Title Page
Contents
ABSTRACT 20
CHAPTER 1. INTRODUCTION 23
Overview 28
Study sites and species 30
References 32
CHAPTER 2. EXCEPTIONAL PROPERTIES OF HYPER-RESISTANT ARMOR OF A HYDROTHERMAL VENT CRAB 39
Abstract 40
Introduction 41
Materials and Methods 43
Sample preparation 43
Properties analysis 44
Statistical analysis 45
Results 46
Mechanical properties 46
Thermal stability 46
Characteristics of structure 47
Component characteristics 48
Discussion 48
Conclusion 52
References 53
Supporting Informations 66
References 70
CHAPTER 3. CONVERGENT EVOLUTION OF HYDROTHERMAL VENT CRUSTACEAN EXOSKELETONS REVEALS THE SPECEIAL PROPERTIES TO RESIST EXTREME TEMPERATURE 75
Abstract 76
Introduction 77
Materials and Methods 79
Sample preparation 79
DNA extraction and Molecular analysis 80
Property analysis 81
Statistical analysis 83
Results 85
Species identification and phylogenetic tree 85
Thermal stability 85
Mechanical properties 86
Structural characteristics 86
Characteristics of components 87
Discussion 88
References 94
Supplementary Materials 108
PCR 108
Molecular analysis 108
References 121
CHAPTER 4. UBIQUITOUS THERMAL RESISTANCE IN THE ARMOR OF THE DEEP-SEA SQUAT LOBSTERS Munidopsis: IS IT PREADAPTATION OR DERIVED FROM ANCESTORS IN HYDROTHERMAL VENTS 127
Abstract 128
Introduction 130
Materials and Methods 133
Sample information 133
Analysis method 133
Statistical analysis 135
Results 136
Thermal stability of exoskeletons 136
Compounds of exoskeleton 136
Discussion 137
References 142
CHAPTER 5. EXOSKELETAL TRADE-OFF BETWEEN CLAWS AND CARAPACE IN DEEP-SEA HYDROTHERMAL VENT DECAPOD 164
Abstract 165
Introduction 167
Materials and Methods 170
Sample Preparation 170
Exoskeleton Analysis 171
Statistical Analysis 172
Results 174
Mechanical properties analysis 174
Components Analysis 175
Structure Analysis 176
Morphology analysis 177
Discussion 178
How to control the mechanical properties of the exoskeleton? 178
Why are carapaces strengthened or claws weakened in hydrothermal species? 180
References 184
CHAPTER 6. CONCLUSION 201
국문 초록 206
Table 1.1. Sample information. 37
Table 4.1. Sampling locations and informations. 159
Table 4.2. FT-IR peaks for exoskeleton of samples. 161
Table 4.3. Raman bands according to references. 162
Table 5.1. Raman peak data / cm⁻¹ of crustacean exoskeleton. 200
Supplementary table 2.1. Indicators related to the TGA results [significant difference is indicated by an asterisk (*), (mean ± SE)]. 68
Supplementary table 2.2. The peaks of the exoskeleton sample with the corresponding substances. 69
Supplementary table 3.1. Substitution model for each gene used for ML and BI analysis. 113
Supplementary table 3.2. Weight loss from TGA (mean ± SE). 114
Supplementary table 3.3. Contents of the elements constituting each layer of the crustacean exoskeleton (mean ± SE). 115
Supplementary table 3.4. Statistical results of comparative analysis of the elements of each layer of the exoskeleton of five species of crustaceans. 118
Supplementary table 3.5. Raman data / cm⁻¹ of five species of the crustacean exoskeleton. 120
Figure 1.1. Sampling site and species. 36
Figure 2.1. Crab samples and mechanical properties. The samples of the two species [(a) Austinograea sp., (b) C. japonica]. (c, d) The hardness and reduced modulus of the exoskeletons of the two crabs in cross-sections (mean ± SE). (e, f) The hardness and... 62
Figure 2.2. Comparison of the hardness for various species (mean ± SE). 63
Figure 2.3. Thermogravimetric analysis (TGA) curve. The first stage of weight loss was visible from room temperature to 200℃, which indicates the removal of water or volatile... 64
Figure 2.4. Component analysis results using EDX. Comparison of (a) main and (b) minor elements of the exoskeleton between the two crabs [significant difference is indicated by an asterisk (*); mean ± SE]. 65
Figure 3.1. Research species on each sampling site (a) and phylogenetic relationship (b). The phylogenetic relationship was estimated using two... 104
Figure 3.2. Thermal stability analysis results. a TGA curve of crustaceans and each combusting substance on each range. Weight loss for each temperature... 105
Figure 3.3. Mechanical properties and thickness ratio of each layer of the exoskeleton. (a hardness, b reduced modulus, and c thickness ratio). 106
Figure 3.4. Crustacean exoskeleton (endocuticle layer) compound analysis graph through Raman analysis. 107
Figure 4.1. Sampling sites for each species distributed across diverse habitats. 156
Figure 4.2. Analysis of thermal stability of exoskeletons. Total temperature range (a) and each ranges [mean ± SE; the significant difference is indicated by an asterisk (*); b]. 157
Figure 4.3. Raman spectra from samples. 158
Figure 5.1. Comparative mechanical properties of crustacean claws and carapaces. Brachyura (a-d; VC and APC) and Anomura (e-h; VSL and PHC). Vent species... 195
Figure 5.2. Component analysis of elements in the exoskeleton in crustaceans. Brachyura (a, b; VC and APC) and Anomura (c, d; VSL and PHC). Vent species are depicted in red; coastal species are depicted in blue (solid: claw and hollow:... 196
Figure 5.3. Comparative analysis of compounds in the exoskeleton of crustaceans. (a) claw and (b) carapace. 197
Figure 5.4. Structural characteristics of the exoskeleton in crustaceans. (a) Diagram of a single layer of Bouligand structure. Stacking height of one single layer of... 198
Figure 5.5. Morphological characteristics of crustacean claws [(a) VC, (b) APC, (d) VSL, (e) PHC]. Comparative graph for the number of teeth on the cutting-edge in two species of (c) Brachyura and (f) Anomura. Red depicts vent species and blue... 199
Supplementary figure 2.1. Cross-section of the exoskeleton of the (a) vent crab and (f) coastal crab. (b, g) The granular structure of the epicuticle. The Bouligand structure of the (c, h) exocuticle and the (d, i) endocuticle. (e, j) The multilayer... 66
Supplementary figure 2.2. Raman spectra of exoskeleton [(a) vent crab and (b) coastal crab]. 67
Supplementary figure 3.1. Crustacean sample image (a VSL, b BC, and c MS). The part of the analysis is indicated as a dotted line. 110
Supplementary figure 3.2. Microstructure of each layer of the VSL (a-d), BC (e-h), MS (i-l). Epicuticle (a e i), exocuticle (b f j), endocuticle (c g k), and membrane layer (d h l). Yellow arrows indicate epicuticle (a e i) and membrane layer (d h l). 111
Supplementary figure 3.3. Raman analysis graph of epicuticle (a), exocuticle (b), and membrane layer (c). 112