University of Alberta Morphology, function and evolution of the sternum V glands in Amphiesmenoptera by Marie Djernæs A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Systematics and Evolution Department of Biological Sciences ©Marie Djernæs Fall 2010 Edmonton, Alberta Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission. Examining Committee Felix A. H. Sperling, Department of Biological Sciences Bruce S. Heming, Department of Biological Sciences Maya L. Evenden, Department of Biological Sciences John Spence, Department of Renewable Resources Mark V. H. Wilson, Department of Biological Sciences Ralph W. Holzenthal, University of Minnesota Abstract I investigated the paired sternum V glands in thirty-eight trichopteran families and all lepidopteran families possessing the gland or associated structures. Using my morphological data and literature data on sternum V gland secretions, I examined phylogenetic trends in morphology and gland products and reconstructed ancestral states. I investigated correlations between gland products, between morphological traits and between chemistry and morphology. The gland is present in twenty-five trichopteran families. It is generally present in Annulipalpia, except Dipseudopsidae, and in Spicipalpia. It is widespread in Plenitentoria, but is often absent in Brevitentoria, especially in males. In Lepidoptera, I present the first report on the reduced, but functional glands in Neopseustidae and Nepticulidae. The gland is typically an invagination from sternum V with a duct leading to a reservoir surrounded by secretory tissue. An opening muscle inserts just inside the opening. I found two non-homologous opening-muscle types, one in Lepidoptera and some Trichoptera, another in the remaining Trichoptera. Muscle fibres often surround the reservoir, sometimes also the secretory tissue. Exceptions are found in Psychomyiidae (no opening muscle), female Philopotamidae (fenestra with separate glandular complex), Agathiphagidae (several unique features), Neopseustidae and Nepticulidae (gland present without gland opening). Using variations in gland structure, I identified phylogenetically useful characters from the superorder to the species level. The fenestrae in female Philopotamidae, Eriocraniidae, Neopseustidae and Nepticulidae are perforated, and perforated patches are present in female Psychomyiidae. The perforated patches are associated with a reservoir, secretory tissue and a distinctive ‘sunburst’ musculature in both Trichoptera and Lepidoptera. The probable ancestral gland compounds are heptan-2-ol, 4-hepten-2-one, 4-hepten-2-ol, nonan-2-one, 6- nonen-2-one and 6-nonen-2-ol, making pheromone production a plausible ancestral function. The most widespread gland compounds are heptan-2-one, heptan-2-ol, nonan-2-one and nonan-2-ol, but these are absent from Apataniidae + Limnephilidae, which instead produce methylated 3-ketones and -ols, unique within Trichoptera. These compounds all probably function as pheromones. Both large and small glands in females can function in sex pheromone production, while large glands in male Hydropsyche (Hydropsychidae) are likely linked to male aggregation pheromone production. Relative sizes of regular gland reservoirs and fenestral gland reservoirs in female philopotamids suggest a complementary function. Acknowledgements I am grateful to N. P. Kristensen for suggesting this interesting project, and to my supervisor F. A. H. Sperling for encouraging me to expand the project and study not only Lepidoptera, but also Trichoptera. I appreciate having had numerous discussions on the intricacies of the sternum V gland with both. I am also enormously grateful to my husband T. J. Simonsen who has provided companionship, practical assistance above and beyond and been a willing discussion partner. This entire project would not have been possible without the loan of specimens from several collections and the collection efforts of a number of individuals. Thus I am grateful to Clemson University (C. J. Geraci and I. Stocks), Los Angeles County Museum (J. P. Donahue and W. Xie), Natural History Museum of Denmark (O. Karsholt and N. P. Kristensen), Smithsonian Institution (C. J. Geraci and O. Flint) and University of Minnesota Insect Collection (R. Holzenthal and D. Robertson) for the loan of specimens. Futhermore, I thank R. Brown, J. Dombroskie, G. Gibbs, O. Karsholt, J.-F. Landry, E. van Nieukerken, G. Pohl, G. S. Robinson and F. A. H. Sperling for collecting or providing specimens. I thank Gatineau Park, Québec, for a permit to collect and conduct experiments on Epimartyria auricrinella . I am grateful to N. P. Kristensen for supplying prepared slides of various Lepidoptera as well as TEM micrographs of Eriocrania semipurpurella . In addition to specimens, a PhD project cannot be completed without the help of a large number of people. I thank C. Löfstedt for supplying unpublished data on gland compounds and T. Garland for advice on phylogenetically independent contrasts. I am grateful to I. Lusebrink who went above and beyond to get me a copy of Ansteeg’s thesis. I gratefully acknowledge B. S. Heming and D. A. Craig for help with microscopy. I thank A. Nimmo for help in identifying specimens, G. Braybrook for assistance with SEM and M. Eberhard for preparing the 1 µm sections of Synempora andesae and Agathiphaga vitiensis . I thank the members of the extended Sperling lab for being good company, and making me feel at home in Edmonton. I thank the members of my PhD committee, B. Heming, M. Evenden and J. Spence, for being willing to serve, and for many useful discussions and pieces of advice over the years. Finally, a project like this requires funding. This study was funded by an NSERC grant to Felix A. H. Sperling, an Alberta Ingenuity Scholarship to M. Djernæs, and operation money from the Danish Council for Independent Research/Natural Sciences (grant no 272-08-0480). Further support for travel to conferences came from FGSR (Marie Louise Imrie award), from the Department of Biological Sciences and from the Graduate Students’ Association. Biography I was born on December 12, 1978, and grew up on a dairy farm in the western part of Denmark. I was an extremely curious child. At around age two I started asking ‘why’ about the world around me: “Why are ducks brown?”, “why are only female ducks brown?” I appreciate the patience of my parents in answering my questions, but also think they were relieved when I learned to read well enough that I could look up the answers myself. But my curiosity often outstripped my reading ability or the literature in the school library. Growing up on a dairy farm, I knew cows, and I had a passion for horses. I could see that cows and horses were in many respects similar, but did wonder how cows got cloven hooves and horses did not. Looking at cow and horse feet, I concluded that if the two halves of a cow foot were to fuse, the result would look very like a horse’s hoof. Soon after that I found a book explaining the evolution of feet in hoofed mammals, and I had to reject my theory. Eventually looking up the answers wasn’t really enough anymore, and I started looking at a career in research. I began studying biology at the University of Copenhagen in 1999, knowing that I wanted to study evolution, but thinking that I would be working with vertebrates. My first introduction to the wonderful world of entomology came in the third year at university when we had to do undergraduate research projects. We were offered several standard and mainly literature based projects, but Klaus- Dieter Klass and Rudolf Meier from the Natural History Museum of Denmark had two small research projects suitable for students, one on flies and one on cockroaches. With a fellow student I went for the cockroach project, not attracted by the cockroaches, but by the promise of being allowed to do real research. This was also the time when I met my future husband Thomas J. Simonsen, truly a time for starting long term relationships. The cockroach project resulted in a talk that I gave at the 22 nd International Congress on Entomology in Brisbane, Australia, my first conference talk ever! I spent a year (2002/2003) as an exchange student at University of California, Riverside where I took my first entomology courses and continued my studies on cockroaches in Daphne Fairbairn’s lab. Back at the University of Copenhagen I took a course in molecular evolution. After the course we students were offered small relevant research projects. This was when I was confirmed as an entomologist: Among the different projects offered were tracing paternity in elephants or trying to place the newly described insect order Mantophasmatodea. I went for Mantophasmatodea, supervised by Jakob Damgaard, and never really considered working on vertebrates after that. The Mantophasmatodea project ended up being about intron positions in EF-1α in the entire Hexapoda, but I still learned valuable new skills about sequencing, data mining, and aligning and analysing gene sequences. In early 2004, I started my Master’s thesis, a molecular phylogeny of cockroaches, at the Natural History Museum of Denmark. Originally I was supervised by Nils Møller Andersen and Jakob Damgaard. Upon the untimely death of Nils Andersen, Niels Peder Kristensen and Jakob Damgaard supervised my thesis work. In the fall of 2004, I married Thomas Simonsen.