Single-Pollen Genotyping (eBook)

Preface 6
Contents 8
Contributors 10
Chapter 1: Significance of Single-Pollen Genotyping in Ecological Research 14
1.1 Introduction 15
1.2 Pollination Methods Have Diversified During the Past 130 Million Years 15
1.3 Importance of Studying Pollen Movement 16
1.4 Traditional and Novel Methods to Infer Pollen Movement 16
References 18
Chapter 2: Procedure for Single-Pollen Genotyping 19
2.1 Introduction 20
2.2 Materials 20
2.2.1 Clean Workspace 20
2.2.2 Stereomicroscope 21
2.2.3 Laboratory Tools and Equipment 21
2.3 Methods 21
2.3.1 Sample Handling 21
2.3.2 DNA Extraction 22
2.3.3 PCR 24
2.4 Notes 26
2.5 Conclusions 26
Box 2.1 Airborne-Pollen Pool Compositions Revealed by Single-Pollen PCR Technique 24
References 27
Chapter 3: Pollination Efficiencies of Insects Visiting Magnolia obovata, as Determined by Single-Pollen Genotyping 28
3.1 Introduction 29
3.2 Traditional Approaches to the Study of Pollination Ecology 29
3.3 Target System and Genetic Analysis 30
3.3.1 Study Species 30
3.3.2 Sample Collection and Counting of Adhered Pollen 31
3.3.3 Genotyping of Transported Pollen Grains 33
3.4 Origin of Pollen Grains: Where was the Source of the Pollen? 34
3.4.1 Proportion of Self-Pollen in the Transported Pollen 34
3.4.2 Composition of Pollen Donor of Adherent Pollen Grains 35
3.5 Pollen Transfer Characteristics of Flower-Visiting Insects 35
3.5.1 Flower Beetles and Small Beetles 35
3.5.2 Bumblebees 39
3.5.3 Conclusions and Future Directions 39
References 40
Chapter 4: Difference in Pollen Donor Composition During the Early Phases of Reproduction as Revealed by DNA Analysis of Pollen 44
4.1 Introduction 45
4.2 Study Species 46
4.3 Study Site 46
4.4 Methods 49
4.4.1 Sampling Design 49
4.4.2 DNA Extraction and Amplification 49
4.4.3 Paternity Analysis 50
4.4.4 Statistical Analysis 50
4.5 Pollen Donor Composition at the Pollination Stage 50
4.6 Pollen Selection 52
4.7 Conclusion 54
References 55
Chapter 5: Expanded Home Range of Pollinator Birds Facilitates Greater Pollen Flow of Camellia .japonica in a Forest Heavily 58
5.1 Introduction 59
5.2 Materials and Methods 60
5.2.1 Study Areas 60
5.2.2 Study Species 61
5.2.3 Flowering Conditions of C. japonica and Home Ranges of Z. japonica 62
5.2.4 Pollen Grain Sampling and Haplotyping 63
5.2.5 Mature Tree Sampling and Genotyping 63
5.2.6 Analysis of Genetic Diversity 64
5.3 Results 64
5.3.1 Flowering Conditions and Home Range of Zosterops japonica 64
5.3.2 Microsatellite Haplotyping from Single Pollen Grains 67
5.3.3 Genetic Diversity 67
5.4 Discussion 69
5.4.1 Resilience of the Camellia japonica–Zosterops japonica System Against Environmental Perturbation 69
5.4.2 Conclusions and Future Research 70
References 71
Chapter 6: Can Tiny Thrips Provide Sufficient Pollination Service During a General Flowering Period in Tropical Rainforest? 74
6.1 Introduction 75
6.2 Materials and Methods 77
6.2.1 Study Site, Target Plant Species, and Mast Flowering in 2005 77
6.2.2 Collection of Insect Visitors to Flower and Tree 78
6.2.3 Genotype Determination of Pollen Grains Adhering to Thrips 79
6.2.4 Proportions of Self-Pollen and Genetic Diversity in Pollen Grains Adhering to Thrips 79
6.2.5 Selfing Rates of Seeds 80
6.2.6 The Contribution of Thrips to Seed Production 80
6.3 Results 81
6.3.1 Flower-Visiting Insects and Their Seasonal Variation 81
6.3.2 Microsatellite Genotyping from Single Pollen Grains 83
6.3.3 Proportions of Self-Pollen and Genetic Diversity in Pollen Grains Adhering to Flower Thrips 83
6.3.4 Selfing Rates of Seeds and the Contribution of Thrips to Seed Production 85
6.4 Discussion 85
6.4.1 Pollination Efficiency of Flower Thrips 85
6.4.2 Pollination System of S. acuminata 87
References 90
Chapter 7: Alien Dandelions Displace a Native Related Species Through Interspecific Pollen Transfer 93
7.1 Introduction 94
7.2 Materials and Methods 95
7.2.1 Study Sites 95
7.2.1.1 Materials 96
7.2.1.2 Sampling Methods 96
7.2.1.3 Molecular Methods 97
7.2.1.4 Statistical Analysis 100
7.3 Results 100
7.4 Discussion 104
References 106
Chapter 8: Single-Pollen Genotyping of Holocene Lake Sediments 110
8.1 Ancient DNA Analysis 111
8.2 Why Fossil Pollen? 111
8.3 Pollen Treatment and DNA Extraction 113
8.4 Choice of Markers and Amplification 114
8.5 Contamination 115
8.6 Validation of Results 116
8.7 Conclusion 117
References 117
Chapter 9: Potential Application of Pollen Genotyping for Evolutionary Genetic and Genomic Studies: Linkage/Recombination An 119
9.1 Previous Studies of Recombination and Linkage Analysis Using Haploid Cells 120
9.2 Potential Application of Pollen Genotyping 121
9.2.1 Recombination Analysis and Construction of Linkage Map 121
9.2.2 Haplotype Sequence Analysis Using a Single Pollen Grain 127
9.3 Conclusions: Application of Pollen Genotyping to Evolutionary Genetic and Genomic Studies 128
Box 9.1 Structure of a Pollen Grain 121
Box 9.2 Haplotype Determination Strategies 124
References 129
Index 132