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In Plant Gene Transfer and Expression Protocols, leading experts present the latest step-by-step recipes for introducing genes into model and agricultural plants and for studying gene activity using reporter enzyme assays, RNA techniques, and immunological methods. The book contains not only a complete collection of mainstream methods, but also methods for studying gene expression and genetic manipulation in chloroplasts and mitochondria. There are also methods for introducing and analyzing plant genes in Xenopus and heterologous yeast hosts. Plant Gene Transfer and Expression Protocols is an indispensable tool for scientists working in both fundamental (biochemical, physiological, and genetic) and applied aspects of plant biotechnology, plant breeding, agriculture, and horticulture. It provides concise practical help that produces the desired results even for beginners and opens the doors to new research possibilities.
Assists policymakers in evaluating the appropriate scientific methods for detecting unintended changes in food and assessing the potential for adverse health effects from genetically modified products. In this book, the committee recommended that greater scrutiny should be given to foods containing new compounds or unusual amounts of naturally occurring substances, regardless of the method used to create them. The book offers a framework to guide federal agencies in selecting the route of safety assessment. It identifies and recommends several pre- and post-market approaches to guide the assessment of unintended compositional changes that could result from genetically modified foods and research avenues to fill the knowledge gaps.
"The book...is, in fact, a short text on the many practical problems...associated with translating the explosion in basic biotechnological research into the next Green Revolution," explains Economic Botany. The book is "a concise and accurate narrative, that also manages to be interesting and personal...a splendid little book." Biotechnology states, "Because of the clarity with which it is written, this thin volume makes a major contribution to improving public understanding of genetic engineering's potential for enlarging the world's food supply...and can be profitably read by practically anyone interested in application of molecular biology to improvement of productivity in agriculture."
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.
Authored by an integrated committee of plant and animal scientists, this review of newer molecular genetic techniques and traditional research methods is presented as a compilation of high-reward opportunities for agricultural research. Directed to the Agricultural Research Service and the agricultural research community at large, the volume discusses biosciences research in genetic engineering, animal science, plant science, and plant diseases and insect pests. An optimal climate for productive research is discussed.
Understanding the physical and genetic structure of cereal genomes and how defined coding and non-coding regions interact with the environment to determine a phenotype are key to the future of plant breeding and agriculture. The production and characteri- tion of transgenic plants is a powerful reverse genetic strategy increasingly used in cereals research to ascribe function to defined DNA sequences. However, the techniques and resources required to conduct these investigations have, until recently, been difficult to achieve or totally lacking in wheat, barley and oat. This book brings together the l- est protocols for the transformation, regeneration and selection using both biolistic and Agrobacterium tumefaciens appropriate for these three species. It includes two chapters describing in vitro Agrobacterium co-cultivation, one leading to germ line transformation with no need for tissue culture-based regeneration. In addition, it has several chapters dedicated to the manipulation of gene expression and characterisation of the recombinant locus and transgenic plants. Finally, it tackles the issues of GM risk assessment, field trials and substantial equivalence in terms of transcriptomics, proteomics and metabolomics. Although this book is dedicated to the temperate small grain cereals wheat, barley and oats, many of the techniques described could be readily adapted for other cereals or plants generally. We thank all the contributing authors for their timely and informative chapters, the staff of Humana Press, especially John Walker for their guidance, and Helen Jenkins for her proof-reading, word processing and administrative support. v Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix PART I.
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.
This volume details protocols for the use of the biolistic DNA delivery method in different plant species. Chapters guide readers through non-protocol chapters that cover relevant topics of interest, a broad overview of the field, exciting modifications of the system, and reliable plant transformation procedures in different 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, Biolistic DNA Delivery: Methods and Protocols aims to provide a comprehensive collection of protocols to intended to be a practical guide for the novice as well as the advanced user in the field of plant genetic transformation.
"Plant transformation technology has played a critical role in advancing biotechnology and fundamental research and evolved as a science. This book describes the breakthrough technologies in all aspects of plant transformation in the last 27 years, which "