Preface to the Second Edition Preface to the First Edition AbbreviatIOns PART I：PLANT TISSUE CUL.TURE 1.INTRODUCTION New technologies Origin of biotechnology History 2 LABORATORY ORGANIZTION Washing facility General laboratory and media preparation area Transfer area Culturing facilities Light units Green houses Laboratory and pemonal safety 3.NUTRITlON MEDIUM Facilities and equipment Units for solution preparation Media composition Inorganic nutrients Carbon and energy source Vitamins Growth regulators Organic supplements Gelling agent pH Protocol：General methodology for medium preparation 4.STERILIZATION TECHNIQUES Preparation of sterile media，containers and small instruments Steam sterilization Units of energy and pressure Dry sterilization Filter sterilization Ultra violet sterilization Maintenance of aseptic conditions AIcohoI sterilization Flame sterilization Sterilization of explant ChemicaI sterilization Protocols Sterilization of seeds Sterilization Of buds，leaf，stem，roots，tubers，scales，etc. Sterilization of tissue for immature embryos，ovules and flower buds for anther culture 5.TYPES OF CULTURE CytodifferentIation Organogenic differentiation Types of culture Seed culture Embryo culture Mature embryo culture Immature embryo culture／embryo rescue Applications of embryo culture Callus culture Organ culture Nucellus culture Application Endosperm culture Application Cell culture Protoplast culture Protocols Protocol for seed germination（Nicotiana） Protocol for embryo culture（Cereals-wheat，maize，barley rice，etc.） Protocol for embryo culture（Legumes-green gram，black gram， french bean，soybean，etc.） Protocol for callus induction（Nicotiana tabacum） Protocol for callus induction（Cereals-wheat，rice，maize，barley,etc.） 6.MICROPROPAGATlON Axillary bud proliferation approach Meristem and shoot tip cultu re Bud culture Single node culture Axillary bud method OrganOgenesis Organogenesis via callus formation Direct adventitious organ formation Embryogenesis Advantages Of micropropagation Problems associated with micrOprOpagatiOn Protocols Meristem and node culture Of potato（Solanum tuberosum） Proliferation of axillary buds（Strawberry-Fragaria chiloensis） OrganOgenesis-adventitious shoot formation Organogenesis via callus formation（Nicotiana） Organogenesis via callus formation（Cereals：wheat，barley, maize，rice，etc.） Embryogenesis（Carrot） 7.CELL SUSPENSION AND SECONDARY MEl-ABOLITES Ivpes of suspension cultures Batch culture Continuous culture Open continuous culture CIosed continuous culture Semi continuous culture G rowth measu rements Synchronization of suspension culture cells Technique for single cell culture-Bergmann cell plating technique Applications Production of secondary metabolites Morphological and chemicaI differentiation Medium composition for secondary product formation G rowth production patterns EnvironmentaI factors Selection of cell lines producing high amounts of a useful metabolite Product analysis Application Problems associated with secondary metabolite production Immobilized cell systems Polymers for immobilization Product release Biotransformation Protocols Protocol for cell suspension culture（Nicotiana tabacum） Protocol for cell suspension culture（Cereals-wheat，rice， maize，barley,etc.） 8.IN VITRO PRODUCTION OF HAPLOIDS Androgenic methods Anther cultu re Microspore culture The various factors governing the success of androgenesis Genotype Physiological statuS of the donor plants Stage of pollen Pretreatment of anthers Culture media Process of androgenesis The ploidy Ievel and chromosome doubling Diploidization Significance and uses of haploids Problems Gynogenic haploids Factors affecting gynogenesis Chromosome elimination technique for production Of haploids in cereals（Barley and wheat） Protocol Protocol for anther culture of cereals（Rice，barley,wheat，etc.） 9.PROTOPLAST ISOLATION AND FUSION Protoplast isolation MechanicaI method Enzymatic method Physiological state of tissue and cell material Enzymes Osmoticum Protoplast purification Protoplast viability and density Culture techniques Culture medium Environmental factors Protoplast development Cell walI formation Growth，division and plant regeneration Somatic hybridization Protoplast fusion Spontaneous fusion Induced fusion methods Treatment with sodium nitrate Calcium ions at high pH Polyethylene glycol method Electrofusion Mechanism of fusion Identification and selection of hybrid cells Chlorophyll deficiency complementation Auxotroph complementation Complementation of resistance markers Use of metabolic inhibitors Use Of visuaI characte ristics Compound selection system 10.SoMACLONAL VARLATION 11.GERMPLASM STORAGE AND CRYOPRESERVATION 12.GENETIC MArERlAL 13.ORGANIZATION OF DNA AND GENE EXPRESSION 14.BASIC TECHNIQUES 15.GENE CLONING：Cutting and Joining DNA Molecules 16.GENE CLONING：Vectors 17.GENE CLONING：cDNA and Genomic Cloning and Analysis of Cloned DNA Sequences 18. POLYMERASE CHAIN REACTlON 19.IN VITRO MUTAGENESlS 20.TRANSPOSON GENETIC ELEMENTS AND GENE TAGGING 21.GENE ISOLATION 22.MOLECULAR MARKERS AND MARKER.ASSlSTED SELECTION 23. GENE TRANSFER IN PLANTS 24. TRANSGENICS IN CROP lMPROVEMENT 25. GENOMICS 26.BIOINFORMATICS 27.INTELLECTUAL PROPERTY RIGHTS Appendix References Glossary Author index Subject index
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