Plant Transformation via Agrobacterium Tumefaciens compiles fundamental and specific information and procedures involving in vitro soybean transformation, which forms the basis for the Agrobacterium-mediated genetic manipulation of soybean using plant tissue culture. This method serves as one of the most preferred, reliable and cost-effective mechanism of transgene expression in both leguminous recalcitrant species and non-legume crops. The technology is favoured due to its simplicity, feasibility and high transformation rates that are so far achieved mostly in monocot plants and a few dicot genotypes.
This book provides a comprehensive review of plant transformation which remains necessary for many researchers who are still facing protocol-related hurdles. Among some of the major topics covered in Plant Transformation via Agrobacterium Tumefaciens are the history and discovery of Agrobacterium bacterium, longstanding challenges causing transformation inefficiencies, types and conditions of explants, development of transgenic plants for stress resistance, and the role of transgenic plants on animal/human health, including the environment.
Plant Transformation via Agrobacterium Tumefaciens helps the reader to understand how soybean, like many other orphan legume crops, faces the risk of overexploitation which may render the currently available varieties redundant and extinct should its narrow gene pool not improve. Plant transformation serves as a key technique in improving the gene pool, while developing varieties that are drought tolerant, have enhanced nutritional value, pest resistant and reduce the destruction by disease causing microorganims. This book is an essential foundation tool that is available for researchers and students to reinforce the application of Agrobacterium-mediated genetic transformation in soybean.
Author(s): Phetole Mangena
Publisher: CRC Press
Year: 2022
Language: English
Pages: 244
City: Boca Raton
Cover
Half Title
Title Page
Copyright Page
Dedication
Table of Contents
Preface
Acknowledgements
Author
Chapter 1 An Overview of Genetic Transformation in Plants
1.1 Introduction
1.2 Molecular Mechanism for DNA Introgression
1.3 Tissue Targeting for Plant Transformation
1.4 Screening and Selection of Transformants
1.5 Transfer to Natural Environment
1.6 Summary
1.7 Abbreviations
References
Chapter 2 Plant Transformation History
2.1 Discovery of Agrobacterium tumefaciens
2.2 Tumour-Inducing Principle
2.3 Structure and Modifications of the Agrobacterium Circular DNA
2.4 Genetic Transformation in Plants
2.5 The First Transgenic Plant
2.6 Role of Plant Tissue Culture
2.7 Summary
2.8 Abbreviations
References
Chapter 3 Agrobacterium tumefaciens
3.1 Introduction
3.2 Classification
3.3 General Characteristics of Agrobacterium
3.4 Size and Morphology
3.5 Nutrition
3.6 Reproduction
3.7 Cell Structure
3.8 Protoplasmic Materials
3.9 Chromosomal DNA and Plasmids
3.10 Summary
3.11 Abbreviations
References
Chapter 4 Getting Started with in Vitro Soybean Transformation
4.1 Introduction
4.2 Bacterial Strains
4.2.1 Bacterial Culture
4.3 Establishing Contamination-Free Cultures
4.3.1 Aseptic Seed Cultures
4.3.2 Culture Medium
4.4 Type and Competency of Explants
4.5 Co-Cultivation of Explants with A. tumefaciens
4.6 Shoot Induction and Elongation
4.7 Rooting of Elongated Shoots
4.8 Acclimatisation
4.9 Screening of Transgenic Plants
4.10 Other Considerations
4.10.1 Equipment and Laboratory Supplies
4.10.2 Surfactants
4.10.3 Antinecrotic Treatments
4.11 Summary
4.12 Abbreviations
References
Chapter 5 In Vitro Cultures Commonly Used for Plant Transformation
5.1 Introduction
5.2 Callus Culture
5.2.1 Purpose of Callus Culture
5.2.2 Requirements for Establishing Callus Culture
5.2.3 Application in Plant Transformation
5.3 Shoot Culture
5.3.1 Purpose of Shoot Culture
5.3.2 Culture Requirements for Efficient Shoot Multiplication
5.3.3 Application of Shoot Culture in Plant Transformation
5.4 Meristem Culture
5.4.1 Purpose of Meristem Culture
5.4.2 Meristem Culture Requirements
5.4.3 Use of Meristem Culture for Plant Improvement
5.5 Protoplast Culture
5.5.1 Purpose of Protoplast Culture
5.5.2 Requirements for Protoplast Culture
5.5.3 Applications in Plant Transformation
5.6 Summary
5.7 Abbreviations
References
Chapter 6 Current and Longstanding Challenges Facing Soybean Transformation
6.1 Introduction
6.2 Longstanding Factors Influencing Soybean Transformation
6.2.1 Genotype Competency and Regenerability
6.2.2 Choice of Explants
6.2.3 Co-Cultivation Conditions
6.2.4 Selection System
6.2.5 Genetic or Chimeric Effects
6.3 Emerging Factors Affecting Soybean Transformation
6.3.1 Seed Quality
6.3.2 Seasonality
6.3.3 Sterilisation Techniques
6.3.4 Plant Growth Regulators
6.3.5 Antibiotics
6.3.6 Oxidative Stress
6.4 Summary
6.5 Abbreviations
References
Chapter 7 Alternative Techniques for Genetic Manipulations in Soybean
7.1 Introduction
7.2 Alternative Techniques for Genetic Improvement
7.2.1 Particle Bombardment/Biolistic Method
7.2.2 Liposome-Mediated Transfection
7.2.3 Fibre-Mediated DNA Delivery
7.2.4 Laser-Induced Genetic Improvement
7.2.5 In-Planta Transformations
7.2.6 Mutagenesis
7.2.7 Marker-Assisted Selection
7.2.8 Qualitative Trait Loci
7.2.9 CRISPR-Cas
7.3 Other Uncommon Methods
7.3.1 Transfection via Calcium Phosphate Precipitation
7.3.2 Diethyl Amino Ethyl (DEAE) Dextran-Mediated DNA Transfer
7.3.3 Polyethylene Glycol (PEG)-Mediated DNA Transfer
7.4 Status of A. rhizogenes in Plant Transformation
7.5 Pros and Cons of Artificial Gene Transfer Methods
7.6 Summary
7.7 Abbreviations
References
Chapter 8 Molecular Aspects of Indirect Gene Transfer via A. tumefaciens
8.1 Introduction
8.2 Molecular Mechanism of Agrobacterium-Mediated Gene Transfer
8.2.1 Pathogenesis
8.2.2 T-DNA
8.2.3 Gene of Interest
8.3 Co-Resident Ti- and At-Plasmid Vectors
8.4 Gene Transfer and Expression
8.5 Infection and Co-Cultivation for Efficient Transgenesis
8.6 Molecular Role of Organic Supplements
8.7 DNA Repair for Efficient Integration
8.8 Immunity to Plant Transformation
8.9 Summary
8.10 Abbreviations
References
Chapter 9 Genetic Transformation in Other Leguminous Crops
9.1 Introduction
9.2 Status of Genetic Transformation in Other Legume Crops
9.2.1 Chickpea – Cicer arietinum
9.2.2 Pigeon Pea – Cajanus cajan
9.2.3 Lentil – Lens culinaris
9.2.4 Common Bean – Phaseolus vulgaris
9.2.5 Peas – Pisum sativum
9.2.6 Faba Bean – Vicia faba
9.2.7 Cowpea – Vigna unguiculata
9.2.8 Mung Bean – Vigna radiata
9.3 Summary
9.4 Abbreviations
References
Chapter 10 Transgenic Plants for Biotic Stress Resistance in Soybean
10.1 Introduction
10.2 Defining Biotic Stress
10.3 Plant-Pathogen Interactions
10.4 Soybean-Pathogen Specificity Genes
10.5 Soybean Defence against Biotic Stress
10.6 Transgenics Conferring Biotic Stress Resistance
10.7 Summary
10.8 Abbreviations
References
Chapter 11 Transgenic Plants for Abiotic Stress Resistance in Soybean
11.1 Introduction
11.2 Defining Abiotic Stress
11.3 Abiotic Stress Impact on Soybean Plants
11.3.1 Salinity
11.3.2 Heat Stress
11.3.3 Chilling Stress
11.3.4 Drought Stress
11.4 General Defence
11.5 Resistance Genes for Abiotic Stress Tolerance
11.6 Abiotic Stress-Tolerant Varieties
11.7 Summary
11.8 Abbreviations
References
Chapter 12 Potential Health Safety Concerns and Environmental Risks
12.1 Introduction
12.2 Evidence of Biosafety and Risks Involving Agrobacterium tumefaciens
12.3 Unintended T-DNA and Selectable Markers Transfer
12.4 Gene Transfer to Non-Targeted Hosts
12.5 Undesirable Gene Flow to Closely Related Species
12.6 Potential Risks to Human and Animal Health
12.7 Environmental Risk Assessment and Contamination
12.8 Microbial Competition with Transformed Agrobacterium Strains
12.9 Impact of Agrobacterium on the Environment
12.10 Summary
12.11 Abbreviations
References
Chapter 13 The Regulatory Management of Transgenic Plants
13.1 Introduction
13.2 Global Approach to GMO Regulations
13.3 Risk Analysis and Management
13.3.1 Risk Assessment of GM-Crops
13.3.2 Risk Communication of GM-Crop Analysis
13.3.3 Risk Management of Potential Transgenic Hazards
13.4 Regulation and Sanctions of Transgenic Products
13.5 Summary
13.6 Abbreviations
References
Chapter 14 Current and Future Prospects in Genetic Engineering
14.1 Introduction
14.2 Current Challenges in Plant Transformation
14.3 Role of GM-Crops in Agriculture
14.4 The Importance of GMOs for Food Security
14.5 Other Plant Breeding Methods
14.5.1 Mutation Breeding and Plant Mutagenesis
14.5.2 Microprojectile Bombardment
14.5.3 Genomic Editing
14.6 Improvement of Crop Traits
14.7 Development of Stress-Resistant Crops
14.8 Legislation and Social Acceptance
14.9 Future Prospects
14.10 Summary
14.11 Abbreviations
References
Chapter 15 Photographic Index of Soybean Transformation Cultures
15.1 Introduction
15.2 The Soybean Plant
15.3 Seed Sterilisation and Germination
15.4 Cotyledonary Node Explants
15.5 Agrobacterium tumefaciens and Co-Cultivation of Explants
15.6 Shoot Induction
15.6.1 In Vitro Elongation and Rooting
15.7 Plant Hardening and Acclimatisation
15.8 Green Fluorescent Protein (GFP)
15.9 Summary
15.10 Abbreviations
References
Glossary
Index
