印度梨形孢对天宝蕉幼苗生长及致病相关类群基因表达的影响
发布时间：2020-11-05 03:48
香蕉是世界上重要的粮食作物。然而,香蕉在全球范围内受到几种病原菌的威胁,其中最严重的病原体之一是引起巴拿马病的古巴专化型尖孢镰刀菌(Fusarium oxysporum f.sp.Cubense,Foc)。受胁迫后的生理响应受基因表达的控制。遗憾的是,表现出抗性的野生型香蕉却不具有市场需求的性状,使用常规育种手段改良香蕉也往往需要耗费大量时间和高昂的成本。因此,深入了解香蕉受此类胁迫应激反应的遗传调控,对香蕉产业的健康发展大有裨益。本研究利用有益微生物印度梨形孢(Piriformospora indica,P.indica)提出促进植物生长的同时又能有效防控枯萎病的技术方案。在现代园艺研究中,该真菌已被证明具有多种显著生物学“促生增抗”的潜力。本研究鉴定了香蕉病程相关(Pathogenesis-related,PR)基因家族。构建了香蕉PR家族基因的系统发育树,结果显示香蕉中最大的PR基因家族PR 9亚家族,有117名成员。进一步研究P.indica对香蕉组培苗和组培移栽苗生长的影响。依据株高、叶片数和生根数等生长指标评估显示,P.indica对两组生长条件下的香蕉苗均具有促生作用。接种P.indica的组培和盆栽香蕉的平均叶片数、生根数、株高、总鲜重和地下部鲜重均显著高于未接种的处理组。说明P.indica能够促进香蕉的生长发育。P.indica的存在,即使在病原真菌的存在下,其生长也明显增强,P.indica处理后的生长率相对优于未经治疗的此外,多种研究表明真菌可以提高植物抗氧化能力,这对植物防御至关重要。香蕉感染Foc的后期,发现植物的过氧化物酶、过氧化氢酶和超氧化物歧化酶的活性水平相对较高。CK组、接种P.indica的处理组、Foc 1处理组、定殖有P.indica的Foc 1处理组(Foc 1-P.indica)、Foc TR4处理组以及定殖有P.indica的FocTR4处理组(Foc TR4-P.indica)的SOD活性峰值分别出现在51、171、123、51、171和171 h;POD活性峰值分别在27、51、171、123、171和51 h;CAT活性峰值分别在51、171、3、123、51和3 h。真菌对试验植物抗氧化活性的逐渐影响表明其参与了对Foc病原体感染的拮抗作用。qRT-PCR表达分析发现PR17受激素和Foc影响较大。FocTR4、ABA和MeJA分别在5、48和12 h达到最大值。GA_3处理在4 h和8 h达到两次峰值,但这两组表达无显著差异。在实验的早期阶段,P。indica和Foc处理的样品的表达水平相对较高,除了PR 5的情况相反。随着暴露时间的延长,P。indica和Foc处理的样品显示各感兴趣基因(PR1,PR2,PR5)的表达水平降低至其在P.indica和Foc处理下的表达低于对照的程度。检测到的表达变化表明,P.indica参与了香蕉病原真菌的早期防御反应本研究说明了菌根真菌印度梨形孢对香蕉生长和枯萎病防控的作用,可为进一步研究香蕉抗逆遗传改良奠定一定基础。
【学位单位】：福建农林大学
【学位级别】：硕士
【学位年份】：2019
【中图分类】：S436.68;S476.1
【文章目录】：
Appendix List of Signs and Abbreviations
摘要
Abstract
CHAPTER 1 Introduction
1.2 Banana （Musa L.）
1.3 Piriformospora indica （P. indica）
1.4 Nature of plant-fungi （P. indica） symbiotic interaction
1.5 Interaction of P. indica with other rhizobacteria in the soil matrix
1.6 Areas for imminent research exploits using P. indica
1.6.1 The mass production of plant secondary metabolites for industrial purposes
1.6.2 Improvement of plant tissue culture and mass propagation systems
1.6.3 Improvement of production and growth of economically important plant biomass
1.6.4 Improvement of germination rate and vigor in seedling production systems
1.6.5 Development of superior and improved breeds in rootstock breeding
1.6.6 Improvement of tolerance in plants under stress duress
1.6.7 Molecular regulator in the expression of genes associated with biotic resistance
1.6.8 Nourishing plants and soil rhizosphere as an eco-friendly bio-fertilizer
1.7 Safety of the use of P. indica as a bio-fertilizer
1.8 Fusarium Wilt
1.9 Pathogenesis-related proteins
1.10 PHYTOHORMONES
1.10.1 Salicylic acid （SA）, Jasmonic acid （JA）, Ethylene （ET） and Auxins
1.11 The Significance of Study
1.12 Impact of Research
1.13 Research Aims and Objectives
1.13.1 General objective
1.13.2 Specific objectives
CHAPTER 2 Analysis of influence of Piriformaspora indica on physiology and growth ofbanana
2.2 Materials and methods
2.2.1 Cultivation and preparation of P. indica
2.2.2 Plant material and in-vitro inoculation
2.2.3 Preparation of Foc TR4 filtrate
2.2.4 In-vitro inoculation
2.2.5 In soil inoculation
2.2.6 Effects of P. indica on the growth of Tianbaojiao banana
2.3 Results
2.3.1 Development patterns of Tianbaojiao banana upon P. indica treatment
2.3.1.1 Effects under tissue culture conditions
2.3.1.2 Effects under potted/soil conditions
2.3.2 Banana plantlet survival rate
2.4 Discussion
CHAPTER 3 Antioxidant enzyme activity analysis
3.2 Materials and methods
3.2.1 Chemicals and reagents
3.2.2 Plant material
3.2.3 Extraction of Antioxidant assay
3.2.4 Measurement of Antioxidant enzyme activity
3.2.4.1 Catalase
3.2.4.2 Peroxidase
3.2.4.3 Superoxide dismutase
3.3 Results
3.4 Discussion
CHAPTER 4 Bioinformatics analysis of pathogenesis-related genes
4.1 Materials and methods
4.2 RESULTS
4.2.1 PR gene BLAST
4.2.2 Phylogeny relationships of Banana PR1, PR2, PR5 and PR 17 gene families
4.2.3 Conserved motif composition analysis
4.2.4 Promoter cis-acting element analysis
4.2.5 Protein-protein interaction （PPI） network construction
4.2.6 Codon bias analysis plant Basic Secretory Protease （PR 17）
4.3 Discussion
4.4 Conclusion
CHAPTER 5 Gene expression analysis
5.2 Materials and methods
5.2.1 Plant material
5.2.2 Preparation of Foc TR4 filtrate
5.2.3 Treatment application
5.2.4 RNA extraction and c DNA synthesis
5.2.5 Quantitative real-time PCR （q RT-PCR）
5.2.6 Gene expression analysis
5.3 Results
5.3.1 Gene expression of PR 17
5.3.2 Gene expression levels PR 1, PR 2, and PR 5
5.4 Discussion
CHAPTER 6 Summary and future prospects
References
Acknowledgeement
Appendix
【相似文献】
本文编号：2871080
【学位单位】：福建农林大学
【学位级别】：硕士
【学位年份】：2019
【中图分类】：S436.68;S476.1
【文章目录】：
Appendix List of Signs and Abbreviations
摘要
Abstract
CHAPTER 1 Introduction
1.2 Banana （Musa L.）
1.3 Piriformospora indica （P. indica）
1.4 Nature of plant-fungi （P. indica） symbiotic interaction
1.5 Interaction of P. indica with other rhizobacteria in the soil matrix
1.6 Areas for imminent research exploits using P. indica
1.6.1 The mass production of plant secondary metabolites for industrial purposes
1.6.2 Improvement of plant tissue culture and mass propagation systems
1.6.3 Improvement of production and growth of economically important plant biomass
1.6.4 Improvement of germination rate and vigor in seedling production systems
1.6.5 Development of superior and improved breeds in rootstock breeding
1.6.6 Improvement of tolerance in plants under stress duress
1.6.7 Molecular regulator in the expression of genes associated with biotic resistance
1.6.8 Nourishing plants and soil rhizosphere as an eco-friendly bio-fertilizer
1.7 Safety of the use of P. indica as a bio-fertilizer
1.8 Fusarium Wilt
1.9 Pathogenesis-related proteins
1.10 PHYTOHORMONES
1.10.1 Salicylic acid （SA）, Jasmonic acid （JA）, Ethylene （ET） and Auxins
1.11 The Significance of Study
1.12 Impact of Research
1.13 Research Aims and Objectives
1.13.1 General objective
1.13.2 Specific objectives
CHAPTER 2 Analysis of influence of Piriformaspora indica on physiology and growth ofbanana
2.2 Materials and methods
2.2.1 Cultivation and preparation of P. indica
2.2.2 Plant material and in-vitro inoculation
2.2.3 Preparation of Foc TR4 filtrate
2.2.4 In-vitro inoculation
2.2.5 In soil inoculation
2.2.6 Effects of P. indica on the growth of Tianbaojiao banana
2.3 Results
2.3.1 Development patterns of Tianbaojiao banana upon P. indica treatment
2.3.1.1 Effects under tissue culture conditions
2.3.1.2 Effects under potted/soil conditions
2.3.2 Banana plantlet survival rate
2.4 Discussion
CHAPTER 3 Antioxidant enzyme activity analysis
3.2 Materials and methods
3.2.1 Chemicals and reagents
3.2.2 Plant material
3.2.3 Extraction of Antioxidant assay
3.2.4 Measurement of Antioxidant enzyme activity
3.2.4.1 Catalase
3.2.4.2 Peroxidase
3.2.4.3 Superoxide dismutase
3.3 Results
3.4 Discussion
CHAPTER 4 Bioinformatics analysis of pathogenesis-related genes
4.1 Materials and methods
4.2 RESULTS
4.2.1 PR gene BLAST
4.2.2 Phylogeny relationships of Banana PR1, PR2, PR5 and PR 17 gene families
4.2.3 Conserved motif composition analysis
4.2.4 Promoter cis-acting element analysis
4.2.5 Protein-protein interaction （PPI） network construction
4.2.6 Codon bias analysis plant Basic Secretory Protease （PR 17）
4.3 Discussion
4.4 Conclusion
CHAPTER 5 Gene expression analysis
5.2 Materials and methods
5.2.1 Plant material
5.2.2 Preparation of Foc TR4 filtrate
5.2.3 Treatment application
5.2.4 RNA extraction and c DNA synthesis
5.2.5 Quantitative real-time PCR （q RT-PCR）
5.2.6 Gene expression analysis
5.3 Results
5.3.1 Gene expression of PR 17
5.3.2 Gene expression levels PR 1, PR 2, and PR 5
5.4 Discussion
CHAPTER 6 Summary and future prospects
References
Acknowledgeement
Appendix
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