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The aim of Transgenic Plants: Methods and Protocols is to provide a source of information to guide the reader through a wide range of frequently used, broadly applicable, and easily reproducible techniques involved in the gene- tion of transgenic plants. Its step-by-step approach covers a series of methods for genetically transforming plant cells and tissues, and for recovering whole transgenic plants from them. The volume then moves on to the use of sele- able and reporter markers, positive selection, marker elimination after rec- ery of transgenic plants, and the analysis of transgene integration, expression, and localization in the plant genome. Although contributors usually refer to model plants in most chapters, the protocols described herein should be widely applicable to many plant species. The last two sections are devoted to me- ods of risk assessment and to exploring the current and future applications of transgenic technology in agriculture and its social implications in a case study. Transgenic Plants: Methods and Protocols is divided into six major s- tions plus an introduction, comprising 27 chapters. Part I, the Introduction, is a review of the past, present, and perspectives of the transgenic plants, from the discovery of Agrobacterium tumefaciens as a feasible transformation vector, to its use as a tool to study gene expression and function, and the current and possible future applications of this technology in agriculture, industry, and medicine.
List of contents
Transgenic Plants: An Historical Perspective.- Transformation.- Plant Transformation: Agrobacterium-Mediated Gene Transfer.- Production of Hairy Root Cultures and Transgenic Plants by Agrobacterium rhizogenes-Mediated Transformation.- Stable Transformation of Plant Cells by Particle Bombardment/Biolistics.- Electroporation: Introduction and Expression of Transgenes in Plant Protoplasts.- Floral Dip: Agrobacterium-Mediated Germ Line Transformation.- Production of Transgenic Crops by the Floral-Dip Method.- Chloroplast Genetic Engineering to Improve Agronomic Traits.- Regeneration.- Organogenesis From Transformed Tomato Explants.- Genetic Transformation of Conifers Utilizing Somatic Embryogenesis.- Regeneration of Transgenic Cassava From Transformed Embryogenic Tissues.- Genetic Transformation of Mature Citrus Plants.- Selection.- Selectable Markers: Antibiotic and Herbicide Resistance.- Histochemical and Fluorometric Assays for uidA (GUS) Gene Detection.- Green Fluorescent Protein Quantification in Whole Plants.- Positive Selection.- Elimination of Marker Genes From Transgenic Plants Using MAT Vector Systems.- Simple and Efficient Removal of Marker Genes From Plastids by Homologous Recombination.- Transgene Integration, Expression, and Localization.- The Study of Transgene Copy Number and Organization.- Analysis of Gene Expression in Transgenic Plants.- Transgene Integration: Use of Matrix Attachment Regions.- Fluorescence In Situ Hybridization to Localize Transgenes in Plant Chromosomes.- Amplification of Genomic Sequences Flanking T-DNA Insertions by Thermal Asymmetric Interlaced Polymerase Chain Reaction.- Risk Assessment.- Agrobacterium Persistence in Plant Tissues After Transformation.- Transgene Dispersal Through Pollen.- Transgenic Crops.- Transgenic Crops: The Current and Next Generations.- Comparative Development and Impact of Transgenic Papayas in Hawaii, Jamaica, and Venezuela.
Summary
The aim of Transgenic Plants: Methods and Protocols is to provide a source of information to guide the reader through a wide range of frequently used, broadly applicable, and easily reproducible techniques involved in the gene- tion of transgenic plants. Its step-by-step approach covers a series of methods for genetically transforming plant cells and tissues, and for recovering whole transgenic plants from them. The volume then moves on to the use of sele- able and reporter markers, positive selection, marker elimination after rec- ery of transgenic plants, and the analysis of transgene integration, expression, and localization in the plant genome. Although contributors usually refer to model plants in most chapters, the protocols described herein should be widely applicable to many plant species. The last two sections are devoted to me- ods of risk assessment and to exploring the current and future applications of transgenic technology in agriculture and its social implications in a case study. Transgenic Plants: Methods and Protocols is divided into six major s- tions plus an introduction, comprising 27 chapters. Part I, the Introduction, is a review of the past, present, and perspectives of the transgenic plants, from the discovery of Agrobacterium tumefaciens as a feasible transformation vector, to its use as a tool to study gene expression and function, and the current and possible future applications of this technology in agriculture, industry, and medicine.
