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.

Inhalt

I Agrobacterium tumefaciens-Mediated Gene Transfer.- 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).- II Agrobacterium rhizogenes-Mediated Gene Transfer.- 6 Agrobacterium rhizogenes-Mediated Transformation: Transformed Roots to Transformed Plants.- III Direct Gene Transfer to Protoplasts.- 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.- IV Biolistic Plant Transformation.- 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.- V Microinjection and Fiber-Mediated Transformation.- 21 Microinjection into Tobacco Protoplasts and Regeneration of Transgenic Plants.- 22 Silicon Carbide Whisker-Mediated Transformation: Regeneration of Transgenic Maize Plants.- VI Tissue Electroporation.- 23 Transient Expression Following Electroporation into Immature Zygotic Embryos of Wheat.- 24 Transgenic Indica Rice Plants by Electroporation of Seed Embryo Cells.- VII Analysis of Transgenic Plants.- 25 Standard Molecular Techniques for the Analysis of Transgenic Plants.- 26 In Situ Hybridization on Plant Metaphase Chromosomes I. Farbos and A. Mouras.- 27 Localisation of RNA Transcripts in Plant Tissue by in Situ Hybridisation.- 28 In Situ Immunolocalisation of Transgene Products in Transgenic Plants.- VIII Establishment and Maintenance of Embryogenic Cultures.- 29 Establishment of and Plant Regeneration from Embryogenic Cell Suspensions and Their Protoplasts in Forage Grasses.- IX Genetic Markers and Expression Signals.- 30 Expression Signals and Vectors.- 31 Selectable Marker and Reporter Genes.- X Guidelines and Biosafety Considerations.- 32 Release of Transgenic Plants: Regulations and Developments.- 33 Risk Assessment.