| Acknowledgements | 第9-21页 |
| List of Abbreviations | 第21-23页 |
| ABSTRACT | 第23-27页 |
| 摘要 | 第28-32页 |
| Chapter 1 General Introduction | 第32-36页 |
| 1.1 Background information | 第32-34页 |
| 1.2 Project goals | 第34页 |
| 1.3 Specific objectives | 第34-36页 |
| Chapter 2 Literature Review | 第36-58页 |
| 2.1 Nomenclature and classification of the pathogen | 第37-38页 |
| 2.2 Pathogenicity determinants of V.dahliae | 第38-41页 |
| 2.2.1 Infectious structures | 第38页 |
| 2.2.2 Cell wall degrading and hydrolytic enzymes | 第38-39页 |
| 2.2.3 Phytotoxins and elicitors | 第39-41页 |
| 2.3 Plant defense responses | 第41-48页 |
| 2.3.1 Phenylpropanoid metabolism and cell wall fortification | 第41-43页 |
| 2.3.2 Carbohydrate metabolism and osmoregulation | 第43-45页 |
| 2.3.3 Induction of pathogenesis-related(PR)proteins | 第45页 |
| 2.3.4 ROS in signal transduction | 第45-46页 |
| 2.3.5 Nitric oxide (NO)in signal transduction | 第46-47页 |
| 2.3.6 Network and interplay of signaling pathways | 第47-48页 |
| 2.4 Management of Verticillium wilt | 第48-51页 |
| 2.4.1 Biological control | 第48-49页 |
| 2.4.2 Organic amendments | 第49-50页 |
| 2.4.3 Resistant germplasm | 第50-51页 |
| 2.5 Brassinosteroids(BRs) | 第51-58页 |
| 2.5.1 BR signaling in plants | 第52-53页 |
| 2.5.2 Ameliorative role of BRs in biotic stresses | 第53-54页 |
| 2.5.3 Mechanism of brassinosteroid mediated stress tolerance | 第54-58页 |
| Chapter 3 Effect of Verticillium dahliae toxin(Vd toxin)on cell structure,plantmorphology and biochemical characteristics of upland cotton | 第58-80页 |
| 3.1 Materials and methods | 第59-64页 |
| 3.1.1 Fungus growth and toxin preparation | 第59页 |
| 3.1.2 Plant growth,treatment conditions and measurement of basic nutrition solution (BNS)uptake | 第59-61页 |
| 3.1.3 Assessment of root dehydrogenase activity | 第61页 |
| 3.1.4 Measurement of chlorophyll fluorescence | 第61页 |
| 3.1.5 Determination of lipid peroxidation,SOD activity and contents of reduced ascorbate(ASA)and reduced glutathione(GSH) | 第61-63页 |
| 3.1.6 Determining the plant growth parameters and root architecture | 第63页 |
| 3.1.7 Scanning electron microscopy(SEM)and transmission electron microscopy (TEM) | 第63-64页 |
| 3.1.8 Stati stical analysis | 第64页 |
| 3.2 Results | 第64-74页 |
| 3.2.1 Plant growth and root dehydrogenase activity | 第64页 |
| 3.2.2 Ultrastructure changes in leaf and root cells | 第64-65页 |
| 3.2.3 SEM observations of root tips and leaf epidermis | 第65-68页 |
| 3.2.4 Basic nutrition solution(BNS)uptake and chlorophyll fluorescence(Fv/Fm) | 第68-69页 |
| 3.2.5 Lipid peroxidation and SOD activity | 第69-71页 |
| 3.2.6 Contents of reduced glutathione(GSH)and reduced ascorbate(ASA) | 第71-72页 |
| 3.2.7 Effect of Vd toxin on root architecture and plant growth parameters | 第72-74页 |
| 3.3 Discussion | 第74-79页 |
| 3.4 Conclusion | 第79-80页 |
| Chapter 4 Role of brassinosteroids to alleviate the negative effects of Vd toxin oncotton callus growth | 第80-99页 |
| 4.1 Materials and methods | 第81-86页 |
| 4.1.1 Plant material,toxin preperation and treatment conditions | 第81-82页 |
| 4.1.2 Determination of chlorophyll and carotenoids contents | 第82页 |
| 4.1.3 Measurement of secondary metabolism-related enzymes activities | 第82-84页 |
| 4.1.4 Quantification of flavonoids,total phenols,soluble sugars and protein | 第84-85页 |
| 4.1.5 Measurement of DNA damage through single cell gel electrophoresis assay (Comet assay) | 第85页 |
| 4.1.6 Tissue processing for transmission electron microscope (TEM) | 第85-86页 |
| 4.1.7 Statistic analysis | 第86页 |
| 4.2 Results | 第86-95页 |
| 4.2.1 Effect of EBR on callus growth | 第86页 |
| 4.2.2 EBR induced tolerance to Vd toxin | 第86-87页 |
| 4.2.3 Chlorophyll and carotenoids contents | 第87-88页 |
| 4.2.4 Activities of secondary metabolism related enzymes under Vd toxin stress | 第88-90页 |
| 4.2.5 Flavonoids,total phenols,soluble sugars and protein | 第90页 |
| 4.2.6 DNA damage in cotton callus | 第90-92页 |
| 4.2.7 Ultrastructural changes under Vd toxin and ameliorative effects of EBR | 第92-95页 |
| 4.3 Discussion | 第95-98页 |
| 4.4 Conclusion | 第98-99页 |
| Chapter 5 Brassinosteroids attenuated Verticillium wilt in upland cotton bymodulating the carbohydrates metabolism, plasma membrane ATPases andintracellular osmolytes | 第99-115页 |
| 5.1 Materials and methods | 第100-104页 |
| 5.1.1 Plant growth,toxin preparation and treatment conditions | 第100-101页 |
| 5.1.2 Evaluation of plant wikting, photosynthesis, transpiration, SPAD and chlorophyll fluorescence | 第101页 |
| 5.1.3 Determination of proline,glycine-betaine and soluble sugar content | 第101-102页 |
| 5.1.4 Determination of the enzymes activities involved in carbohydrate metabolism | 第102-103页 |
| 5.1.5 Measurement of different ATPases activity | 第103页 |
| 5.1.6 Statistical analysis | 第103-104页 |
| 5.2 Results | 第104-110页 |
| 5.2.1 EBR application attenuated plant wilting | 第104-105页 |
| 5.2.2 Activity of different ATPase in cotton leaves | 第105页 |
| 5.2.3 Accumulation of intracellular osmolytes | 第105-107页 |
| 5.2.4 Effect of Vd toxin and EBR on activities of enzymes involved in carbohydrate metabolism | 第107-108页 |
| 5.2.5 Photosynthetic parameters,chlorophyll content and fluorescence | 第108页 |
| 5.2.6 Root and shoot biomass | 第108-110页 |
| 5.3 Discussion | 第110-114页 |
| 5.4 Conclusion | 第114-115页 |
| Chapter 6 Understanding the physiological and biochemical basis of innate andbrassinosteroid induced tolerance to Verticillium wilt in cotton leaves | 第115-135页 |
| 6.1 Materials and methods | 第116-120页 |
| 6.1.1 Plant material,growth and treatment conditions | 第116页 |
| 6.1.2 Measurement of superoxide radical(O_2~-),hydrogen peroxide(H_2O_2),their histochemical detection and electrolyte leakage in cotton leaves | 第116-117页 |
| 6.1.3 Determination of leaf osmotic potential and relative water contents | 第117-118页 |
| 6.1.4 Measurement of the enzymes activities involved in carbohydrate and secondary metabolism in cotton leaves | 第118页 |
| 6.1.5 Determination of callose and lignin content | 第118页 |
| 6.1.6 Analysis of the activities of antioxidative enzymes | 第118-120页 |
| 6.1.7 Dynamics of glutathione-S-transferase(GST),dehydroascorbate reductase (DHAR) and contents of non enzymatic antioxidants | 第120页 |
| 6.1.8 Quantification of total phenol,DPPH activity,flavonoids and total soluble proteins | 第120页 |
| 6.1.9 Statistical analysis | 第120页 |
| 6.2 Results | 第120-131页 |
| 6.2.1 H_2O_2/O_2~- accumulation and electrolyte leakage in cotton leaves | 第120-124页 |
| 6.2.2 Changes in leaf osmotic potential(OP) and relative water content(RWC) | 第124-125页 |
| 6.2.3 Stem lignin and callose deposition | 第125-126页 |
| 6.2.4 Changes in the activities of enzymes involved in carbohydrate metabolism(AI, SuSy,SPS)in cotton leaves | 第126-127页 |
| 6.2.5 Activities of enzymes involved in secondary metabolism in cotton leaves | 第127-128页 |
| 6.2.6 Glutathione-S-transferase (GST), dehydroascorbate reductase (DHAR) and contents of phenols, flavonoids, soluble protein and non enzymatic antioxidants | 第128-130页 |
| 6.2.7 Dynamics of antioxidative enzymes in cotton leaves | 第130-131页 |
| 6.3 Discussion | 第131-134页 |
| 6.4 Conclusion | 第134-135页 |
| Chapter 7 Genotypic differences in carbohydrate, secondary metabolism andantioxidative capacities in cotton root and amino acid contents in root exudates | 第135-159页 |
| 7.1 Materials and Methods | 第136-138页 |
| (A) Analysis in cotton root | 第136页 |
| 7.1.1 Measurement of O_2~- and H_2O_2 contents, enzymes activities of carbohydrate, secondary metabolism and antioxidants, phenols, flavonoids and callose in cotton root | 第136页 |
| (B) Analysis in cotton root exudates | 第136-138页 |
| 7.1.2 Plant material, growth, inoculum preparation and treatment conditions | 第136-137页 |
| 7.1.3 Evaluation of disease resistance | 第137页 |
| 7.1.4 Collection of root exudates and analysis of amino acids | 第137页 |
| 7.1.5 In vitro activities of amino acid on the growth of V. dahliae | 第137-138页 |
| 7.1.6 Statistical analysis | 第138页 |
| 7.2 Results | 第138-152页 |
| 7.2.1 H_2O_2 and O_2~- accumulation in cotton roots | 第138-139页 |
| 7.2.2 Root dehydrogenase activity and lipid peroxidation | 第139-141页 |
| 7.2.3 Enzyme activities of carbohydrate metabolism in cotton roots | 第141页 |
| 7.2.4 Activities of enzymes involved in phenylpropanoid pathway (PAL, PPO, CAD, and SKDH) | 第141-143页 |
| 7.2.5 Changes in the contents of DPPH, flavonoids and callose in cotton root | 第143-145页 |
| 7.2.6 Activities of antioxidative enzymes in cotton root | 第145页 |
| 7.2.7 Glutathione dependent detoxification | 第145-146页 |
| 7.2.8 Disease resistance evaluation and plant growth | 第146-148页 |
| 7.2.9 Changes in the contents of amino acids in cotton root exudates | 第148-151页 |
| 7.2.10 In vitro effects of amino acids on V. dahliae growth | 第151-152页 |
| 7.3 Discussion | 第152-158页 |
| 7.4 Conclusion | 第158-159页 |
| Chapter 8 Liquid chromatography mass spectroscopy (LC-MS) analyses reveal thatphenolic compounds and endogenous plant hormones contribute to thebrassinosteroid mediated disease resistance | 第159-184页 |
| 8.1 Materials and methods | 第160-163页 |
| 8.1.1 Plant growth, inoculum preparation and treatment conditions | 第160页 |
| 8.1.2 Evaluation of disease resistance, H_2O_2 accumulation, electrolyte leakage and fungal colonization | 第160-161页 |
| 8.1.3 Measurement of enzyme activities of phenylpropanoid pathway | 第161页 |
| 8.1.4 Extraction of phenolic compounds and endogenous plant hormones | 第161页 |
| 8.1.5 Liquid chromatography mass spectroscopy(LC-MS/MS) | 第161-163页 |
| 8.1.6 Real-time quantitative PCR(qPCR)analysis | 第163页 |
| 8.1.7 Statistical analysis | 第163页 |
| 8.2 Results | 第163-178页 |
| 8.2.1 Dynamics of PAL and CAD | 第163-166页 |
| 8.2.2 Expression analysis of PAL and CAD through quantitative real time(qPCR) | 第166页 |
| 8.2.3 Patterns of phenolic compounds in cotton leaf and Vd toxin | 第166-171页 |
| 8.2.4 Changes in the levels of endogenous plant hormones | 第171-173页 |
| 8.2.5 Expression of genes involved in hormone synthesis | 第173-175页 |
| 8.2.6 H_2O_2 accumulation and electrolyte leakage of detached cotton leaves | 第175-178页 |
| 8.2.7 Effect of V.dahliae inoculation and EBR application on disease development,plant growth and fungus colonization | 第178页 |
| 8.3 Discussion | 第178-183页 |
| 8.4 Conclusion | 第183-184页 |
| References | 第184-202页 |
| List of Publications | 第202-203页 |
