1. TRACING THE EVOLUTIONARILY DISTINCTIVENESS OF PEROMYSCUS MANICULATUS IN WYOMING Valorie Lyman, Dr. Hayley Lanier University of Wyoming at Casper

2. Acknowledgements I would like to thank: Yellowstone folks: Zachary Roehrs Meredith Roehrs Scott Seville Ashkia Campbell Andy Kulikowski Kevin Crouch-Jacobs Samantha Haller Dominique Schlumpf Kayla Wilson Jennifer Forrester for collecting Yellowstone samples UW-Casper Conservation Biology Course for skinning Casper samples and providing tissue samples National Park Service (NPS) Hayley Lanier for all her guidance, expertise, and advice in doing this project Wyoming INBRE program Nathan Stack for providing assistance in the lab

3. Acknowledgements I would like to thank: Freya and Sparky, Dr. Lanier’s cats for collecting the samples on Casper Mountain

4. Distribution  Peromyscus maniculatus is one of the most widely distributed mammals in North America  Found in many habitats and across the widest elevation range of any mammal

5. Disease Transmission Across their distribution Peromyscus maniculatus is the host of at least two strains of Hantavirus, the Sin Nombre Virus in Western range and Monongahela Fever in Eastern range Annual number of cases (bars) and case-fatality (line) for hantavirus pulmonary syndrome, United States, 1993-2013 Year Number of cases or % fatality rate

6. Peromyscus as a model species  Evolution of coloration on different substrates in the Peromyscus maniculatus species complex  Evolution of Hemoglobin oxygen affinity in high mountains

7. P. polionotus P. melanotis P. keeni P. arcticus P. maniculatus

8. Natarajan, Hoffmann, Lanier, et al. 2015 ?

9. Research in Wyoming  16 samples collected at this site  5 samples collected from this site Casper Mountain SamplesYellowstone National Park Samples

10. H 0 : Wyoming samples will fall within Clade 1 or Clade 2 due to their location H 1 : These populations will be in a new clade not yet described Hypotheses: Natarajan, Hoffmann, Lanier, et al. 2015

11. Methods- Lab Extract DNA from tissue samples using Qiagen DNeasy Blood and Tissue kit Polymerase Chain Reaction (PCR) to amplify mtDNA and gel electrophoresis to run out PCR on agarose gel Visual DNA on gels using BioRad Transilluminator Send successful samples off for sequencing at Macrogen

12. Polymerase Chain Reaction Internal primersExternal Primers

13. Methods-Analysis Troublesome sequence Clean sequence Red-Adenine Green-Thymine Blue-Cytosine Yellow-Guanine

14. Methods- Compiling data

15. Wyoming tree Casper 1 Yellowstone1 Casper 2 Yellowstone 3 Yellowstone 2 Yellowstone 4 Yellowstone 5 Casper 3 Casper 4 Casper 5 Casper 6 Casper 7 Casper 9 Casper 8 Casper 10 Casper 11 Casper 12 0.02

16. Greater genetic diversity in Casper Mountain population than in Yellowstone population Clade 1 has undergone more recent expansion than the other clades All Wyoming samples fall within Clade 1 Results: Clade 1 Red- Casper Mountain Green- Yellowstone Black- Previous samples sites

17. Hypothesis supported Natarajan, Hoffmann, Lanier, et al. 2015