Diplomarbeit vorgelegt zur Erlangung des Grades eines Diplom-Biologen an der Fakultät für Biologie der Ruhr-Universität Bochum Molecular genetic, chemotaxonomic, and autecological investigations of European Sericostomatidae (Insecta: Trichoptera) Molekulargenetische, chemotaxonomische und autökologische Unter- suchungen europäischer Sericostomatidae (Insecta: Trichoptera) von Florian Leese aus Lauterbach (Hess.) angefertigt an der Fluss-Station des Max-Planck-Instituts für Limnologie Schlitz, sowie der Arbeitsgruppe Spezielle Zoologie Bochum Betreuer: Prof. Dr. R. Wagner Referent: Prof. Dr. J.-W. Wägele Korreferent: Prof. Dr. H. Haeupler F. Leese (2004) – Table of contents 1 TABLE OF CONTENTS 1. Preface................................................................................................................................... 3 2. Zusammenfassung................................................................................................................ 4 3. Summary............................................................................................................................... 6 4. Introduction .......................................................................................................................... 8 4.1 An overview of the order ‘Trichoptera’ ................................................................... 8 4.2 The family Sericostomatidae Stephens, 1836 ........................................................ 10 4.3 The Sericostoma species complex.......................................................................... 11 4.4 Aims of this study................................................................................................... 13 5. Autecological investigations .............................................................................................. 14 5.1 Introduction ................................................................................................................ 14 5.1.1 Ecological research among the Sericostoma species.............................. 14 5.1.2 Aims of autecological investigations ....................................................... 19 5.2 Material and methods................................................................................................. 20 5.2.1 Collection sites of specimens................................................................... 20 5.2.2 Recirculating laboratory streams............................................................ 21 5.2.3 Rearing of larvae and pupae ................................................................... 21 5.2.4 Mating experiments ................................................................................. 22 5.2.5 Thermostat-controlled water-bath system ............................................... 22 5.2.6 Egg mass swelling and embryonic development ..................................... 24 5.3 Results ........................................................................................................................ 26 5.3.1 Emergence ............................................................................................... 26 5.3.2 Diurnal activity........................................................................................ 27 5.3.3 Aspects of reproduction ........................................................................... 28 5.4. Discussion ................................................................................................................. 32 5.4.1 Emergence ............................................................................................... 32 5.4.2 Diurnal activity........................................................................................ 32 5.4.3 Aspects of reproduction ........................................................................... 33 5.4.4 Conclusions.............................................................................................. 35 6. Chemotaxonomic investigations........................................................................................ 36 6.1 Introduction ................................................................................................................ 36 6.1.1 Pheromones in Trichoptera ..................................................................... 36 6.1.2 Aims of chemotaxonomic investigations.................................................. 37 6.2 Material and methods................................................................................................. 39 F. Leese (2004) – Table of contents 2 6.2.1 Material ................................................................................................... 39 6.2.2 Methods.................................................................................................... 39 6.3 Results ........................................................................................................................ 42 6.3.1 Morphology.............................................................................................. 42 6.3.2 Qualitative and quantitative composition phermononal glands.............. 42 6.4 Discussion .................................................................................................................. 47 7. Molecular Genetic Investigations ..................................................................................... 48 7.1 Introduction ................................................................................................................ 48 7.1.1 The Sericostoma species complex............................................................ 49 7.1.2. Phylogenetic relationship between European sericostomatid genera.... 50 7.2 Material and Methods................................................................................................. 51 7.2.1 Material and collection sites .................................................................. 51 7.2.2 Laboratory methods................................................................................. 53 7.2.3 Analysis methods ..................................................................................... 64 7.2.4 Performing analyses ................................................................................ 75 7.3 Results ........................................................................................................................ 79 7.3.1 Descriptive sequence characteristics ...................................................... 79 7.3.2 Phylogenetic analyses.............................................................................. 84 7.4 Discussion ................................................................................................................ 102 7.4.1 Methodological aspects ......................................................................... 102 7.4.2 Phylogenetic analyses............................................................................ 105 7.4.3 Conclusions............................................................................................ 107 8. Final conclusions .............................................................................................................. 111 9. Danksagung....................................................................................................................... 114 10. Literature ........................................................................................................................ 115 11. Appendix ......................................................................................................................... 128 11.1 Morphological variation ..................................................................................... 128 11.2 16S rDNA sequence alignment .......................................................................... 129 11.3 COI sequence alignment..................................................................................... 135 F. Leese (2004) – Preface 3 1. PREFACE Several methods employed in this work are rather complex. Therefore in the ‘material and methods’ chapters basic principles of these methods are explained for better understanding of the results. Maps were created using the program PanMap (Diepenbroek et al. 2000). Illustra- tions (graphs, map design) were prepared with Photoshop 7.0 and Corel Draw 9.0. If not cre- ated by myself, the source of the graphics is mentioned in the text. Sequence data can be found in the appendix. F. Leese (2004) – Zusammenfassung 4 2. ZUSAMMENFASSUNG Über den taxonomischen Status der zahlreichen beschriebenen westpaläarktischen Arten von Sericostoma Latereille, 1825 (Trichoptera: Sericostomatidae), sowie deren Verbreitung wird seit Mitte des 19. Jahrhunderts kontrovers diskutiert. Artunterscheidungen basieren zumeist ausschließlich auf der Morphologie der ♂ Genitalien, mit häufig nur geringen Unterschieden. Zur Überprüfung der zurzeit als gültig angesehenen Arten, speziell der beiden mitteleuropäi- schen Morphospezies S. personatum und S. schneideri, wurden autökologische, chemotaxo- nomische, sowie molekulargenetische Untersuchungsmethoden gewählt. Autökologische Untersuchungen konzentrierten sich auf Unterschiede hinsichtlich der Schlupfdynamik, diurnalen Aktivität, sowie Quellung und temperaturabhängiger Entwicklung der Ei-Gelege. Der chemotaxonomische Ansatz beschränkte sich auf die Identifizierung von Pheromonen der beiden mitteleuropäischen Sericostoma ‚Arten’. Für die genetischen Unter- suchungen wurden zwei mitochondriale Gene (16S rDNA, Cytochromoxidase