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Hybridization in <I>Diabrotica Barberi</I> Smith and Lawrence University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Dissertations and Student Research in Entomology Entomology, Department of 12-2009 Hybridization in Diabrotica barberi Smith and Lawrence and Diabrotica longicornis (Say) (Coleoptera: Chrysomelidae): Biology, behavior, field introgression, and a reevaluation of taxonomic status Laura A. Campbell University of Nebraska at Lincoln Follow this and additional works at: https://digitalcommons.unl.edu/entomologydiss Part of the Entomology Commons Campbell, Laura A., "Hybridization in Diabrotica barberi Smith and Lawrence and Diabrotica longicornis (Say) (Coleoptera: Chrysomelidae): Biology, behavior, field introgression, and a reevaluation of taxonomic status" (2009). Dissertations and Student Research in Entomology. 3. https://digitalcommons.unl.edu/entomologydiss/3 This Article is brought to you for free and open access by the Entomology, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Dissertations and Student Research in Entomology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. HYBRIDIZATION IN DIABROTICA BARBERI SMITH AND LAWRENCE AND DIABROTICA LONGICORNIS (SAY) (COLEOPTERA: CHRYSOMELIDAE): BIOLOGY, BEHAVIOR, FIELD INTROGRESSION, AND A REEVALUATION OF TAXONOMIC STATUS By Laura A. Campbell A DISSERTATION Presented to the Faculty of The Graduate College at the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Doctor of Philosophy Major: Entomology Under the Supervision of Professor Lance J. Meinke Lincoln, Nebraska December, 2009 HYBRIDIZATION IN DIABROTICA BARBERI SMITH AND LAWRENCE AND DIABROTICA LONGICORNIS (SAY) (COLEOPTERA: CHRYSOMELIDAE): BIOLOGY, BEHAVIOR, FIELD INTROGRESSION, AND A REEVALUATION OF TAXONOMIC STATUS Laura A. Campbell, Ph.D. University of Nebraska, 2009 Advisor: Lance J. Meinke Diabrotica barberi Smith and Lawrence, the northern corn rootworm, and D. longicornis (Say) are currently recognized as closely related chrysomelid species. Multiple studies of morphological and genetic data have strongly supported that the two species are sister taxa, and hybridization has been proposed to occur between them. The objectives of this work were to: 1) assess life history parameters of the two taxa and their hybrids, 2) document and compare mating behavior, pheromone response, and spermatophore transfer of both taxa and their hybrids, and 3) examine genetic (amplified fragment length polymorphism, AFLP), and morphological (color and head capsule width) data of both taxa for evidence of introgression. Using lab-reared D. barberi and D. longicornis, fitness parameters and potential for population growth were assessed. D. barberi and D. longicornis were very similar for most life history traits but differed in longevity and fecundity. Hybrids of a D. longicornis mother and D. barberi father were as viable as individuals of either parental species, but hybrids of a D. barberi mother and D. longicornis father demonstrated consistently poor fitness. Studies of pheromone response, mating behavior, and spermatophore size demonstrated that F1 hybrids (D. longicornis mother, D. barberi father) are reproductively fit and also that the two parental taxa exhibit numerous small differences in reproduction. The genetic and morphological data provided evidence for field introgression between D. barberi and D. longicornis. Gene flow between the two taxa was relatively high, which prevents strong differentiation of populations. In total, this research demonstrates that hybridization most likely occurs under field conditions, and that the relative fitness of hybrids of a D. longicornis mother and D. barberi father is comparable to the parental taxa. Currently available data support a reevaluation of the current taxonomic status of the taxa. Redesignation of the taxa as subspecies may be most appropriate. Greater work on the ecology of both taxa is needed to assess the degree of ecological separation and how gene flow occurs. i Acknowledgements I would like to thank my advisor, Lance Meinke, for all of his advice and support throughout my M.S. and Ph.D. Lance has been generous with his time and is always curious about the natural world, and I have been lucky to have him as my advisor. I will truly miss his discussions of science and pest management. I would also like to thank my committee, Tiffany Heng-Moss, Svata Louda, Guillermo Ortí, and Blair Siegfried. They have all provided wonderful advice and have always challenged me to improve. I would also like to thank Erin Blankenship for many discussions on statistics and Kent Eskridge for help with multivariate statistics. I would also like to thank Claudia Rauter for serving as my Preparing Future Faculty mentor, Johanne and Mahlon Fairchild for first interesting me in entomology, and Karen Schaffer for challenging me in biology. My work would not have been possible without a great deal of help. Jim Brown has always assisted with his practical knowledge of corn rootworms. Katie Becker, Sabine Becker, Jason Brewer, Scott Garvin, Sarah Holdt, Kevin Jergen, Crystal Ramm, and Zach Smith all assisted my work at some point and contributed so much to it. Tom Clark, Pete Clark, and John Foster graciously allowed me to work with their genetic data. Eli Levine, Elson Shields, Tom Clark, Tim Nowatzki, Mark Boetel, Tom Hunt, and Bruce Eisley all helped with D. barberi collections. Tom Weissling lent a great deal of his time in advising me on Y-tube setup and Diabrotica behavior. I would also like to thank my many friends in the Entomology Department and in the School of Biological Sciences, who have provided a great deal of support through friendship and scientific ideas. These include, in no particular order: Adela Roa-Varon ii and Naikoa Aguilar-Amuchastegui, Michael Mellon, Tomomi Suwa, Stuart Willis, Jeremy Brozek, Roger and Kate Santer, Analiza Alves, Andre Crespo, Haichuan Wang, Leonardo Magalhaes, Sasi Maliphan, Eliseu Pereira, and Gabe and Melanie Langford. I would especially like to acknowledge Natalie West, who has always shared with me her great love of thistles, science, western Nebraska, life, politics, and sparkly things. Many other people have supported me throughout this process. My family is small, but has always been there for me. In addition, I am lucky to have friends who act like family, including Carol Schreiber, Allie Giffin, Josh Smith, and Julie Williams. Bob and Wanda Jones deserve special mention for always encouraging me and teaching me so much about life; I could not have done this without their support over the years. Lastly, but not at all least, I would like to thank Agustín Jiménez-Ruiz for his love, good humor, and constant encouragement for the progress of science. iii TABLE OF CONTENTS List of Figures…………………………………………………………………………....vi List of Tables…………………………………………………………………………… vii Chapter 1: Literature Review………………………………………………………… ..1 Diabrotica…………………………………………………………………………1 D. barberi and D. longicornis (Say)………………………………………………2 Habitat and Host Preferences…………….………………………………………..5 Species Origins……………………………………………………………………6 Pheromone Response and Mating Behavior in Diabrotica……………………….8 Species Concept and Its Application to D. barberi and D. longicornis…………11 Justification………………………………………………………………………13 Research Goals…………………………………………………………………...14 References Cited…………………………………………………………………15 Figures……………………………………………………………………………22 iv Chapter 2: Fitness of Diabrotica barberi Smith and Lawrence, Diabrotica longicornis (Say), and their hybrids (Coleoptera: Chrysomelidae)…………………23 Introduction……………………………………………………………………...23 Materials and Methods…………………………………………………………..27 D. barberi, D. longicornis, and hybrid colonies…………………………27 Rearing, mating, and oviposition………………………………………..28 Fitness Parameters……………………………………………………….29 Population Growth……………………………………….………………30 Color-Scoring…………………………………………………………….32 Data Analyses……………………………………………………………32 Results……………………………………………………………………………33 Life History Parameters………………………………………………….33 Population Growth……………………………………….………………37 Color Scores..…………………………………………………………….37 Discussion………………………………………………………………………..37 Life History Traits……………………………………………………….37 Color Clines……………………………………………………………...42 Physiological Effects of Mating…………….……………………………43 Future Research………………………………………………………….44 Conclusions………………………………………………………………46 References Cited…………………………………………………………………47 Tables…………………………………………………………………………….55 v Chapter 3: Pheromone response, mating behavior, and spermatophore transfer in Diabrotica barberi, Diabrotica longicornis, and their hybrids (Coleoptera: Chrysomelidae)………………………………………………………………………… 66 Introduction……………………………………………………………………...66 Materials and Methods…………………………………………………………..69 D. barberi, D. longicornis, and hybrid colonies…………………………69 Pheromone Response…………………………………………………….70 Courtship and Mating Behavior…………………………………………71 Spermatophore Transfer…………………………………………………73 Data Analyses……………………………………………………………75 Results……………………………………………………………………………76 Pheromone Response…………………………………………………….76 Courtship and Mating Behavior…………………………………………77 Spermatophore Transfer…………………………………………………82 Discussion………………………………………………………………………..84 Pheromone Response………….…………………………………………84 Courtship Behavior………………………………………………………85 Mating Behavior…………………………………………………………87 Spermatophore Transfer…………………………………………………88 Hybridization between D. barberi and D. longicornis…………………..90 Future Work……………………………………………………………...91 Conclusions………………………………………………………………92
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