With a detailed account of basic techniques and important applications alike, this book provides a general introduction as well as a selected survey of key advances in the fascinating field of plant cell and tissue culture as a tool in biotechnology.

The adv antages of those systems are counterbalanced by some important dis- vantages. For one, in heterotrophic and mixotrophic systems high concentrations of organic ingredients are required in the nutrient medium (particularly sugar at 2% or more), associated with a high risk of microbial contamination. How, and to which extent this can be avoided will be dealt with in Chapter 3. Other disadvantages are the difficulties and limitations of extrapolating results based on tissue or cell c- tures, to interpreting phenomena occurring in an intact plant during its development. It has always to be kept in mind that tissue cultures are only model systems, with all positive and negative characteristics inherent of such experimental setups. To be realistic, a direct duplication of in situ conditions in tissue culture systems is still not possible even today in the 21st century, and probably never will be. The organization of the genetic system and of basic cell structures is, however, essentially the same, and therefore tissue cultures of higher plants should be better suited as model s- tems than, e.g., cultures of algae, often employed as model systems in physiological or biochemical investigations. The domain cell and tissue culture is rather broad, and necessarily unspecif ic. In terms of practical aspects, basically five areas can be distinguished (see Figs. 1.1 , 1.2 ), which here shall be briefly surveyed before being discussed later at length.

Presents detailed descriptions of basic cell culture techniques Covers important applications Of interest to both beginners and experienced scientists

Contenu
1 Introduction 2 Historical Developments of Cell and Tissue Culture Techniques 3 Callus Cultures 3.1 Establishment of a Primary Culture from Explants of the Secondary Phloem of the Carrot Root 3.2 Fermenter Cultures 3.3 Immobilized Cell Cultures 3.4 Nutrient Media 3.5 Evaluation of Experiments 3.6 Maintenance of Strains, Cryopreservation 3.7 Some Physiological, Biochemical, and Histological Aspects 4 Cell Suspension Cultures 4.1 Methods to Establish a Cell Suspension 4.2 Cell Population Dynamics 5 Protoplast Cultures 5.1 Production of Protoplasts 5.2 Protoplast Fusion 6 Haploid Techniques 6.1 Application Possibilities 6.2 Physiological and Histological Background 6.3 Methods for Practical Application 6.4 Haploid Plants 7 Plant PropagationMeristem Cultures, Somatic Embryogenesis 7.1 General Remarks, and Meristem Cultures 7.2 Protocols of Some Propagation Systems 7.2.1 In Vitro Propagation of Cymbidium 7.2.2 Meristem Cultures of Raspberries 7.2.3 In Vitro Propagation of Anthurium 7.3 Somatic Embryogenesis 7.3.1 Basics of Somatic Embryogenesis 7.3.2 Ontogenesis of Competent Cells 7.3.3 Genetic AspectsDNA Organization 7.3.4 The Phytohormone System 7.3.5 The Protein System 7.3.6 Cell Cycle Studies 7.4 Practical Application of Somatic Embryogenesis 7.5 Artificial Seeds 7.6 Embryo Rescue 8 Some Endogenous and Exogenous Factors in Cell Culture Systems 8.1 Endogenous Factors 8.1.1 Genetic Influences 8.1.2 Physiological Status of 'Mother Tissue' 8.1.3 Growth Conditions of the 'Mother Plant' 8.2 Exogenous Factors 8.2.1 Growth Regulators 8.2.2 Nutritional Factors 8.2.2.1 Improvement of Nutrient Uptake by Transgenic Carrot Cultures 8.3 Physical Factors 9 Primary Metabolism 9.1 Carbon Metabolism 9.2Nitrogen Metabolism 10 Secondary Metabolism 10.1 Introduction 10.2 Mechanism of Production of Secondary Metabolites 10.3 Historical Background 10.4 Plant Cell Cultures and Pharmaceuticals, and Other Biologically Active Compounds 10.4.1 Antitumor Compounds 10.4.2 Anthocyanin Production 10.5 Strategies for Improvement of Metabolite Production 10.5.1 Addition of Precursors, and Biotransformations 10.5.2 Immobilization of Cells 10.5.3 Differentiation and Secondary Metabolite Production 10.5.4 Elicitation 10.5.4.1 Jasmonic Acid 10.5.4.2 Effect of UV on Production of Secondary Metabolites in Cultured Tissues 10.6 Organ Cultures 10.6.1 Shoot Cultures 10.6.2 Root Cultures 10.7 Genetic Engineering of Secondary Metabolites 10.8 Membrane Transport and Accumulation of Secondary Metabolites 10.9 Bioreactors 10.9.1 Technical Aspects of Bioreactor Systems 10.10 Prospects 11 Phytohormones and Growth Regulators 12 Cell Division, Cell Growth, Cell Differentiation 13 Genetic Problems and Gene Technology 13.1 Somaclonal Variations 13.1.1 Ploidy Stability 13.1.2 Some More Somaclonal Variations 13.2 Gene Technology 13.2.1 Transformation Techniques 13.2.1.1 Direct Gene Transfer 13.2.1.2 Agrobacterium-Mediated Gene Transformation 13.2.2 Selectable Marker Genes 13.2.2.1 Reporter Genes 13.2.2.2 Variants of GFP 13.2.3 ß-Glucuronidase (GUS) 13.2.3.1 Procedures for Assay of GUS Gene Expression 13.2.4 Antibiotics Resistance Genes 13.2.5 Elimination of Marker Genes 13.2.5.1 Cre-lox Recombination-Based Systems 13.2.5.2 Ac/Ds System 13.2.5.3 Double Cassette System 13.2.6 Agrobacterium-Mediated Transformation in Dicotyledonous Plants 13.2.6.1 Transgenic Carrot: Potential Source of Edible Vaccines 13.2.6.2 Cell Culture and Transformation Procedures 13.2.6.3 Uses of Transgenes to Increase Host Plant