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Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agricultural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate a single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Agrobacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent developments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.
Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agric- tural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Ag- bacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent devel- ments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.
Rapid changes and significant progress have been made in the Agrobacterium field, such as genetically transforming plants for both basic research purposes and agricultural development. In Agrobacterium Protocols, Third Edition, Volumes 1 and 2, a team of leading experts and veteran researchers describe in detail techniques for delivering DNA to plant cells and permanently altering their genomes. This edition emphasizes agricultural crops and plant species with economic values, with updated protocols on 32 plant species and protocols involving 19 new species. Together with the 1st and 2nd editions, these two volumes offer Agrobacterium-mediated genetic transformation protocols for a total of 76 plant species. For a number of important plants such as rice, barley, wheat and citrus, multiple protocols using different starting plant materials for transformation are included. Volume 1 details updated techniques available for 18 plant species drawn from cereal crops, legume plants, vegetable plants, and three model plant species: Brachypodium distachyon, Medicago truncatula, and Setaria viridis. It also updates a chapter for vector construction, a step critical to a successful plant transformation process. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Agrobacterium Protocols, Third Edition facilitates the transfer of this rapidly developing technology to all researchers for use in both fundamental and applied biology.
A collection of protocols for the expression of agrobacterium genes in higher plants that aim to provide a profound effect on research in plant development.
Rapid changes and significant progress have been made in the Agrobacterium field, such as genetically transforming plants for both basic research purposes and agricultural development. In Agrobacterium Protocols, Third Edition, Volumes 1 and 2, a team of leading experts and veteran researchers describe in detail techniques for delivering DNA to plant cells and permanently altering their genomes. This edition emphasizes agricultural crops and plant species with economic values, with updated protocols on 32 plant species and protocols involving 19 new species. Together with the 1st and 2nd editions, these two volumes offer Agrobacterium-mediated genetic transformation protocols for a total of 76 plant species. For a number of important plants such as rice, barley, wheat and citrus, multiple protocols using different starting plant materials for transformation are included. Volume 2 contains 29 chapters with updated techniques for industrial plants, root plants, nuts and fruits, tropic plants, and other important plant species. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Agrobacterium Protocols, Third Edition facilitates the transfer of this rapidly developing technology to all researchers in both fundamental and applied biology.
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.
A comprehensive state-of-the-art collection of the most frequently used techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods range from general methodologies, such as culture induction, growth and viability evaluation, and contamination control, to such highly specialized techniques as chloroplast transformation involving the laborious process of protoplast isolation and culture. Most of the protocols are currently used in the research programs of the authors or represent important parts of business projects aimed at the generation of improved plant materials. Two new appendices explain the principles for formulating culture media and the composition of the eight most commonly used media formulations, and list more than 100 very useful internet sites.
Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agric- tural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Ag- bacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent devel- ments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.
Methods in Plant Molecular Biology and Biotechnology emphasizes a variety of well-tested methods in plant molecular biology and biotechnology. For each detailed and tested protocol presented, a brief overview of the methodology is provided. This overview considers why the protocol is used, what other comparable methods are available, and what limitations can be expected with the protocol. Other chapters in the book present overviews regarding how to approach particular problems and introduce unique methods - such as how to use computer methodology to study isolated genes. The book will be a practical reference for plant physiologists, plant molecular biologists, phytopathologists, and microbiologists.