Ecological and Evolutionary Relationships between Bees and their Bacterial Gut Microbiota Item Type text; Electronic Dissertation Authors Martinson, Vincent G. Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 23/09/2021 12:57:33 Link to Item http://hdl.handle.net/10150/223362 ECOLOGICAL AND EVOLUTIONARY RELATIONSHIPS BETWEEN BEES AND THEIR BACTERIAL GUT MICROBIOTA by Vincent G. Martinson _____________________ A Dissertation Submitted to the Faculty of the GRADUATE INTERDISCIPLINARY PROGRAM IN INSECT SCIENCE In Partial Fulfillment of the Requirements For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College THE UNIVERSITY OF ARIZONA 2012 2 THE UNIVERSITY OF ARIZONA GRADUATE COLLEGE As members of the Dissertation Committee, we certify that we have read the dissertation prepared by Vincent G. Martinson entitled ECOLOGICAL AND EVOLUTIONARY RELATIONSHIPS BETWEEN BEES AND THEIR BACTERIAL GUT MICROBIOTA and recommend that it be accepted as fulfilling the dissertation requirement for the Degree of Doctor of Philosophy _______________________________________________________________________ Date: 03/08/12 Nancy A. Moran _______________________________________________________________________ Date: 03/08/12 Howard Ochman _______________________________________________________________________ Date: 03/08/12 Michael J. Sanderson _______________________________________________________________________ Date: 03/08/12 Diana E. Wheeler _______________________________________________________________________ Date: 03/08/12 Noah K. Whiteman Final approval and acceptance of this dissertation is contingent upon the candidate’s submission of the final copies of the dissertation to the Graduate College. I hereby certify that I have read this dissertation prepared under my direction and recommend that it be accepted as fulfilling the dissertation requirement. ________________________________________________ Date: 03/08/12 Dissertation Director: Nancy A. Moran 3 STATEMENT BY AUTHOR This dissertation has been submitted in partial fulfillment of requirements for an advanced degree at the University of Arizona and is deposited in the University Library to be made available to borrowers under rules of the Library. Brief quotations from this dissertation are allowable without special permission, provided that accurate acknowledgment of source is made. Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the head of the major department or the Dean of the Graduate College when in his or her judgment the proposed use of the material is in the interests of scholarship. In all other instances, however, permission must be obtained from the author. SIGNED: _______________________________ Vincent G. Martinson 4 ACKNOWLEDGEMENTS Nancy Moran is an excellent scientist and advisor that has an unparalleled enthusiasm for science. I thank Nancy for giving me the opportunity to help develop a new research program within her lab. Not only did she provide her expert advise on the direction of my research and presenting my work, but generously provided resources that allowed this work to happen. Thank you to Mike Sanderson, Diana Wheeler, and Noah Whiteman for contributing their time to be on my dissertation committee, and providing helpful feedback. I have learned much from Howard Ochman through classes, lab meetings, and as a committee member, and thank him for his honest discussions. I thank Molly Hunter for filling in at my oral examination and for her interest in my research. The Moran and Ochman lab members (Nicole Gerardo, Mark van Passel, John Stavrinides, Pradeep Reddi Marri, Hema Narra, Renyi Liu, Chi-Horng Kuo, Baoyu Tian, Kerry Oliver, Eva Nováková, John McCutcheon, Allison Hansen, Rahul Raghavan, Zakee Sabree, Philipp Engel, Walden Kwong, Patrick Degnan, Kevin Vogel, and Gaelen Burke) have been greatly influential in my scientific development. Thank you to the many lab technicians that have given me support over the years (Helen Dunbar, Heather McLaughlin, Tyler Jarvik, Eli Powell, and Kim Hammond). Especially Kim and Eli for wrangling irritable bee colonies and keeping the rest of the colonies happy. Thank you to the members of the Carl Hayden Bee Research Center (Gloria DeGrandi-Hoffman, Thomas Deeby, Mark Carroll, Bruce Eckholm) for access to hives and training me how to tend bees, without which I could not have performed my work. James Nieh and Jay Evans contributed critical specimens for my research. My co-authors Bryan Danforth, Robert Minckley, Olav Rueppell, Salim Tingek, Tanja Magoc, Steven Salzberg gave valuable feedback on manuscripts that became part of this dissertation. Undergraduate Jamie Moy helped to collect data. Thank you to Becky Nankivell, Sharon Richards, Teresa Kudrna, Ryan Nystrom, Ming Beckwith, Pennie Liebig, and the EEB office staff for their administrative support (and encouragement). Thank you to Andrea Grantham, Tony Day, Carl Boswell for teaching me microscopy techniques that aided in my research. Thank you to the NSF IGERT Program in Comparative Genomics and the director Michael Nachman, which not only funded much of my graduate research, but also taught me invaluable techniques, critical thinking, and about surviving in academia. Thank you to the Center for Insect Science and the NSF (through funding awards to Nancy Moran) for funding my research and travel to scientific conferences. Thank you to the Department of Ecology and Evolutionary Biology for the platform to share my data, feedback from faculty, and free food. Thank you to all my family and friends (you know who you are) for being there through thick and thin. 5 DEDICATION To Ellen for your love, and to my family, Mom (Joanne) and Dad (Wynn) and my brothers Philip and Jonathan for all their support, encouragement, and positive thinking. 6 TABLE OF CONTENTS ABSTRACT.........................................................................................................................7 INTRODUCTION...............................................................................................................8 PRESENT STUDY............................................................................................................19 REFERENCES..................................................................................................................23 APPENDIX A: A SIMPLE AND DISTINCTIVE MICROBIOTA ASSOCIATED WITH HONEY BEES AND BUMBLE BEES.............................................................................29 APPENDIX B: ESTABLISHMENT OF CHARACTERISTIC GUT BACTERIA DURING DEVELOPMENT OF THE HONEY BEE WORKER.....................................73 APPENDIX C: HAPPENSTANCE SEQUENCING OF THE NEAR-COMPLETE GENOME OF A BUMBLE BEE GUT SYMBIONT.....................................................123 7 ABSTRACT Gut microbial communities exist in the vast majority of animals, and often form complex symbioses with their hosts that affect their host’s biology in numerous ways. To date, the majority of studies of these complex interactions have focused on the nutritional benefits provided by the microbiota; however, the natural microbiota can also influence development, immunity, and the metabolism of its host. Apis mellifera, the honey bee, harbors a distinctive bacterial community that is present in individuals from distant locations around the world; however, the basis of the bee-microbiota association is unknown. This dissertation explores properties of the bacterial microbiota within bees, including its persistence of this association, mechanisms of transmission, localization through host ontogeny, and basic metabolic capabilities that define and maintain the symbiotic relationship. Apis and Bombus species (honey and bumble bees) share a distinct bacterial microbiota that is not present in other bees and wasps. Close analysis of the A. mellifera microbiota revealed consistent communities in adult worker gut organs and a general lack of bacteria in larvae. Contact between workers and with hive materials were identified as major routes of transmission for bacterial communities, showing the importance of social behavior in this association. Genomic analysis of a gut bacterium co-sequenced with the Bombus impatiens genome revealed it as a divergent lineage of Gammaproteobacteria, and deletions of conserved metabolic pathways, reduction in genome size, and its low GC content all suggest that the bacterial species has had a long association with its host. 8 INTRODUCTION Explanation of the problem and its context The evolution and physiology of animals are greatly influenced by the microbial communities residing within them (Lee & Mazmanian 2010; Ley et al. 2008; Ley et al. 2006; Wostmann 1996). Many host-microbe interactions occur within the gut, which is not only the chief area of food digestion and nutrient absorption, but also a main location for immune system activity (Chapman 1998; Hooper et al. 2002; Lee & Mazmanian 2010; Tellam 1996; Vallet-Gely et al. 2008). Gut microbial communities exist in the vast majority of animals and have formed complex symbioses with their hosts, affecting their host’s biology in abundant ways (Dehority 1997; Dillon & Dillon 2004; Mackie 1997). Advances in