水稻細(xì)菌性穗枯病是由穎殼伯克氏菌（Burkholderia glumae）和唐菖蒲伯克氏菌（B.gladioli）引起,穎殼伯克氏菌是其主要病原。對細(xì)菌性穗枯病病原菌的遺傳和表觀遺傳學(xué)特征進(jìn)行研究,有助于制定其可持續(xù)防控方案。因此,我們在國內(nèi)首次報道了一種伯克氏菌引起的水稻病害,同時對B.glumae的DNA甲基化和基因表達(dá)進(jìn)行了研究。采用生物化學(xué)和分子生物學(xué)方法,從我國具有典型穗部枯萎病癥狀的病株中分離得到唐菖蒲伯克氏菌。DNA甲基化由DNA甲基化轉(zhuǎn)移酶催化完成,是原核和真核生物基因組中常見的表觀修飾形式,然而甲基化修飾在多數(shù)植物病原細(xì)菌與寄主互作過程中的功能還有待研究。B.glumae的DNA甲基化作用尚不明確。本項目旨在利用SMRT測序技術(shù)研究B.glumae菌株LMG2196的全基因組DNA甲基化模式。我們檢測了B.glumae基因組中6mA和4mC兩種甲基化形式,結(jié)果共檢測到11,450預(yù)測的6mA殘基和12,049預(yù)測的4mC殘基,明確了6mA殘基4mC殘基在基因組中的位點;共同和菌株特異的DNA甲基化基序的分析結(jié)果表明DNA甲基化呈現(xiàn)高度的分歧;這些研究結(jié)果有助于推進(jìn)細(xì)菌... 

【文章來源】：中國農(nóng)業(yè)科學(xué)院北京市

【文章頁數(shù)】：78 頁

【學(xué)位級別】：博士

【文章目錄】：
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


【參考文獻(xiàn)】：
期刊論文
[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)



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