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Chemical Ecology of Pollination in Deceptive Ceropegia

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Chemical Ecology of Pollination in Deceptive Ceropegia Chemical Ecology of Pollination in Deceptive Ceropegia CHEMICAL ECOLOGY OF POLLINATION IN DECEPTIVE CEROPEGIA DISSERTATION zur Erlangung des Doktorgrades Dr. rer. nat. an der Bayreuther Graduiertenschule für Mathematik und Naturwissenschaften (BayNAT) der Universität Bayreuth vorgelegt von Annemarie Heiduk Bayreuth, Januar 2017 Die vorliegende Arbeit wurde in der Zeit von Februar 2012 bis Dezember 2016 in Bayreuth am Lehrstuhl Pflanzensystematik unter der Betreuung von Herrn Univ.-Prof. Dr. Stefan Dötterl (Erst-Mentor) und Herrn PD Dr. Ulrich Meve (Zweit-Mentor) angefertigt. Gefördert wurde die Arbeit von Februar bis April 2012 durch den ‛Feuerwehrfond’ zur Doktorandenförderung der Universität Bayreuth, von Mai 2012 bis April 2015 durch ein Stipendium nach dem Bayerischen Eliteförderungsgesetzt (BayEFG), und von Mai bis Juli 2015 durch ein Stipendium des Bayerischen Programms zur Förderung der Chancengleichheit für Frauen in Forschung und Lehre. Vollständiger Abdruck der von der Bayreuther Graduiertenschule für Mathematik und Naturwissenschaften (BayNAT) der Universität Bayreuth genehmigten Dissertation zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften (Dr. rer. nat.). Dissertation eingereicht am: 02.02.2017 Zulassung durch das Leitungsgremium: 10.02.2017 Wissenschaftliches Kolloquium: 31.05.2017 Amtierender Direktor: Prof. Dr. Stephan Kümmel Prüfungsausschuss: Prof. Dr. Stefan Dötterl (Erstgutachter) Prof. Dr. Konrad Dettner (Zweitgutachter) Prof. Dr. Heike Feldhaar (Vorsitz) Prof. Dr. Bettina Engelbrecht Declaration of self-contribution This dissertation is submitted as a “Cumulative Thesis“ and contains a general synopsis (Part I) and three manuscripts (Part II) about the chemical ecology and pollination biology of Ceropegia . The major part of the research presented here was accomplished by myself under supervision of Univ.-Prof. Dr. Stefan Dötterl (Universities of Bayreuth and Salzburg) and PD Dr. Ulrich Meve (University of Bayreuth). In collaboration with my supervisors, I developed the methods, collected most of the data, and drafted the manuscripts. All co-authors discussed the results and supported preparation of manuscripts. 1st manuscript: Authors: Annemarie Heiduk, Hanghui Kong, Irina Brake, Michael von Tschirnhaus, Till Tolasch, Armin G. Tröger, Elisabeth Wittenberg, Wittko Francke, Ulrich Meve, and Stefan Dötterl Title: Deceptive Ceropegia dolichophylla fools its kleptoparasitic fly pollinators with exceptional floral scent Status: Published on 03 July 2015 in Frontiers of Ecology and Evolution 3: 66. doi: 10.3389/fevo.2015.00066 Own contribution: concept and study design 60%, data acquisition 70%, analyses of scent samples 95%, data analyses 90%, preparation of figures and tables 100%, discussion of results 70%, manuscript writing 90%. The study was designed by AH, SD, and UM. Bioassays in China in 2012 were performed by HK. AH collected all other field data. Electrophysiological measurements were performed by SD. Identification of pollinators was carried out by IB and MV. Identification and/or synthesis of some of the compounds were done by WF, AT, EW, and TT. AH analyzed the data and wrote the first draft of the manuscript. WF wrote the part on the chemical synthesis. All authors contributed to interpretation of the findings, and edited and approved the manuscript. i 2nd manuscript: Authors: Annemarie Heiduk, Irina Brake, Michael von Tschirnhaus, Matthias Göhl, Andreas Jürgens, Steven D. Johnson, Ulrich Meve, and Stefan Dötterl Title: Ceropegia sandersonii mimics attacked honeybees to attract kleptoparasitic flies for pollination Status: Published on 24 October 2016 in Current Biology 26, 2787-2793. Own contribution: concept and study design 50%, data acquisition 70%, analyses of scent samples 90%, data analyses 90%, preparation of figures and tables 95%, discussion of results 70%, manuscript writing 85%. The study was conceived by AH, SD, and UM. The experiments were designed by AH, AJ, and SD. Data were collected by AH, AJ, SD, and UM. Statistical analyses were performed by AH, SDJ, and SD. Flies were identified by IB and MvT. ( E)-2- octen-1-yl acetate was synthesized by MG. Lab facilities and equipment at UKZN, South Africa was provided by SDJ. The manuscript was drafted by AH and all authors contributed valuable discussions. 3rd manuscript: Authors: Annemarie Heiduk, Irina Brake, Michael von Tschirnhaus, Jean-Paul Haenni, Raymond Miller, John Hash, Samuel Prieto-Benítez, Andreas Jürgens, Steven D. Johnson, Stefan Schulz, Sigrid Liede-Schumann, Ulrich Meve, and Stefan Dötterl Title: Floral scent and pollinators of Ceropegia trap flowers Status: Published on 01 July 2017 in Flora 232, 169-182 (invited contribution). Own contribution: concept and study design 60%, data acquisition 70%, analyses of scent samples and data 90%, preparations of figures and tables 100%, discussion of results 80%, manuscript writing 95%. The study was conceived by AH, SD, and UM. Data were collected by AH, UM, AJ . Genetic analyses were performed by SLS and PBS. Statistical analyses were performed by AH. Flies were identified by IB, MvT, JPH, RM, and JH. Floral compounds of C. stenantha were identified and synthesized by SS. Lab facilities and equipment at UKZN, South Africa was provided by SDJ. The manuscript was drafted by AH and all authors contributed valuable discussions. ii CONTENT Abstract ............................................................................................................................ 1 Zusammenfassung ............................................................................................................ 3 PART I - Synopsis ............................................................................................................... 6 General introduction ................................................................................................................ 7 Pollination in general ..................................................................................................................................... 7 Mutualistic pollination and unidirectional exploitation ................................................................................ 9 Trap flowers ................................................................................................................................................. 10 The genus Ceropegia L. ................................................................................................................................ 11 Flower morphology ................................................................................................................................. 12 Flower scent ............................................................................................................................................ 13 Pollinating flies ........................................................................................................................................ 13 Aims of my research .................................................................................................................................... 14 Materials and Methods .......................................................................................................... 16 Plant material and study sites (Publications 1, 2, 3) ................................................................................... 16 Determination of pollinators (Publications 1, 2, 3) ..................................................................................... 18 Pollen transfer efficiency (PTE) (Publications 1, 3) ..................................................................................... 18 Dynamic headspace (Publications 1, 2, 3) ................................................................................................... 18 Gas chromatography/mass spectrometry (GC/MS) (Publications 1, 2, 3) .................................................. 20 Electrophysiological analyses (GC/EAD) (Publications 1, 2) ........................................................................ 21 Behavioral studies (Publications 1, 2) ......................................................................................................... 21 Genetic relatedness of study plants (Publication 3) ................................................................................... 22 Statistical analyses ....................................................................................................................................... 22 Floral scent (Publications 1, 2, 3) ........................................................................................................... 22 Relationships between genetic relatedness, floral scent, and flower visiting/pollinating flies (Publication 3) ........................................................................................................................................ 23 Bioassays (Publication 2) ........................................................................................................................ 23 Results and Discussion ............................................................................................................ 24 Pollen transfer efficiency, flower visitors and pollinators (Publications 1, 3) ............................................. 24 Floral scent (Publication 3) .......................................................................................................................... 28 Pollinator specificity through floral scent chemistry
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