Transgenic Plants: Methods and Protocols

PENA, L ed.
cover image
Publication
Totowa, NJ : Humana Press, 2004.
ISBN
9781588292636 / eISBN:9781592598274
Printed edition: 9781588292636
Physical Description
XIV, 437 p. online resource.
Series
Methods in Molecular Biology™ Vol 286
URL
Language
eng
 
 
 
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$a 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.
520
$a Transgenic plants serve as essential tools for research in plant biology and are the basis of modern agricultural biotechnology. In Transgenic Plants: Methods and Protocols, leading scientists describe readily reproducible techniques they have perfected for the genetic transformation and regeneration of plants. The authors cover the most commonly used transformation systems, including Agrobacterium tumefaciens, Agrobacterium rhizogenes, particle bombardment, electroporation, floral-dip, and chloroplast transformation, as well as the regeneration of whole transgenic plants by organogenesis and somatic embryogenesis from different explant cells/tissues and from such diverse plant species as tomato, cassava, conifers, and citrus. Also described are the use of selectable and reporter markers, positive selection (for maize transformation), and the controlled excision and removal of marker genes from both nuclei and plastids once transgenic shoots have been efficiently generated. Additional techniques employ quantitative real-time PCR, reverse transcription PCR, fluorescence in situ hybridization, and thermal asymmetric interlaced PCR. Risk assessment methods offer the opportunity to study Agrobacterium persistence in plant tissues and to investigate the possibility of transgene dispersal. The protocols follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and easy to use, Transgenic Plants: Methods and Protocols offers both novice and experienced researchers a cutting-edge laboratory manual covering all aspects of plant genetic transformation and transgenic crops.
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$a PLANT SCIENCE
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$a BOTANY
650
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$a TRANSGENIC ORGANISMS
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$a METHODS AND PROTOCOLS
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Subject
LIFE SCIENCES
PLANT SCIENCE
BOTANY
TRANSGENIC ORGANISMS
METHODS AND PROTOCOLS
E-BOOKS
Summary
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.
Table 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.
Notes
Springer Protocols
Methods in Molecular Biology™, 1064-3745 ; 286