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水稻白叶枯病菌3个GGDEF结构域蛋白的功能鉴定

摘要第5-6页
ABSTRACT第6-7页
LIST OF ABBREVIATIONS, ACRONYMS AND SYMBOLS第8-16页
CHAPTER I GENERAL INTRODUCTION第16-36页
    1.1 Research survey of the rice pathogen Xanthomonas oryzae pv. oryzae第16-22页
        1.1.1 Genomic study on Xanthomonas oryzae pv. oryzae第16-18页
        1.1.2 Pathogenic traits from Xanthomonas oryzae pv. oryzae contributing host-plant interaction第18-22页
            1.1.2.1 Surface-associated virulence factors第18-19页
                1.1.2.1.1 Extracellular polysaccharides (EPSs)第18-19页
                1.1.2.1.2 Lipopolysaccharides (LPS)第19页
            1.1.2.2 Type II secretion system第19-20页
            1.1.2.3 Type III secretion system第20-21页
            1.1.2.4 Other factors involved in virulence of Xanthomonas oryzae pv. oryzae第21-22页
    1.2 Cyclic di-GMP signaling in the virulence and environmental adaptation of Xanthomonas第22-25页
        1.2.1 Cyclic di-GMP generation第24页
        1.2.2 Cyclic di-GMP dissolution第24-25页
    1.3 Cyclic di-GMP receptor types第25-28页
        1.3.1 Pli Z domain containing proteins第25-26页
        1.3.2 Degenerate GGDEF, EAL domains as c-di-GMP receptors第26-27页
        1.3.3 Transcription factors第27页
        1.3.4 Riboswitches第27-28页
    1.4 Cyclic di-GMP regulated cell functions第28-32页
        1.4.1 Regulation of motility by c-di-GMP第28-29页
        1.4.2 Regulation of biofilm formation by c-di-GMP第29-30页
        1.4.3 Regulation of virulence by c-di-GMP第30-31页
        1.4.4 C-di-GMP regulates cell cycle and cell differentiation第31-32页
    1.5 Cyclic di-GMP signaling system in bacterial leaf blight disease of rice第32-34页
    1.6 Significance of research study and objectives第34-35页
    1.7 Thesis outline第35-36页
CHAPTER II GENERATION OF SINGLE GENE-KNOCKOUT MUTANTS第36-50页
    2.1 Experimental materials第37-38页
        2.1.1 Bacterial strains and plasmids第37页
        2.1.2 Reagents and equipment第37页
        2.1.3 Culture media and growth conditions第37-38页
        2.1.4 Maintenance of bacterial strains and plasmids第38页
    2.2 Experimental methods第38-41页
        2.2.1 Genomic DNA extraction第38-39页
        2.2.2 Generation of PCR fragments from genomic DNA第39页
        2.2.3 Cloning of gene fragments with middle vector第39-40页
        2.2.4 Construction of vector plasmids第40页
        2.2.5 Preparation of Xoo PXO99A competent cells第40页
        2.2.6 Generation of single gene-knockout mutants第40页
        2.2.7 Preservation第40-41页
    2.3 Results第41-49页
        2.3.1 Bioinformatics and homology alignments of GGDEF-domain proteins in the genome of X. oryzae pv. oryzae第41-44页
        2.3.2 Construction of p KMS1 vector plasmids第44-47页
        2.3.3 Generation of single gene-knockout mutants第47-49页
    2.4 Discussion第49-50页
CHAPTER III GENERATION OF MULTIPLE GENE-KNOCKOUT MUTANTS第50-57页
    3.1 Experimental materials第50-52页
        3.1.1 Bacterial strains and plasmids第50页
        3.1.2 Reagents and equipment第50-51页
        3.1.3 Culture media and culture condition第51-52页
        3.1.4 Maintenance of bacterial strains and plasmids第52页
    3.2 Experimental methods第52-53页
        3.2.1 Preparation of competent cells第52页
        3.2.2 Generation of double gene deletion mutants第52-53页
        3.2.3 Generation of triple gene deletion mutant第53页
        3.2.4 Preservation第53页
    3.3 Results第53-56页
        3.3.1 Construction of p KMS1 vector plasmids第53页
        3.3.2 Generation of double and triple-gene knockout mutants第53-56页
    3.4 Discussion第56-57页
CHAPTER IV PHENOTYPIC ANALYSIS OF SINGLE AND MULTIPLE GENE-KNOCKOUT MUTANTS第57-69页
    4.1 Experimental materials第58页
        4.1.1 Bacterial strains and plasmids第58页
        4.1.2 Reagents and equipment第58页
        4.1.3 Culture media and culture condition第58页
    4.2 Experimental methods第58-59页
        4.2.1 Virulence assay第58-59页
        4.2.2 Motility assay第59页
        4.2.3 EPS production第59页
        4.2.4 Biofilm formation第59页
    4.3 Statistical analyses第59页
    4.4 Results第59-66页
        4.4.1 GGDEF-domain proteins modulate X. oryzae pv. oryzae motility第59-61页
        4.4.2 GGDEF protein regulates EPS production第61页
        4.4.3 GGDEF-domain influence biofilm formation第61页
        4.4.4 GGDEF-domain proteins regulate X. oryzae pv. oryzae virulence in rice第61-66页
    4.5 Discussion第66-69页
CHAPTER V SUMMARY第69-70页
REFERENCES CITED第70-93页
APPENDICES第93-97页
ACKNOWLEDGEMENT第97-99页
CURRICULUM VITAE第99-100页

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