Plant Transformation via Agrobacterium tumefaciens: Culture Conditions, Recalcitrance and Advances in Soybean

245 Pages·2022·22.437 MB·English

by  Phetole Mangena

Plant Transformation via
Agrobacterium tumefaciens
This book presents information from authentic and highly regarded sources in
plant biotechnology. It provides a comprehensive review of plant transformation
in soybean. The book discusses protocol ineffciencies, the type and conditions of
explants, culture conditions, complexity in transformation processes, host-specifcity
and Agrobacterium-genotype interactions. The book provides insights promoting
soybean use in the manufacturing of health-promoting supplements, feed production,
food products and biodiesel production. It helps the reader in achieving successful
soybean transformation and outlining the possibilities that could help reduce
recalcitrance in soybean.

Plant Transformation via
Agrobacterium tumefaciens
Culture Conditions, Recalcitrance
and Advances in Soybean
Phetole Mangena
First edition published 2023
by CRC Press
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Library of Congress Cataloging‑in‑Publication Data
Names: Mangena, Phetole, author.
Title: Plant transformation via agrobacterium tumefaciens : culture
conditions, recalcitrance and advances in soybean / Phetole Mangena.
Other titles: Culture conditions, recalcitrance and advances in soybean
Description: First edition. | Boca Raton, FL : CRC Press, 2023. | Includes
bibliographical references and index.
Identifers: LCCN 2022009578 (print) | LCCN 2022009579 (ebook) | ISBN
9781032250373 (hardback) | ISBN 9781032250380 (paperback) | ISBN
9781003281245 (ebook)
Subjects: LCSH: Agrobacterium tumefaciens. | Plant biotechnology. |
Soybean.
Classifcation: LCC QR82.R45 M36 2023 (print) | LCC QR82.R45 (ebook) |
DDC 577.5/7--dc23/eng/20220330
LC record available at https://lccn.loc.gov/2022009578
LC ebook record available at https://lccn.loc.gov/2022009579
ISBN: 9781032250373 (hbk)
ISBN: 9781032250380 (pbk)
ISBN: 9781003281245 (ebk)
DOI: 10.1201/b22829
Typeset in Times
by Deanta Global Publishing Services, Chennai, India
This book is affectionately dedicated to Relebogile Ida,
Lethabo and Lerato Mahlare Mahlogonolo Mangena.

Contents
Preface.................................................................................................................... xiii
Acknowledgements ..................................................................................................xv
Author ....................................................................................................................xvii
Chapter 1 An Overview of Genetic Transformation in Plants ..............................1
1.1 Introduction ...............................................................................1
1.2 Molecular Mechanism for DNA Introgression ..........................3
1.3 Tissue Targeting for Plant Transformation ................................4
1.4 Screening and Selection of Transformants ................................5
1.5 Transfer to Natural Environment...............................................7
1.6 Summary ...................................................................................9
1.7 Abbreviations .............................................................................9
References .......................................................................................... 10
Chapter 2 Plant Transformation History ............................................................. 11
2.1 Discovery of Agrobacterium tumefaciens ............................... 11
2.2 Tumour-Inducing Principle ......................................................12
2.3 Structure and Modifcations of the Agrobacterium
Circular DNA .......................................................................... 14
2.4 Genetic Transformation in Plants ............................................16
2.5 The First Transgenic Plant ....................................................... 17
2.6 Role of Plant Tissue Culture .................................................... 19
2.7 Summary ................................................................................. 21
2.8 Abbreviations ...........................................................................22
References ..........................................................................................22
Chapter 3 Agrobacterium tumefaciens ...............................................................25
3.1 Introduction .............................................................................25
3.2 Classifcation ...........................................................................25
3.3 General Characteristics of Agrobacterium ..............................27
3.4 Size and Morphology ..............................................................29
3.5 Nutrition ..................................................................................30
3.6 Reproduction ........................................................................... 31
3.7 Cell Structure ..........................................................................32
3.8 Protoplasmic Materials ............................................................ 33
3.9 Chromosomal DNA and Plasmids ..........................................34
3.10 Summary ................................................................................. 35
3.11 Abbreviations ........................................................................... 35
References ..........................................................................................36
vii
viii Contents
Chapter 4 Getting Started with in Vitro Soybean Transformation .....................39
4.1 Introduction .............................................................................39
4.2 Bacterial Strains ......................................................................40
4.2.1 Bacterial Culture ........................................................ 41
4.3 Establishing Contamination-Free Cultures .............................42
4.3.1 Aseptic Seed Cultures ................................................ 43
4.3.2 Culture Medium .........................................................45
4.4 Type and Competency of Explants ..........................................46
4.5 Co-Cultivation of Explants with A. tumefaciens ..................... 47
4.6 Shoot Induction and Elongation ..............................................48
4.7 Rooting of Elongated Shoots ...................................................49
4.8 Acclimatisation ........................................................................49
4.9 Screening of Transgenic Plants ...............................................50
4.10 Other Considerations ...............................................................51
4.10.1 Equipment and Laboratory Supplies ..........................52
4.10.2 Surfactants .................................................................. 52
4.10.3 Antinecrotic Treatments ............................................. 53
4.11 Summary ................................................................................. 53
4.12 Abbreviations ...........................................................................54
References .......................................................................................... 55
Chapter 5 In Vitro Cultures Commonly Used for Plant Transformation ............59
5.1 Introduction .............................................................................59
5.2 Callus Culture ..........................................................................60
5.2.1 Purpose of Callus Culture ..........................................60
5.2.2 Requirements for Establishing Callus Culture ...........62
5.2.3 Application in Plant Transformation ..........................63
5.3 Shoot Culture ...........................................................................63
5.3.1 Purpose of Shoot Culture ...........................................64
5.3.2 Culture Requirements for Effcient Shoot
Multiplication .............................................................65
5.3.3 Application of Shoot Culture in Plant
Transformation ...........................................................66
5.4 Meristem Culture .....................................................................66
5.4.1 Purpose of Meristem Culture .....................................68
5.4.2 Meristem Culture Requirements ................................69
5.4.3 Use of Meristem Culture for Plant Improvement .......69
5.5 Protoplast Culture ....................................................................70
5.5.1 Purpose of Protoplast Culture ....................................71
5.5.2 Requirements for Protoplast Culture ..........................72
5.5.3 Applications in Plant Transformation ........................73
5.6 Summary .................................................................................73
5.7 Abbreviations ........................................................................... 74
References .......................................................................................... 75
Contents ix
Chapter 6 Current and Longstanding Challenges Facing Soybean
Transformation ...................................................................................79
6.1 Introduction .............................................................................79
6.2 Longstanding Factors Infuencing Soybean Transformation .......80
6.2.1 Genotype Competency and Regenerability ................80
6.2.2 Choice of Explants ..................................................... 81
6.2.3 Co-Cultivation Conditions .........................................82
6.2.4 Selection System.........................................................83
6.2.5 Genetic or Chimeric Effects .......................................84
6.3 Emerging Factors Affecting Soybean Transformation ............85
6.3.1 Seed Quality ...............................................................85
6.3.2 Seasonality .................................................................86
6.3.3 Sterilisation Techniques .............................................87
6.3.4 Plant Growth Regulators ............................................88
6.3.5 Antibiotics ..................................................................88
6.3.6 Oxidative Stress ..........................................................89
6.4 Summary .................................................................................90
6.5 Abbreviations ........................................................................... 91
References .......................................................................................... 91
Chapter 7 Alternative Techniques for Genetic Manipulations in Soybean ........95
7.1 Introduction .............................................................................95
7.2 Alternative Techniques for Genetic Improvement ..................95
7.2.1 Particle Bombardment/Biolistic Method ...................96
7.2.2 Liposome-Mediated Transfection ..............................97
7.2.3 Fibre-Mediated DNA Delivery ..................................98
7.2.4 Laser-Induced Genetic Improvement .........................99
7.2.5 In-Planta Transformations .........................................99
7.2.6 Mutagenesis ..............................................................100
7.2.7 Marker-Assisted Selection ........................................100
7.2.8 Qualitative Trait Loci ............................................... 101
7.2.9 CRISPR-Cas9 ...........................................................102
7.3 Other Uncommon Methods ...................................................102
7.3.1 Transfection via Calcium Phosphate Precipitation ....... 103
7.3.2 Diethyl Amino Ethyl (DEAE) Dextran-Mediated
DNA Transfer ........................................................... 103
7.3.3 Polyethylene Glycol (PEG)-Mediated
DNA Transfer ........................................................... 104
7.4 Status of A. rhizogenes in Plant Transformation ...................104
7.5 Pros and Cons of Artifcial Gene Transfer Methods .............105
7.6 Summary ............................................................................... 106
7.7 Abbreviations ......................................................................... 107
References ........................................................................................ 108