印度梨形孢對天寶蕉幼苗生長及致病相關類群基因表達的影響
發(fā)布時間：2020-11-05 03:48
香蕉是世界上重要的糧食作物。然而,香蕉在全球范圍內受到幾種病原菌的威脅,其中最嚴重的病原體之一是引起巴拿馬病的古巴�；图怄哏牭毒�(Fusarium oxysporum f.sp.Cubense,Foc)。受脅迫后的生理響應受基因表達的控制。遺憾的是,表現(xiàn)出抗性的野生型香蕉卻不具有市場需求的性狀,使用常規(guī)育種手段改良香蕉也往往需要耗費大量時間和高昂的成本。因此,深入了解香蕉受此類脅迫應激反應的遺傳調控,對香蕉產業(yè)的健康發(fā)展大有裨益。本研究利用有益微生物印度梨形孢(Piriformospora indica,P.indica)提出促進植物生長的同時又能有效防控枯萎病的技術方案。在現(xiàn)代園藝研究中,該真菌已被證明具有多種顯著生物學“促生增抗”的潛力。本研究鑒定了香蕉病程相關(Pathogenesis-related,PR)基因家族。構建了香蕉PR家族基因的系統(tǒng)發(fā)育樹,結果顯示香蕉中最大的PR基因家族PR 9亞家族,有117名成員。進一步研究P.indica對香蕉組培苗和組培移栽苗生長的影響。依據(jù)株高、葉片數(shù)和生根數(shù)等生長指標評估顯示,P.indica對兩組生長條件下的香蕉苗均具有促生作用。接種P.indica的組培和盆栽香蕉的平均葉片數(shù)、生根數(shù)、株高、總鮮重和地下部鮮重均顯著高于未接種的處理組。說明P.indica能夠促進香蕉的生長發(fā)育。P.indica的存在,即使在病原真菌的存在下,其生長也明顯增強,P.indica處理后的生長率相對優(yōu)于未經治療的此外,多種研究表明真菌可以提高植物抗氧化能力,這對植物防御至關重要。香蕉感染Foc的后期,發(fā)現(xiàn)植物的過氧化物酶、過氧化氫酶和超氧化物歧化酶的活性水平相對較高。CK組、接種P.indica的處理組、Foc 1處理組、定殖有P.indica的Foc 1處理組(Foc 1-P.indica)、Foc TR4處理組以及定殖有P.indica的FocTR4處理組(Foc TR4-P.indica)的SOD活性峰值分別出現(xiàn)在51、171、123、51、171和171 h;POD活性峰值分別在27、51、171、123、171和51 h;CAT活性峰值分別在51、171、3、123、51和3 h。真菌對試驗植物抗氧化活性的逐漸影響表明其參與了對Foc病原體感染的拮抗作用。qRT-PCR表達分析發(fā)現(xiàn)PR17受激素和Foc影響較大。FocTR4、ABA和MeJA分別在5、48和12 h達到最大值。GA_3處理在4 h和8 h達到兩次峰值,但這兩組表達無顯著差異。在實驗的早期階段,P。indica和Foc處理的樣品的表達水平相對較高,除了PR 5的情況相反。隨著暴露時間的延長,P。indica和Foc處理的樣品顯示各感興趣基因(PR1,PR2,PR5)的表達水平降低至其在P.indica和Foc處理下的表達低于對照的程度。檢測到的表達變化表明,P.indica參與了香蕉病原真菌的早期防御反應本研究說明了菌根真菌印度梨形孢對香蕉生長和枯萎病防控的作用,可為進一步研究香蕉抗逆遺傳改良奠定一定基礎。
【學位單位】：福建農林大學
【學位級別】：碩士
【學位年份】：2019
【中圖分類】：S436.68;S476.1
【文章目錄】：
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
【相似文獻】
本文編號：2871080
【學位單位】：福建農林大學
【學位級別】：碩士
【學位年份】：2019
【中圖分類】：S436.68;S476.1
【文章目錄】：
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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