Beschreibung

For several decades, Arabidopsis thaliana has been the organism of choice in the laboratories of many plant geneticists, physiologists, developmental biologists, and biochemists around the world. During this time, a huge amount of knowledge has been acquired on the biology of this plant species, which has resulted in the development of molecular tools that account for much more efficient research. The significance that Arabidopsis would attain in biological research may have been difficult to foresee in the 1980s, when its use in the laboratory started. In the meantime, it has become the model plant organism, much the same way as Drosophila, Caenorhabditis, or mouse have for animal systems. Today, it is difficult to envision research at the cutting edge of plant biology without the use of Arabidopsis. Since the first edition of Arabidopsis Protocols appeared, new developments have fostered an impressive advance in plant biology that prompted us to prepare Arabidopsis Protocols, Second Edition. Completion of the Arabidopsis genome sequence offered for the first time the opportunity to have in hand all of the genetic information required for studying plant function. In addition, the development of whole systems approaches that allow global analysis of gene expression and protein and metabolite dynamics has encouraged scientists to explore new scenarios that are extending the limits of our knowledge.

This collection of readily reproducible Arabidopsis protocols has been updated to reflect recent advances in plant biology, the completion of the Arabidopsis genome sequence-essential for studying plant function, and the development of whole systems approaches that allow global analysis of gene expression and protein and metabolite dynamics. The authors have included nearly all techniques developed in Arabidopsis, others recently adapted from the traditional work in crop species, and the most recent ones using Arabidopsis as a model system.

Klappentext
Emulating the much-praised first edition of Arabidopsis Protocols, leading scientists have generated an up-to-date work that reflects recent advances in plant biology, the completion of the Arabidopsis genome sequence-essential for studying plant function-and the development of whole systems approaches that allow global analysis of gene expression, as well as protein and metabolite dynamics. The authors have included nearly all techniques developed in Arabidopsis, others recently adapted from more traditional work in crop species, and the latest using Arabidopsis as a model system. Highlights include the most recent methods-transcriptomics, proteomics, and metabolomics-and their novel applications (phosphoproteomics, DNA microarray-based genotyping, high throughput metabolite profiling, and single-cell RNA). Traditional protocols from the agricultural sciences and others developed in crop species (grafting and chloroplast transformation) have been adapted to exploit the advantages of the Arabidopsis model. The protocols themselves follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Indispensable and highly practical, Arabidopsis Protocols, Second Edition offers both novice and experienced plant biologists cutting-edge tools to explore new scenarios and gain an understanding of how this complex, multicellular organism works, how it copes with a sessile life style, and how these strategies compare with those developed in other organisms.

Inhalt
Part I. Growing Arabidopsis Growth of Plants and Preservation of Seeds Luz Rivero-Lepinckas, Deborah Crist, and Randy Scholl Prevention and Control of Pests and Diseases Jenifer Bush, Georg Jander, and Frederick M. Ausubel Growth of Other Species Related to Arabidopsis thaliana Krithika Yogeeswaran and Mikhail Elia Nasrallah. Grafting, Katherine Bainbridge, Tom Bennett, Colin Turnbull, and Ottoline Leyser Synchronization, Transformation, and Cryopreservation of Suspension-Cultured Cells Margit Menges and James A. H. Murray Part II. Genetic Analyses Genetic Analysis Maarten Koornneef, Carlos Alonso-Blanco, and Piet Stam QTL Analysis Carlos Alonso-Blanco, Maarten Koornneef, and Johan W. van Ooijen EMS Mutagenesis of Arabidopsis YongSig Kim, Karen S. Schumaker, and Jian-Kang Zhu Low-Resolution Mapping of Untagged Mutations M. Rosa Ponce, Pedro Robles, Francisca M. Lozano, M. Asunción Brotóns, and José L. Micol Gene Identification and Cloning by Molecular Marker Mapping Georg Jander High-Throughput TILLING for Arabidopsis Bradley J. Till, Trenton Colbert, Christine Codomo, Linda Enns, Jessica Johnson, Steven H. Reynolds, Jorja G. Henikoff, Elizabeth A. Greene, Michael N. Steine, Luca Comai, and Steven Henikoff Genotyping and Mapping With High-Density Oligonucleotide Arrays Justin Borevitz Forward Genetic Screening of Insertional Mutants Anja Schneider and Dario Leister PCR-Based Screening for Insertional Mutants Anna N. Stepanova and Jose M. Alonso Cytogenetic Analysis of Arabidopsis Martin Lysak, Paul Fransz, and Ingo Schubert Using Information From Public Arabidopsis Databases to Aid Research Margarita Garcia-Hernandez and Leonore Reiser Part III. Transient and Stable Transformation In PlantaAgrobacterium-Mediated Transformation by Vacuum Infiltration Brian W. Tague and Joanna Mantis Transient Expression Assay by Agroinfiltration of Leaves Min Woo Lee and Yinong Yang Functional Analysis of Transcription Factors by Microparticle Bombardment Ida Ruberti, Giovanna Sessa, and Giorgio Morelli Transfection Assays With Protoplasts Containing Integrated Reporter Genes Shiv Tiwari, Shucai Wang, Gretchen Hagen, and Tom J. Guilfoyle Chloroplast Genetic Engineering Via Organogenesis or Somatic Embryogenesis Amit Dhingra and Henry Daniell b-Glucuronidase as Reporter Gene: Advantages and Limitations Kye-Won Kim, Vincent R. Franceschi, Laurence B. Davin, and Norman G. Lewis Use of Fluorescent Proteins as Reporters Guido Jach Luciferases as Reporter Genes Megan M. Southern, Paul E. Brown, and Anthony Hall Aequorin as a Reporter Gene Christoph Plieth Applications of Chemical-Inducible Expression Systems in Functional Genomics and Biotechnology Jianru Zuo, Peter D. Hare, and Nam-Hai Chua Part IV. Transcriptomics RNA Extraction Huazhong Shi and Ray Bressan Target Preparation for DNA Microarray Hybridization Tong Zhu, Sherman H. Chang, and Pedro Gil Statistical Issues in Microarray Data Analysis Willem A. Rensink and Samuel P. Hazen Preparation and Quality Assesment of RNA From Cell-Specific Samples Obtained by Laser Microdissection Regine Kleber and Julia Kehr Part V. Proteomics Protein Isolation and Second-Dimension Electrophoretic Separation Bernhard Schlesier and Hans-Peter Mock Isolation of Nuclear Proteins Tomasz T. Calikowski and Iris Meier Purification and Fractionation of Membranes for Proteomic Analysis Anne Marmagne, Daniel Salvi, Norbert Rolland, Geneviève Ephritikhine, Jacques

Produktinformationen