A phylogeographic, ecological and genomic analysis of the recent range expansion of the wasp spider Argiope bruennichi Dissertation zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen Fakultät der Christian-Albrechts-Universität zu Kiel vorgelegt von Henrik Krehenwinkel Plön, August, 2013 Erstgutachter: Prof. Dr. Diethard Tautz Zweitgutachter: Prof. Dr. Guenther B. Hartl Tag der mündlichen Prüfung: 02.10.2013 Zum Druck genehmigt: 02.10.2013 gez. Prof. Dr. Wolfgang Duschl (Dekan) Table of Contents Summary of the thesis ......................................................................................................... 4 Zusammenfassung der Dissertation .................................................................................... 6 General introduction ........................................................................................................... 9 Species ranges, their limits and expansion in a changing world................................. 9 Adaptations and range expansions - Impediments and possibilities ......................... 10 Hybridization – Destructive or creative force? ......................................................... 11 Phylogenetics and phylogeography - Detecting genetic structure and hybridization 13 Detecting the phenotypic consequences of adaptation ............................................. 13 Genomics of adaptation - Detecting its functional signatures and architecture........ 14 The wasp spider Argiope bruennichi - A new model species for studying adaptation in the face of global change ...................................................................................... 17 Summary ................................................................................................................... 19 Chapter 1: Northern range expansion of European populations of the wasp spider Argiope bruennichi is associated with global-warming-correlated genetic admixture and population specific temperature adaptations ..................................................................... 21 1.1 Introduction ............................................................................................................. 21 1.2 Material and methods .............................................................................................. 23 Sample collection and morphological analysis ......................................................... 23 Molecular analysis .................................................................................................... 24 Phylogeographic and population genetic analysis .................................................... 26 Thermal preference tests and reciprocal transplant experiment ............................... 27 1.3 Results ..................................................................................................................... 29 Mitochondrial data .................................................................................................... 29 Microsatellite data ..................................................................................................... 34 SNP data.................................................................................................................... 35 Body size ................................................................................................................... 38 Thermal preference and overwintering ..................................................................... 39 1.4 Discussion ............................................................................................................... 41 Phylogeography of the species.................................................................................. 41 Introgression, admixture and range expansion ......................................................... 42 Adaptation and phenotypic responses ....................................................................... 44 The role of other factors on the species´ invasion success ....................................... 46 1.5 Conclusion .............................................................................................................. 48 Chapter 2: Phylogeographic surveys of two widely distributed Palearctic spider species highlight the importance of extra-European glacial refugia and reveal a pronounced incongruence between nuclear - and mitochondrial markers ........................................... 49 2.1 Introduction ............................................................................................................. 49 2.2 Material and methods .............................................................................................. 51 Sample collection ...................................................................................................... 51 Molecular analysis .................................................................................................... 52 2.3 Results ..................................................................................................................... 53 Mitochondrial data .................................................................................................... 53 Nuclear microsatellites - and SNP loci ..................................................................... 55 1 Nuclear DNA sequences ........................................................................................... 56 Genetic diversity ....................................................................................................... 57 The nursery web spider Pisaura mirabilis ................................................................ 58 2.4 Discussion ............................................................................................................... 62 Pleistocene isolation and postglacial recolonization in Argiope bruennichi ............ 62 Secondary contact, differential introgression and limits to gene flow in a highly dispersive species ...................................................................................................... 63 The phylogeography of Pisaura mirabilis - differences and similarities with Argiope bruennichi ................................................................................................................. 66 2.5 Summary ................................................................................................................. 68 Chapter 3: An assessment of genotyping success in museum specimens identifies unexpected predictors of DNA integrity and shows an historical increase in genetic diversity in an expansive spider species ........................................................................... 69 3.1 Introduction ............................................................................................................. 69 3.2 Material and methods .............................................................................................. 70 Sample preparation and tests for genotyping success ............................................... 70 Changes of genetic diversity during a range expansion............................................ 72 3.3 Results ..................................................................................................................... 73 Factors, which affect DNA integrity in wet preserved spiders ................................. 73 Changes of genetic diversity during a range expansion............................................ 77 3.4 Discussion ............................................................................................................... 77 Factors, which affect DNA integrity in wet preserved spiders ................................. 77 Changes of genetic diversity during a range expansion............................................ 79 3.5 Conclusion .............................................................................................................. 79 Chapter 4: Whole genome sequencing reveals genetic signatures of ecological differentiation during a contemporary range expansion of a spider species ..................... 80 4.1 Introduction ............................................................................................................. 80 4.2 Material and methods .............................................................................................. 82 Generation of a reference transcriptome from an ontogenetic series ....................... 82 Generation of a wasp spider draft genome ............................................................... 83 Estimation of genomic divergence during a range expansion .................................. 85 A PCR based screen for range-wide genetic differentiation in genomic candidate loci............................................................................................................................. 87 4.3 Results ..................................................................................................................... 88 Reference transcriptome and - genome ..................................................................... 88 Estimation of genomic divergence during a range expansion .................................. 90 A PCR based screen for range-wide genetic differentiation in genomic candidate loci............................................................................................................................. 94 4.4 Discussion ..............................................................................................................