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Genetic Transformation of Wheat for Salt Tolerance

ACKNOWLEDGEMENTS第4-5页
Preface第5-10页
List of Tables第10-11页
List of Figures第11-13页
1- INTRODUCTION第13-17页
2- LITERATURE REVIEW第17-43页
    2.1 Genetic Transformation of Wheat第17-18页
    2.2 Genetic Engineering of Wheat for Abiotic Stresses第18-21页
    2.3 Transformation Methods Used for Wheat第21-35页
        2.3.1 Direct Methods第21-27页
            2.3.1.1 Simple direct methods第21-23页
            2.3.1.2 Bilolistic/microprojectile bombardment第23-27页
        2.3.2 Indirect Methods第27-35页
            2.3.2.1 Agrobacterium-Mediated Transformation第27-32页
            2.3.2.2 Agrobacterium-Mediated inplanta Transformation第32-35页
    2.4 Genetic Markers第35-40页
        2.4.1 Scorable/Screenable Markers第35-37页
        2.4.2 Selectable Markers第37-40页
            2.4.2.1 Antibiotic Resistance Markers第37-39页
            2.4.2.2 Herbicide Resistance Markers第39-40页
            2.4.2.3 Metabolic/Auxotrophic Marker Genes第40页
    2.5 Promoters第40-43页
3 GENE GUN TRANSFORMATION OF WHEAT第43-61页
    3.1 MATERIALS AND METHODS第43-49页
        3.1.1 Callus Induction第43页
        3.1.2 Osmotic Treatment第43-44页
        3.1.3 Plasmid Vector第44页
        3.1.4 Extraction of Plasmid DNA第44-45页
        3.1.5 Preparing Micro-carriers第45-46页
        3.1.6 Coating plasmid DNA onto Micro-Carriers第46页
        3.1.7 Bombardment of Callus第46页
        3.1.8 Selection and regeneration第46-47页
        3.1.9 Extraction of genomic DNA from leaves第47页
        3.1.10 PCR (Polymerase Chain Reaction) Analysis第47-48页
        3.1.11 Phosphinothricin Leaf Paint Assay第48-49页
        3.1.12 Salt tolerance第49页
    3.2 RESULTS第49-56页
        3.2.1 Selection第49-50页
        3.2.2 Regeneration第50-51页
        3.2.3 Screening of Transformants第51-56页
            3.2.3.1 PCR test第51页
            3.2.3.2 PPT Leaf-painting第51-56页
            3.2.3.3 Salt tolerance第56页
    3.3 DISCUSSION第56-59页
    3.5 CONCLUSION第59-61页
4 AGROBACTERIUM-MEDIATED TRANSFORMATION OF WHEAT第61-76页
    4.1 MATERIALS AND METHODS第61-64页
        4.1.1 Callus Induction第61-62页
        4.1.2 Agrobacterium Culture第62页
        4.1.3 Inoculation and co-cultivation第62页
        4.1.4 Selection and Regeneration第62-63页
        4.1.5 Molecular Characterization of Putative Transformants第63页
        4.1.6 Extraction of Plasmid DNA第63-64页
        4.1.7 Determination of osmotic stress resistance第64页
    4.2 RESULTS第64-71页
        4.2.1 Selection第64-66页
        4.2.2 Regeneration第66页
        4.2.3 PCR test for transformed status of plants第66-70页
        4.2.4 Salt tolerance of transformed plants第70-71页
    4.3 DISCUSSION第71-74页
    4.4 CONCLUSION第74-76页
5 IN PLANTA APICAL MERISTEM TRANSFORMATION OF WHEAT第76-96页
    5.1 MATERIALS AND METHODS第76-81页
        5.1.1 Plant material, Inoculation, Co-cultivation and Selection第76-80页
            5.1.1.1 Transient GUS expression experiments第76-77页
            5.1.1.2 Transformation of apical meristem after germination第77-79页
            5.1.1.3 Transformation of apical meristem before germination第79-80页
        5.1.2. GUS Assay第80页
        5.1.3 Molecular Evaluation of Putative Transformants第80-81页
    5.2 RESULTS第81-89页
        5.2.1 Transient GUS expression第81-82页
        5.2.2 GUS expression in inflorescences from meristems inoculation after germination第82-84页
        5.2.3 GUS expression in inflorescences after inoculation of meristems before seed germination第84-86页
        5.2.4 PCR Analysis of T1 generation from GUS positive plants第86页
        5.2.5 Transformation of apical meristems with BADH and nptII第86-88页
        5.2.6 PCR test of kanamycim resistant plants第88-89页
    5.3 DISCUSSION第89-94页
    5.4 CONCLUSION第94-96页
6 BADH TRANSGENE AND SALT TOLERANCE第96-99页
7 SUMMARY第99页
8 SUMMARY IN CHINESE第102-105页
9 REFERENCES第105-122页

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