Melvin J. Oliver is a Supervisory Research Geneticist with the United States Department of Agriculture’s Agricultural Research Service and Adjunct Professor in the Division of Plant Sciences at the University of Missouri.Yi Li is Professor of Horticultural Plant Biotechnology at the University of Connecticut and Director of the New England Center for Invasive Plants.

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• List of Contributors ixForeword xiJ. Perry Gustafson and Peter H. RavenSection 1 Perspectives of Gene Flow 1Chapter 1 Potential Environmental Impacts of Transgene Flow in Rice with a Particular View on Herbicide Resistance 3Bao-Rong Lu and Wei WangIntroduction 3Transgene Escape and Its Potential Environmental Impacts 4Herbicide-Resistant Rice 8Gene Flow from Herbicide-Resistant Rice and Its Potential Impacts 12Managing Potential Environmental Impacts from Transgene Flow 17Conclusion 19Acknowledgments 20References 20Chapter 2 Assessment and Detection of Gene Flow 27Wei Wei, Charles Kwit, Reginald J. Millwood, Hong S. Moon, and C. Neal Stewart Jr.Importance of Assessment and Detection of Gene Flow 27Detection Methodologies 29Applications in Assessment of Gene Flow 35Perspectives 36Acknowledgments 37References 37Chapter 3 Bt Resistance Monitoring in European Corn Borers and Western Corn Rootworms 43Blair D. Siegfried and Terence SpencerIntroduction 43Resistance Monitoring 44Bioassay Methods and Baseline Susceptibility 47Recent Developments 49Conclusions 51Acknowledgments 52References 52Chapter 4 Commercialization of GMOs: the Company Perspective on Risks and Containment 57Kater HakeCommercialization of Traits Derived from Plant Biotechnology (GMOs and Alien Chromosome Introgression) 57Gene Containment During the Commercial Utilization of Crops Derived from Biotechnology 60Gene Containment During Research, Development, and Seed Production 62Stewardship and Gene Containment 63References 65Section 2 Strategies for Gene Containment 67Chapter 5 Trait Control (Gene Switch Technology) 69M. Isabel Ordiz, Jaemo Yang, and Roger N. BeachyComponents of Gene Switch Systems 69Application of Gene Switch Technologies 75Future Perspectives 79References 80Chapter 6 Male Sterility and Hybrid Plant Systems for Gene Confinement 85Albert P. Kausch, Joel Hague, Adam Deresienski, Michael Tilelli, and Kimberly NelsonIntroduction 85Cytoplasmic Male Sterility 86Transgenic Nuclear Male Sterility 87Transgenic Nuclear Male Sterility for Gene Confinement 89Male Sterility for Recovery of Wide Crosses with Gene Confinement 90Hybrid Plant Systems 92Seed-Based Hybrid Plant Systems 92Gene-Deletor System 92Male and Female Sterile Lines 94Total Sterility 95Stable Knockout Mutations 96References 96Chapter 7 Gene-Deletor Technology and Its Potential Applications in Addressing Gene Flow and Food Safety Concerns over Transgenic Plants 101Yi Li, Hui Duan, Yongqin Chen, and Richard McAvoyIntroduction 101The Principle of Gene-Deletor Technology and Supporting Experimental Data 102Characteristics and Requirements of Gene-Deletor Technology 105Potential Applications of Gene-Deletor Technology 107Conclusions and Perspectives 109Acknowledgments 110References 110Chapter 8 Seed-Based Gene Containment Strategies 113Melvin J. Oliver and Kater HakeSeedless Crops 114Seed-Based Containment 115GeneSafe (Technology Protection System (TPS)) Technology 117Expression Cascades for Gene Containment 120References 123Chapter 9 Transgenic Mitigation of Transgene Dispersal by Pollen and Seed 125Jonathan Gressel and Hani Al-AhmadIntroduction—Containers Leak 125Mitigation—The Concept of Conferring a Selective Disadvantage Outside of Cultivation 128Evidence That TM Mitigates Transgene Establishment 133Tailoring Mitigator Genes to Crops and Cropping Situation 137Concluding Remarks 142References 142Chapter 10 Tapping RNA Silencing for Transgene Containment through the Engineering of Sterility in Plants 147Zongrang Liu and Stacy D. SingerIntroduction 147References 166Chapter 11 Control of Cleistogamy and Seed Dormancy for Biological Gene Containment in Oilseed Rape (Brassica napus L.) 175Sabine Gruber and Alexandra H¨uskenIntroduction 175Pathways of Gene Flow in Oilseed Rape 176Managing Gene Flow in Oilseed Rape 184Combining Biological Measures Package for Gene Containment 191Conclusion 192Acknowledgments 192References 192Index 199A color plate section appears between pages 114 and 115

Melvin J. Oliver is a Supervisory Research Geneticist with the United States Department of Agriculture’s Agricultural Research Service and Adjunct Professor in the Division of Plant Sciences at the University of Missouri.Yi Li is Professor of Horticultural Plant Biotechnology at the University of Connecticut and Director of the New England Center for Invasive Plants.

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• List of Contributors ixForeword xiJ. Perry Gustafson and Peter H. RavenSection 1 Perspectives of Gene Flow 1Chapter 1 Potential Environmental Impacts of Transgene Flow in Rice with a Particular View on Herbicide Resistance 3Bao-Rong Lu and Wei WangIntroduction 3Transgene Escape and Its Potential Environmental Impacts 4Herbicide-Resistant Rice 8Gene Flow from Herbicide-Resistant Rice and Its Potential Impacts 12Managing Potential Environmental Impacts from Transgene Flow 17Conclusion 19Acknowledgments 20References 20Chapter 2 Assessment and Detection of Gene Flow 27Wei Wei, Charles Kwit, Reginald J. Millwood, Hong S. Moon, and C. Neal Stewart Jr.Importance of Assessment and Detection of Gene Flow 27Detection Methodologies 29Applications in Assessment of Gene Flow 35Perspectives 36Acknowledgments 37References 37Chapter 3 Bt Resistance Monitoring in European Corn Borers and Western Corn Rootworms 43Blair D. Siegfried and Terence SpencerIntroduction 43Resistance Monitoring 44Bioassay Methods and Baseline Susceptibility 47Recent Developments 49Conclusions 51Acknowledgments 52References 52Chapter 4 Commercialization of GMOs: the Company Perspective on Risks and Containment 57Kater HakeCommercialization of Traits Derived from Plant Biotechnology (GMOs and Alien Chromosome Introgression) 57Gene Containment During the Commercial Utilization of Crops Derived from Biotechnology 60Gene Containment During Research, Development, and Seed Production 62Stewardship and Gene Containment 63References 65Section 2 Strategies for Gene Containment 67Chapter 5 Trait Control (Gene Switch Technology) 69M. Isabel Ordiz, Jaemo Yang, and Roger N. BeachyComponents of Gene Switch Systems 69Application of Gene Switch Technologies 75Future Perspectives 79References 80Chapter 6 Male Sterility and Hybrid Plant Systems for Gene Confinement 85Albert P. Kausch, Joel Hague, Adam Deresienski, Michael Tilelli, and Kimberly NelsonIntroduction 85Cytoplasmic Male Sterility 86Transgenic Nuclear Male Sterility 87Transgenic Nuclear Male Sterility for Gene Confinement 89Male Sterility for Recovery of Wide Crosses with Gene Confinement 90Hybrid Plant Systems 92Seed-Based Hybrid Plant Systems 92Gene-Deletor System 92Male and Female Sterile Lines 94Total Sterility 95Stable Knockout Mutations 96References 96Chapter 7 Gene-Deletor Technology and Its Potential Applications in Addressing Gene Flow and Food Safety Concerns over Transgenic Plants 101Yi Li, Hui Duan, Yongqin Chen, and Richard McAvoyIntroduction 101The Principle of Gene-Deletor Technology and Supporting Experimental Data 102Characteristics and Requirements of Gene-Deletor Technology 105Potential Applications of Gene-Deletor Technology 107Conclusions and Perspectives 109Acknowledgments 110References 110Chapter 8 Seed-Based Gene Containment Strategies 113Melvin J. Oliver and Kater HakeSeedless Crops 114Seed-Based Containment 115GeneSafe (Technology Protection System (TPS)) Technology 117Expression Cascades for Gene Containment 120References 123Chapter 9 Transgenic Mitigation of Transgene Dispersal by Pollen and Seed 125Jonathan Gressel and Hani Al-AhmadIntroduction—Containers Leak 125Mitigation—The Concept of Conferring a Selective Disadvantage Outside of Cultivation 128Evidence That TM Mitigates Transgene Establishment 133Tailoring Mitigator Genes to Crops and Cropping Situation 137Concluding Remarks 142References 142Chapter 10 Tapping RNA Silencing for Transgene Containment through the Engineering of Sterility in Plants 147Zongrang Liu and Stacy D. SingerIntroduction 147References 166Chapter 11 Control of Cleistogamy and Seed Dormancy for Biological Gene Containment in Oilseed Rape (Brassica napus L.) 175Sabine Gruber and Alexandra H¨uskenIntroduction 175Pathways of Gene Flow in Oilseed Rape 176Managing Gene Flow in Oilseed Rape 184Combining Biological Measures Package for Gene Containment 191Conclusion 192Acknowledgments 192References 192Index 199A color plate section appears between pages 114 and 115

Melvin J. Oliver is a Supervisory Research Geneticist with the United States Department of Agriculture’s Agricultural Research Service and Adjunct Professor in the Division of Plant Sciences at the University of Missouri.Yi Li is Professor of Horticultural Plant Biotechnology at the University of Connecticut and Director of the New England Center for Invasive Plants.

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• List of Contributors ixForeword xiJ. Perry Gustafson and Peter H. RavenSection 1 Perspectives of Gene Flow 1Chapter 1 Potential Environmental Impacts of Transgene Flow in Rice with a Particular View on Herbicide Resistance 3Bao-Rong Lu and Wei WangIntroduction 3Transgene Escape and Its Potential Environmental Impacts 4Herbicide-Resistant Rice 8Gene Flow from Herbicide-Resistant Rice and Its Potential Impacts 12Managing Potential Environmental Impacts from Transgene Flow 17Conclusion 19Acknowledgments 20References 20Chapter 2 Assessment and Detection of Gene Flow 27Wei Wei, Charles Kwit, Reginald J. Millwood, Hong S. Moon, and C. Neal Stewart Jr.Importance of Assessment and Detection of Gene Flow 27Detection Methodologies 29Applications in Assessment of Gene Flow 35Perspectives 36Acknowledgments 37References 37Chapter 3 Bt Resistance Monitoring in European Corn Borers and Western Corn Rootworms 43Blair D. Siegfried and Terence SpencerIntroduction 43Resistance Monitoring 44Bioassay Methods and Baseline Susceptibility 47Recent Developments 49Conclusions 51Acknowledgments 52References 52Chapter 4 Commercialization of GMOs: the Company Perspective on Risks and Containment 57Kater HakeCommercialization of Traits Derived from Plant Biotechnology (GMOs and Alien Chromosome Introgression) 57Gene Containment During the Commercial Utilization of Crops Derived from Biotechnology 60Gene Containment During Research, Development, and Seed Production 62Stewardship and Gene Containment 63References 65Section 2 Strategies for Gene Containment 67Chapter 5 Trait Control (Gene Switch Technology) 69M. Isabel Ordiz, Jaemo Yang, and Roger N. BeachyComponents of Gene Switch Systems 69Application of Gene Switch Technologies 75Future Perspectives 79References 80Chapter 6 Male Sterility and Hybrid Plant Systems for Gene Confinement 85Albert P. Kausch, Joel Hague, Adam Deresienski, Michael Tilelli, and Kimberly NelsonIntroduction 85Cytoplasmic Male Sterility 86Transgenic Nuclear Male Sterility 87Transgenic Nuclear Male Sterility for Gene Confinement 89Male Sterility for Recovery of Wide Crosses with Gene Confinement 90Hybrid Plant Systems 92Seed-Based Hybrid Plant Systems 92Gene-Deletor System 92Male and Female Sterile Lines 94Total Sterility 95Stable Knockout Mutations 96References 96Chapter 7 Gene-Deletor Technology and Its Potential Applications in Addressing Gene Flow and Food Safety Concerns over Transgenic Plants 101Yi Li, Hui Duan, Yongqin Chen, and Richard McAvoyIntroduction 101The Principle of Gene-Deletor Technology and Supporting Experimental Data 102Characteristics and Requirements of Gene-Deletor Technology 105Potential Applications of Gene-Deletor Technology 107Conclusions and Perspectives 109Acknowledgments 110References 110Chapter 8 Seed-Based Gene Containment Strategies 113Melvin J. Oliver and Kater HakeSeedless Crops 114Seed-Based Containment 115GeneSafe (Technology Protection System (TPS)) Technology 117Expression Cascades for Gene Containment 120References 123Chapter 9 Transgenic Mitigation of Transgene Dispersal by Pollen and Seed 125Jonathan Gressel and Hani Al-AhmadIntroduction—Containers Leak 125Mitigation—The Concept of Conferring a Selective Disadvantage Outside of Cultivation 128Evidence That TM Mitigates Transgene Establishment 133Tailoring Mitigator Genes to Crops and Cropping Situation 137Concluding Remarks 142References 142Chapter 10 Tapping RNA Silencing for Transgene Containment through the Engineering of Sterility in Plants 147Zongrang Liu and Stacy D. SingerIntroduction 147References 166Chapter 11 Control of Cleistogamy and Seed Dormancy for Biological Gene Containment in Oilseed Rape (Brassica napus L.) 175Sabine Gruber and Alexandra H¨uskenIntroduction 175Pathways of Gene Flow in Oilseed Rape 176Managing Gene Flow in Oilseed Rape 184Combining Biological Measures Package for Gene Containment 191Conclusion 192Acknowledgments 192References 192Index 199A color plate section appears between pages 114 and 115