马铃薯立枯丝核菌毒素的分离鉴定及对茎溃疡的致病机理研究
发布时间：2021-11-04 03:50
近年来随着马铃薯产业的迅猛发展,我国马铃薯主产区土传病害日趋加重,尤其是由立枯丝核菌（Rhizoctonia solani）侵染引起的马铃薯茎溃疡病（也称马铃薯立枯病,Potato Rhizoctonia canker,黑痣病,black scurf）已成为影响马铃薯产业发展的限制因子,不仅严重影响马铃薯块茎的产量和品质,降低其商品性和贮藏性,也给种植户造成了巨大的经济损失。立枯丝核菌对马铃薯植株的致病机理十分复杂,产生的毒素可能是其致病的关键因子,探明立枯丝核菌毒素在植株发病过程中的作用,对防治马铃薯黑痣病具有重要意义。本研究拟采用高效液相色谱（HPLC）、薄层色谱（TLC）、柱层析、红外光谱（FTIR）、质谱和核磁（NMR）分析等手段分离和鉴定病原菌毒素的活性成分,同时用毒素处理马铃薯茎基部,综合采用植物病理学、生理学和细胞生物学等多学科的方法,揭示病原菌毒素对马铃薯茎溃疡的致病机理,为更有效地防治马铃薯茎溃疡病提供理论依据。主要试验结果如下:1.立枯丝核菌AG-3毒素的分离和鉴定（1）将分离到的38个样品进行TLC分析,在254nm紫外光下可以看见清晰的斑点,制备TLC的最佳展开...
【文章来源】：甘肃农业大学甘肃省
【文章页数】：116 页
【学位级别】：博士
【文章目录】：
摘要
Summary
Abbreviations
Chapter 1 Introduction
1.1 Economic importance of potato
1.2 Phytopathogenic fungi attacking potato
1.3 Rhizoctonia diseases of potato
1.3.1 Rhizoctonia disease cycle and disease symptoms
1.3.2 Anastomosis groups of R. solani
1.3.3 Epidemiology of R. solani
1.4 The management of Rhizoctonia potato disease
1.4.1 Biocontrol
1.4.2 Cultural practices
1.4.3 Chemical control
1.4.4 Preformed defense systems
1.4.5 Organic matter amendments
1.5 Fungal phytotoxins
1.5.1 Definitions and chemical structures
1.5.2 Phytotoxins from Rhizoctonia solani
1.6 Objectives and significance of the study
Chapter 2 Isolation, purification and characterization of phytotoxins produced byRhizoctonia solani AG-3, the caused agent of potato stem canker
2.1 Introduction
2.2 Materials and methods
2.2.1 Collection of sample
2.2.2 Maintenance and storage of fungal cultures
2.2.3 Culture media
2.2.4 Isolation and identification of fungal pathogens
2.2.5 Fungal cultures and toxin production
2.2.6 Biological activity and stem canker assay
2.2.7 Bioactivity of soluble macromolecular compounds obtained from culturefiltrates
2.2.8 Solvent extraction
2.2.9 Isolation and purification of the phytotoxins from culture filtrates
2.2.10 Characterization of purified compound
2.2.11 Potato stem canker response to MTPA and MTAA
2.2.12 Statistical analysis
2.3 Results
2.3.1 Isolation and identification of fungi isolates
2.3.2 Preliminary in vitro characterization
2.3.3 Isolation and purification of R. solani toxin
2.3.4 Potato stem canker response to toxin fractions
2.3.5 IR spectra of the purified compounds
2.3.6 HPLC of the purified compounds
2.3.7 NMR spectra of the purified compounds
2.3.8 Potato stem canker response to MTPA and MTAA
2.4 Discussion
Chapter 3 Investigation of 3-methylthiopropionic acid of Rhizoctonia solani AG-3 and itsrole in the pathogenicity of the fungus
3.1 Introduction
3.2 Materials and methods
3.2.1 Soil sterilization, source of seed and MTPA
3.2.2 Potato stems canker response to MTPA
3.2.3 Rhizoctonia growth response to MTPA
3.2.4 Fungal culture filtrates and toxin production
3.2.5 Bioassay for phytotoxicity
3.2.6 Influence of pH on toxin activity
3.2.7 Experimental design
3.2.8 Data collection
3.2.9 Statistical analyses
3.3 Results
3.3.1 The effect of pH on phytotoxin activity
3.3.2 Plant height, stem diameter, number of stolon and symptomless stolon
3.3.3 Canopy development
3.3.4 Effect of MTPA phytotoxin on yield of potato plants
3.3.5 Biomass of potato plants
3.3.6 Chlorophyll content
3.3.7 Potato seedling root response to MTPA
3.3.8 Assessment of disease of potato plants
3.4 Discussion
Chapter 4 Investigation of mechanisms of Rhizoctonia solani phytotoxins in causingpotato stem canker and antioxidant enzyme activities
4.1 Introduction
4.2 Materials and methods
4.2.1 Soil sterilization, potato cultivars, fungal isolates and MTPA
4.2.2 Experimental design and treatment description
4.2.3 Data Collected
4.2.4 Enzymatic assays
4.2.5 Statistical analysis
4.3 Results
4.3.1 Soil physicochemical properties
4.3.2 Effect of R. solani phytotoxin on stem canker
4.3.3 Influence of R. solani phytotoxin on black scurf of potato
4.3.4 Effect of MTPA phytotoxin on MDA content
4.3.5 Effect of MTPA phytotoxin on SOD activity
4.3.6 Effect of MTPA phytotoxin on POD activity
4.3.7 Effect of MTPA phytotoxin on CAT activity
4.3.8 Effect of MTPA phytotoxin on chitinase activity
4.3.9 Relationship between the antioxidant enzyme activities and stem canker ofpotato
4.4 Discussion
Chapter 5 General conclusions and future works
5.1 General conclusions
5.2 Future works
References
Acknowledgements
Personal Resume
Publications originating from this dissertation
Supervisor’s Brief Introduction
本文编号：3474940
【文章来源】：甘肃农业大学甘肃省
【文章页数】：116 页
【学位级别】：博士
【文章目录】：
摘要
Summary
Abbreviations
Chapter 1 Introduction
1.1 Economic importance of potato
1.2 Phytopathogenic fungi attacking potato
1.3 Rhizoctonia diseases of potato
1.3.1 Rhizoctonia disease cycle and disease symptoms
1.3.2 Anastomosis groups of R. solani
1.3.3 Epidemiology of R. solani
1.4 The management of Rhizoctonia potato disease
1.4.1 Biocontrol
1.4.2 Cultural practices
1.4.3 Chemical control
1.4.4 Preformed defense systems
1.4.5 Organic matter amendments
1.5 Fungal phytotoxins
1.5.1 Definitions and chemical structures
1.5.2 Phytotoxins from Rhizoctonia solani
1.6 Objectives and significance of the study
Chapter 2 Isolation, purification and characterization of phytotoxins produced byRhizoctonia solani AG-3, the caused agent of potato stem canker
2.1 Introduction
2.2 Materials and methods
2.2.1 Collection of sample
2.2.2 Maintenance and storage of fungal cultures
2.2.3 Culture media
2.2.4 Isolation and identification of fungal pathogens
2.2.5 Fungal cultures and toxin production
2.2.6 Biological activity and stem canker assay
2.2.7 Bioactivity of soluble macromolecular compounds obtained from culturefiltrates
2.2.8 Solvent extraction
2.2.9 Isolation and purification of the phytotoxins from culture filtrates
2.2.10 Characterization of purified compound
2.2.11 Potato stem canker response to MTPA and MTAA
2.2.12 Statistical analysis
2.3 Results
2.3.1 Isolation and identification of fungi isolates
2.3.2 Preliminary in vitro characterization
2.3.3 Isolation and purification of R. solani toxin
2.3.4 Potato stem canker response to toxin fractions
2.3.5 IR spectra of the purified compounds
2.3.6 HPLC of the purified compounds
2.3.7 NMR spectra of the purified compounds
2.3.8 Potato stem canker response to MTPA and MTAA
2.4 Discussion
Chapter 3 Investigation of 3-methylthiopropionic acid of Rhizoctonia solani AG-3 and itsrole in the pathogenicity of the fungus
3.1 Introduction
3.2 Materials and methods
3.2.1 Soil sterilization, source of seed and MTPA
3.2.2 Potato stems canker response to MTPA
3.2.3 Rhizoctonia growth response to MTPA
3.2.4 Fungal culture filtrates and toxin production
3.2.5 Bioassay for phytotoxicity
3.2.6 Influence of pH on toxin activity
3.2.7 Experimental design
3.2.8 Data collection
3.2.9 Statistical analyses
3.3 Results
3.3.1 The effect of pH on phytotoxin activity
3.3.2 Plant height, stem diameter, number of stolon and symptomless stolon
3.3.3 Canopy development
3.3.4 Effect of MTPA phytotoxin on yield of potato plants
3.3.5 Biomass of potato plants
3.3.6 Chlorophyll content
3.3.7 Potato seedling root response to MTPA
3.3.8 Assessment of disease of potato plants
3.4 Discussion
Chapter 4 Investigation of mechanisms of Rhizoctonia solani phytotoxins in causingpotato stem canker and antioxidant enzyme activities
4.1 Introduction
4.2 Materials and methods
4.2.1 Soil sterilization, potato cultivars, fungal isolates and MTPA
4.2.2 Experimental design and treatment description
4.2.3 Data Collected
4.2.4 Enzymatic assays
4.2.5 Statistical analysis
4.3 Results
4.3.1 Soil physicochemical properties
4.3.2 Effect of R. solani phytotoxin on stem canker
4.3.3 Influence of R. solani phytotoxin on black scurf of potato
4.3.4 Effect of MTPA phytotoxin on MDA content
4.3.5 Effect of MTPA phytotoxin on SOD activity
4.3.6 Effect of MTPA phytotoxin on POD activity
4.3.7 Effect of MTPA phytotoxin on CAT activity
4.3.8 Effect of MTPA phytotoxin on chitinase activity
4.3.9 Relationship between the antioxidant enzyme activities and stem canker ofpotato
4.4 Discussion
Chapter 5 General conclusions and future works
5.1 General conclusions
5.2 Future works
References
Acknowledgements
Personal Resume
Publications originating from this dissertation
Supervisor’s Brief Introduction
本文编号：3474940
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