Barley: Methods and Protocols

Description

This detailed volume explores barley as both a crop and a model, with practical techniques such as crossing barley, a range of tissue culture methods, the preparation of barley tissues for different forms of microscopy, and the assessment of sensitivity to abiotic stresses. Efficient protocols are provided for transformation, TILLING, virus-induced gene silencing and genome editing. There is also particular emphasis on a range of protocols for genotyping and for the analysis of gene expression. Written for the highly successful Methods in Molecular Biology series, chapters include introductions on 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 easy-to-use, Barley: Methods and Protocols serves as a valuable reference volume for cereal researchers and breeders by providing detailed protocols covering important traditional skills such as crossing and tissue culture through to the latest technologies for genotyping, expression analysis, and genome editing. Front Matter ....Pages i-xi An Introduction to Barley: The Crop and the Model (Wendy A. Harwood)....Pages 1-5 Mutation Breeding in Barley: Historical Overview (Ludmila Ohnoutkova)....Pages 7-19 A Practical Guide to Barley Crossing (William T. B. Thomas, Hazel Bull, Allan Booth, Ruth Hamilton, Brian P. Forster, Jerome D. Franckowiak)....Pages 21-36 Barley Anther Culture (Ludmila Ohnoutkova, Tomas Vlcko, Mentewab Ayalew)....Pages 37-52 Isolated Microspore Culture in Barley (Patricio Esteves, François J. Belzile)....Pages 53-71 TILLING in Barley (Matthias Jost, Miriam Szurman-Zubrzycka, Katarzyna Gajek, Iwona Szarejko, Nils Stein)....Pages 73-94 Virus-Induced Gene Silencing (VIGS) for Functional Characterization of Disease Resistance Genes in Barley Seedlings (Lokanadha R. Gunupuru, Alexandre Perochon, Shahin S. Ali, Steven R. Scofield, Fiona M. Doohan)....Pages 95-114 Agrobacterium-Mediated Transformation of Barley Immature Embryos (Alison Hinchliffe, Wendy A. Harwood)....Pages 115-126 Methods for the Simple and Reliable Assessment of Barley Sensitivity to Abiotic Stresses During Early Development (Agata Daszkowska-Golec, Anna Skubacz, Marzena Kurowska, Michał Słota, Dorota Swiergolik, Iwona Szarejko)....Pages 127-151 Preparation of Barley Roots for Histological, Structural, and Immunolocalization Studies Using Light and Electron Microscopy (Marek Marzec, Michael Melzer)....Pages 153-166 Preparation of Barley Pollen Mother Cells for Confocal and Super Resolution Microscopy (Sybille Mittmann, Mikel Arrieta, Luke Ramsay, Robbie Waugh, Isabelle Colas)....Pages 167-179 Microarrays for High-Throughput Gene Expression Analysis of Barley (Jenny A. Morris, Pete E. Hedley)....Pages 181-194 Genome Engineering Using TALENs (Goetz Hensel, Jochen Kumlehn)....Pages 195-215 Creating Targeted Gene Knockouts in Barley Using CRISPR/Cas9 (Tom Lawrenson, Wendy A. Harwood)....Pages 217-232 Genotyping-by-Sequencing on the Ion Torrent Platform in Barley (Amina Abed, Gaétan Légaré, Sonia Pomerleau, Jérôme St-Cyr, Brian Boyle, François J. Belzile)....Pages 233-252 DNA Methylation Analysis in Barley and Other Species with Large Genomes (Karolina Chwialkowska, Urszula Korotko, Miroslaw Kwasniewski)....Pages 253-268 High-Resolution RT-PCR Analysis of Alternative Barley Transcripts (Craig G. Simpson, John Fuller, Paulo Rapazote-Flores, Claus-Dieter Mayer, Cristiane P. G. Calixto, Linda Milne et al.)....Pages 269-281 Exome Capture for Variant Discovery and Analysis in Barley (Micha Bayer, Jenny A. Morris, Clare Booth, Allan Booth, Niki Uzrek, Joanne R. Russell et al.)....Pages 283-310 Back Matter ....Pages 311-313