... Preface... xix. Biological Control 4 Parasitoids 5 Predators 5. Baculoviruses. Entomopathogenic

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1 Foreword... xvii Preface... xix 1. Pest Management and the Environment 1 Introduction 1 Pest-Associated Crop Losses and the Need for Pest Management What Is Available in the Basket and What Can We Do? 2 3 Pest Management Components... L1 Economic Thresholds... L1 Biological Control 4 Parasitoids 5 Predators 5 Entomopathogenic Baculoviruses Bacteria 6 6 Entomopathogenic Fungi 7 Entomopathogenic Cultural Control Nematodes 7 8 Date of Sowing and Planting Density 8 Nutrient Management 8 Intercropping and Crop Rotations 9 Field Sanitation and Tillage Chemical Control 9 9 Development of Resistance to Insecticides and Strategies for Resistance Management... Pest Resurgence Pesticide Residues in Food and Food Products Contamination of Soil and Water... Pesticides of Plant Origin... Host Plant Resistance... Integrated Pest Management... Mating Disruption and Mass Trapping

2 vi Population Prediction Models and Early Warning Systems 15 The IPM practice 16 Is Genetic Engineering of Plants and Biocontrol Agents an Answer? References Applications Introduction of Biotechnology in Agriculture: The Prospects The Genomics Revolution 24 Marker-Assisted Selection 25 Gene Sequence and Function 27 Metabolic Pathways 27 Trait Analysis 28 Genetic Transformation 28 Resistance to Insect Pests, Diseases, and Herbicides 29 Tolerance to Abiotic Stresses 29 Increased Starch and Sugar Production y 30 Altering Senescence and Drought Resistance 30 Increased Photosynthetic Efficiency, Crop Growth, and Yield 30 Improved Nutrition 31 Production of Pharmaceuticals and Vaccines 32 Production of Antibodies 32 Genetic Improvement of Entomopathogenic Microorganisms 33 Genetic Improvement of Natural Enemies 33 Application of Biotechnology in Biosystematics and Diagnostics 34 Exploitation of Male-sterility and Apomixis References Evaluation of Transgenic Plants and Mapping Populations for Resistance to Insect Pests 41 Introduction 41 Techniques to Screen for Resistance to Insects under Natural Infestation 42 Use of Hot-Spot Locations 42 Adjusting Planting Date 44 Manipulation of Cultural Practices 44 Planting Infester Rows 44 Grouping the Material According to Maturity and Height 45 Sequential Plantings 46 Selective Control of Nontarget Insects 46 Augmentation of Insect Populations 46 Labelling the Plants or Inflorescences Flowering at the Same Time 47 Techniques to Screen for Resistance to Insects under Artificial Infestation 47 Mass Rearing : 47 Infestation Techniques 48 Planning the Rearing Schedule and Egg and Pupal Storage 51 Caging the Plants with Insects 51 Greenhouse Screening Use of Excised Plant Parts Detached Leaf Assay 55

3 vii Leaf Disc or Pod/Boll Assay 58 Oviposition Nonpreference 58 Diet Impregnation Assay to Assess Antibiosis 59 Bioassay of Transgenic Plants for Resistance to Insects 60 Phenotyping Mapping Populations for Resistance to Insects 62 Measurement of Host Plant Resistance to Insects 62 Visual Damage Rating 63 Indirect Feeding Injury 63 Simulated Feeding Injury 64 Association of Physico-Chemical Characteristics and Molecular Markers with Insect Resistance 64 Sampling Insect Populations 64 Measurements of Yield and Quality 68 Measurements of Insect Survival and Development 68 Consumption and Utilization of Food 68 Measurements of Insect Behavior 69 Selection Indices 70 Tolerance Index 70 Loss in Grain Yield 70 Relative Efficiency Index 71 Fischer and Maurer's Stress Susceptibility Index Fernandez Stress Tolerance Index References ; Host Plant Resistance to Insects: Potential and Limitations 83 Introduction 83 Identification and Utilization of Resistance 85 Wild Relatives of Crops as Sources of Resistance to Insects 87 Inducible Resistance 89 Factors Affecting Expression of Resistance to Insects 91 Soil Moisture 91 Plant Nutrition 91 Temperature 91 Photoperiod 92 Insect Biotypes 92 Influence of HPR on Pest Population Dynamics and Economic Injury Levels 92 Host Plant Resistance in Integrated Pest Management 94 HPR as a Principal Method of Insect Control 94 HPR and Biological Control 98 Tritrophic Interactions 99 Compatibility of Plant Resistance and Biological Control 100 Incompatibility of Plant Resistance and Biological Control 101 Plant Resistance-Insect Pathogen Interaction 102 Manipulation of Plant Characteristics to Increase the Effectiveness of Natural Enemies 102 HPR and Cultural Control 102 Asynchrony Between Plant Growth and Insect Populations 103 Genetic Diversity 103

4 viii Multilines/Synthetics Trap Crops Nutrient Application and Plant Resistance HPR and Chemical Control Interaction between Antixenotic Mechanism of Resistance and Chemical Control Interaction between Antibiosis Mechanism of Resistance and Chemical Control Moderate Levels of Plant Resistance and Chemical Control... High Levels of Plant Resistance and Chemical Control... Advantages of HPR... Limitations of HPR References Mechanisms and Inheritance of Resistance to Insect Pests... Introduction,... Mechanisms of Resistance to Insects... Antixenosis... Antibiosis... Tolerance... Escape... Breeding for Resistance to Insect Pests... Mass Selection... Recurrent Selection... Pedigree Breeding... Backcross Breeding... Development of Fl Hybrids Using Cytoplasmic Genetic Basis of Resistance Male-sterility Oligogenic Resistance... Polygenic Resistance... Cytoplasmic Effects... Genetics and Inheritance of Resistance to Insect Pests... Rice... Wheat and Barley... Maize... Sorghum... Cotton Oilseeds Alfalfa Potato Grain Legumes... Vegetables... Fruits References Physico-Chemical and Molecular Markers for Resistance to Insect Pests Introduction Mapping Populations

5 ix Physico-chemical Markers Associated with Resistance to Insects Morphological Markers Visual Stimuli Phenological Traits Leaf Hairs Trichomes Plant Growth Responses Biochemical Markers Attractants Repellents Phagostimulants Antifeedants Growth Inhibitors Nonprotein Amino Acids Nutritional Factors Enzymes Molecular Markers Restriction Fragment Length Polymorphisms Sequence-Tagged Sites Expressed Sequence Tags Single Strand Conformation Polymorphisms Microsatellites Simple Sequence Repeats Randomly Amplified Polymorphic DNA Inter Simple Sequence Repeat Sequence Characterized Amplified Regions Amplified Fragment Length Polymorphisms Single Nucleotide Polymorphisms Diversity Array Technology Molecular'Markers Linked to Insect Resistance in Different Crops Cotton Rice Wheat Maize Sorghum Potato Tomato Chickpea Pigeonpea Cowpea Common Bean Greengram Soybean Groundnut Gene Synteny Molecular Markers and Metabolic Pathways The Transgenic Approach and Gene Pyramiding through MAS Marker-Assisted versus Phenotypic Selection References

6 x 7. Genetic Transformation of Crops for Resistance to Insect Pests Introduction Genetic Transformation: The Protocols Agrobacterium-Mediated Gene Transfer Microprojectile Bombardment with DNA or Biolistics Direct DNA Transfer into Isolated Protoplasts Gene Expression Genetic Transformation of Crop Plants for Resistance to Insects Toxin Proteins from Bacillus thuringiensis Cotton Maize Rice Sorghum Sugarcane Oilseed Crops Grain Legumes... Tobacco,... Potato Vegetables Fruits and Forest Trees Ornamentals Vegetative Insecticidal Proteins Toxin Proteins from Photorhabdus luminescens Secondary Plant Metabolites Pro tease Inhibitors Tobacco Potato Cotton Maize Rice Wheat Sugarcane Vegetables Chrysanthemum Alpha Amylase Inhibitors Enzymes Plant Lectins Tobacco Potato Cotton Cereals Sugarcane Vegetables Viruses, Neurotoxins, and Insect Hormones Neurotoxins Viruses Neuropeptides and Peptidic Hormones Biotin-Binding Proteins Antibodies Inducible Resistance

7 xi Gene Pyramiding References Genetic Engineering of Entomopathogenic Microbes for Pest Management 255 Introduction 255 Natural versus Engineered Microbes 256 Genetic Engineering of Microbes 257 Entomopathogenic Virus es Baculoviruses with Neurotoxins 259 Baculoviruses Expressing Insect Diuretic Hormones 262 Expression of Entomopoxvirus in Bacteria 262 Effect of Insecticides on Biological Activity of Baculoviruses Expressing Neurotoxins 262 Role of Baculoviruses in Pest Management 262 Entomopathogenic Bacteria 263 Bacillus thuringiensis 263 Gene Expression and Cry Structure 264 Mode of Action 265 Biopesticides Based on Bacillus thuringiensis 267 Genetic Engineering of Bacteria 268 The Alternative Delivery Systems for Bt Toxins 269 Entomopathogenic Fungi 272 Entomopathogenic Protozoa 275 Entomopathogenic Nematodes 275 Biosafety Considerations for Using Genetically Engineered Microbes References Genetic Ehgineering Introduction of Natural Enemies for Integrated Pest Management Techniques for Genetic Engineering of Arthropods 295 DNA Injection into Eggs 295 Maternal Microinjection 296 Sperm-Mediated Transfer 296 Paratransgenesis 297 Vectors for Genetic Engineering of Arthropods 297 Transposable Elements 297 Viral Vectors 298 Baculoviruses 298 Genetic Improvement of Beneficial Arthropods 299 Cryopreservation 299 Altering Biological Attributes 301 Quality Control of Insect Cultures and Mass Production 301 Adaptation to Extreme Environmental Conditions 301 Improving Resistance to Insecticides 301 Dominant Repressible Lethal Genetic System 303 Markers and Promoters 304 Environmental Release and Potential Risks 305 Genetic Exchange with Natural Populations 306

8 xii Horizontal Gene Flow Transgene Instability References Deployment of Insect-Resistant Transgenic Crops for Pest Management: Potential and Limitations... Introduction... Progress in Development of Insect-Resistant Transgenic Crops... Effectiveness of Transgenic Crops for Controlling Insect Pests... Cotton... Cereals... Grain Legumes Potato Vegetables... Deployment of Transgenic Plants for Pest Management... Transgenic Crops and Chemical Control "j'... Transgenic Crops and Biological Control... Transgenic Crops and Cultural Control... Advantages and Limitations of Insect-Resistant Transgenic Crops... Stability of Transgene Expression... Performance Limitations... Secondary Pest Problems... Insect Sensitivity... Evolution of Insect Biotypes... Environmental Influence on Gene Expression References Transgenic Resistance to Insects: Interactions with Nontarget Organisms... Introduction... Bt Sprays, Transgenic Plants, and Nontarget Organisms... Interaction of Transgenic Crops with Nontarget Organisms: Protocols for Ecotoxicological Evaluation of Transgenic Plants... Influence of Transgenic Crops on Diversity of Nontarget Organisms... Influence of Transgenic Crops on Activity and Abundance of Pollinators... Interaction of Transgenic Crops with Predators... Synergistic/Neutral Interactions... Bt Toxins... Lectins... Antagonistic Interactions... Bt Toxins... Protease Inhibitors... Lectins... Interaction of Transgenic Crops with Parasitoids... Synergistic/Neutral Interactions... Bt Toxins... Lectins... Antagonistic Interactions... Bt Toxins

9 xiii Protease Inhibitors... Lectins... Interactions of Transgenic Plants with Fauna and Flora in the Rhizosphere References Development of Resistance to Transgenic Plants and Strategies for Resistance Introduction Management Factors Influencing Insect Susceptibility to Bt Toxins... Variation in Insect Populations for Susceptibility to Bt Toxins... Effect of Host Plant on Insect Susceptibility to Cry Proteins... Detection, Development, and Monitoring of Resistance... Detection of Resistance... Development of Resistance under Laboratory Conditions... Development of Resistance under Field Conditions... Mechanisms of Resistance... Reduced Protoxin-Toxin Conversion... Reduced Binding to Receptor Proteins... Feeding Behavior Cross Resistance Genetics of Resistance... Frequency of Resistance Alleles... Inheritance of Resistance... Strategies for Resistance Management... Stable Transgene Expression... High Level of Transgene Expression... Gene Pyramiding... Pyramiding Two or More Bt Genes... Pyrarniding Bt with Protease Inhibitor and Lectin Genes... Pyramiding Bt Genes with Conventional Host Plant Resistance... Pyramiding Genes for Resistance to Multiple Pests... Regulation of Gene Expression and Gene Deployment... Planting Refuge Crops... Refuge Types... Role of Alternate Hosts as Refugia... Removal of Alternate Hosts and Destruction of Carryover Population... Use of a Planting Window... Crop Rotations... Integrated Pest Management... Simulation Models for Resistance Management References Transgenic Resistance to Insects: Gene Flow Introduction Role of Pollinators in Gene Flow Role of Viruses in Gene Flow Gene Flow into the Wild Relatives of Crops: Vertical Gene Flow Cotton

10 xiv Cereals... Brassicas... Soybean... Sunflower... Vegetables... Beets... Development of Resistance to Antibiotics: Horizontal Gene Flow... Gene Flow, Selection Pressure, and Enhanced Fitness of Herbivores... Gene Flow and Selection Pressure from Herbivores... Gene Flow and Enhanced Fitness of Herbivores... Gene Flow and Genetic Purity of Crops... Gene Flow and the Centers of Genetic Diversity... Gene Flow and Aquatic Environments... Management of Gene Flow... Use of Taxonomic Information... Geographic Isolation... Use of Border Rows... Use of Cytoplasmic Male-sterility ' References Transgenic Resistance to Insects: Nature of Risk and Risk Management... Introduction... Assessment of Risk to Agriculture... Selection for Weedy Traits... Invasiveness... Assessment of Nature of Risk to the Environment... Risk Assessment... Probability of Harm... Hazard to the Environment... Probability of Horizontal and Vertical Gene Flow... Risk Management... General Information... DNA Donor and Receiving Species... Conditions of Release and the Target Environment... Interaction Between the Transgenic Plants and the Environment... Control, Monitoring, and Waste Treatment... Post Release Monitoring of the Transgene References Biosafety of Food from GeneticalIy Modified Crops... Introduction... Biosafety Assessment of Food from Genetically Modified Crops... Comparative Effects of Traditional Breeding and Genetic Engineering on Food Quality... Substantial Equivalence to the Nontransgenic Food... Nutritional Quality

11 xv Toxicological Effects and Metabolism of the Transgene Products... Biosafety of Transgene Products... Bt Cry Proteins..... Protease Inhibitors... Lectins... Changes in Chemical Profile/Secondary Metabolites... Biosafety of Transgenic Feed/Forage for Animals... DNA Transfer from Transgenic Food to Microorganisms/Humans... Potential Effects on Human Health Resulting from the Use of Viral DNA in Human Food... Transposable Elements... Fate of Genetically Modified Plant DNA in the Digestive System... Allergenic Effects... Public Attitude to Food fram Transgenic Plants... Introduction of Food fram Genetically Modified Craps to the General Public... Consumer Response to Food from Genetically Modified Crops... Role of the Scientific Community, NGOs, and the Media References Detection and Monitoring of Food and Food Products Derived from GeneticalIy Modified Crops... Introduction... Sampling for Detection of Genetically Modified Food... Detection of Genetically Modified Foods... DNA-Based Methods... Qualitative PCR Analysis... Multiplex PCR-Based Detection Methods... Quantitative PCR... Microarray Technology... RNA-Based Methods... Protein-Based Methods... Enzyme-Linked Immunosorbent Assay... Lateral Flow Sticks... Phenotypic Characterization... Mass Spectrometry... Surface Plasmon Resonance... Monitoring of Genetically Modified Food and Food Labeling References Molecular Markers for Diagnosis of Insect Pests and Their Natural Enemies Introduction... Molecular Tools for Diagnosis of Insect Pests... Polymerase Chain Reaction... Random Amplified Polymorphic DNA

12 xvi Expressed Sequence Tags 478 Restriction Fragment Length Polymorphisms 479 Inter-Simple Sequence Repeats 479 Application of Molecular Markers for Insect Diagnosis 479 Diagnosis of Insect Pests and Their Natural Enemies 479 Detection of Insect Biotypes 480 Phylogenetics and Population Structure 481 Application of Molecular Markers for Studying Population Genetics 482 Application of Molecular Markers for Studying Molecular Basis of Insect-Plant Interactions Social Behavior of Insects Application of Molecular Markers to Understand of Insects Functional Genomics 484 Detection of Natural Enemy-Insect Host Interactions References Molecular Techniques for Developing New Insecticide Molecules I and Monitoring Introduction Insect Resistance to Insecticides Development of New Insecticide Molecules 493 Molecular Markers for Monitoring Insect Resistance to Insecticides 494 Comparative Genomics and Divergent Evolution of Detoxification Genes 495 Microarrays and Regulatory Mutations in Cytochrome P450s 496 Quantitative Trait Loci, Positional Cloning, and Multiple Resistance to Bt Toxins 496 Selective Sweeps and Genomic Consequences References Biotechnology, Introduction Pest Management, and the Environment: The Future Genetic Engineering of Crops for Resistance to Insect Pests 501 Genetic Improvement of Natural Enemies 504 Genetic Improvement of Entomopathogenic Microorganisms 504 Molecular Markers, Biosystematics, and Diagnostics 505 Development of New Insecticide Resistance to Insecticides Molecules and Monitoring Insect 506 Marker-Assisted Selection and the Genomics Revolution 506 Transgenic Crops and the Environment 507 Biosafety of Food from Transgenic Crops References 509 Species Index 513 Subject Index 521