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Abstract and EpOR6 are the most closely related E. postvittana ORs to other moth pheromone receptors. However, EpOR1 binds plant semiochemicals and not pheromone components and EpOR6 remains to be functionally annotated. Tissue expression analysis of 26 of the ORs revealed that three (EpOR30, 33 and 34) are more than 600 times more highly expressed in male than female antennae, making them good candidates for being the pheromone receptors of E. postvittana. Functional analysis of these ORs will reveal their role(s) in E. postvittana olfaction. ii
Acknowledgements Acknowledgements I would like to extend my sincere thanks for the guidance and support I received from my supervisors: Richard Newcomb, Andrew Kralicek and David Christie. This PhD would not have been possible without your encouragement and open door policy for us students. Thank you for always being available for discussions from the commencement through to the completion of my research. No work is complete without the efforts of all the members of a team. My heartfelt thanks to all the members of the Molecular Sensing Team. Thank you Melissa for doing the groundwork on EpOR1; to Cyril for your expertise in recombinant protein expression and purification; to Colm and Aidan for sharing your expertise of Sf9 cell culture; to Nadeesha for your guidance in the lab and for your friendship and to Edwige for your valuable time and guidance in dealing with the ghosts of qRT-PCR, and your friendly support and encouragement in the highs and lows of my life. A big thank you to Pablo, Andy, Sylvia, Jeremy, Leah and Caroline for your company in the lab and during the many lunch breaks. The time spent as part of the Molecular Sensing team would not have been enjoyable without you all! A very big thank you to Anne Barrington of the insect rearing facility at Plant and Food Research Ltd, for rearing the thousands of sacrificial moths without whom a major part of this thesis would have been incomplete. Thank you to the members of the microarray facility at Plant and Food Research, especially Luke Luo for setting up the hybridisations and Bart Jenssen for your expertise on data analysis. I would also like to show my gratitude to the Bioinformatics department at Plant and Food Research, especially Ross Crowhurst for doing all the raw data analysis of the tons of sequencing with a smile! Thank you a lot Ross! I would also like to thank the Allan Wilson Centre Genome Service, especially Lorraine Berry for your advice on sample preparations and genomic sequencing; and to the team at the University of Otago high-throughput DNA sequencing unit. iii
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Functional characterisation of Epip
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3.1 Introduction 3 The roles of Epi
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The roles of Epiphyas postvittana G
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4.1 Introduction 4 Identification o
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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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