Mechanisms of Olfaction in Insects - ResearchSpace@Auckland ...

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Acknowledgements I owe my deepest gratitude to my family, without whose support and encouragement this achievement would never have been possible. Thank you mum and dad for always encouraging me in my studies; to my siblings Shaireen, Imtiyaz, Imraan and Rizwaan for sharing in my life. I would also like to thank my parents-in-law and Arisha for your support, and to Sohail and Shayaan for being the brightest stars in my life. A very big thank you to my husband Aznain for your understanding and sacrifices during the course of my project, especially for your patience and words of guidance during the times when writing seemed like an unattainable feat. It just would not have been possible without all your love, support and encouragement. Finally, I would like to thank School of Biological Sciences and The University of Auckland for the travel grants for supporting my attendance to the Queenstown Molecular Biology Meeting (2006), to the International Symposium of Taste and Olfaction (2008) and to the Genetics Society of Australasia (2010). This research has been made possible through grants from Foundation of Research, Science and Technology and the Ministry of Research, Science and Technology of New Zealand. iv
Contents v Contents Abstract……………………………………………………………………...…...…...i Acknowledgements…………………………………………………………….……iii List of Figures……………………………………………………………………..…ix List of Tables…………………………………………………………………...……xi List of Abbreviations…………………………………………………………...…..xii 1 General Introduction ........................................................................................... 1 1.1 General overview ............................................................................................ 1 1.2 Olfaction in insects .......................................................................................... 2 1.3 Olfaction in moths ........................................................................................... 2 1.3.1 Sex pheromones ....................................................................................... 2 1.3.2 Other odorants detected by moths............................................................ 3 1.4 The olfactory system ....................................................................................... 4 1.5 Components of the olfactory system ............................................................... 7 1.5.1 Proteins found in the sensillum lymph..................................................... 7 1.5.2 Membrane proteins ................................................................................ 12 1.5.2.1 Odorant receptor identification in moths ........................................... 13 1.5.2.2 Insect odorant receptor activation ...................................................... 18 1.6 Insect pest control strategies ......................................................................... 21 1.7 Light brown apple moth ................................................................................ 24 1.8 Aims .............................................................................................................. 32 2 Functional Characterisation of Epiphyas postvittana Odorant Receptor 1 .. 34 2.1 Introduction ................................................................................................... 34 2.1.1 Aims ....................................................................................................... 39 2.2 Materials and methods .................................................................................. 39 2.2.1 Materials ................................................................................................ 39 2.2.2 Insect cell culture ................................................................................... 40 2.2.3 Transfection of cells ............................................................................... 40 2.2.4 Detection of mRNA encoding pIB-EpOR1 by RT-PCR ....................... 41 2.2.5 Membrane Fraction Isolation and Western Blot .................................... 41 2.2.6 EpOR1 functional assay ......................................................................... 43 2.2.7 Data analysis .......................................................................................... 44
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Page 3 and 4: Abstract Abstract Olfaction or the
Page 5: Acknowledgements Acknowledgements I
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
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Page 37 and 38: General Introduction 21 et al., 200
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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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