the research ........................................................... 14
iv
3.2. Materials .............................................................................................. 14
3.3. Methods ............................................................................................... 15
3.3.1. Samples collection method .......................................................................... 15
3.3.2. Isolating and subculturing method ......................................................... 16
3.3.3. Pathogenicity test .................................................................................. 17
3.3.4. Studying on some biological characteristics of the selected fungi strain... 17
3.3.4.1. Studying on the morphological characteristics of conidia, mycelium and
conidiophore of the selected fungi strain................................................... 17
3.3.4.2. Studying on the culturing features of the pathogenic fungi in different medium
............................................................................................................... 18
3.3.4.3. Screening the ability to produce extracellular enzyme cellulase of the selected
fungi strain .............................................................................................. 18
3.3.5. Indentification of the selected fungi strain ............................................... 18
3.3.5.1. DNA extraction method ......................................................................... 18
3.3.5.2. Amplification, sequencing and analyzing ITS gene sequence of isolated fungi
strain method .......................................................................................... 20
3.3.6. Screening, detecting toxic genes of fungal strains ..................................... 21
IV. RESULTS AND DISCUSSION ..................................................................... 23
4.1. Results of samples collection, isolation and subculturing the pathogenic
pathogenic fungi strain. ........................................................................... 23
4.2. Pathogenicity test ................................................................................... 25
4.3. Classification of pathogenic fungi strain HN 5.2. ..................................... 26
4.3.1. Amplification, sequencing and analyzing ITS gene sequence of fungi strain
HN 5.2. ................................................................................................... 26
4.3.2. Clg2P gene amplification ........................................................................ 29
4.4. Screening the development of HN 5.2 strain in different medium ............. 29
4.5. Testing the ability to extract cellulase of HN 5.2 strain .............................. 32
v
4.6. Screening for the genes encoding virulence factors biosynthesis in pathogenic
HN 5.2 fungi ........................................................................................... 33
V. CONCLUSION AND PROPOSAL ................................................................. 36
5.1. Conclusion...................................................................................................... 36
5.2. Proposal ......................................................................................................... 36
REFERENCES .................................................................................................... 37
APPENDIX .......................................................................................................... 45
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