Incluye bibliografía

Part I Agrobacterium tumefaciens-Mediated Gene Transfer.. Introduction I Agrobacterium tumefaciens, a Natural Vector System.. Protocols.. 1 Transformation of Arabidopsis thaliana Using Root Explants.. 2 Seed Transformation in Arabidopsis thaliana.. 3 In Planta Agrobacterium-Mediated Transformation of Adult Arabidopsis thaliana Plants by Vacuum-Infiltration.. 4 Agrobacterium-Mediated Transformation of Potato (Solanum tuberosum.. 5 Agrobacterium-Mediated Transformation of Rapeseed (Brassica napus.. Part II Agrobacterium rhizogenes-Mediated Gene Transfer.. Introduction II Agrobacterium rhizogenes, a Natural Transformation System.. Protocol.. 6 Agrobacterium rhizogenes-Mediated Transformation: Transformed Roots to Transformed Plants.. Part III Direct Gene Transfer to Protoplasts.. Introduction III Direct Gene Transfer to Protoplasts.. Protocols.. 7 Polyethylene Glycol-Mediated Direct Gene Transfer to Tobacco Protoplasts and Regeneration of Transgenic Plants.. 8 Polyethylene Glycol-Mediated Direct Gene Transfer to Indica Rice Protoplasts and Regeneration of Transgenic Plants.. 9 Plastid Transformation by Polyethylene Glycol Treatment of Protoplasts and Regeneration of Transplastomic Tobacco Plants.. 10 Electroporation-Mediated Gene Transfer to Protoplasts and Regeneration of Transgenic Plants.. 11 Electroporation Mediated Gene Transfer to Japonica Rice Protoplasts and Regeneration of Transgenic Plants.. 12 Regeneration of Transgenic Maize Plants from Embryogenic Protoplasts After Polyethylene Glycol-Mediated DNA Uptake.. 13 Gene Transfer to Protoplasts: Transient Gene Expression Analysis.. Part IV Biolistic Plant Transformation.. Introduction IV The Biolistic Transformation System.. Protocols.. 14 Transient Gene Expression Studies with the Biolistic System.. 15 Maize Transformation by Microprojectile Bombardment of Immature Embryos

16 Biolistic Transformation of Microspore-Derived and Immature Zygotic Embryos and Regeneration of Fertile Transgenic Barley Plants.. 17 Transformation of Soybean (Glycine max Through Electric Discharge Particle Acceleration.. 18 Ballistic Microtargeting of Visible Marker Genes to the Shoot Meristem of Wheat.. 19 Stable Transformation of Wheat by Microprojectile Bombardment.. 20 Biolistic-Based Analysis of Chloroplast Gene Expression.. Part V Microinjection and Fiber-Mediated Transformation.. Introduction V Microinjection into Plant Cells: Methodology and Applications.. Protocols.. 21 Microinjection into Tobacco Protoplasts and Regeneration of Transgenic Plants.. 22 Silicon Carbide Whisker-Mediated Transformation: Regeneration of Transgenic Maize Plants.. Part VI Tissue Electroporation.. Introduction VI Transformation by Tissue-Electroporation.. Protocols.. 23 Transient Expression Following Electroporation into Immature Zygotic Embryos of Wheat.. 24 Transgenic Indica Rice Plants by Electroporation of Seed Embryo Cells.. Part VII Analysis of Transgenic Plants.. Introduction VII Analysis of Transgenic Plants.. Protocols.. 25 Standard Molecular Techniques for the Analysis of Transgenic Plants.. 25.1 Isolation of Genomic DNA from Plants.. 25.2 Isolation of Total RNA from Plant Tissues.. 25.3 Southern Blotting Technique with Genomic DNA from Plants.. 25.4 Southern Blot Hybridization with Radioactive Probes 2.. 25.5 Southern Blot Hybridization with Nonradioactive Probes and Detection by Chemiluminescence.. 25.6 RNA Analysis by Northern Blot Hybridization.. 25.7 Western Blot Analysis.. 25.8 PCR Amplification of Plant Genomic DNA.. 25.9 Enzyme Assay for Phosphinotricin Acetyltransferase.. 25.10 Enzyme Assay for Hygromycin Phosphotransferase.. 25.11 Enzyme Assay for Neomycin Phosphotransferase II.. 25.12 Enzyme Assay for Cloramphenicol Acetyltransferase.. 25.13 Enzyme Assay for ß-Glucuronidase

25.14 Firefly Luciferase Assay.. 26 In Situ Hybridization on Plant Metaphase Chromosomes.. 27 Localisation of RNA Transcripts in Plant Tissue by in Situ Hybridisation.. 28 In Situ Immunolocalisation of Transgene Products in Transgenic Plants.. Part VIII Establishment and Maintenance of Embryogenic Cultures.. Introduction VIII Embryogenic Cultures: a Source of Target Cells for Gene Transfer.. Protocol.. 29 Establishment of and Plant Regeneration from Embryogenic Cell Suspensions and Their Protoplasts in Forage Grasses.. Part IX Genetic Markers and Expression Signals.. Introduction IX Genetic Markers and Expression Signals.. 30 Expression Signals and Vectors.. 31 Selectable Marker and Reporter Genes.. Part X Guidelines and Biosafety Considerations.. Introduction X Gene Technology with Plants and the Public.. 32 Release of Transgenic Plants: Regulations and Developments.. 33 Risk Assessment

Using the clearly structured protocols given in this manual, it will be easy to apply the latest techniques in plant biotechnology in order to create new plant varieties or varieties with altered and optimized characteristics. Direct gene transfer into plant cells or protoplasts by the use of microinjection, electroporation or biolistic systems, or mediated by Agrobacteria is described in detail for various plant species, including relevant crops and cereals. Also included are protocols of the standard molecular techniques for the analysis of transgenic plants as well as a section on biosafety considerations and regulations. Inglés