Plant biotechnology : the genetic manipulation of plants

The purpose of this book is to provide an overview of the production of GM crops, highlighting the key scientific and technical advances that underpin their development. The text begins with a summary of current knowledge about plant genome organisation and gene expression, followed by an introduction to the techniques of plant tissue culture and genetic transformation and their application to crop plants. A consideration of the design of constructs for plant genetic manipulation precedes a series of chapters covering specific targets for GM crops. These include the genetic manipulation of herbicide resistance, pest resistance and disease resistance. Strategies for engineering stress tolerance and the improvement of crop yield and quality are discussed, and the prospects for "molecular farming" are considered. Key themes and strategies are developed using appropriate case studies, which place the science in its broader agricultural/commercial context. The text concentrates on the core molecular biological issues, while the accompanying Online Resource Centre encourages an exploration of the wider implications and concerns about GM crops. Online Resource Centre All the figures from the book are available to download free from the Online Resource Centre. It also features weblinks to relevant sites with additional information or discussion including those hosted by the protagonists of the GM debate.

Author Notes

Dr Adrian Slater is the Deputy Director of the Norman Borlaug Institute for Plant Science Research at De Montfort University, Leicester
Dr Nigel Scott is a Principal Lecturer in plant molecular biology in the Norman Borlaug Institute
Dr Mark Fowler is a Senior Lecturer in plant molecular biology in the Norman Borlaug Institute

Reviews 1

Choice Review

As the world's population increases, we need to produce more food in a low-cost, environmentally friendly way. Scientists are improving crops by adding herbicides, disease, and pest resistances. This is the first book to cover the scientific aspects of genetically modified crops as well as the social and ethical debate. Slater, Scott, and Fowler all are senior scientists at the Norman Borlaug Institute for Plant Science Research, De Montfort University, UK. The first four chapters explain the laboratory techniques used to transform plants. Four chapters discuss the introduction of resistance traits into agricultural plants and three chapters treat stress tolerance, crop yields, and the use of plants to produce important molecules. The final chapter provides a balanced discussion about genetically modified crops of the present day and what might be possible in the future. Information is also provided about government regulations in Europe and the US and about the concerns of environmentalists. Excellent tables and figures; comprehensive, accessible information. The book's Web page should be very useful. ^BSumming Up: Recommended. Lower-division undergraduates through professionals; two-year technical program students. P. M. Bradley Worcester State College

Preface	p. v
List of Abbreviations	p. xiii
Foreword	p. xxi
1 Plant genomes: the organisation and expression of plant genes	p. 1
Introduction	p. 1
DNA, chromatin and chromosome structure	p. 1
An introduction to gene structure and gene expression	p. 6
Gene structure and expression in a eukaryotic protein-coding gene	p. 6
Translation	p. 7
Regulation of gene expression	p. 12
Chromatin conformation	p. 14
Gene transcription	p. 14
RNA modification, splicing, turnover and transport	p. 15
Translation	p. 18
Post-translational modification	p. 18
Localisation	p. 19
Protein turnover	p. 19
Conclusions	p. 20
Implications for plant transformation	p. 20
Examples of promoter elements used to drive transgene expression	p. 24
Protein targeting	p. 24
Heterologous promoters	p. 24
Genome size and organisation	p. 25
Arabidopsis and the new technologies	p. 25
Genome-sequencing projects--technology, findings and applications	p. 26
Biotechnological implications of the 'Arabidopsis Genome Sequencing Initiative'	p. 30
Crop plant genome sequencing	p. 30
Summary	p. 32
Further reading	p. 33
2 Plant tissue culture	p. 35
Introduction	p. 35
Plant tissue culture	p. 35
Plasticity and totipotency	p. 35
The culture environment	p. 36
Plant cell culture media	p. 36
Plant growth regulators	p. 40
Culture types	p. 43
Callus	p. 43
Cell-suspension cultures	p. 43
Protoplasts	p. 44
Root cultures	p. 45
Shoot tip and meristem culture	p. 45
Embryo culture	p. 45
Microspore culture	p. 45
Plant regeneration	p. 46
Somatic embryogenesis	p. 47
Organogenesis	p. 50
Integration of plant tissue culture into plant transformation protocols	p. 51
Summary	p. 52
Further reading	p. 52
3 Techniques for plant transformation	p. 55
Introduction	p. 55
Agrobacterium-mediated gene transfer	p. 55
The biology of Agrobacterium	p. 55
The Ti plasmid	p. 56
Ti-plasmid features	p. 57
The process of T-DNA transfer and integration	p. 59
Practical applications of Agrobacterium-mediated plant transformation	p. 63
Transformation in planta	p. 66
Direct gene transfer methods	p. 66
Particle bombardment	p. 67
Polyethylene glycol (PEG)-mediated transformation	p. 72
Electroporation	p. 72
Silicon carbide fibres--WHISKERS	p. 75
Summary	p. 75
Further reading	p. 77
4 Binary vectors for plant transformation	p. 79
Introduction	p. 79
Desirable features of any plasmid vector	p. 79
Development of plant transformation vectors	p. 80
Basic features of vectors for plant transformation	p. 80
Promoters and terminators	p. 82
Selectable markers	p. 87
Reporter genes	p. 88
Origins of replication	p. 93
Co-integrative and binary vectors	p. 93
Families of binary vectors	p. 93
Optimisation	p. 94
Arrangement of genes in the vector	p. 94
Transgene copy number	p. 96
Transgene position	p. 96
Transgene features	p. 97
Clean gene technology	p. 97
Summary	p. 99
Further reading	p. 100
5 The genetic manipulation of herbicide resistance	p. 103
Introduction	p. 103
The use of herbicides in modern agriculture	p. 104
What types of compounds are herbicides?	p. 105
Strategies for engineering herbicide resistance	p. 107
Prospects for plant detoxification systems	p. 121
Commercialisation of herbicide-resistant plants to date	p. 122
The environmental impact of herbicide-resistant crops	p. 125
The development of 'super weeds'	p. 126
Summary	p. 129
Further reading	p. 129
6 The genetic manipulation of pest resistance	p. 131
Introduction	p. 131
The nature and scale of insect pest damage to crops	p. 131
GM strategies for insect resistance: The Bacillus thuringiensis approach to insect resistance	p. 132
The use of 'Bt' as a biopesticide	p. 136
Bt-based genetic modification of plants	p. 137
The problem of insect resistance to Bt	p. 139
The environmental impact of Bt crops	p. 143
The 'Copy Nature' strategy	p. 145
Insect resistant crops and food safety	p. 152
Summary	p. 153
Further reading	p. 153
7 Plant disease resistance	p. 157
Introduction	p. 157
Existing non-GM approaches	p. 157
Plant-pathogen interactions	p. 159
Prokaryotes	p. 160
Fungi and water moulds	p. 160
Viruses	p. 161
Natural disease resistance pathways--overlap between pests and diseases	p. 162
Anatomical defences	p. 163
Pre-existing protein and chemical protection	p. 163
Inducible systems	p. 163
Systemic responses	p. 170
Biotechnological approaches to disease resistance	p. 170
Protection against fungal pathogens	p. 171
Antimicrobial proteins	p. 174
Induction of HR and SAR in transgenic plants	p. 175
Summary	p. 176
Further reading	p. 177
8 Reducing the effects of viral disease	p. 179
Introduction	p. 179
Types of plant viruses	p. 179
RNA viruses	p. 181
Entry and replication--points of inhibition	p. 183
How has industry dealt with viruses?	p. 184
The transgenic approach--PDR	p. 187
Interactions involving viral proteins	p. 188
RNA effects	p. 192
What has been commercialised in the West?	p. 198
Yellow squash and zucchini	p. 198
Papaya	p. 198
Potato	p. 199
Risk	p. 199
Summary	p. 201
Further reading	p. 202
9 Strategies for engineering stress tolerance	p. 205
Introduction	p. 205
The nature of abiotic stress	p. 206
The nature of water-deficit stress	p. 207
Different abiotic stresses create a water deficit	p. 208
Targeted approaches towards the manipulation of tolerance to specific water-deficit stresses	p. 215
Alternative approaches to salt stress	p. 215
Alternative approaches to cold stress	p. 218
Tolerance to heat stress	p. 220
Secondary effects of abiotic stress--the production of reactive oxygen species	p. 222
Strategy 1 Expression of enzymes involved in scavenging ROS	p. 223
Strategy 2 Production of antioxidants	p. 227
Summary	p. 229
Further reading	p. 229
10 The improvement of crop yield and quality	p. 231
Introduction	p. 231
The genetic manipulation of fruit ripening	p. 232
Engineering plant protein composition for improved nutrition	p. 251
The genetic manipulation of crop yield by enhancement of photosynthesis	p. 252
Manipulation of light harvesting and the assimilate distribution--phytochromes	p. 253
Direct manipulation of photosynthesis--enhancement of dark reactions	p. 256
Summary	p. 257
Further reading	p. 258
11 Molecular farming/'pharming'	p. 261
Introduction	p. 261
Carbohydrates and lipids	p. 261
Carbohydrate production	p. 263
Metabolic engineering of lipids	p. 270
Molecular farming of proteins	p. 279
Production systems	p. 280
Medically related proteins	p. 289
Economic considerations for molecular farming	p. 298
Summary	p. 301
Further reading	p. 302
12 Future prospects for GM crops	p. 305
Introduction	p. 305
The current state of transgenic crops	p. 305
Who has benefited from these first-generation crops?	p. 307
What will drive the development of the future generations of GM crops?	p. 309
Concerns about GM crops	p. 309
Antibiotic resistance genes	p. 309
Herbicide resistance and 'super-weeds'	p. 311
Gene containment	p. 311
Big business	p. 312
The regulation of GM crops and products	p. 316
The Euopean Union (EU)	p. 316
The USA	p. 320
Future developments in the science of plant biotechnology	p. 324
'Greener' genetic engineering	p. 324
Summary	p. 329
Further reading	p. 330
Index	p. 335

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