Transgenic Wheat, Barley and Oats: Production and Characterization Protocols

ISBN-13: 9781588299611 / Angielski / Twarda / 2008 / 349 str.

Peter Shewry; Huw D. Jones

Opis

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.

Spis treści

Preface Contributors Part I: Introduction 1. Transgenic Wheat, Barley and Oats: Production and Characterization Paul A. Lazzeri and Huw D. Jones Part II: Transformation and Regeneration 2. Selection of Transformed Plants Huw D. Jones and Caroline A. Sparks 3. Reporter Genes Alison Huttly 4. Biolistics Transformation of Wheat Caroline A. Sparks and Huw D. Jones 5. Agrobacterium-Mediated Transformation of Bread and Durum Wheat Using Freshly Isolated Immature Embryos Huixia Wu, Angela Doherty and Huw D. Jones 6. Floral Transformation of Wheat Sujata Agarwal, Star Loar, Camille Steber and Janice Zale 7. Highly Efficient Agrobacterium-Mediated Transformation of Wheat via In Planta Inoculation Thierry Risacher, Melanie Craze, Sarah Bowden, Wyatt Paul and Tina Barsby 8. Barley Transformation Using Biolistic Techniques Wendy A. Harwood and Mark A. Smedley 9. Barley Transformation Using Agrobacterium-Mediated Techniques Wendy A. Harwood, Joanne G. Bartlett, Silvia C. Alves, Matthew Perry, Mark A. Smedley, Nicola Leyland and John W. Snape 10. Transformation of Oats and its Application to Improving Osmotic Stress Tolerance Shahina B. Maqbool, Heng Zhong, Hesham F. Oraby and Mariam B. Sticklen Part III: Gene and Protein Expression 11. Promoter Sequences for Defining Transgene Expression Huw D. Jones and Caroline A. Sparks 12. Down-Regulationof Gene Expression by RNA-Induced Gene Silencing Silvia Travella and Beat Keller Part IV: Characterisation of Transgenic Plants 13. Gene Insertion Patterns and Sites Phillippe Vain and Vera Thole 14. Fluorescent In Situ Hybridization to Detect Transgene Integration Into Plant Genomes Trude Schwartzacher 15. Establishing Substantial Equivalence: Transcriptomics María Marcela Baudo, Stephen J. Powers, Rowan A.C. Mitchell and Peter R. Shewry 16. Establishing Substantial Equivalence: Proteomics Alison Lovegrove, Louise Salt and Peter R. Shewry 17. Establishing Substantial Equivalence: Metabolomics Michael H. Beale, Jane L. Ward and John M. Baker 18. Design and Management of Field Trials of Transgenic Cereals Zoltán Bedo, Mariann Rakszegi and László Láng 19. GM Risk Assessment Penelope A.C. Sparrow Part V: Conclusions 20. Transgenic Wheat, Barley and Oats: Future Prospects Jim M. Dunwell

Blurb

The understanding of the physical and genetic structure of plant genomes and the determination of phenotypes via interactions of coding and non-coding regions with the environment has quickly become the key to the future of plant breeding and agriculture.  In Transgenic Wheat, Barley and Oats: Production and Characterization Protocols, expert investigators contribute the latest protocols for the transformation, regeneration and selection of these three species, using both biolistics and Agrobacterium tumefaciens, in order to fill the need for techniques and resources in this vital area of research.  Covering such topics as in vitro Agrobacterium co-cultivation, the manipulation of gene expression, and GM risk assessment, this versatile collection focuses on the study of temperate small grain cereals while many of its readily reproducible procedures could be easily adapted to accommodate other cereals and plants.  The in-depth chapters adhere to the highly successful Methods in Molecular Biology™ series format, featuring brief introductions to the subject, lists of the necessary materials and reagents, step-by-step laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Transgenic Wheat, Barley and Oats: Production and Characterization Protocols is an ideal guide for scientists pursuing this powerful, forward-thinking genetic strategy.