水稻细菌性穗枯病病原鉴定及颖壳伯克氏菌（Burkholderia glumae）基因组甲基化研究

发布时间：2021-09-01 19:35

　　水稻细菌性穗枯病是由颖壳伯克氏菌（Burkholderia glumae）和唐菖蒲伯克氏菌（B.gladioli）引起,颖壳伯克氏菌是其主要病原。对细菌性穗枯病病原菌的遗传和表观遗传学特征进行研究,有助于制定其可持续防控方案。因此,我们在国内首次报道了一种伯克氏菌引起的水稻病害,同时对B.glumae的DNA甲基化和基因表达进行了研究。采用生物化学和分子生物学方法,从我国具有典型穗部枯萎病症状的病株中分离得到唐菖蒲伯克氏菌。DNA甲基化由DNA甲基化转移酶催化完成,是原核和真核生物基因组中常见的表观修饰形式,然而甲基化修饰在多数植物病原细菌与寄主互作过程中的功能还有待研究。B.glumae的DNA甲基化作用尚不明确。本项目旨在利用SMRT测序技术研究B.glumae菌株LMG2196的全基因组DNA甲基化模式。我们检测了B.glumae基因组中6mA和4mC两种甲基化形式,结果共检测到11,450预测的6mA残基和12,049预测的4mC残基,明确了6mA残基4mC残基在基因组中的位点;共同和菌株特异的DNA甲基化基序的分析结果表明DNA甲基化呈现高度的分歧;这些研究结果有助于推进细菌... 

【文章来源】：中国农业科学院北京市

【文章页数】：78 页

【学位级别】：博士

【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATION
CHAPTER 1 INTRODUCTION
CHAPTER 2 Identification and Diagnosis of Plant Pathogenic Bacteria Causing Rice Diseases
    2.1 Introduction
    2.2 First Report of Burkholderia gladioli Causing Rice Panicle Blight and Grain Discoloration in China
CHAPTER 3 Precision Genome wide DNA Methylations Characterization of Burkholderia glumae Using Single Molecule Real Time (SMRT) Sequencing
    3.1 Introduction
        3.1.1 Rice Bacterial Panicle Blight
        3.1.2 Epigenetic
        3.1.3 DNA Methylation
        3.1.4 Restriction-Modification Systems
        3.1.5 Third Generation Sequencing Technology
    3.2 Materials and Methods
        3.2.1 Strain growth condition and Genomic DNA Extraction
        3.2.2 DNA Dot blot assay
        3.2.3 DNA library preparation and sequencing
        3.2.4 Bioinformatic analyses of SMRT sequencing
        3.2.5 Functional enrichment analysis of the methylated promoter operons
        3.2.6 Metabolic Pathway Analysis
    3.3 Results
        3.3.1 Preliminary Analysis
        3.3.2 Deep Sequencing Analysis
        3.3.3 SMRT sequencing determines divergent DNA modification profiles
        3.3.4 Base Modification Site Annotation
        3.3.5 Motif Analysis
        3.3.6 KEGG and GO annotation analysis
        3.3.7 Dot Blot Assay
    3.4 Discussion
CHAPTER 4 A System to Study the Expression of Phytopathogenic Genes Encoded by Burkholderia glumae
    4.1 Introduction
    4.2 Materials and Methods
        4.2.1 Plant material and collection of root exudates
        4.2.2 Bacterial strain and Growth Condition
        4.2.3 RNA extraction and Quantitative Real-Time PCR
        4.2.4 Pathogen Assays
    4.3 Results
        4.3.1 Pathogenicity test
        4.3.2 Expression Patterns of Pathogenicity Genes
    4.4 Discussion
CHAPTER 5 MAJOR FINDINGS AND FUTURE PERSPECTIVES
    5.1 Major Findings
    5.2 Future Perspectives
REFERENCES
LIST OF PUBLICATIONS
ACKNOWLEDGEMENT
AUTHOR RESUME


【参考文献】：
期刊论文
[1]Research Status and Prospect of Burkholderia glumae, the Pathogen Causing Bacterial Panicle Blight[J]. CUI Zhou-qi,ZHU Bo,XIE Guan-lin,LI Bin,HUANG Shi-wen.  Rice Science. 2016(03)



本文编号：3377564