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甜高粱遗传转化体系优化及烟草果胶质多糖修饰研究

摘要第7-8页
abstract第8-9页
Abbreviation第17-20页
Chapter Ⅰ Optimization of callus induction and plant regeneration from different explants in sorghum(Sorghum bicolor)第20-68页
    1.0 Introduction第20-24页
    1.1 Transformation methods第24-28页
        1.1.1 Micro projectile transformation第24-26页
        1.1.2 Agrobacterium-mediated transformation第26-27页
        1.1.3 Electroporation-mediated transformation第27页
        1.1.4 Pollen-mediated transformation第27-28页
    1.2 Factors influencing the sorghum transformation第28-45页
        1.2.1 Genotypes第28-29页
        1.2.2 Sources of explants第29-30页
        1.2.3 Agrobacterium strains and vectors第30-31页
        1.2.4 Marker selection第31-32页
        1.2.5 Reporter genes第32-33页
        1.2.6 Agrobacterium concentration第33-34页
        1.2.7 In-vitro culture media第34-35页
        1.2.8 Osmotic treatment第35-36页
        1.2.9 Antioxidants第36页
        1.2.10 Antibiotics第36-37页
        1.2.11 Phenolic compounds第37-38页
        1.2.12 Temperature第38-45页
    1.3 Materials and Methods第45-48页
        1.3.1 Plant materials第45页
        1.3.2 Regeneration第45-46页
        1.3.3 Agrobacterium strains and binary vectors第46页
        1.3.4 Agrobacterium transformation第46-48页
    1.4 Results第48-54页
        1.4.1 Callus induction from immature embryos第48-49页
        1.4.2 Callus induction from mature embryos第49-50页
        1.4.3 Callus induction and regeneration from immature inflorescence第50-51页
        1.4.4 Transformation through peircing of mature embryos第51-52页
        1.4.5 Production of phenolic compounds第52-53页
        1.4.6 Excessive growth of Agrobacterium during co-cultivation第53-54页
    1.5 Discussion第54-58页
    1.6 References第58-68页
CHAPTER Ⅱ CRISPR/Cas9 and virus-induced gene silencing(VIGS)-mediated functional characterization of pectin and lignin related genes第68-152页
    2.0 Introduction第68-70页
    2.1 Lignins第70-76页
        2.1.1 Structure and synthesis of lignin第70-73页
        2.1.2 Monolignin biosynthesis第73-74页
        2.1.3 Role of CCoAMT,COMT and CCRT in lignin synthesis第74-76页
    2.2 Pectin第76-79页
        2.2.1 Structure of homogalacturonan(HG)第76页
        2.2.2 Structure of xylogalacturonan第76-77页
        2.2.3 Structure of rhamnogalacturonan II第77-78页
        2.2.4 Structure of rhamnogalacturonan I第78-79页
    2.3 Synthesis of Monosaccharides第79-82页
        2.3.1 Synthesis of UDP-D-glucose and UDP-D-galactose第79-80页
        2.3.2 Synthesis of UDP-L-rhamnose第80页
        2.3.3 Synthesis of UDP-D-glucuronic acid第80-81页
        2.3.4 Synthesis of UDP-D-galacturonic acid,UDP-D-xylose and UDP-D-apiose第81页
        2.3.5 Synthesis of UDP-L-arabinose第81页
        2.3.6 Synthesis of GDP-D-mannose and GDP-L-fucose第81-82页
        2.3.7 SYNTHESIS OF GDP-D-RHAMNOSE AND GDP-L-GALACTOSE第82页
    2.4 UDP-D-glucuronic acid4-epimerase involved in synthesis of pectin第82-84页
    2.5 GAlactUronosylTransferase(GAUT)involved in synthesis of pectin第84页
    2.6 Role of homogalacturonan in biological processes第84-88页
        2.6.1 Role of homogalacturonan in plant development第85页
        2.6.2 HG modulation and its impact on the physical properties to the end-product第85-86页
        2.6.3 Role of HG modifications in plant responses to biotic stress第86-87页
        2.6.4 Roles of HG modulation in structural resistance to plants第87-88页
    2.7 Virus-induced gene silencing for functional characterization第88-91页
    2.8 CRISPR/Cas9 as a tool of genome editing第91-94页
        2.8.1 CRISPR/Cas9 vectors for gene editing in plants第93-94页
    2.9 Materials and Methods第94-109页
        2.9.1 Plasmid construction for VIGS第94-95页
        2.9.2 Phylogenetic analysis第95页
        2.9.3 Gene structure analysis第95页
        2.9.4 Statistical analysis第95页
        2.9.5 Cell wall isolation and extraction第95-96页
        2.9.6 Estimation of monosaccharaides第96页
        2.9.7 Biphenyl assay第96页
        2.9.8 RNA extraction protocol第96-98页
        2.9.9 PCR product purification protocol第98-99页
        2.9.10 Plasmid extraction第99-100页
        2.9.11 DNA ligation第100页
        2.9.12 E.coli transformation第100-101页
        2.9.13 Analyzing positive clones第101-102页
        2.9.14 Analyzing positive clones by restriction digestion第102页
        2.9.15 Synthesis of cDNA第102-103页
        2.9.16 Amplification of PCR product第103页
        2.9.17 Vector digestion第103-104页
        2.9.18 Agarose gel electrophoresis第104页
        2.9.19 Semi-quantitative reverse transcriptase PCR(RT-PCR)第104-105页
        2.9.20 GV3101 Electro-competent cells transformation第105页
        2.9.21 Virus gene inducing(VIGs)protocol第105-106页
        2.9.22 Agrobacterium-mediated leaf disc transformation第106-107页
        2.9.23 Quantitative real-time PCR protocol第107-109页
    2.10 Results第109-120页
        2.10.1 The identification of NbGAE Gene family in tobacco第109-112页
        2.10.2 Phylogenetic analysis of the NbGAE genes in tobacco and other species第112页
        2.10.3 Gene Structure analysis of the NbGAE gene family in the tobacco第112-114页
        2.10.4 Sequence alignments of NbGAE family第114页
        2.10.5 Down regulation in expression of NbGAE6 homologous第114-117页
        2.10.6 Estimation of GalA content through biphenyl assay第117-118页
        2.10.7 Monosaccharide composition in silenced NbGAE6第118-119页
        2.10.8 Relative expression of NbGAE6 in tobacco after VIGS第119-120页
    2.11 Results for CRISPR/Cas9第120-130页
        2.11.1 Vector and gene construction for CRISPR/Cas第121页
        2.11.2 Identification of Protospacer Adjacent Motif(PAM)of pectin related genes第121-122页
        2.11.3 Identification of Protospacer Adjacent Motif(PAM)of lignin related genes第122-123页
        2.11.4 Phylogenetic analysis of CCoAMT第123-124页
        2.11.5 Semi-quantitative(RT-PCR)analysis for CCoAMT第124-125页
        2.11.6 Semi-quantitative(RT-PCR)analysis of NtabGAE6 and NtabGAE第125-126页
        2.11.7 Establishment of optimized CRISPR/Cas9 system and mutation analysis第126-130页
    2.12 Discussion第130-133页
    2.13 Conclusion第133-138页
    2.14 REFERENCES第138-152页
CHAPTER Ⅲ Conclusion and recommendations第152-154页
ACKNOWLEDGEMENT第154-155页
CURRICULUM VITAE第155页

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