Plant Tissue Culture : Techniques and Experiments

Plant Tissue Culture: Techniques and Experiments, Fourth Edition, builds on the classroom tested, audience proven manual that has guided users through successful plant culturing for almost 30 years. The book's experiments demonstrate major concepts and can be conducted with a variety of plant materials readily available throughout the year. This fully updated edition describes the principles of the newest technologies, including CRISPR/Cas9 gene editing and RNAi technology with plant cell and tissue cultures and their applications. Bridging the gap between theory and practice, this book contains detailed methodology supported by comprehensive illustrations, giving users a diverse learning experience for both university students and plant scientists. Provides fundamental principles, methods and techniques in plant cell, tissue and organ culture that can be applied to all crop plants, including agronomic crops, horticulture and forestry crops for germplasm improvement Guides readers from lab setup to supplies, stock solution and media preparation, explant selection and disinfestations, and experimental observations and measurement Contains the latest advances and updates since the previous edition published in 2012 Front Cover Plant Tissue Culture Copyright Page Contents Preface Acknowledgments Scheduling and interrelationships of exercises 1 History of plant cell culture Introduction The early years The era of techniques development The recent past Cell behavior Plant modification and improvement Pathogen-free plants and germplasm storage Clonal propagation Product formation The present era References 2 Setup of a tissue culture laboratory Work areas Media preparation/culture evaluation/record-keeping area Aseptic transfer area Environmentally controlled culture area Reference Further reading Books Journals Organizations 3 Media components and preparation Inorganic salts Plant growth regulators Vitamins Carbohydrates Hexitols Gelling agent Amino acids Antibiotics Natural complexes Media pH Medium preparation References 4 Explant preparation Explant age Season Explant size Plant quality Goal Genotype Aseptic technique Aseptic germination of seeds Carrot, sunflower, broccoli, cotton, or any seed of choice Medium preparation: 1-L equivalent, seed germination medium Explant preparation Carrot, sunflower, and broccoli Cotton Douglas Fir Medium preparation: sterile vermiculite Explant preparation Observations Reference Further reading 5 Contamination Explant Disinfectant agents Explant source Internal microbial contamination Explant pretreatment to control microbial contaminants Personnel Laminar airflow hood Insects Control of insect contamination Media Instruments Room air handling system References 6 Callus induction Callus initiation Medium preparation Explant preparation Carrot seedlings Lemon Broccoli Carrot root tissue Culture Observations Data and questions Explant orientation Medium preparation Explants Observations Establishment of competent cereal cell cultures Medium preparation Explants Data and questions Salt selection in vitro Medium preparation Explants Data and questions Growth curves Cotton Medium preparation Explant preparation Disinfestation of explant source Culture Tobacco Explant preparation Medium preparation Disinfestation of expant source and culture Observations Data and questions Cellular variation from callus cultures Medium preparation Explants Data and questions References Further reading 7 Regeneration and morphogenesis Controlled morphogenesis: potato tuberization Explants Data and questions Somatic embryogenesis Explant preparation Procedure Data and questions Regeneration of rice Regeneration Data and questions Dormancy requirements of explants Explant preparation Questions Primary morphogenesis: Douglas fir Explant preparation Data and questions References Further reading 8 Woody shrubs and trees Rhododendrons and azaleas Explants Advanced studies Roses Explant Advanced studies Birch trees Explant Advanced studies White cedar (arborvitae) Explant References 9 Haploid plants from anther culture Datura anther culture Explant preparation Questions African violet Explant preparation Observations Root tip chromosome squash technique Questions Tobacco Explant preparation Observations Questions Anther squash technique References Further reading 10 Embryo rescue Sweet corn embryo culture Preparation and culture of corn embryos Questions Crabapple and pear Explant preparation Observations Questions References Further reading 11 Meristem culture for virus-free plants Isolation of the shoot apical meristem Explant preparation Questions Establishing virus- and bacteria-free plants Explant preparation Garlic propagation from bulb scales Explant preparation Data References Further reading 12 In vitro propagation for commercial production of ornamentals Boston fern Stage I Explant preparation Observations Stage II Explant preparation Observations Stage III Explant preparation Transplantation Staghorn fern Stage I Explant preparation Observations Stage II Explant preparation Observations Stage III Observations Data and questions Ficus Stage I Explant preparation Stage II Stage III Explant preparation Questions Kalanchoe Explant preparation Questions African violet Explant preparation Data and questions Endangered species Pitcher plants Explant preparation Cactus Explant preparation References Further reading 13 Protoplast isolation and fusion Corn Supplies Plant material preparation (2 weeks) Protoplast liberation Protoplast purification Protoplast transformation Tomato Supplies Plant material preparation (4 weeks) Protoplast liberation Protoplast purification Protoplast transformation Other plant protoplast isolation and fusion Supplies Protoplast liberation Alternative protocol Protoplast purification Alternative protocol Vital staining Protoplast fusion Yield determination Developing a protoplast isolation protocol References Further reading 14 Agrobacterium-mediated transformation of plants Petunia or tobacco leaf disk Sequence Preculture Explant preparation Cocultivation with Agrobacterium Agrobacterium preparation Transgenic shoot selection and elimination of Agrobacterium Antibiotic stocks Procedure Rooting and pretesting for transformed shoots Procedure Tomato cotyledon/hypocotyl or lettuce leaf disk Sequence Preculture Explant preparation Cocultivation with Agrobacterium Transgenic shoot selection and elimination of Agrobacterium Antibiotic stocks Procedure Rooting and pretesting for transformed shoots Procedure Petunia shoot apex Sequence Seed germination Shoot induction Shoot apex transformation Rooting Mature rice seed-derived callus Sequence Seed preparation and callus induction Cocultivation with Agrobacterium Agrobacterium preparation Transformed callus selection and elimination of Agrobacterium Shoot regeneration Rooting and pretesting for transformed shoots Arabidopsis floral dip Sequence Grow Arabidopsis Grow Agrobacterium Culture Agrobacterium Harvest Agrobacterium Procedure Dip Arabidopsis Seed screening Seed preparation Tobacco leaf infiltration Sequence Grow Agrobacterium Culture Agrobacterium Harvest Agrobacterium Procedure Infiltration tobacco leaves References Further reading 15 Genome editing and RNA interference technologies in plants Genome editing using a CRISPR/Cas9 or TALEN system Introduction Design CRISPR–Cas9 expression vectors Sequence of targeted mutagenesis in tomato Rooting and pretesting of targeted mutagenesis in tomato Principle of TALEN-mediated genome editing RNA interference technology to suppress gene expression Introduction Mechanism of RNAi Applications of RNAi in agriculture Design RNAi expression vectors Sequence of targeted gene silencing in lettuce Pretesting of targeted gene silencing in lettuce References Further reading Appendix I: Useful measurements Length Temperature Concentration Weight Appendix II: Review of solution preparation Percentage solutions Examples Problems Answers Molar solutions Problems Answers Dilution of a stock solution Problems Answers Appendix III: List of suppliers Appendix IV: Common plant tissue culture terms Index Back Cover __Plant Tissue Culture: Techniques and Experiments, Fourth Edition,__