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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.
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.
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.
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.
Agrobacterium is a plant pathogen which causes the “crown-gall” disease, a neoplastic growth that results from the transfer of a well-defined DNA segment (“transferred DNA”, or “T-DNA”) from the bacterial Ti (tumor-inducing) plasmid to the host cell, its integration into the host genome, and the expression of oncogenes contained on the T-DNA. The molecular machinery, needed for T-DNA generation and transport into the host cell and encoded by a series of chromosomal (chv) and Ti-plasmid virulence (vir) genes, has been the subject of numerous studies over the past several decades. Today, Agrobacterium is the tool of choice for plant genetic engineering with an ever expanding host range that includes many commercially important crops, flowers, and tree species. Furthermore, its recent application for the genetic transformation of non-plant species, from yeast to cultivated mushrooms and even to human cells, promises this bacterium a unique place in the future of biotechnological applications. The book is a comprehensive volume describing Agrobacterium's biology, interactions with host species, and uses for genetic engineering.
The broad host range pathogenic bacterium Agrobacterium tumefaciens has been widely studied as a model system to understand horizontal gene flow, secretion of effector proteins into host cells, and plant-pathogen interactions. Agrobacterium-mediated plant transformation also is the major method for generating transgenic plants for research and biotechnology purposes. Agrobacterium species have the natural ability to conduct interkingdom genetic transfer from bacteria to eukaryotes, including most plant species, yeast, fungi, and even animal cells. In nature, A. tumefaciens causes crown gall disease resulting from expression in plants of auxin and cytokinin biosynthesis genes encoded by the transferred (T-) DNA. Gene transfer from A. tumefaciens to host cells requires virulence (vir) genes that reside on the resident tumor-inducing (Ti) plasmid. In addition to T-DNA, several Virulence (Vir) effector proteins are also translocated to host cells through a bacterial type IV secretion system. These proteins aid in T-DNA trafficking through the host cell cytoplasm, nuclear targeting, and T-DNA integration. Genes within native T-DNAs can be replaced by any gene of interest, making Agrobacterium species important tools for plant research and genetic engineering. In this research topic, we provided updated information on several important areas of Agrobacterium biology and its use for biotechnology purposes.