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An Examination of the DNA Content, Taxonomy and Phylogeny of Penstemon (Plantaginaceae)

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An Examination of the DNA Content, Taxonomy and Phylogeny of Penstemon (Plantaginaceae) Brigham Young University BYU ScholarsArchive Theses and Dissertations 2010-03-19 An Examination of the DNA Content, Taxonomy and Phylogeny of Penstemon (Plantaginaceae) Shaun R. Broderick Brigham Young University - Provo Follow this and additional works at: https://scholarsarchive.byu.edu/etd Part of the Animal Sciences Commons BYU ScholarsArchive Citation Broderick, Shaun R., "An Examination of the DNA Content, Taxonomy and Phylogeny of Penstemon (Plantaginaceae)" (2010). Theses and Dissertations. 2044. https://scholarsarchive.byu.edu/etd/2044 This Thesis is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected], [email protected]. An examination of the DNA content, taxonomy and phylogeny of Penstemon (Plantaginaceae) Shaun R. Broderick A thesis submitted to the faculty of Brigham Young University in partial fulfillment of the requirements for the degree of Master of Science Mikel R. Stevens Dale T. Lindgren Brad Geary Eric N. Jellen Department of Plant and Wildlife Sciences Brigham Young University April 2010 Copyright © 2010 Shaun R. Broderick All Rights Reserved ABSTRACT An examination of the DNA content, taxonomy and phylogeny of Penstemon (Plantaginaceae) Shaun R. Broderick Department of Plant and Wildlife Sciences Master of Science Penstemon is the largest genus in North America with more than 270 reported species. However, little is known about the genome size of this genus and how this information may be useful in selecting species in developing hybrids for landscape use. Using flow cytometry, we estimated the genome size of approximately 40% of the genus (117 specimens from 104 different species.) Genome sizes for the putative diploids ranged from 2C = 0.94 – 1.89 pg (1C = 462 – 924 Mbp) and the putative polyploids ranged from 2.57 – 6.54 pg (1C = 1,257 – 3,156 Mbp). Chromosome counts were compiled and compared with the flow cytometry results for the species within this publication. Ploidy within the genus ranged from diploid to dodecaploid. These data were compared and contrasted with the current taxonomy of Penstemon and previously published ITS and cpDNA phylogenetic work. Based on genome size, reassigning P. montanus, P. cardinalis, and P. uintahensis to the subgenus Penstemon and P. personatus to the subgenus Dasanthera, would better reflect the phylogeny of the genus. Both auto- and allo- polyploidization are plausible mechanisms for increasing ploidy within the genus. The diploid species within the subgenus Saccanthera contain on average 1.09 pg (1C = 532 Mbp); however, two species within this subgenus are tetraploid and octaploid. The DNA content of subgenus Penstemon exhibits high plasticity and spans a six-fold increase. Our study found flow cytometry to be useful in species identification and verification. This represents the first published work on the genome size of Penstemon. This research will aid in future DNA sequencing experiments and breeding programs. Keywords: Penstemon, flow cytometry, DNA content, polyploid, C value, phylogeny ACKNOWLEDGEMENTS I would like to thank Dr. Mikel Stevens for his continual encouragement over these several months. He was always there to provide great support, corrections on my writing, and his friendship. I would also like to thank Dr. Dale T. Lindgren for his expertise in Penstemon and for having him on my committee, Dr. Brad Geary for providing a great lab environment, and Dr. Eric N. Jellen for instructing and coaching me with cytogenetics. I would also like to thank all of those students that helped me in the greenhouse, in the field, and in sample preparation. I really could not have completed this work without their assistance. Work aside, I really appreciated their friendships. My acknowledgements would be incomplete without mentioning my wife. Her support was continual throughout every frustrating and stressful day. My parents are also in need of thanking as they are the ones that instilled the need of continual education in me. I would have stopped my education long ago without them. CONTENTS TITLE PAGE ..................................................................................................................................................................i ABSTRACT ................................................................................................................................................................. ii ACKNOWLEDGEMENTS ......................................................................................................................................... iii CHAPTER 1: A SURVEY OF PENSTEMON’S GENOME SIZE ............................................................................... 1 Introduction ....................................................................................................................................................................... 2 Materials and Methods ...................................................................................................................................................... 4 Penstemon diversity and identification ................................................................................................................... 4 Tissue sampling ...................................................................................................................................................... 4 Isolation of nuclei and genome size analysis .......................................................................................................... 5 Results and discussion ...................................................................................................................................................... 6 Ploidy within the Penstemon genus ........................................................................................................................ 7 Subgenus Cryptostemon ......................................................................................................................................... 9 Subgenus Dasanthera ............................................................................................................................................. 9 Subgenus Dissecti ................................................................................................................................................. 10 Subgenus Habroanthus ........................................................................................................................................ 10 Subgenus Penstemon ............................................................................................................................................ 11 Subgenus Saccanthera .......................................................................................................................................... 12 Hybrids ................................................................................................................................................................. 13 Flow cytometry and Penstemon classification ...................................................................................................... 13 Conclusion ...................................................................................................................................................................... 14 Acknowledgements ......................................................................................................................................................... 16 References ....................................................................................................................................................................... 17 CHAPTER 2: LITERATURE REVIEW ..................................................................................................................... 24 Introduction ..................................................................................................................................................................... 25 Penstemon taxonomy............................................................................................................................................ 26 Marker development and phylogenetic studies in Penstemon .............................................................................. 28 Ploidy in Penstemon ............................................................................................................................................. 29 iv Flow cytometry..................................................................................................................................................... 31 Development of flow cytometry ...................................................................................................................... 32 Sample preparation .......................................................................................................................................... 33 Secondary metabolites interfere with flow cytometry ..................................................................................... 33 Cell division .................................................................................................................................................... 34 Conclusion .....................................................................................................................................................................
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