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Crocanthemum Canadense (L.) Britt MIXED MATING AND ITS ECOLOGICAL IMPACT ON AN ENDANGERED PLANT: CROCANTHEMUM CANADENSE (L.) BRITT. by Emily Evans Thesis submitted in partial fulfillment of the requirements for the Degree of Bachelor of Science with Honours in Biology Acadia University April, 2017 © Copyright by Emily Evans, 2017 This thesis by Emily Evans is accepted in its present form by the Department of Biology as satisfying the thesis requirements for the degree of Bachelor of Science with Honours Approved by the Thesis Supervisors __________________________ ____________________ Dr. Rodger Evans Date __________________________ ____________________ Dr. Kirk Hillier Date Approved by the Head of the Department __________________________ ____________________ Dr. Brian Wilson Date Approved by the Honours Committee __________________________ ____________________ Dr. Jun Yang Date ii I, Emily Evans, grant permission to the University Librarian at Acadia University to reproduce, loan or distribute copies of my thesis in microform, paper or electronic formats on a non-profit basis. I, however, retain the copyright in my thesis. _________________________________ Signature of Author _________________________________ Date iii Acknowledgements I would like to thank my supervisors, Dr. Rodger Evans and Dr. Kirk Hillier, for providing me with support and guidance throughout my research experience. Through their passion for their work, both were huge motivators throughout the summer and academic year. Rodger’s positivity and sense of humour always brightened the lab space, while Kirk’s curiosity greatly inspired me to discuss and pursue questions of my own. A big thanks to Victoria Brown for lending a helping hand during the many hours we spent in the field, baking in the hot sun. Also, thank you Erica Gillis for your support and for introducing your awesome music to our lab space. In the K.C. Irving Environmental Science Centre, Phyllis Essex-Fraser, Dr. David Kristie, and Dr. Robin Browne provided critical guidance and feedback on my project, especially regarding germination procedures and statistical analyses. Phyllis, your dedication to helping students and hilarious sense of humour was a highlight of my honours experience! Finally, I would like to thank my fellow honours students and the faculty in Acadia’s Biology Department for providing such a supportive and welcoming environment, especially during the summer months. This project was made possible by the cooperation of the Canadian Forces Base 14 Wing Greenwood and on-site support by Alan Ng and Alyssa Walker, who helped locate plant populations. Funding from both the Arthur Irving Academy Foundation and NSERC through an Undergraduate Student Research Award was critical for the progression of the project. iv Table of Contents Acknowledgements ........................................................................................................... iv List of Tables ................................................................................................................... vii List of Appendices .............................................................................................................. x Abstract .............................................................................................................................. xi 1. Introduction .................................................................................................................... 1 1.1 Studying Endangered Plants ................................................................................. 1 1.2 Crocanthemum canadense in Nova Scotia ........................................................... 1 1.3 Reproductive Biology ........................................................................................... 3 1.4 Florivory by Mompha capella .............................................................................. 7 1.5 Objective ............................................................................................................... 8 2. Materials and Methods ................................................................................................ 11 2.1 Sample Collections of C. canadense .................................................................. 11 2.2 Differentiating Between Flower Types ............................................................... 11 2.3 Quantifying Floral Characteristics ...................................................................... 12 2.3.1 Dissection ....................................................................................................... 12 2.3.2 Pollen Counts and Viability Testing .............................................................. 13 2.4 Floral Morphology .............................................................................................. 14 2.4.1 Scanning Electron Microscopy ...................................................................... 14 2.4.2 Spurr’s Resin Embedding .............................................................................. 15 2.5 Pollinator Exclusion Trial ................................................................................... 16 2.6 Seed Studies ........................................................................................................ 17 2.6.1 Germination Trial ........................................................................................... 17 2.6.2 Viability Testing ............................................................................................. 18 v 2.7 Infestation Rate by Mompha capella .................................................................. 19 3. Results ........................................................................................................................... 21 3.1 Quantifying Floral Characteristics ...................................................................... 21 3.2 Floral Morphology .............................................................................................. 24 3.3 Pollinator Exclusion Trial ................................................................................... 29 3.4 Seed Studies ........................................................................................................ 30 3.5 Infestation Rates by Mompha capella ................................................................. 31 4. Discussion ..................................................................................................................... 35 4.1 Seasonal Transition in Floral Morphology and Mating Strategy ........................ 35 4.2 Floral Contributions to Genetic Diversity and Fitness ....................................... 37 4.3 Implications of Mompha capella on Future Populations .................................... 38 5. References ..................................................................................................................... 41 Appendix A ....................................................................................................................... 45 Appendix B ....................................................................................................................... 49 vi List of Tables Table 1. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in the mean number of stamens between C. canadense flower types .................. 45 Table 2. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in the mean number of pollen grains per anther between C. canadense flower types ........................................................................................................................................ 45 Table 3. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in the estimated mean number of pollen grains per flower between C. canadense flower types. ............................................................................................................................ 46 Table 4. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in the mean number of ovules between C. canadense flower types .................... 46 Table 5. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in the mean number of seeds between C. canadense flower types ...................... 47 Table 6. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in the estimated mean inviable-to-total pollen grains per anther of seasonal flower types of C. canadense ............................................................................................................. 47 Table 7. Data under comparison in a post hoc Tukey’s multiple comparisons test to assess differences in seed production between flower types of C. canadense treated under open (un- bagged) and closed (bagged) pollinator conditions ................................................................ 48 Table 8. Data under comparison in a post hock Tukey’s multiple comparisons test to assess differences in seed germination percentage between flower types of C. canadense after a two-week incubation period .................................................................................................... 48 Table 9. Summary of estimated contribution per plant to the genetic diversity of subsequent generations based on mean number of flowers per plant, seeds produced per flower, and germination % of seeds for each flower type .........................................................................
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