Testing for genetic manipulation in plants /
The debate over genetic manipulation and its use in plant improvement and protection has led to an increased demand for developing methods for detecting and characterizing genetic manipulation in plants and plant products such as seeds and foods. This book is unique in presenting all relevant method...
| Other Authors: | , , |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
Berlin ; New York :
Springer,
[2002], ©2002
Berlin ; New York : Springer-Verlag Berlin Heidelberg, c2002 New York : 2002 Berlin ; New York : [2002] |
| Series: | Molecular methods of plant analysis ;
v. 22 Molecular methods of plant analysis ; v. 22 |
| Subjects: |
Table of Contents:
- 1 Selectable and Screenable Markers for Rice Transformation / R.M. Twyman, E. Stoger, A. Kohli, T. Capell, P. Christou
- 1.2 Dominant Selectable Markers for Rice 3
- 1.2.1 Aminoglycoside 3'-Phosphotransferase (Neomycin Phosphotransferase) 3
- 1.2.2 Hygromycin Phosphotransferase 4
- 1.2.3 Phosphinothricin Acetyltransferase 4
- 1.2.4 Other Dominant Selectable Markers 5
- 1.3 Novel Selectable Markers 7
- 1.3.1 Innocuous Markers 7
- 1.3.2 Counterselectable Markers 7
- 1.4 Screenable Marker Genes 8
- 1.4.1 [beta]-Glucuronidase (gusA) 9
- 1.4.2 Firefly Luciferase (luc) 11
- 1.4.3 Green Fluorescent Protein (gfp) 11
- 1.5 Strategies for Marker-Gene Delivery 13
- 2 Use of Green Fluorescent Protein to Detect Transformed Shoots / J. Molinier, G. Hahne
- 2.2 GFP: Suitable as a Visually Selectable Marker In Planta? 20
- 2.2.1 Important Properties of the Protein 20
- 2.2.2 Properties of a Useful Selectable Marker in Plant Transformation Technology 21
- 2.3 GFP Expression and Detection in Primary Transformed Tissues 22
- 2.3.1 Transient Expression and GFP Detection 22
- 2.3.2 Detection Equipment and Troubleshooting 22
- 2.3.3 Stable Expression and GFP Detection in Primary Transformed Tissues 23
- 2.4 GFP for Screening of Segregating Populations 28
- 3 Luciferase Gene Expressed in Plants, Not in Agrobacterium / S.L. Mankin
- 3.2 Preventing Bacterial Expression 31
- 3.3 Imaging Luciferase Activity In Planta 32
- 3.4 Measuring Luciferase Activity in Plant Extracts 34
- 4 Use of [beta]-Glucuronidase (GUS) To Show Dehydration and High-Salt Gene Expression / K. Nakashima, K. Yamaguchi-Shinozaki
- 4.2 What Is GUS? 39
- 4.3 Trasgenic Plants Carrying Promoter-GUS Constructs 41
- 4.3.1 Construction of Promoter-GUS Fusion Genes 44
- 4.3.2 Introduction of Promoter-GUS Constructs into Agrobacterium 45
- 4.3.3 Transformation of Plants with Agrobacterium 46
- 4.3.3.1 Transformation of Arabidopsis Plants 46
- 4.4 Fluorometric Assay 49
- 4.4.2 Stress Conditions 49
- 4.4.2.1 Plant Preparation 49
- 4.4.2.2 Dehydration 49
- 4.4.2.3 High Salinity 49
- 4.4.2.4 ABA Treatment 50
- 4.4.2.5 Other Treatments 50
- 4.4.3 Protein Assay 50
- 4.4.4 Sample Preparation 50
- 4.4.5 Fluorometric Assay 51
- 4.5 Histochemistry 51
- 4.5.2 Histochemistry 52
- 4.6 Northern Analysis of GUS 52
- 4.6.2 RNA Extraction 53
- 4.6.3 RNA Blotting 54
- 4.6.4 Northern Hybridization 55
- 4.7 Application of the GUS System 56
- 4.7.1 Transient Assay 56
- 4.7.2 Transactivation Experiment 57
- 4.7.3 Promoter Tagging (Enhancer Trap) 57
- 5 Methods for Detecting Genetic Manipulation in Grain Legumes / H.-J. Jacobsen, R. Greiner
- 5.2 Detection at the DNA Level 64
- 5.3 PCR Analysis 65
- 5.4 Control PCR and Specific PCR Systems 66
- 5.5 Quantitative Approach 69
- 5.6 Competitive PCR 69
- 5.7 Real-Time PCR Systems 70
- 6 Elimination of Selectable Marker Genes from Transgenic Crops / A.P. Gleave
- 6.2 Co-transformation 74
- 6.3 Transposon-Mediated Approaches 78
- 6.3.1 Transposon-Mediated Repositioning 78
- 6.3.2 Transposon-Mediated Elimination 80
- 6.4 Site-Specific Recombination 81
- 6.4.1 The Cre/loxP System 82
- 6.4.2 The FLP/frt System 86
- 6.4.3 The R/RS System 88
- 6.5 Intrachromosomal Homologous Recombination 89
- 6.6 Conclusions and Future Prospects 90
- 7 GST-MAT Vector for the Efficient and Practical Removal of Marker Genes from Transgenic Plants / H. Ebinuma, K. Sugita, E. Matsunaga, S. Endo, K. Yamada
- 7.2 ipt-Type MAT Vectors 96
- 7.2.1 Transposable Element 96
- 7.2.2 Site-Specific Recombination System 97
- 7.2.3 Advantages of the ipt Gene 101
- 7.3 Two-Step Transformation 103
- 7.3.1 Promoter of the R Gene 103
- 7.3.2 Promoter of the ipt Gene 105
- 7.3.3 Combination of the ipt and iaaM/H Genes 108
- 7.3.4 Transgene Stacking 110
- 7.4 Single-Step Transformation 112
- 7.5 Cloning Vector for Desired Genes 114
- 8 Safety Assessment of Insect Protected Crops: Testing the Feeding Value of Bt Corn and Cotton Varieties in Poultry, Swine and Cattle / B. Hammond, E. Stanisiewski, R. Fuchs, J. Astwood, G. Hartnell
- 8.1.1 Food Safety Standards 119
- 8.1.2 Testing for Food and Feed Safety 120
- 8.2 Insect Protection Traits 122
- 8.3 Benefits 123
- 8.4 Safety Assessment of the Cry Insect-Control Proteins 124
- 8.5 Mode of Action 125
- 8.6 Substantial Equivalence Based on Compositional Analysis 126
- 8.7 Current Products 127
- 8.8 Grower Acceptance 127
- 8.9 Future Products 127
- 8.10 Farm-Animal Studies 128
- 8.10.1 Bt Corn 128
- 8.10.1.1 Poultry 128
- 8.10.1.2 Lactating Cows 131
- 8.10.1.3 Beef and Sheep 131
- 8.10.1.4 Swine 132
- 8.11 Cottonseed 133
- 9 Safety Assessment of Genetically Modified Rice and Potatoes with Soybean Glycinin / K. Momma, W. Hashimoto, S. Utsumi, K. Murata
- 9.2 Safety Assessment of Genetically Modified Crops 140
- 9.2.1 Genetically Modified Rice 141
- 9.2.2 Genetically Modified Potatoes 144
- 10 Chromosomal and Genetic Aberrations in Transgenic Soybean / R.J. Singh
- 10.2 Times in Culture with 2,4-D Prior to Transformation 154
- 10.3 Genetic Background of the Explants 159
- 10.4 Seed Fertility in Transgenic Soybean 160
- 10.5 Cytological Basis of Gene Silencing 163
- 11 Transgenic Barley (Hordeum vulgare L.) and Chromosomal Variation / M.-J. Cho, H.W. Choi, P. Bregitzer, S. Zhang, P.G. Lemaux
- 11.2 Chromosomal Variation in Nontransgenic Barley Plants 170
- 11.3 Chromosomal Variation in Transgenic Barley Plants 172
- 11.4 Fidelity and Quality of Transgenic Barley Plants 177
- 11.4.1 Comparative Analysis of Genomic Stability in Plants Derived from Tissues Generated Using Different in Vitro Proliferation Processes 177
- 11.4.2 Somaclonal Variation and Field Performance of Transgenic Plants Derived from Embryogenic Callus 179
- 11.4.3 Stability of Transgenes and Transgene Expression 180
- 11.5 Conclusions and Future Perspectives 184