The geographic scale of genetic variation in common plant species – Implications for genebanks and restoration DISSERTATION ZUR ERLANGUNG DES DOKTORGRADES DER NATURWISSENSCHAFTEN (DR. RER. NAT.) DER FAKULTÄT FÜR BIOLOGIE UND VORKLINISCHE MEDIZIN DER UNIVERSITÄT REGENSBURG vorgelegt von Daniela Maria Listl aus Regensburg im Jahr 2016 Das Promotionsgesuch wurde eingereicht am: 01.07.2016 Die Arbeit wurde angeleitet von: Prof. Dr. Christoph Reisch Unterschrift: ii Für meine zwei Deifln und meine Eltern iii iv TABLE OF CONTENTS Acknowledgements………………………………………………………………………… vii Summary…………………………………………………………………………………….. ix CHAPTER 1 GENERAL INTRODUCTION……………………...…………………………..…………………… 1 1 Biodiversity……………………………………………………………………….................. 1 > Excursus: Genebanks worldwide – a short overview…………………………………… 4 2 Genebank standards……………………………………………………………………...... 8 4 Thesis outline……………………………………………………………………….............. 11 CHAPTER 2 PHYLOGEOGRAPHY OF A TOUGH ROCK SURVIVOR IN EUROPEAN DRY GRASSLANDS……… 15 Abstract…………………………………………………………………………....................... 15 Materials and Methods……………………………………………………………………….. 18 Results ………………………………………………………………………………………… 25 Discussion………………………………………………………………………...................... 33 CHAPTER 3 THE GEOGRAPHIC SCALE OF GENETIC VARIATION IN A COMMON SPECIES – A CASE STUDY ON LATHYRUS PRATENSIS L. ………………………………………...…..... 38 Abstract……………………………………………………………………….......................... 38 Introduction ………………………………………………………………………................... 39 Material and Methods ………………………………………………………………………. 41 Results………………………………………………………………………............................ 50 Discussion ………………………………………………………………………..................... 56 CHAPTER 4 GENETIC DIVERSITY IN POPULATIONS OF HEPATICA NOBILIS SCHREB. IN BAVARIA AGAINST THE BACKGROUND OF SEED TRANSFER GUIDELINES IN FORESTRY AND RESTORATION....... 60 Abstract ……………………………………………………………........................................ 60 Introduction……………………………………………………………................................... 61 Material and Methods ………………………………………………….……………......... 65 Results………………………………………………………………...................................... 70 Discussion …..…………………………………………………………….............................. 74 v CHAPTER 5 GENETIC ADAPTATION AND ENVIRONMENTAL INFLUENCES ON SEED QUALITY AND LONGEVITY……………………………………………………………………………………… 80 Abstract ……………………………………………………………...................................... 80 Introduction ………………………………………………………………............................ 82 Material and Methods ……………………………………………………………………..... 84 Results ...……………………………………………………………….................................. 93 Discussion………………………………………………………………………...................... 99 CHAPTER 6 GENERAL DISCUSSION AND CONCLUSION………………………………………................... 103 History of plant populations ………………………………………………………………… 103 Plant traits……………………………………………………………..................................... 104 Intraspecific differences in seeds…………………………………………………………... 105 Strengths and limitations ………………………………………………………………........ 105 Conclusion and Perspectives …………………………………........................................... 107 References……………………………………………………………………...…………….. 109 Appendix………………………………………………………………………………………. 130 vi ACKNOWLEDGEMENTS General Acknowledgements First, I would like to thank Christoph Reisch and Peter Poschlod. Christoph was there, even before start, over up and downs, to the end of this thesis – he was always supportive and with some new ideas and background information – and almost always positive ;-) I thank you very much! Peter Poschlod was trustful and supportive all the time – even at the beginning, when he did not really know me. I appreciate this very much! His knowledge is impressive and all his suggestions for the thesis were very helpful! Petra Schitko was always contact person number one in the lab: next to her knowledge and experience, she has this kind of steadiness and confidence – and she brightens up lab work even in moments when nothing is working. For very pleasant co-working in the lab – and for tolerating my small nervous breakdowns I also want thank Eva very much. I want to thank Peter Hummel, Manuela Bog, Salvatore Tomasello, Roland Greiner, Sven Himmelreich and the whole working group Oberprieler for help, when there was help necessary! Special thanks also go to the partners of the Genbank WEL and WIPs in Osnabrück, Karlsruhe (KIT und PH), Berlin, esp. Peter Borgmann, Annemarie Radkowitsch, Joachim Daumann, Elke Zippel. I am deeply grateful to Judith Lang, Philipp Glaab and Conny Straubinger - they were doing a very good job at the genebanks in Regensburg. It is a pleasure to work with you! A big thanks also goes to Sabine for a lot of help and instructions to ArcGIS! Thanks to the whole Regensburg group for their pleasant mix of spirits that was inspiring and very enjoyable: esp. Annika Sezi, Annisa Satyanti, Bille Bauer, Christina Meindl, Christina Putz, Christina Gleixner, Christine Gutmann, Christoph Schmid, Ellen Pagel, Eva Bauer, Eva-Rosa Wagner, Günther Kolb, Hannah Gutheil, Heike Pfaffenzeller, Irina Weinberger, Juliane Drobnik, Katrin Binder, Maik Bartelhaimer, Maxi Bleicher, Mehdi Abedi, Sabine Fischer, Sabrina Senger, Sergey Rosbakh, Shuang Li, Steffi Meier, Tobi Maul, Tobi Windmaisser, Verena Busch – and many, many more! And a special thanks to my wonderful roommates Franzi Kaulfuß and Josef Simmel! I want to thank my friends: Susi, Gudrun, Babsi, Bine, Susanne, Carola, Christiane, Birgit, Andi H., Andi N., Axel, Falk, Francis, Geimi, Karsten, Lugge, Lukas, Markus, Michi, Olla, Roli and all the “Sinzinger” and “Prüfeninger” that made it possible to think about something else – and they even gave me company at collection trips and gave me bed & breakfast all over Bavaria and Europe. vii My family has supported me during the long time of preparing this thesis. My parents Helga and Paul have always been very supportive - financially and morally, “Oma Maria” und “Oma Helga” have been there for us each day for three years now. Of course, I am very grateful to my partner Basti: he accompanied collections trips for all of the chapters during his holidays – without him I would not have been able to start this neither to bring this to an end! Finally, thanks to Theo - you have been very patient with your confused mother! Chapter 2 A lot of people collected plant material for this chapter: Andreas Hilpold, Andreas Renner, Annemarie Radkowitsch, Anton Meier, Camilla Wellstein, Carla Pinto Cruz, Christiane Sonnleitner, John Dickie, Helga Listl, Maria Hanauer, Melanie Harze, Nicola Schoenenberger, Paul Listl, Petr Karlik, Roland Kittel, Salvatore Tomasello, Sven Himmelreich, Zsolt Molnár, - thanks a lot to everyone! Special thanks go to Simone Tausch and Martin Leipold for collections in Serbia and Spain, and the organization of several other collections! For helpful tips on the manuscript we thank two anonymous reviewers. Chapter 3 Thanks to Sebastian Teufel for the help at several collection trips all over Bavaria. For help with statistics I want to thank Christina Meindl. Prof. Christoph Oberprieler gave me very helpful hints on ploidy levels of Lathyrus pratensis - thank you a lot! Further, Kathrin Kiehl and three anonymous reviewers made helpful comments on the manuscript. Chapter 4 Sincere thanks goes to Katrin Binder for collection of plants and lab collaboration that was very pleasant. For plant collection I also want to thank Sebastian Teufel; and of course a big “Thanks” to Shuang Li for her very, very ambitioned work in the lab! Chapter 5 I thank Sergey Rosbakh for collaboration of the projects and for taking me with him to Berchtesgaden several times. Thanks to Simone Tausch and Martin Leipold for the collaboration in obtaining information and material (Martins brother for help with the boxes!). Josef Simmel, Juliane Drobnik and Philipp Glaab helped with the collection of seeds – thank you a lot. I further want to say “Thank you” to Volker Debus and the gardeners for fostering the plants - and to famous Inge Lauer for her help in the lab and a lot of washing! Last but not least, I want to thank Christoph Schmid for his generous help with statistical analysis in R! viii SUMMARY Genebanks for plant seeds are a necessary tool to support the conservation of biodiversity. In the last decades also the importance of intraspecific diversity within the genebank collections was emphasised. Even though background information on the distribution of genetic diversity within and between populations was scarce for common plant species, seed provenances have been delineated for seed collection and production. Seeds have to be used in the same provenance, where they were collected for restoration projects, agriculture and forestry. The present study aims to elucidate distribution of intraspecific genetic diversity in three common plant species over different geographic scales. In the last part, further intraspecific differences in seed traits were investigated. Chapter 2 dealt with the distribution of genetic diversity within and between populations of Sedum album in its distribution range in Europe and the history of Central European populations. With a combination of neutral and chloroplast specific markers, we reconstructed postglacial processes that lead to the present distribution pattern in S. album. Distinct haplotypes could be detected on the Iberian Peninsula and in Eastern and Southern Europe. Central European populations have their origin in Southern Europe and show meanwhile high influence of Eastern lineages. A clear East/West subdivision