Testing for Genetic Manipulation in Plants

by Jackson, John Flex; Linskens, H. F.; Inman, R. B.

• ISBN13: 9783540431534
• ISBN10: 3540431535
• Format: Hardcover
• Copyright: 2002-12-01
• Publisher: Springer Verlag

Summary

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 methods together in one volume: those for using and determining markers retained in genetically manipulated products as well as methods for eliminating marker genes and procedures for characterizing chromosomal aberrations in genetically manipulated plants.

Table of Contents

Selectable and Screenable Markers for Rice Transformation
R.M. Twyman
E. Stoger
A. Kohli
T. Capell
P. Christou
Introduction	1	(2)
Dominant Selectable Markers for Rice	3	(4)
Aminoglycoside 3'-Phosphotransferase (Neomycin Phosphotransferase)	3	(1)
Hygromycin Phosphotransferase	4	(1)
Phosphinothricin Acetyltransferase	4	(1)
Other Dominant Selectable Markers	5	(2)
Novel Selectable Markers	7	(1)
Innocuous Markers	7	(1)
Counterselectable Markers	7	(1)
Screenable Marker Genes	8	(5)
β-Glucuronidase (gusA)	9	(2)
Firefly Luciferase (luc)	11	(1)
Green Fluorescent Protein (gfp)	11	(2)
Strategies for Marker-Gene Delivery	13	(6)
References	14	(5)
Use of Green Fluorescent Protein to Detect Transformed Shoots
J. Molinier
G. Hahne
Introduction	19	(1)
GFP: Suitable as a Visually Selectable Marker In Planta?	20	(2)
Important Properties of the Protein	20	(1)
Properties of a Useful Selectable Marker in Plant Transformation Technology	21	(1)
GFP Expression and Detection in Primary Transformed Tissues	22	(6)
Transient Expression and GFP Detection	22	(1)
Detection Equipment and Troubleshooting	22	(1)
Stable Expression and GFP Detection in Primary Transformed Tissues	23	(5)
GFP for Screening of Segregating Populations	28	(1)
Conclusion	29	(2)
References	29	(2)
Luciferase Gene Expressed in Plants, Not in Agrobacterium
S.L. Mankin
Introduction	31	(1)
Preventing Bacterial Expression	31	(1)
Imaging Luciferase Activity In Planta	32	(2)
Measuring Luciferase Activity in Plant Extracts	34	(3)
References	35	(2)
Use of β-Glucuronidase (GUS) To Show Dehydration and High-Salt Gene Expression
K. Nakashima
K. Yamaguchi-Shinozaki
Introduction	37	(2)
What Is GUS?	39	(2)
Trasgenic Plants Carrying Promoter-GUS Constructs	41	(8)
Construction of Promoter-GUS Fusion Genes	44	(1)
Introduction of Promoter-GUS Constructs into Agrobacterium	45	(1)
Transformation of Plants with Agrobacterium	46	(1)
Transformation of Arabidopsis Plants	46	(3)
Fluorometric Assay	49	(2)
Introduction	49	(1)
Stress Conditions	49	(1)
Plant Preparation	49	(1)
Dehydration	49	(1)
High Salinity	49	(1)
ABA Treatment	50	(1)
Other Treatments	50	(1)
Protein Assay	50	(1)
Sample Preparation	50	(1)
Fluorometric Assay	51	(1)
Histochemistry	51	(1)
Introduction	51	(1)
Histochemistry	52	(1)
Northern Analysis of GUS	52	(4)
Introduction	52	(1)
RNA Extraction	53	(1)
RNA Blotting	54	(1)
Northern Hybridization	55	(1)
Application of the GUS System	56	(1)
Transient Assay	56	(1)
Transactivation Experiment	57	(1)
Promoter Tagging (Enhancer Trap)	57	(1)
Conclusion	57	(6)
References	59	(4)
Methods for Detecting Genetic Manipulation in Grain Legumes
H.-J. Jacobsen
R. Greiner
Introduction	63	(1)
Detection at the DNA Level	64	(1)
PCR Analysis	65	(1)
Control PCR and Specific PCR Systems	66	(3)
Quantitative Approach	69	(1)
Competitive PCR	69	(1)
Real-Time PCR Systems	70	(1)
Concluding Remarks	70	(3)
References	71	(2)
Elimination of Selectable Marker Genes from Transgenic Crops
A.P. Gleave
Introduction	73	(1)
Co-transformation	74	(4)
Transposon-Mediated Approaches	78	(3)
Transposon-Mediated Repositioning	78	(2)
Transposon-Mediated Elimination	80	(1)
Site-Specific Recombination	81	(8)
The Cre/loxP System	82	(4)
The FLP/frt System	86	(2)
The R/RS System	88	(1)
Intrachromosomal Homologous Recombination	89	(1)
Conclusions and Future Prospects	90	(5)
References	91	(4)
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
Introduction	95	(1)
ipt-Type MAT Vectors	96	(7)
Transposable Element	96	(1)
Site-Specific Recombination System	97	(4)
Advantages of the ipt Gene	101	(2)
Two-Step Transformation	103	(9)
Promoter of the R Gene	103	(2)
Promoter of the ipt Gene	105	(3)
Combination of the ipt and iaaM/H Genes	108	(2)
Transgene Stacking	110	(2)
Single-Step Transformation	112	(2)
Cloning Vector for Desired Genes	114	(1)
Concluding Remarks	115	(4)
References	116	(3)
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
Introduction	119	(3)
Food Safety Standards	119	(1)
Testing for Food and Feed Safety	120	(2)
Insect Protection Traits	122	(1)
Benefits	123	(1)
Safety Assessment of the Cry Insect-Control Proteins	124	(1)
Mode of Action	125	(1)
Substantial Equivalence Based on Compositional Analysis	126	(1)
Current Products	127	(1)
Grower Acceptance	127	(1)
Future Products	127	(1)
Farm-Animal Studies	128	(5)
Bt Corn	128	(1)
Poultry	128	(3)
Lactating Cows	131	(1)
Beef and Sheep	131	(1)
Swine	132	(1)
Cottonseed	133	(1)
Conclusions	134	(5)
References	135	(4)
Safety Assessment of Genetically Modified Rice and Potatoes with Soybean Glycinin
K. Momma
W. Hashimoto
S. Utsumi
K. Murata
Introduction	139	(1)
Safety Assessment of Genetically Modified Crops	140	(6)
Genetically Modified Rice	141	(3)
Genetically Modified Potatoes	144	(2)
Concluding Remarks	146	(7)
References	149	(4)
Chromosomal and Genetic Aberrations in Transgenic Soybean
R.J. Singh
Introduction	153	(1)
Times in Culture with 2,4-D Prior to Transformation	154	(5)
Genetic Background of the Explants	159	(1)
Seed Fertility in Transgenic Soybean	160	(3)
Cytological Basis of Gene Silencing	163	(1)
Conclusions	164	(5)
References	165	(4)
Transgenic Barley (Hordeum vulgare L.) and Chromosomal Variation
M.-J. Cho
H.W. Choi
P. Bregitzer
S. Zhang
P.G. Lemaux
Introduction	169	(1)
Chromosomal Variation in Nontransgenic Barley Plants	170	(2)
Chromosomal Variation in Transgenic Barley Plants	172	(5)
Fidelity and Quality of Transgenic Barley Plants	177	(7)
Comparative Analysis of Genomic Stability in Plants Derived from Tissues Generated Using Different in Vitro Proliferation Processes	177	(2)
Somaclonal Variation and Field Performance of Transgenic Plants Derived from Embryogenic Callus	179	(1)
Stability of Transgenes and Transgene Expression	180	(4)
Conclusions and Future Perspectives	184	(5)
References	185	(4)
Subject Index	189