Genetic Transformation of Plants

by Jackson, J. F.; Linskens, H. F.

• ISBN13: 9783540002925
• ISBN10: 3540002928
• Format: Hardcover
• Copyright: 2003-09-01
• Publisher: Springer Verlag

Summary

Whilst genetic transformation of plants is commonly viewed as a means of bringing about plant improvement, it has not so readily been recognised as a tool for analysing the function of plant genes. This book is unusual in that it focuses on the genetic transformation of a range of plants using a number of different methods. Many plants have been found to be quite difficult to transform, and so various techniques were developed. These techniques include: Agrobacterium suspension drops, electroporation, PEG, "whiskers", and various biolistic methods. A chapter on intellectual and property rights is included.

Table of Contents

1 Exclusive Rights in Life: Biotechnology, Genetic Manipulation, and Intellectual Property Rights
E.R. GOLD
1.1 Introduction	1	(1)
1.2 Biotechnological Innovation	2	(4)
1.2.1 Physical Innovations	3	(1)
1.2.1.1 DNA and Protein Molecules	3	(1)
1.2.1.2 Cells	3	(1)
1.2.1.3 Whole Organisms	4	(1)
1.2.2 Information and Other Intangibles	4	(2)
1.2.2.1 DNA Sequences and Cells	4	(1)
1.2.2.2 Processes Using Biological Matter	5	(1)
1.2.2.3 Bioinformatics	5	(1)
1.2.3 Summary	6	(1)
1.3 Introduction to Intellectual Property Rights	6	(11)
1.3.1 Exclusive Rights vs. Rights to Things	6	(1)
1.3.2 Property and Intellectual Property Rights	7	(1)
1.3.3 Trade Secrets	7	(1)
1.3.3.1 Subject Matter	8	(1)
1.3.3.2 Requirements	8	(1)
1.3.4 Patents	8	(6)
1.3.4.1 Subject Matter	9	(1)
1.3.4.1.1 Invention vs. Discovery	10	(1)
1.3.4.1.2 Exclusions	10	(1)
1.3.4.2 Requirements	11	(1)
1.3.4.2.1 Substantive Criteria	11	(1)
1.3.4.2.1.1 Novelty	12	(1)
1.3.4.2.1.2 Inventive Step (Nonobviousness)	12	(1)
1.3.4.2.1.3 Industrial Application (Utility)	12	(1)
1.3.4.2.2 Procedural Criterion: Disclosure	13	(1)
1.3.4.3 Remedies	14	(1)
1.3.5 Copyright and Database Protection	14	(2)
1.3.5.1 Subject Matter	15	(1)
1.3.5.2 Requirements	15	(1)
1.3.5.3 Remedies	16	(1)
1.3.6 Plant Variety Protection	16	(1)
1.3.6.1 Subject Matter	16	(1)
1.3.6.2 Requirements	17	(1)
1.4 Challenges	17	(4)
1.4.1 Incentive vs. Access	18	(2)
1.4.1.1 Justification for Property Rights	18	(1)
1.4.1.2 Economic Reality	19	(1)
1.4.2 Fairness to Providers of Biological Matter	20	(6)
1.4.2.1 Rights to Biological Matter	20	(1)
1.5 Conclusion	21	(1)
References	21	(2)
2 Agrobacterium rhizogenes-Mediated Transformation of Plants
W. VAN DE VELDE, M. KARIMI, G. DEN HERDER, M. VAN MONTAGU, M. HOLSTERS, and S. GOORMAGHTIG
2.1 Introduction	23	(3)
2.2 Aspects Influencing A. rhizogenes Transformation Efficiency	26	(5)
2.2.1 Choice of A. rhizogenes Strain	26	(2)
2.2.2 Choice of Explant	28	(1)
2.2.3 Preparation of Bacterial Inoculum and Infection of Explants	29	(1)
2.2.4 Co-cultivation	29	(2)
2.3 Establishing the Transformed Nature of Hairy Roots	31	(1)
2.4 Cotransformation of Binary T-DNA	31	(2)
2.5 Propagation of Hairy Root Lines in Liquid Cultures	33	(1)
2.5.1 The Clonal Status of Hairy Roots	33	(1)
2.5.2 Stability of Long-Term Hairy Root Cultures	34	(1)
2.6 Regeneration of Plants from Hairy Roots	34	(1)
2.7 The Multi-Auto-Transformation (MAT) Vector System	35	(2)
2.8 Conclusions	37	(1)
Protocol 1: Production of Transformed Hairy Roots	37	(1)
Protocol 2: Plant Regeneration from Hairy Roots	38	(1)
Protocol 3: Hairy Root Liquid Culture	39	(1)
References	39	(6)
3 Transformation of Petunia hybrida by the Agrobacterium Suspension Drop Method
S.J. WYLIE, D. TJOKROKUSUMO, and J.A. McC0MB
3.1 Introduction	45	(2)
3.2 Transformation	47	(1)
3.3 Analysis of Transformants	47	(2)
3.3.1 Screening Petunia Seedlings for Herbicide Resistance	47	(1)
3.3.2 Transmission of Basta Resistance Phenotype to T2 Progeny	48	(1)
3.3.3 β-Glucuronidase Assay	48	(1)
3.3.4 DNA Analysis	49	(1)
3.4 Conclusion	49	(1)
References	49	(4)
4 Onion, Leek and Garlic Transformation by Co-cultivation with Agrobacterium
C.C. EADY
4.1 Introduction	53	(2)
4.1.1 Current Applications of Allium Transformation Technology	53	(2)
4.1.1.1 Physiological Studies	53	(1)
4.1.1.2 Herbicide Resistance	54	(1)
4.1.1.3 Antimicrobial Resistance	54	(1)
4.1.1.4 Insect Resistance	55	(1)
4.2 Onion Transformation Protocols	55	(7)
4.2.1 Transformation Using Antibiotic and Visual Selection	56	(1)
4.2.1.1 Bacterial Strain and Plasmids	56	(1)
4.2.1.2 Transformation Procedure (Modified from Eady et al. 2000	56	(1)
4.2.2 Transformation Using Herbicide Selection	57	(1)
4.2.2.1 Bacterial Strain and Plasmids	58	(1)
4.2.2.2 Transformation Procedure	58	(1)
4.2.3 Ex-Flasking and Growth in Containment	58	(1)
4.2.4 Transgene Detection	59	(1)
4.2.5 Transgene Expression and Stability	60	(3)
4.2.5.1 Visual Reporter Genes	60	(1)
4.2.5.2 Expression of Herbicide Resistance	61	(1)
4.2.5.3 Antisense Alliinase Gene Expression	61	(1)
4.3 Leek Transformation	62	(1)
4.4 Garlic Transformation Protocol	63	(1)
4.4.1 Bacterial Strain and Plasmids	63	(1)
4.4.2 Transformation Procedure	63	(1)
4.5 Concluding Remarks	64	(1)
References	65	(4)
5 Electroporation Transformation of Barley
E. GUREL and N. GÖZÜKIRMIZI
5.1 Introduction	69	(2)
5.2 Background of Electroporation Procedures	71	(3)
5.2.1 Pre- and Post-Electroporation Period	71	(2)
5.2.2 Electrical Variables	73	(1)
5.3 Culture and Electroporation of Barley Explants	74	(9)
5.3.1 Protoplasts	74	(2)
5.3.2 Microspores	76	(1)
5.3.3 Intact Tissues	77	(15)
5.3.3.1 Analysis and Inheritance of Transgenes in Electroporated Tissues	81	(2)
5.4 Conclusions and Future Perspectives	83	(1)
References	84	(7)
6 Sorghum Transformation
Z. ZHAO and D. TOMES
6.1 Introduction	91	(1)
6.2 Sorghum Transformation Process and Optimization	92	(4)
6.2.1 Plant Materials and Transformation Systems	92	(1)
6.2.2 Transformation Via Microprojectile Bombardment	93	(1)
6.2.3 Agrobacterium-Mediated Transformation	94	(2)
6.3 Analysis of Transgenic Plants and the Progeny	96	(3)
6.3.1 Molecular Analysis of To Plants	97	(1)
6.3.2 Foreign Gene Expression in To Plants	98	(1)
6.3.3 Genetic and Molecular Analysis of the Progeny	99	(1)
6.4 Marker-Free Sorghum Transgenic Plants	99	(3)
6.4.1 Importance of Marker-Free Transgenics in Sorghum	100	(1)
6.4.2 Methods to Eliminate Markers from Transgenic Plants	100	(1)
6.4.3 Agrobacterium 2 T-DNA Co-Transformation System	101	(1)
References	102	(7)
7 Transgenic Sunflower: PEG-Mediated Gene Transfer
P.C. BINSFELD
7.1 Introduction	109	(1)
7.2 Genetic Variability and Transgenic Breeding	109	(1)
7.3 Gene Transfer Systems	110	(4)
7.3.1 PEG-Mediated Gene Transfer	111	(4)
7.3.1.1 Short DNA Molecule Uptake	111	(1)
7.3.1.2 Large DNA Molecule Uptake	112	(2)
7.4 Plant Regeneration	114	(1)
7.5 General Analytical Considerations	115	(9)
7.5.1 Molecular Analysis	115	(3)
7.5.1.1 DNA Extraction	116	(1)
7.5.1.2 Southern Hybridization	116	(1)
7.5.1.3 Polymerase Chain Reaction	116	(1)
7.5.1.4 Random Amplified Polymorphic DNA	117	(1)
7.5.2 Biochemical Analysis	118	(2)
7.5.2.1 Multiple Molecular Forms of Enzymes	118	(1)
7.5.2.2 Enzymatic Assay	119	(1)
7.5.3 Cytogenetic Analysis	120	(3)
7.5.3.1 Flow Cytometric Analysis	120	(2)
7.5.3.2 Mitotic and Meiotic Cell Analysis	122	(1)
7.5.3.3 In Situ Hybridization	122	(1)
7.5.4 Morphological Analysis	123	(1)
7.6 Conclusions and Future Perspectives	124	(1)
References	124	(3)
8 Transformation of Norway Spruce (Picea abies) by Particle Bombardment
D.H. CLAPHAM, H. HÄGGMAN, M. ELFSTRAND, T. ARONEN, and S. VON ARNOLD
8.1 Introduction	127	(1)
8.2 Types of Particle Accelerator	127	(1)
8.3 Transformation of Embryogenic Cultures	128	(5)
8.3.1 Transient Expression in Embryogenic Cultures	128	(1)
8.3.2 Production of Stably Transformed Cell Cultures and Transgenic Plants	129	(2)
8.3.3 Stability of Transgene Expression	131	(1)
8.3.4 Trends in Transgenic Plant Production	131	(2)
8.4 Transformation of Pollen	133	(3)
8.4.1 The Reproductive Biology of Norway Spruce	133	(1)
8.4.2 Transient Expression in Pollen	134	(1)
8.4.3 Development of Controlled Pollination Techniques for Bombarded Pollen	135	(1)
8.5 Applications of Transgenic Norway Spruce in Research	136	(3)
8.5.1 Genes Regulating Embryogenesis	136	(1)
8.5.2 Genes with Similarity to Defense Genes	137	(2)
8.6 Prospects for Transgenic Norway Spruce in Practical Forestry	139	(4)
References	143	(4)
9 WHISKERS-Mediated Transformation of Maize
J.F. PETOLINO, M. WELTER, and C. QIHUA CAI
9.1 Introduction	147	(1)
9.2 Preparation of Purified DNA Fragments	147	(3)
9.3 Establishment and Maintenance of Embryogenic Suspension Cultures	150	(2)
9.4 DNA Delivery via WHISKERS	152	(1)
9.5 Transgene Copy Number Estimation	153	(4)
9.6 Regeneration of Transgenic Plants and Progeny	157	(1)
9.7 Conclusions and Future Perspectives	157	(1)
References	158	(1)
10 Genetic Transformation of Soybean with Biolistics
D. SIMMONDS
10.1 Introduction	159	(1)
10.2 Tissue Culture and Plant Regeneration	160	(4)
10.2.1 Genotype Specificity	160	(1)
10.2.2 Initiation and Repetitive Proliferation of Somatic Embryogenic Cultures	161	(2)
10.2.3 Embryo Histodifferentiation and Maturation	163	(1)
10.2.4 Germination, Conversion and Plant Fertility	163	(1)
10.3 Transformation	164	(3)
10.3.1 Gene Delivery	164	(1)
10.3.2 Target Tissue Optimization	165	(1)
10.3.3 Selection	165	(1)
10.3.4 Transgenic Plant Recovery	166	(1)
10.4 Conclusions	167	(1)
10.5 Protocol	168	(2)
10.5.1 Induction and Maintenance of Proliferative Embryogenic Cultures	168	(1)
10.5.2 Transformation	168	(1)
10.5.3 Selection	169	(1)
10.5.4 Plant Regeneration	169	(1)
References	170	(5)
11 Genotoxic Effects of Tungsten Microparticles Under Conditions of Biolistic Transformation
J. BUCHOWICZ and C. KRYSIAK
11.1 Introduction	175	(1)
11.2 Biological Significance of Tungsten	175	(3)
11.2.1 Early Observations on Biological Effects of Tungsten	175	(1)
11.2.2 Catalytic Activity of Simple Tungsten Compounds	176	(1)
11.2.3 Tungstoenzymes	176	(1)
11.2.4 Tungsten-DNA Interaction	177	(1)
11.3 Tungsten Microparticles in Biotechnological Applications	178	(4)
11.3.1 Biolistic Transformation	178	(3)
11.3.1.1 An Overview	178	(2)
11.3.1.2 Technical Details	180	(1)
11.3.2 Biolistic Inoculation and Related Applications of Tungsten Particles	181	(1)
11.4 Assessment of Tungsten-Induced DNA Lesions	182	(4)
11.4.1 Electrophoretic Analysis of Tungsten-Damaged Plasmid DNA	182	(2)
11.4.2 A Modified TUNEL Method for Detection of Cellular DNA Fragmentation	184	(2)
11.5 Post-Bombardment Inhibition of Somatic Embryogenesis	186	(2)
11.6 Concluding Remarks	188	(1)
References	188	(7)
Subject Index	195