Genetically Engineered Foods...v Contents List of Contributors..... xv Foreword ..... xvii Series Preface..... xix

Genetically Engineered Foods

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Genetically Engineered FoodsHandbook of Food Bioengineering,

Volume 6

Edited by

Alina Maria HolbanAlexandru Mihai Grumezescu

Academic Press is an imprint of Elsevier125 London Wall, London EC2Y 5AS, United Kingdom525 B Street, Suite 1800, San Diego, CA 92101-4495, United States50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United StatesThe Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom

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v

Contents

List of Contributors .............................................................................................xvForeword .........................................................................................................xviiSeries Preface ....................................................................................................xixPreface for Volume 6: Genetically Engineered Foods ............................................ xxiii

Chapter 1: Genetic Engineered Organisms (Plants and Animals) ...............................1Sana Khalid, Zubaida Yousaf, Ahmad A. Shahid1 Genetic Engineering ................................................................................................ 12 Purpose of Genetically Engineered Organisms ...................................................... 13 Genetic Engineering Versus Hybridization Techniques .......................................... 24 Genetically Modified Plants .................................................................................... 3

4.1 Bacillus thuringiensis (Bt) Berliner Crops: A Blueprint to the GM Plantation ...................................................................................................6

5 Techniques to Develop Genetically Modified Plants .............................................. 65.1 Recombinant DNA Technology ....................................................................... 75.2 RNAi (Silencing) ............................................................................................. 75.3 Tissue Culture and Transformation .................................................................. 85.4 Infiltration ........................................................................................................ 85.5 Silicon Carbide-Mediated Transformation ...................................................... 95.6 The Pollen-Tube Pathway Method ................................................................ 105.7 Liposomes ...................................................................................................... 105.8 Microinjection ................................................................................................ 10

6 Genetically Modified Animals .............................................................................. 117 Molecular Advancements in the Development of GM Animals ........................... 118 Significance of GM Livestock and Fishery ........................................................... 12

8.1 Effect of Bt Crops on Nontarget Insects ........................................................ 128.2 Pollinator Insects ........................................................................................... 138.3 Phytophilous Insects ...................................................................................... 148.4 Effect on Soil Organisms ............................................................................... 158.5 Effects on Vertebrates .................................................................................... 158.6 Effect on Aquatic Organisms ......................................................................... 168.7 Effect on Human Cells ................................................................................... 16

9 Other Unrevealed and Impulsive Effects of GMOs .............................................. 16

Contents

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10 Risk Assessment.................................................................................................. 1711 Biosafety Regulations in Pakistan ....................................................................... 1912 Cartagena Protocol on Biosafety ........................................................................ 2113 Conclusions ......................................................................................................... 2214 Future Prospects .................................................................................................. 22References ................................................................................................................. 23

Chapter 2 : Sugar and Sugar Alcohol Production in Genetically Modified Cyanobacteria ....................................................................31

Niels-Ulrik Frigaard1 Introduction ........................................................................................................... 312 Cyanobacteria ....................................................................................................... 34

2.1 Description of Cyanobacteria ........................................................................ 342.2 Large-Scale Production With Cyanobacteria ................................................. 352.3 Synthetic Biology Using Cyanobacteria ........................................................ 36

3 Production of Sugars and Sugar Alcohols ............................................................ 363.1 Strategies to Produce Valuable Sugars in Cyanobacteria ............................... 363.2 Sucrose Production ........................................................................................ 383.3 Glucose and Fructose Production .................................................................. 403.4 Glycerol Production ....................................................................................... 403.5 Erythritol Production ..................................................................................... 413.6 Mannitol Production ...................................................................................... 42

4 Human Health Concerns About Cyanobacteria .................................................... 435 Conclusions ........................................................................................................... 44References ................................................................................................................. 45

Chapter 3 : Genetically Modified Plants as Sustainable and Economic Sources for RUTFs............................................................................49

Soleyman Dayani, Mohammad R. Sabzalian 1 Introduction ......................................................................................................... 49

1.1 Food and Human Nutrition .......................................................................... 491.2 Malnutrition ................................................................................................. 501.3 Global Demand for More Value in Foods: A Health Need or

a Social Trend? ..............................................................................................521.4 Functional Foods .......................................................................................... 53

2 Ready-To-Use Therapeutic Foods ....................................................................... 54 3 Perspectives for Alternative Sources ................................................................... 56 4 Crop Plants .......................................................................................................... 56 5 Herbs and Noncrop Plants .................................................................................. 58 6 Cultivated and Wild Edible Mushrooms ............................................................. 59 7 Genetic Modifications to Enhance the Nutritional Quality of Plants ................. 62 8 Micronutrients ..................................................................................................... 66 9 Antinutritional and Promoter Factors .................................................................. 6710 Essential Amino Acids ........................................................................................ 6911 Ethical Issues ...................................................................................................... 71

Contents

vii

12 In Silico Tools for RUTF Development .............................................................. 7213 Food Processing Techniques ............................................................................... 7214 Conclusions and Future Perspectives .................................................................. 73References ................................................................................................................. 74

Chapter 4 : Approved Genetically Engineered Foods: Types, Properties, and Economic Concerns .....................................................................85

Grigore M. Elena, Biscu E. Ramona, Alina M. Holban1 Introduction ........................................................................................................... 852 Advantages of Genetically Modified Foods .......................................................... 89

2.1 Herbicide Tolerance ....................................................................................... 902.2 Insect and Pest Resistance ............................................................................. 902.3 Disease Resistance ......................................................................................... 912.4 Temperature Tolerance ................................................................................... 912.5 Salinity Tolerance .......................................................................................... 922.6 Nutrition ......................................................................................................... 93

3 Common Genetically Modified Foods .................................................................. 933.1 Genetically Modified Plants (GMPs) ............................................................. 933.2 Genetically Modified Animals ....................................................................... 96

4 Risks Associated With Genetically Modified Foods ............................................. 984.1 Unintended Harm to Other Organisms .......................................................... 994.2 Reduced Effectiveness of Pesticides ............................................................ 1004.3 Gene Transfer to Nontarget Species ............................................................ 1014.4 Human Health Related Risks ....................................................................... 101

5 Economic Concerns ............................................................................................ 1026 Conclusions and Perspectives ............................................................................. 104References ............................................................................................................... 104

Chapter 5 : Influence of Feed From Genetically Modified Plants on the Composition and Quality of Foods of Animal Origin ...........................109

Gerhard Flachowsky1 Introduction ......................................................................................................... 1092 Global Food Situation ......................................................................................... 1113 Feeding Studies With GM Plants of the 1st Generation ..................................... 115

3.1 Recombinant DNA and Newly Expressed Proteins ..................................... 1153.2 Product Composition and Quality ............................................................... 116

4 Feeding Studies With GM Plants of the 2nd Generation .................................... 1224.1 Increased Content of Amino Acids .............................................................. 1254.2 Expression and Uses of Stearidonic Acid in Soybeans (C18:4 n−3;SDA) .... 1264.3 Conjugated Linoleic Acids .......................................................................... 1284.4 Increasing Concentrations of Micronutrients

(the Vitamin A Precursor β-Carotene) ..........................................................1284.5 Lower Content of Undesirable Ingredients .................................................. 129

5 Conclusions ......................................................................................................... 130References ............................................................................................................... 131

Contents

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Chapter 6 : Genetically Modified Microorganisms: Harmful or Helpful? .................143Ortansa Csutak, Ionela Sarbu1 Introduction ......................................................................................................... 1432 Incidence of Microorganisms in Foods ............................................................... 144

2.1 Lactic Acid Bacteria .................................................................................... 1442.2 Yeasts ........................................................................................................... 145

3 Strategies for the Improvement of Microorganisms in Foods and Beverages .... 1473.1 Baking Products ........................................................................................... 1493.2 Beer .............................................................................................................. 1513.3 Wine ............................................................................................................. 1533.4 Dairy Products and Nutritional Additives for the Dairy Industry ................ 1553.5 Probiotics ..................................................................................................... 1593.6 Meat Products .............................................................................................. 1603.7 Fermented Vegetable Products ..................................................................... 1603.8 Other Industrial Enzymes and Food Additives ............................................ 1613.9 GMMs: Harmful or Helpful? ....................................................................... 165


Chapter 7 : Techniques for Production and Quality Assessment of Genetically Modified Foods ..........................................................177

Sapna Jain, Mukesh K. Yadav, Ajay Kumar1 Introduction ......................................................................................................... 1772 Types of GM Foods ............................................................................................. 178

2.1 Cotton .......................................................................................................... 1792.2 Rice .............................................................................................................. 1802.3 Brinjal .......................................................................................................... 1802.4 Milk and Dairy Products .............................................................................. 1812.5 Rapeseed ...................................................................................................... 1812.6 Soybean ........................................................................................................ 1812.7 Maize ........................................................................................................... 1822.8 Honey ........................................................................................................... 1822.9 Papaya .......................................................................................................... 1822.10 Other Genetically Modified Foods ............................................................ 182

3 Production of GM Foods .................................................................................... 1823.1 Modification of Plants .................................................................................. 1833.2 Modification of Animals .............................................................................. 187

4 Risks Related to Genetically Modified Foods .................................................... 1894.1 Environmental Hazards ................................................................................ 1894.2 Human Health Risks .................................................................................... 1904.3 Economic Concerns ..................................................................................... 190

5 Methods for Assessing the Safety Profiles .......................................................... 1915.1 Targeted Quantitative Analysis .................................................................... 1915.2 Profiling Methods ........................................................................................ 192

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6 Testing and Labeling of Genetically Modified Foods ......................................... 1936.1 qPCR Technology for Detection of Genetically Modified Organisms ........ 1956.2 PCR Capillary Gel Electrophoresis Technology ......................................... 1976.3 Microarray Technology for the Detection of Genetically

Modified Organisms .....................................................................................1976.4 Luminex Technology ................................................................................... 1986.5 Digital PCR Technology .............................................................................. 1986.6 Loop-Mediated Isothermal Amplification (LAMP) ..................................... 1986.7 DNA Walking .............................................................................................. 199


Chapter 8 : Awareness and Utilization of Genetically Modified Foods in Nigeria .....203Brilliant O. Agaviezor 1 Introduction ....................................................................................................... 203 2 Nigeria as a Country ......................................................................................... 204 3 Agriculture in Nigeria ....................................................................................... 204 4 Animal Rearing in Nigeria ................................................................................ 205 5 Problems and Proffered Solutions of Agriculture in Nigeria ............................ 206 6 Awareness and Utilization of Genetically Modified Foods in Nigeria ............. 209 7 Some Genetically Modified Foods in Nigeria .................................................. 212 8 Advantages of Genetically Modified Foods ...................................................... 213 9 Challenges Related to Genetically Modified Foods .......................................... 21410 Controversies About Genetically Modified Foods in Nigeria ........................... 214

10.1 The Dangers of Genetically Modified Crops in Nigeria .......................... 21410.2 Nigeria’s Desperate Need for Export Earnings ........................................ 21410.3 Monsanto, Genetically Modified Organisms, and Nigeria’s

Bleak Future ..............................................................................................21510.4 Revisiting Genetically Modified Foods in Nigeria .................................. 21510.5 Reasons Why Nigeria Must not Accept Genetically

Modified Organisms ..................................................................................21711 Conclusions ....................................................................................................... 217References ............................................................................................................... 218

Chapter 9 : Biosensors as Advanced Device for the Transgenic Plants and Food and Detection...................................................................221

Juliana B. Sousa, Joilson Ramos-Jesus, Rosana Anita S. Fonseca, Cristina Delerue-Matos, Maria Fátima Barroso, José Ribeiro Santos, Jr.1 Introduction ......................................................................................................... 2212 Conventional Analytical Methods for the Assessment of GMOs ....................... 223

2.1 Protein-Based Methods for the Detection of GMOs ................................... 2232.2 DNA-Based Methods for the Detection and Quantification of GMOs .........225

3 Biosensor Development for the Assessment of GMOs ....................................... 2273.1 Development of Immunosensors for the Detection of GMOs ..................... 2283.2 Development of Genosensors for the Determination of GMOs .................. 232

Contents

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Chapter 10 : Genetically Engineered Food Crops to Abiotic Stress Tolerance ...........................................................................247

Sávio P. dos Reis, Deyvid N. Marques, Nicolle L. Ferreira Barros, Carinne de Nazaré Monteiro Costa, Cláudia R. Batista de Souza1 Introduction ......................................................................................................... 2472 Genetic Transformation of Plants ....................................................................... 251

2.1 Biolistic-Mediated Genetic Transformation ................................................ 2522.2 Agrobacterium-Mediated Genetic Transformation ...................................... 253

3 Proteins With Protective Roles Against Abiotic Stress ....................................... 2543.1 Late Embryogenesis Abundant Proteins ...................................................... 2543.2 Heat Shock Proteins ..................................................................................... 2563.3 Aquaporins ................................................................................................... 258

4 Signaling Molecules\Transcription Factors ........................................................ 2594.1 Calcineurin B-Like Proteins ........................................................................ 2594.2 Basic Leucine Zipper Proteins ..................................................................... 2614.3 AP2/ERF Proteins ........................................................................................ 262

5 Osmoprotectant Metabolites ............................................................................... 2645.1 Trehalose-6-Phosphate Synthases (TPS) and Phosphatases (TPP) ............. 2645.2 Pyrroline-5-Carboxylate Synthases (P5CS) and Reductases (P5CR) ......... 266


Chapter 11 : Detection of Prevented DNA Damage by Therapeutic Foods ........................................................................281

Burcu Dogan-Topal, Bengi Uslu, Sibel A. Ozkan1 Introduction ......................................................................................................... 2812 DNA Damage and Aging .................................................................................... 282

2.1 Types of DNA Damage ................................................................................ 2842.2 Detection of DNA Damage .......................................................................... 2852.3 Repair of DNA Damage............................................................................... 287

3 Therapeutic Foods (Nutraceuticals) .................................................................... 2894 Detection of Prevented DNA Damage ................................................................ 290

4.1 Phenolic Compounds ................................................................................... 2934.2 β-Glucan ...................................................................................................... 2984.3 Vitamin E ..................................................................................................... 2994.4 Fermented Papaya ........................................................................................ 3004.5 Vitamin C ..................................................................................................... 3004.6 Carotenoid .................................................................................................... 3014.7 Multicomponent ........................................................................................... 302


Contents

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Chapter 12 : Genetically Engineered Crops: Opportunities, Constraints, and Food Security at a Glance of Human Health, Environmental Impact, and Food Quality .........................................311

Vishal V. Dawkar, Ashok D. Chougale, Vitthal Barvkar, Rahul S. Tanpure, Ashok P. Giri 1 Introduction ....................................................................................................... 311 2 Green Revolution .............................................................................................. 313 3 GM Crops: An Overview of Human Health, Their Environmental Impact,

and Food Quality ................................................................................................3153.1 Food Quality and Nutritional Content ....................................................... 315

4 Fulfill Food Needs by Increasing Food Production .......................................... 317 5 Safety Issues ...................................................................................................... 318

5.1 Potential Effects of GM Plants on Human Health and the Environment ..........................................................................................318

5.2 Allergy Concerns ....................................................................................... 319 6 Gene Flow and Landrace Contamination .......................................................... 319 7 Swapping Insect Pests for Targeted Attacks ..................................................... 321 8 BT Crop Resistance .......................................................................................... 322 9 Superweeds ....................................................................................................... 32210 Bioethical and Social Issues of GM Crops ....................................................... 32211 If Anything is Wrong With GM Technology There is no

Turning Back! .............................................................................................. 32412 Food Security .................................................................................................... 32513 Conventional Breeding, Biopesticides, and GM Crops: What to Choose

for Food Security? ..............................................................................................32814 Conclusions ....................................................................................................... 330References ............................................................................................................... 330

Chapter 13 : Safety and Risk Assessment of Food From Genetically Engineered Crops and Animals: The Challenges ...............................335

Wilna J. van Rijssen, E. Jane Morris1 Introduction ......................................................................................................... 3352 Safety and Risk Assessment of Food From GE Plants (Crops) .......................... 336

2.1 Food Safety and Risks ................................................................................. 3363 Approaches to Safety and Risk Assessment ........................................................ 339

3.1 The Targeted Approach ................................................................................ 3393.2 A Nontarget Approach ................................................................................. 3413.3 Hierarchical, Multistep, Risk-Based, and Weight-of-Evidence-Based

Approach .......................................................................................................3424 Problem Formulation .......................................................................................... 3425 Procedures for the Initial Risk and Safety Assessment of GE Plants

Described by Codex Alimentarius .......................................................................3435.1 Molecular Characterization ......................................................................... 343

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5.2 Plant Breeding ............................................................................................. 3445.3 Compositional Analysis ............................................................................... 3455.4 Natural Variation of Food Component Concentrations................................ 346

6 The Novel Protein(s) ......................................................................................... 3466.1 Toxicity ...................................................................................................... 3466.2 Allergenicity ............................................................................................... 3476.3 Endogenous Allergens and Toxicants ........................................................ 3486.4 Antibiotic Marker Genes ............................................................................ 348

7 Animal Studies .................................................................................................. 3487.1 Nutrition Evaluation ................................................................................... 3487.2 GE “Output Traits” .................................................................................... 3497.3 Quality Control and Quality Assurance ..................................................... 3497.4 Long-Term Animal Feeding Studies .......................................................... 3497.5 Limitations of Animal Feeding Studies ..................................................... 3507.6 Defective Studies ....................................................................................... 352

8 Unintended Effects ............................................................................................ 352 9 New Molecular Modification Techniques ......................................................... 353

9.1 New Plant Breeding Techniques ................................................................ 3539.2 Gene Modulation ....................................................................................... 3549.3 Critical Points in Food Safety Assessment ................................................ 354

10 GE Stacked Events ............................................................................................ 35611 GE Animals ....................................................................................................... 356

11.1 General ..................................................................................................... 35611.2 Atlantic Salmon ........................................................................................ 358

12 The Precautionary Approach (Principle)........................................................... 36113 Societal Decision Making ................................................................................. 36114 Benefit Assessment ........................................................................................... 36115 Conclusions ....................................................................................................... 361References ............................................................................................................... 362

Chapter 14 : Identification of Genetically Modified Foods .....................................369Florin Iordache, Horia Maniu, Carmen Curutiu, Petruta Cornea, Aneta Pop1 Introduction ......................................................................................................... 3692 Sampling ............................................................................................................. 3703 GMO Testing Methods ........................................................................................ 371

3.1 Indirect Methods .......................................................................................... 3713.2 Direct Method: DNA-Based Methods ......................................................... 373

4 Validation and Standardization of Analytical Methods ....................................... 3795 Conclusions and Perspectives ............................................................................. 380References ............................................................................................................... 380

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Chapter 15 : Methods for Plant Genetic Modification ..........................................385Coralia Bleotu, Lilia Matei, Laura D. Dragu, Liana Grigorescu, Carmen C. Diaconu, Gabriela Anton1 Introduction ......................................................................................................... 3852 Methods for Genetic Modification ...................................................................... 386

2.1 Nongenetic Engineering Methods ............................................................... 3862.2 Genetic Engineering .................................................................................... 388

3 The Role of Epigenetics in Plant Transformation ............................................... 3944 Potential Risks Related to the Use of Genetically Modified Foods .................... 3955 Conclusions ......................................................................................................... 397References ............................................................................................................... 398

Index ..............................................................................................................403

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Documents

Genetically Engineered Foods...v Contents List of Contributors..... xv Foreword ..... xvii Series Preface..... xix