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阿维菌素降解菌株的分离、鉴定及生物降解途径研究

摘要第10-12页
ABSTRACT第12-14页
LIST OF ABBREVIATIONS第15-17页
LIST OF TABLES第17-18页
LIST OF FIGURES第18-19页
CHAPTER 1 INTRODUCTION第19-21页
CHAPTER 2 REVIEW OF LITERATURE第21-51页
    1. AVERMECTINS第21-22页
    2. ABAMECTIN第22-23页
    3. GENERAL PROPERTIES第23页
    4. PHYSICAL/CHEMICAL PROPERTIES第23-24页
    5. MODE OF ACTION第24-26页
    6. METABOLISM第26-27页
    7. TOXICITY第27-28页
    8. ENVIRONMENTAL FATE第28-41页
        8.1. Concentrations and persistence in environmental compartments第28-32页
        8.2. Impacts on terrestrial fauna第32-36页
            8.2.1. Lethal impacts第32-33页
            8.2.2. Sublethal impacts第33-36页
        8.3. Impacts on aquatic fauna第36-41页
            8.3.1. Impacts on target aquatic animals第37-38页
            8.3.2. Impacts on non-target marine organisms第38-39页
            8.3.3. Impacts on non-target freshwater organisms第39-41页
    9. BIODEGRADATION/BIOREMEDIATION第41-44页
    References第44-51页
CHAPTER 3 ISOLATION AND CHARACTERIZATION OF BACTERIAL STRAIN GB-01第51-78页
    Abstract第51页
    1. INTRODUCTION第51-53页
    2. MATERIALS AND METHODS第53-64页
        2.1 Chemicals第53页
        2.2 Culture media第53-54页
            2.2.1 Washing and sterilization of glassware第53页
            2.2.2 Medium preparation第53-54页
        2.3. Enrichment of the abamectin-degrading consortium第54-55页
        2.4 Isolation of pure culture第55-57页
        2.5 Maintenance of bacterial culture第57页
        2.6 Identification and characterization of strain GB-01第57-60页
            2.6.1 Morphological characterization第57-59页
                2.6.1.1 Transmission electron microscopy第57-58页
                2.6.1.2 Smear preparation and Gram staining第58-59页
                2.6.1.3 Cultural characterization第59页
            2.6.2 Physiological and biochemical characterization第59-60页
                2.6.2.1 Fermentation of sugars第59页
                2.6.2.2 Methyl red and Voges Praskaeur test第59页
                2.6.2.3 Indole test第59页
                2.6.2.4 Citrate utilization test第59-60页
                2.6.2.5 Nitrate reduction test第60页
                2.6.2.6 Catalase test第60页
                2.6.2.7 Liquefication of gelatin第60页
        2.7 Biodegradation assay of abamectin by strain GB-01第60-63页
            2.7.1 Inoculum preparation第60-61页
            2.7.2 Construction of reactors第61页
            2.7.3 Effects of Temperature第61-62页
            2.7.4 Effects of pH第62页
            2.7.5 Effects of initial concentration第62-63页
            2.7.6 Biodegradation kinetics第63页
        2.8 Analytical techniques第63-64页
            2.8.1 Extraction of abamectin for HPLC第63页
            2.8.2 High pressure liquid chromatography (HPLC)第63-64页
            2.8.3 Calibration of HPLC instrument第64页
    3. RESULTS AND DISCUSSION第64-72页
        3.1 Degradability of abamectin through enrichment culture第64-65页
        3.2 Isolation and characterization of the strain GB-01第65-68页
        3.3 Biodegradation of abamectin by strain GB-01 in cell culture第68-69页
        3.4 Degradation at different temperatures第69-70页
        3.5 Degradation at different pH第70-71页
        3.6 Degradation at different initial concentrations of abamectin第71-72页
    4. CONCLUSION第72-74页
    References第74-78页
CHAPTER 4 TAXONOMIC ANALYSIS OF STRAIN GB-01第78-126页
    Abstract第78页
    1. INTRODUCTION第78-86页
        1.1 History and structure of the Burkholderia genus第78-81页
        1.2 Natural diversity and potential application of Burkholderia cepacia complex species第81-83页
        1.3 Identification and Classification of Bcc第83-84页
        1.4 Research aim第84-86页
    2. MATERIALS AND METHODS第86-96页
        2.1 Bacterial strain and culture conditions第86页
        2.2 Genomic DNA extraction第86-87页
        2.3 DNA quantification and purity check第87页
        2.4 Polymerase chain reaction (PCR)第87-88页
        2.5 Agarose gel electrophoresis第88页
        2.6 DNA (PCR product) purification from agarose gel第88-90页
        2.7 Cloning of PCR product第90页
        2.8 Transformation of E. coli第90-91页
        2.9 Plasmid purification from E. coli第91-92页
        2.10 Restriction endonuclease digestion第92页
        2.11 Cycle sequencing of DNA第92页
        2.12 Molecular sequence analysis第92-93页
        2.13 Biochemical and substrate utilization test第93-95页
            2.13.1 API 20 NE第93页
            2.13.2 ID 32 GN第93-94页
            2.13.3 API 50 CH第94页
            2.13.4 Biolog GN2 MicroPlate第94-95页
        2.14 Whole cell fatty acid profiling第95页
        2.15 Whole genome G+C mol% determination第95页
        2.16 Whole genome DNA-DNA hybridization第95-96页
    3. RESULTS AND DISCUSSION第96-116页
        3.1 Biochemical and substrate utilization characteristics of strain GB-01第96-104页
        3.2 Whole cell fatty acid profile of strain GB-01第104-106页
        3.3 G+C mol% of Strain GB-01第106-108页
        3.4 Phylogenetic analysis of 16S rRNA gene第108-111页
        3.5 Amplification of recA gene fragments from strain GB-01第111-113页
        3.6 Phylogenetic analysis of recA gene第113-115页
        3.7 Whole genome DNA-DNA hybridization第115-116页
    4. CONCLUSION第116-117页
    References第117-126页
CHAPTER 5 BIODEGRADATION PATHWAY OF ABAMECTIN BY STRAIN GB-01第126-145页
    Abstract第126页
    1. INTRODUCTION第126-133页
        1.1 High performance liquid chromatography (HPLC)第127-128页
        1.2 Mass spectrometry第128-131页
            1.2.1 Triple quadrupole mass spectrometry第129-131页
        1.3 Electrospray Ionization (ESI)第131-133页
    2. MATERIALS AND METHODS第133-135页
        2.1 Chemicals, bacterium and culture media第133-134页
        2.2 Instrumentation第134页
        2.3 Biodegradation experimental system第134-135页
        2.4 Sample preparation第135页
    3. RESULTS AND DISCUSSION第135-141页
        3.1 Degradation products第135-138页
        3.2 Structural elucidation of the metabolites第138-141页
    4. CONCLUSION第141-142页
    References第142-145页
CHAPTER 6 EVALUATION OF BIOREMEDIATION POTENTIAL OF STRAIN GB-01 IN SOIL MICROCOSMS第145-163页
    Abstract第145页
    1. INTRODUCTION第145-147页
    2. MATERIALS AND METHODS第147-151页
        2.1 Chemicals, organism and culture conditions第147页
        2.2 Effect of different factors on the growth of strain GB-01第147-148页
        2.3 Induction studies第148页
        2.4 Preparation of the inoculum and soil microcosms第148-150页
        2.5 Study of different factors affecting the abamectin degradation in soil microcosms第150-151页
        2.6 Extraction and HPLC analysis第151页
        2.7 Reproducibility第151页
    3. RESULTS AND DISCUSSION第151-158页
        3.1 Growth characterization of strain GB-01 for inoculum preparation第152页
        3.2 Effects of substrate concentration第152-154页
        3.3 Effects of pH and temperature第154-156页
        3.4 Effects of inoculum size, inoculation frequency and flooded conditions第156-158页
    4. CONCLUSION第158-159页
    References第159-163页
SUMMARY AND CONCLUSIONS第163-165页
SIGNIFICANCE OF THE RESEARCH第165-167页
FUTURE PERSPECTIVES第167-169页
PUBLICATIONS第169-171页
APPENDICES第171-173页
ACKNOWLEDGMENT第173-174页

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