PLANT TISSUE CULTURE 10.3. Ovary Culture 140 10.4. Flower Culture 142 11. EMBRYO AND ENDOSPERM CULTURE

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  • PLANT TISSUE CULTURE (THEORY AND PRACTICALS)

    2nd Edition

  • PLANT TISSUE CULTURE (THEORY AND PRACTICALS)

    2nd Edition

    T. Pullaiah Department of Botany, Sri Krishnadevaraya University, Anantapur - 515003, Andhra Pradesh (India)

    M.V. Subba Rao Department of Botany, Andhra University, Visakhapatnam - 530003, Andhra Pradesh (India)

    and

    E. Sreedevi Department of Botany, Government Degree College, Anantapur - 515001, Andhra Pradesh (India)

  • Published by SCIENTIFIC PUBLISHERS (INDIA)

    Jodhpur –

    5 A, New Pali Road P.O. Box 91 Jodhpur - 342 001 INDIA

    © 2017, Authors

    All rights reserved. No part of this publication or the information contained herein may be reproduced, adapted, abridged, translated, stored in a retrieval system, computer system, photographic or other systems or transmitted in any form or by any means, electronic, mechanical, by photocopying, recording or otherwise, without written prior permission from the author. Disclaimer: Whereas every effort has been made to avoid errors and omissions, this publication is being sold on the understanding that neither the editors (or authors) nor the publishers nor the printers would be liable in any manner to any person either for an error or for an omission in this publication, or for any action to be taken on the basis of this work. Any inadvertent discrepancy noted may be brought to the attention of the publisher, for rectifying it in future editions, if published. Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for identification and explanation without intent to infringe.

    ISBN: 978-93-86347-35-0 (PB) 978-93-86347-34-3 (HB)

    eISBN: 978-93-87869-80-6

    Visit the Scientific Publishers (India) website at http://www.scientificpub.com

    Printed in India

  • PREFACE

    Since the publication of the first edition in 2009, several new and exciting developments have taken place in the field of plant tissue culture which pursues us to attempt to provide current information and examples. From this book, faculty in various fields of plant sciences, graduate students, and undergraduates may gain basic and advanced knowledge about application of various aspects of Plant Cell and Tissue Culture. The revised edition presents updated information and will prove to be an excellent text and reference book for theoretical, practical and applied aspects of plant tissue culture with hands on training approach. Each chapter as before, is written in lucid language which has now been thoroughly revised for recent developments to include relevant media protocols, and is profusely illustrated with self- explanatory diagrams. This book includes new additions in every chapter starting from tissue culture history to haploid production, suspension cultures, nucellus culture, protoplast fusion and hybridization and secondary metabolite production and enables to make the readers to know about recent developments in Plant tissue culture. This revised edition includes detailed drawings that complement both knowledge of theory and experiments.

    T. Pullaiah M.V. Subba Rao

    E. Sreedevi

  • LIST OF ABBREVIATIONS

    % Per cent

    2iP 2-Isopentenyl adenine

    ABA Abscisic Acid

    AC Activated Charcoal

    AIDS Acquired Immuno Deficiency Syndrome

    AR Analytical Reagent

    ATP Adenosine Triphosphate

    BB5 Gamborg’s Medium

    BA 6-Benzyladenine

    BAP 6-Benzyl-Aminopurine

    BM Basal Medium

    BuDR BromoDeoxyuridine

    CCC Cycocel

    cDNA complementary DNA

    CH Casein Hydrolysate

    CM Coconut Milk

    CMS Cytoplasmic Male Sterility

    CPPU N-2-chloro-4puridyl-N-phenyl Urea

    CPW Cell-protoplast Washing

    DES Diethyl Sulphate

    DFR Dihydroflavonol-4-reductase

    DH Diploid Homozygous

    DMSO Dimethyl Sulphoxide

    DNA Deoxyribonucleic Acid

  • vi Plant Tissue Culture (Theory and Practicals)

    DPU N,N’-diphenyl Urea

    DRT Decimal Reduction Time

    ECs Embryogenic Clumps

    EDTA EthylenediamineTetraacetic acid

    ELISA Enzyme-linked Immunosorbent Assay

    EMS Ethyl Methane Sulphonate

    F3’5’H Flavanoid 3’, 5’- hydroxylase

    FDA Fluorescein Diacetate

    FudR 5-Fluorodeoxyuridine

    GA3 Gibberellic Acid

    GCMS Gas Chromatography Mass Spectrometer

    GR Guaranteed Reagent

    H2O2 Hydrogen peroxide

    HEPA High Efficiency Particulate Air filter

    HgCl2 Mercuric chloride

    HPLC High Performance Liquid Chromatography

    HU Hydroxy Urea

    IAA Indole-3-Acetic Acid

    IBA Indole-3-Butyric Acid

    IBPGR International Board for Plant Genetic Resources

    IEDCs Induced Embryogenic Determined Cells

    JA Jasmonic Acid

    JAMe methyl Jasmonate

    KN Kinetin

    LN Liquid Nitrogen

    LOS Low-oxygen Storage

    LPS Low-pressure Storage

    LS medium Linsmaier and Skoog medium

    MCO Mitotic Crossing Over

    MCPA 2-methyl-4-chlorophenoxyacetic acid

    mRNA messenger RNA

  • List of Abbreviations vii

    MS Murashige and Skoog’s Medium

    MSO Methionine Sulphoximide

    NAA Naphthalene Acetic Acid

    NOA Naphthoxyacetic Acid

    NR Nitrate Reductase oC Degree Celsius

    OD Optical Density

    ODAP β-oxalyl-L-α,β-diaminopropionic acid

    PCPA 4-Chlorophenoxyacetic acid

    PCR Polymerase Chain Reaction

    PCV Packed Cell Volume

    PE Plating Efficiency

    PEDCs Pre-embryogenic Determined Cells

    PEG Polyethylene Glycol

    PGRs Plant Growth Regulators

    PLRV Potato Leaf-Roll Virus

    ppm Parts per million

    Psi Pounds per square inch

    PVP Polyvinylpyrrolidone

    RBC Red Blood Cell

    RFLP Restriction Fragment Length Polymorphism

    RO Reverse Osmosis

    rRNA Ribosomal Ribonucleic Acid

    T-DNA Transfer Deoxyribonucleic Acid

    TDP Thermal Death Point

    TDT Thermal Death Time

    TDZ Thiadiazuron

    TMV Tobacco Mosaic Virus

    UV Ultraviolet

    WBC White Blood Cell

    WM White’s Medium

  • viii Plant Tissue Culture (Theory and Practicals)

    WPM Woody Plants Medium

    TTC 2,3,5-Triphenyltetrazolium Chloride

    2,4,5-T 2,4,5-Trichloro phenoxy acetic acid

    Picloram 4-Amino, 3,5,6-Trichloropicolinic acid

    B-9 Succinic acid 2, 2-dimethyl hydrazide

    2,4-D 2,4-Dichloro phenoxy acetic acid

  • CONTENTS

    1. HISTORY OF PLANT TISSUE CULTURE 1 1.1. Introduction 1 1.2. Totipotency of the Cells 3 1.3. Morphogenesis 4 1.4. History of Plant Tissue Culture 9 1.5. Applications of Plant Tissue Culture 19 1.6. Terminology 20 1.7. Books for Further Reading 24

    2. LABORATORY REQUIREMENTS FOR PLANT TISSUE CULTURE

    25

    2.1. Laboratory Organization 25

    3. TISSUE CULTURE MEDIA 34

    3.1. Introduction 34 3.2. Media Composition 34 3.3. Media Preparation 48 3.4. Commonly used Concentrations in Tissue Culture 50

    4. METHODS OF STERILIZATION 52

    4.1. Introduction 52 4.2. Sterilization by Heat 53 4.3. Filter Sterilization 55 4.4. Chemicals 55 4.5. Infrared Radiation 56 4.6. Surfactants 56 4.7. Antibiotics for Control of Bacterial and Fungal Contaminants 57 4.8. Maintenance of Aseptic Conditions 58

  • x Plant Tissue Culture (Theory and Practicals)

    4.9. Surface Sterilization of Explant 58

    4.10. Transfer of An Explant 59

    5. CELL AND SUSPENSION CULTURE 62

    5.1. Introduction 62 5.2. Procedure for Cell and Suspension Culture 62 5.3. Establishment of Cell Suspension Culture 62 5.4. Cell Suspension Culture and Cell Growth 64 5.5. Formation of Somatic Embryos and Plant Regeneration 64 5.6. Types of Suspension Cultures 64 5.7. Growth Patterns in Suspension Culture 66 5.8. Measurement of Cell Growth in Suspension Cultures 67 5.9. Synchronization of Suspension Cell Cultures 69 5.10. Technique for Single Cell Culture 69 5.11. Tests for Viability of Cultured Cells 71 5.12. Advantages of Cell and Suspension Culture 72 5.13. Plant Cell Reactors 72 5.14. Immobilized Plant Cell Reactors 77 5.15. Culture of Isolated Single Cells 78

    6. SOMATIC EMBRYOGENESIS 80

    6.1. Introduction 80 6.2. Some Examples of Somatic Embryogenesis 82 6.3. Theories on Embryogenesis 83 6.4. Factors Affecting Somatic Embryogenesis 85 6.5. Anatomy and Cytology to Somatic Embryo Development 88

    7. SYNTHETIC SEEDS 93

    7.1. Introduction 93

    7.2. Encapsulation 93

    7.3. Production of Artificial Seeds 95

    7.4. Applications 97

    7.5. Limitations 98

    7.6. Uses of Artificial Seeds 98

    7.7. Reference 99

  • Contents xi

    8. MICROPROPAGATION OF HIGHER PLANTS 100

    8.1. Introduction 100 8.2. Somatic Cell Embryogenesis 102 8.3. Production of Adventitious Shoots and Related Organs 103 8.4. Enhanced Axillary Branching 103 8.5. Steps Involved in Plant Multiplication 105 8.6. Buds Culture 107 8.7. Factors Affecting Micropropagation 109 8.8. Micropropagation of Ornamental Plants 112 8.9. Advantages of Micropropagation 113

    9. HAPLOID PRODUCTION 115

    9.1. Introduction 115 9.2. Anther Culture 115 9.3. Pollen Culture 122 9.4. Comparison between Anther