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General Introduction 30 this receptor is referred to as BmOR49 but in Tanaka et al. (2009), another OR has been annotated as BmOR49 so in this thesis, the BmOR49 of Anderson et al. (2009) will be referred to as BmOR49J], and tissue expression analysis showed it to be expressed in both male and female antennae at similar levels. Functional characterisation of EpOR3 by heterologous expression in Sf9 cells decoded its ligands to be members of terpenes, alcohols and esters at high concentrations, while at lower concentrations, the receptor bound citral, geraniol, geranial and geranyl acetate. Dose response curves showed EpOR3 has the highest affinity for the oviposition deterrent citral, with an EC50 value of 1.1 x 10 -13 M (Jordan et al., 2009), implicating a role of this receptor as an OR and not PR. Three ORs have so far been identified from E. postvittana; EpOR2 is the Drosophila OR83b homologue, EpOR3 is the receptor for general plant odorants while EpOR1 remains to be functionally characterised. Further analysis of these receptors and the identification of new receptors will lead to a better understanding of the olfactory mechanisms of this moth. This knowledge will aid in the development of pest control strategies targeting the olfactory system of the moth. Two GOBP members have also been identified; however this class of soluble proteins have very little functional data available, therefore an attempt will be made to deduce the role(s), if any, of this class of protein in odour perception in in vitro assay systems.
General Introduction 31 BmOr2 HvOr2 MsepOR2 DiOR2 PxOR2 EpOR2 0.1 100 * 80 HvOr15 HvOr14 MsepOR1 HvOr6 HvOr16 HvOr11 BmOr3 PxOR1 BmOr7 BmOr5 BmOr9 BmOr4 MsepOR3 HvOr13 DiOR3 BmOr1 DiOR1 EpOR1 PxOR3 PxOR4 Male-biased sex pheromone receptor clade 31 77 99 BmOr6 100 BmOr22 BmOr21 HvOr19 41 62 99 100 98 HvOr21 BmOr19 BmOr17 BmOr8 BmOr20 100 43 BmOr38 BmOr35 BmOr37 96 HvOr12 45 41 BmOr24 69 BmOr25 99 100 88 89 HvOr7 BmOr11 HvOr9 BmOr23 BmOr42 54 100 HvOr18 40 100 100 HvOr20 BmOr28 57 95 BmOr14 100 BmOr34 100 BmOr33 BmOr30 BmOr36 34 HvOr3 100 BmOR22J 100 BmOr15 99 BmOr12 30 73 100 100 HvOr8 BmOr13 EpOR3 * EpOR3 BmOR49 BmOR49J 100 100 BmOr48 BmOr46 Female-biased 100 BmOr47 BmOr45 receptor clade 100 BmOR19J 94 97 HvOr10 BmOr10 40 100 BmOr16 89 BmOR23J BmOr26 HvOr17 98 BmOr29 BmOr27 Or83b clade 98 99 48 100 60 25 100 Figure 1.8: Phylogenetic tree constructed from lepidopteran odorant receptors with the three E. postvittana ORs indicated by *. Adapted from Jordan et al. (2009) where the tree was originally constructed using the FITCH method from Jones-Thorton distances and the given bootstrap values calculated using 1000 replicates. 50 62 96 33 100 25 34 91 100 94 100 100 100 *
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Page 3 and 4: Abstract Abstract Olfaction or the
Page 5 and 6: Acknowledgements Acknowledgements I
Page 7 and 8: Contents v Contents Abstract……
Page 9 and 10: Contents 4.2.5 Degenerate PCR .....
Page 11 and 12: List of Figures List of Figures Fig
Page 13 and 14: List of Tables List of Tables Table
Page 15 and 16: List of Abbreviations gDNA Genomic
Page 17 and 18: 1.1 General overview 1 General Intr
Page 19 and 20: General Introduction 3 moth in loca
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Page 23 and 24: General Introduction 7 1.5 Componen
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Page 51 and 52: Functional characterisation of Epip
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Identification of putative odorant
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Microarray OR 454 Transcriptome OR
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5.1 Introduction 5 Concluding Discu
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Concluding Discussion 127 VOBA sett
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Concluding Discussion 129 redundant
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Concluding Discussion 131 against t
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Concluding Discussion 133 sequencin
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Appendix B 135 Buffered TB (Sambroo
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Appendix C 137 C. Appendix C - Clus
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Appendix D 139 D. Appendix D - Solu
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Appendix E 141 I II
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Appendix E 143 V
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Appendix E 145 VII continued Figure
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References 147 Ansebo, L., Ignell,
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References 149 Butenandt, A., Beckm
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References 151 Fedurco, M., Romieu,
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References 153 Große-Wilde, E., St
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References 155 Hrdy, I., F. Kuldova
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References 157 Kaissling, K. E. (19
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References 159 Krieger, J., Große-
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References 161 Lopez-Gomez, R. and
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References 163 Miura, N., Nakagawa,
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References 165 Pell, J. K., Macaula
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References 167 Røstelien, T., Stra
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References 169 Stowers, L. and Loga
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References 171 Turner, C. T., Davy,
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References 173 Vogt, R. G., Riddifo
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References 175 Widmayer, P., Heifet
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