Transgenic Wheat, Barley and Oats

1. Front Matter

   Pages i-x

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2. Introduction

   1. Front Matter

      Pages 1-1

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   2. Transgenic Wheat, Barley and Oats: Production and Characterization

      • Paul A. Lazzeri*, Huw D. Jones

      Pages 3-20

3. Transformation and Regeneration

   1. Front Matter

      Pages 22-22

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   2. Selection of Transformed Plants

      • Huw D. Jones*, Caroline A. Sparks

      Pages 23-37

   3. Reporter Genes

      • Alison Huttly

      Pages 39-69

   4. Biolistics Transformation of Wheat

      • Caroline A. Sparks, Huw D. Jones*

      Pages 71-92

   5. Agrobacterium-Mediated Transformation of Bread and Durum Wheat Using Freshly Isolated Immature Embryos

      • Huixia Wu, Angela Doherty, Huw D. Jones

      Pages 93-103

   6. Floral Transformation of Wheat

      • Sujata Agarwal, Star Loar, Camille Steber, Janice Zale*

      Pages 105-113

   7. Highly Efficient Agrobacterium-Mediated Transformation of Wheat Via In Planta Inoculation

      • Thierry Risacher, Melanie Craze, Sarah Bowden, Wyatt Paul*, Tina Barsby

      Pages 115-124

   8. Barley Transformation Using Biolistic Techniques

      • Wendy A. Harwood*, Mark A. Smedley

      Pages 125-136

   9. Barley Transformation Using Agrobacterium-Mediated Techniques

      • Wendy A. Harwood*, Joanne G. Bartlett, Silvia C. Alves, Matthew Perry, Mark A. Smedley, Nicola Leyl et al.

      Pages 137-147

   10. Transformation of Oats and Its Application to Improving Osmotic Stress Tolerance

       • Shahina B. Maqbool*, Heng Zhong, Hesham F. Oraby, Mariam B. Sticklen

       Pages 149-168

4. Gene and Protein Expression

   1. Front Matter

      Pages 170-170

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   2. Promoter Sequences for Defining Transgene Expression

      • Huw D. Jones*, Caroline A. Sparks

      Pages 171-184

   3. Down-Regulation of Gene Expression by RNA-Induced Gene Silencing

      • Silvia Travella*, Beat Keller

      Pages 185-199

5. Characterisation of Transgenic Plants

   1. Front Matter

      Pages 202-202

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   2. Gene Insertion Patterns and Sites

      • Philippe Vain, Vera Thole

      Pages 203-226

   3. Fluorescent In Situ Hybridization to Detect Transgene Integration into Plant Genomes

      • Trude Schwarzacher

      Pages 227-246

   4. Establishing Substantial Equivalence: Transcriptomics

      • María Marcela Baudo*, Stephen J. Powers, Rowan A. C. Mitchell, Peter R. Shewry

      Pages 247-272

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.

• Agrobacterium
• Biolistics
• Cereal research
• Embryo
• Flora
• Gene and protein expression
• Proteom
• Transformation and regeneration
• Transgenic plants
• genome
• wheat
• genetic engineering
• Transgenic organisms

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