Plant Cell Culture PDF
Preview Plant Cell Culture
Plant Cell Culture MonoclonalAntibodies PCR AnalyzingChromosomes SeparatingCells Biological Centrifugation Plant Cell Culture Forthcoming titles AnalysingGene Expression Animal Cell Culture andTechnology, Second Edition Gene Mapping ReconstructingEvolutionaryTrees Plant Cell Culture D. E. Evans Oxford Brookes University, Oxford, UK J.O.D. Coleman Oxford Brookes University, Oxford, UK and A. Kearns Oxford Brookes University, Oxford, UK @ Taylor&Francis ~ Taylor&FrancisGroup LONDONANDNEWYORK ©Taylor & Francis, 2003 First published 2003 All rights reserved. No part of this book may be reprinted or reproduced or utilised in any form or by any electronic, mechanical, or other means, now known or hereafter invented, including photocopying and recording, or in any information storage or retrieval system, without permission in writing from the publishers. A CIP catalogue record for this book is available from the British Library. ISBN 1 85996 320 X Published by Taylor & Francis 2 Park Square, Milton Park, Abingdon, Oxon, 0Xl4 4RN 52 Vanderbilt Avenue, New York, NY 10017 Transferred to Digital Printing 2007 Typeset by J&L Composition, N Yorks, UK Publisher's Note The publisher has gone to great lengths to ensure the quality of this reprint but points out that some imperfections in the original may be apparent Contents Abbreviations xi Preface xiii Chapter I An introduction to plant cell and tissue culture 1. Plantcellandtissue culture 1 2. Industrial-scaleplantcellculture 3 3. Planttissue culture, plantbreedingand crop improvement 3 4. Planttissue cultureandplantgenetic engineering 4 Chapter 2 Basic plant biology for cell culture 7 1. Tissuesandorgans 7 2. Planttissues 7 3. Meristems 8 4. Plantreproductivetissues 9 5. The embryoandembryogenesis 10 6. Developmentoftissues 11 7. Protoplasts 12 8. Media, nutrientsandrequirementsforgrowth 12 Chapter 3 Tissue culture in agriculture, horticulture and forestry I9 1. Introduction 19 2. Micropropagation 20 3. Plantbreeding 23 4. Biodiversityand conservationofgermplasm 26 Chapter 4 Tissue culture in genetic engineering and biotechnology 3I 1. Introduction 31 2. Genetic engineering 31 3. Applicationsofplantgenetic engineering 39 VI Contents Chapter 5 Culture facilities, sterile technique and media preparation 43 1. Introduction 43 2. Thebasic laboratorylayoutandequipment 43 3. Sterilization 47 4. Media preparation 50 5. Contamination 53 6. Disposingofcontaminatedwaste 53 7. Safetyin the laboratory 54 Protocol5.1: Hygiene in the tissue culturelaboratory 55 Protocol5.2: Preparingbasal mediumfrom commercial formulations orfrom individualcomponents 57 Protocol5.3: Preparationofhormoneadditionsto basal media 60 Protocol5.4: Preparationofcoconutwater supplementfor basal media 62 Chapter 6 Callus cultures 63 1. Introduction 63 2. Originofcallus 63 3. Typesofcallus 63 4. Roleofcallus in embryogenesis, organogenesisand cellculture 64 5. Initiationandestablishmentofcallus cultures 64 6. Monitoringthegrowthofcallus 67 7. Genetictransformationofcallus 68 Protocol 6.1: Preparationofplantmaterialandexplants 70 Protocol 6.2: Callus from adicotroot: Arabidopsis thaliana and Nicotiana tabacum 73 Protocol 6.3: Callus from dicotshoot/leaf- Arabidopsisthaliana andNicotiana tabacum 74 Protocol 6.4: Callus from amonocot(e.g.maize, rice) 75 Protocol 6.5: Callus from gymnosperms (Norwayspruce) 76 Protocol 6.6: Transformingmaizecallus byparticlebombardment 77 Protocol 6.7: PreparationofatransgenicArabidopsisthalianacallus usingAgrobacterium 79 Chapter 7 Cell suspension cultures 81 1. Introduction 81 2. Initiationofcellsuspensioncultures 81 3. Maintenanceofcellsuspensioncultures 82 4. Growthcharacteristicsofcellsuspensioncultures 83 5. Usesofcellsuspensioncultures 87 Protocol 7.1: Ageneric protocolfor initiatingasuspensionculture 88 Protocol 7.2: Monitoringthe growthofsuspensioncultures 90 Protocol 7.3: Measurementofcellviability 92 Protocol 7.4: Genetictransformationofsuspensionculturedcells 94 Contents vii Chapter 8 Protoplast culture 95 1. Introduction 95 2. Isolationofprotoplasts 95 3. Protoplastculture 97 4. Usesofprotoplasts 98 Protocol8.1: Preparationofprotoplastsfrom suspensioncultures (e.g.maize) usingalongincubation 101 Protocol8.2: Isolationofprotoplastsfrom carrotsuspension culturesusingarapid incubation 102 Protocol8.3: Rapid preparationofmesophyllprotoplastsfrom maizeleaves 103 Protocol8.4: Preparationofmesophyllprotoplastsfrom tobacco leavesandpurificationofprotoplastson adensity gradient 104 Protocol8.5: Rapidpreparationofprotoplastsfrom maizeroots 106 Protocol8.6: Preparationofprotoplastsfrom roots (arabidopsis) withpartialpurificationofprotoplastsbyflotation 108 Protocol 8.7: The cultureofprotoplastsusingnurseculture 110 Protocol8.8: The cultureofprotoplastsusingthe hangingdrop method 112 Protocol8.9: Detectingregenerationofcellwall in isolated protoplasts 114 Protocol8.10: Protoplastfusion inducedbypolyethyleneglycol (PEG) 115 Protocol 8.11: Electricalfusion ofprotoplasts 116 Protocol 8.12: Protoplasttransformationbyelectroporation 118 Protocol8.13: Transformationofprotoplastsmediatedby polyethyleneglycol(PEG) 119 Chapter 9 Haploid cultures 121 1. Introduction 121 2. Antherculture 121 3. Microsporeculture 122 4. Anthercultureversus microsporeculture 122 5. Intergenericcrosses andembryorescue 123 6. Proceduresfor the inductionofandrogenesis 124 Protocol 9.1: Anthercultureofwheat (Triticum aestivum L.) 125 Protocol 9.2: Wheatmicrosporeculture 127 Chapter I0 Organ and embryo culture 129 1. Introduction 129 2. Hairy roots 129 3. Embryo culture 129 4. Minitubersandmicrotubers 131 VIII Contents Protocol 10.1: Isolationandcultureofthe primaryseedlingroot ofdicots 133 Protocol 10.2: Isolationandcultureofrootsofmonocots (e.g.maize) 134 Protocol 10.3: Hairy rootcultures (planttransformationwith Agrobacteriumrhizogenes) 135 Protocol 10.4: Sterilecultureofthe ear ofacereal (e.g.maize) 137 Protocol 10.5: Embryo rescue ofadicot- Phaseolus vulgaris 138 Chapter II Regeneration ofplants and micropropagation 141 1. Introduction 141 2. Regenerationvia somaticembryogenesis 141 3. Control ofembryogenesis 144 4. Regenerationvia organogenesis 144 5. Protocols 145 Protocol 11.1: Embryogenesisfrom callus in adicot(e.g.carrot) or monocot(cereals, rice) 148 Protocol 11.2: Plantregenerationbyorganogenesis 149 Protocol 11.3: SomaticembryogenesisofNorway spruce usinga suspensionculturestep 151 Chapter 12 Somaclonal variation IS3 1. Introduction 153 2. Originsandmechanismsofsomaclonalvariability 153 3. The importanceofsomaclonalvariation 155 4. Methodsofassessing somaclonalvariationin regeneratedplants 155 5. Somaclonalvariationasatechniqueforcrop improvement 158 Chapter 13 Bacterial culture in the plant cell culture laboratory IS9 1. Introduction 159 2. Facilitiesfor bacterialculture 159 Protocol 13.1: TocultureE.coliin Luriabroth (LB) 161 Protocol 13.2: Calciumchloride-mediatedtransformationofE.coli 162 Protocol 13.3: Calciumchloride-mediatedtransformationof Agrobacterium 164 Protocol 13.4: CultureofAgrobacterium tumefaciensin culture medium,YEB 166 Protocol 13.5: CultureofAgrobacteriumrhizogenes in culture medium,YMA 167 Contents ix Chapter 14 Industrial usesofplant cell culture 169 1. Introduction 169 2. Plantcellsin culture 169 3. Apparatusfor large-scaleplantcultures 170 4. Examples ofindustrial-scaleproductionusing culturedcells 172 Chapter I5 Prospects and future challenges 175 1. Recentdevelopments 175 2. Thefuture 175 Suppliers ofchemicals, apparatus and cell culture products 181 Glossary 185 Index 189
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