| 摘要 | 第8-10页 |
| ABSTRACT | 第10-11页 |
| LIST OF ABBREVIATIONS | 第12-13页 |
| CHAPTER 1 GENERAL INTRODUCTION | 第13-39页 |
| 1.1 INTRODUCTION OF THIS STUDY | 第13-16页 |
| 1.2 REVIEW OF LITERATURE | 第16-39页 |
| 1.2.1 Oil seed rape | 第16-19页 |
| 1.2.1.1 Crop history | 第16-17页 |
| 1.2.1.2 Production and economic importance | 第17-18页 |
| 1.2.1.3 Important fungal diseases on Oil seed rape | 第18-19页 |
| 1.2.2 Sclerotinia sclerotiorum | 第19-32页 |
| 1.2.2.1 History | 第19页 |
| 1.2.2.2 Economic importance | 第19-20页 |
| 1.2.2.3 Taxonomy and morphology | 第20-22页 |
| 1.2.2.4 Host range | 第22-24页 |
| 1.2.2.5 Symptom | 第24-25页 |
| 1.2.2.6 Disease cycle | 第25-26页 |
| 1.2.2.7 Epidemiology | 第26-27页 |
| 1.2.2.8 Management of Sclerotinia sclerotiorum | 第27-32页 |
| 1.2.2.8.1 Host resistance | 第27页 |
| 1.2.2.8.2 Cultural management | 第27-30页 |
| 1.2.2.8.3 Biological control | 第30页 |
| 1.2.2.8.4 Chemical control | 第30-32页 |
| 1.2.2.8.4.1 Seed treatment | 第30-31页 |
| 1.2.2.8.4.2 Foliar sprays | 第31-32页 |
| 1.2.3 Detection of resistance to different fungicides in Sclerotinia sclerotiorum | 第32-34页 |
| 1.2.4 Molecular mechanisms of resistance to different fungicides in Sclerotiniasclerotiorum | 第34-36页 |
| 1.2.5 Os1/Group-III Histidine kinase (Hk) and its importance | 第36-37页 |
| 1.2.6 c DNA library transfer approach | 第37-39页 |
| CHAPTER 2 EXPLORING MECHANISMS OF RESISTANCE TODIMETHACHLONE IN SCLEROTINIA SCLEROTIORUM | 第39-60页 |
| 2.1 INTRODUCTION | 第39-40页 |
| 2.2 MATERIALS AND METHODS | 第40-47页 |
| 2.2.1 Isolates of Sclerotinia sclerotiorum | 第40-41页 |
| 2.2.2 Sensitivity assay of Sclerotinia sclerotiorum isolates to dimethachlone andother fungicides | 第41-42页 |
| 2.2.3 Determination of mycelial growth and sclerotial production on PDA | 第42页 |
| 2.2.4 Determination of pathogenicity on detached leaves of oilseed rape plants | 第42页 |
| 2.2.5 Determination of sensitivity to osmotic pressure and oxidative stress | 第42-43页 |
| 2.2.6 Measurement of electrical conductivity for mycelial exudates | 第43页 |
| 2.2.7 Amplification and sequencing of key genes in the HOG signalling pathway | 第43-44页 |
| 2.2.8 RNA extraction and c DNA synthesis | 第44页 |
| 2.2.9 Determination of the expression of Ss Os1 and Ss Hog1 genes by real-time PCRand transporter genes in FD47R/S by RT-PCR | 第44-46页 |
| 2.2.10 Data analysis | 第46-47页 |
| 2.3 RESULTS | 第47-56页 |
| 2.3.1 Sensitivity of Sclerotinia sclerotiorum isolates to dimethachlone and otherfungicides | 第47-48页 |
| 2.3.2 Mycelial growth and sclerotial production on PDA | 第48-49页 |
| 2.3.3 Lesion development of Sclerotinia sclerotiorum isolates and mutants on untreated and dimethachlone-treated detached leaves of oilseed rape cultivarHuayouza 5 | 第49-51页 |
| 2.3.4 Influence of osmotic pressure and oxidative stress on mycelial growth | 第51-52页 |
| 2.3.5 Resistant isolates released more electrolytes | 第52-53页 |
| 2.3.6 Investigation of point mutations in the Ss Os1 gene | 第53页 |
| 2.3.7 Expression of Ss Os1 and Ss Hog1 genes | 第53-56页 |
| 2.4 DISCUSSION | 第56-60页 |
| CHAPTER 3 SCREENING DIMETHACHLONE RESISTANCE GENE IN SCLEROTINIA SCLEROTIORUM BY CONSTRUCTING AND TRANSFERRING OF THE CDNA LIBRARY FROM THE DIMETHACHLONE RESISTANT FIELDISOLATE HN4561JBJ TO SENSITIVE ISOLATE HN4042J | 第60-70页 |
| 3.1 INTRODUCTION | 第60页 |
| 3.2 MATERIALS AND METHODS | 第60-65页 |
| 3.2.1 Fungicide, media and isolates of Sclerotinia sclerotiorum | 第60-61页 |
| 3.2.2 Extraction of total RNA from the dimethachlone resistant field isolateHN4561JBJ | 第61页 |
| 3.2.3 Synthesis of ss c DNA | 第61页 |
| 3.2.4 Synthesis of ds c DNA | 第61-62页 |
| 3.2.5 Purification of ds c DNA in removing the fragments below 300bp | 第62-63页 |
| 3.2.6 c DNA library construction and evaluation | 第63页 |
| 3.2.7 Extraction and transfer of c DNA library from E.coli and transfer toAgrobacterium | 第63-64页 |
| 3.2.8 Agrobacterium transformation and evaluation of transformant strains | 第64-65页 |
| 3.2.9 Fungicide screening and identification of resistant mutants | 第65页 |
| 3.2.10 Extraction of genomic DNA from resistant mutants and investigation ofc DNA fragment insertion | 第65页 |
| 3.3 RESULTS | 第65-68页 |
| 3.3.1 Total RNA analysis | 第65-66页 |
| 3.3.2 ds c DNA synthesis | 第66-67页 |
| 3.3.3 c DNA library construction and evaluation | 第67页 |
| 3.3.4 Agrobacterium mediated transformation and identification of resistant mutantsto dimethachlone | 第67页 |
| 3.3.5 Investigation of dimethachlone resistant mutants | 第67-68页 |
| 3.4 DISCUSSION | 第68-70页 |
| CHAPTER 4 NON-SPECIFIC LOCATED POINT MUTATIONS IN SSOS1 GENE OF SCLEROTINIA SCLEROTIORUM MAY CONFER RESISTANCE TODIMETHACHLONE | 第70-80页 |
| 4.1 INTRODUCTION | 第70页 |
| 4.2 MATERIALS AND METHODS | 第70-72页 |
| 4.2.1 Fungal isolate and Media | 第70-71页 |
| 4.2.2 Level and stability of dimethachlone resistance in the mutants of Sclerotiniasclerotiorum | 第71页 |
| 4.2.3 Amplification of the Ss Os1 gene in the Dimethachlone resistant mutants andtheir parental isolate by PCR | 第71-72页 |
| 4.2.4 Sequence analysis of Ss Os1 gene from dimethachlone resistant mutants andtheir parental isolates | 第72页 |
| 4.3 RESULTS | 第72-78页 |
| 4.3.1 Identification of Sclerotinia sclerotiorum mutants resistant to dimethachlone | 第72-73页 |
| 4.3.2 Level and stability of dimethachlone resistant mutants | 第73-75页 |
| 4.3.3 Amplification of Ss Os1 gene in resistant mutants as well as parental isolate byPCR | 第75-76页 |
| 4.3.4 Sequence analysis of Os1 gene in Sclerotinia sclerotiorum | 第76-78页 |
| 4.4 DISCUSSION | 第78-80页 |
| CHAPTER 5 SUMMARY AND OUTLOOK | 第80-82页 |
| 5.1. CONCLUSION | 第80页 |
| 5.2 FUTURE WORK | 第80-82页 |
| REFERENCES | 第82-104页 |
| APPENDIX | 第104-108页 |
| LIST OF PUBLICATION DURING PHD STUDY | 第108-109页 |
| ACKNOWLEDGEMENTS | 第109-111页 |
