Transgenic Plants: Methods and Protocols

ISBN-13: 9781617374616 / Angielski / Miękka / 2010 / 437 str.

Leandro Pena

Opis

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.

Spis treści

Part I. Introduction Transgenic Plants: An Historical Perspective Luis Herrera-Estrella, June Simpson, and Miguel Martínez-Trujillo Part II. Transformation Plant Transformation: Agrobacterium-Mediated Gene Transfer Abhaya M. Dandekar and Henry J. Fisk Production of Hairy Root Cultures and Transgenic Plants by Agrobacterium rhizogenes-Mediated Transformation Mary C. Christey and Robert H. Braun Stable Transformation of Plant Cells by Particle Bombardment/Biolistics Julie R. Kikkert, José R. Vidal, and Bruce I. Reisch Electroporation: Introduction and Expression of Transgenes in Plant Protoplasts Henry J. Fisk and Abhaya M. Dandekar Floral Dip: Agrobacterium-Mediated Germ Line Transformation Steven J. Clough Production of Transgenic Crops by the Floral-Dip Method Ian S. Curtis Chloroplast Genetic Engineering to Improve Agronomic Traits Henry Daniell, Oscar N. Ruiz, and Amit Dhingra Part III. Regeneration Organogenesis From Transformed Tomato Explants Anne Frary and Joyce Van Eck Genetic Transformation of Conifers Utilizing Somatic Embryogenesis Krystyna Klimaszewska, Robert G. Rutledge, and Armand Séguin Regeneration of Transgenic Cassava From Transformed Embryogenic Tissues Peng Zhang and Johanna Puonti-Kaerlas Genetic Transformation of Mature Citrus Plants Magdalena Cervera, José Juárez, Luis Navarro, and Leandro Peña Part IV. Selection Selectable Markers: Antibiotic and Herbicide Resistance Julia L. Goodwin, Gabriela M. Pastori, Michael R. Davey, and Huw D. Jones Histochemical and Fluorometric Assays for uidA (GUS) Gene Detection Magdalena Cervera Green Fluorescent Protein Quantification in Whole Plants Matthew D. Halfhill, Reginald J. Millwood, and C. Neal Stewart, Jr. PositiveSelection Allan Wenck and Geneviève Hansen Elimination of Marker Genes From Transgenic Plants Using MAT Vector Systems Hiroyasu Ebinuma, Koichi Sugita, Saori Endo, Etsuko Matsunaga, and Keiko Yamada Simple and Efficient Removal of Marker Genes From Plastids by Homologous Recombination Anil Day, Vasumathi Kode, Panagiotis Madesis, and Siriluck Iamtham Part V. Transgene Integration, Expression, and Localization The Study of Transgene Copy Number and Organization David J. Ingham Analysis of Gene Expression in Transgenic Plants Andrew F. Page and Subhash C. Minocha Transgene Integration: Use of Matrix Attachment Regions George C. Allen, Steven Spiker, and William F. Thompson Fluorescence In Situ Hybridization to Localize Transgenes in Plant Chromosomes Wendy A. Harwood, Lorelei J. Bilham, Silvia Travella, Haroldo Salvo-Garrido, and John W. Snape Amplification of Genomic Sequences Flanking T-DNA Insertions by Thermal Asymmetric Interlaced Polymerase Chain Reaction Yao-Guang Liu, Yuanlin Chen, and Qunyu Zhang Part VI. Risk Assessment Agrobacterium Persistence in Plant Tissues After Transformation Jaime Cubero and María M. López Transgene Dispersal Through Pollen Laura C. Hudson, Matthew D. Halfhill, and C. Neal Stewart, Jr. Part VII. Transgenic Crops Transgenic Crops: The Current and Next Generations Jim M. Dunwell Comparative Development and Impact of Transgenic Papayas in Hawaii, Jamaica, and Venezuela Gustavo Fermín, Paula Tennant, Carol Gonsalves, David Lee, and Dennis Gonsalves Index

Blurb

Transgenic plants serve as essential tools for research in plant biology and are the basis of modern agricultural biotechnology. In Transgenic Plants: Methods and Protocols, leading scientists describe readily reproducible techniques they have perfected for the genetic transformation and regeneration of plants. The authors cover the most commonly used transformation systems, including Agrobacterium tumefaciens, Agrobacterium rhizogenes, particle bombardment, electroporation, floral-dip, and chloroplast transformation, as well as the regeneration of whole transgenic plants by organogenesis and somatic embryogenesis from different explant cells/tissues and from such diverse plant species as tomato, cassava, conifers, and citrus. Also described are the use of selectable and reporter markers, positive selection (for maize transformation), and the controlled excision and removal of marker genes from both nuclei and plastids once transgenic shoots have been efficiently generated. Additional techniques employ quantitative real-time PCR, reverse transcription PCR, fluorescence in situ hybridization, and thermal asymmetric interlaced PCR. Risk assessment methods offer the opportunity to study Agrobacterium persistence in plant tissues and to investigate the possibility of transgene dispersal. The protocols follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and easy to use, Transgenic Plants: Methods and Protocols offers both novice and experienced researchers a cutting-edge laboratory manual covering all aspects of plant genetic transformation and transgenic crops.