Multi-Intron Analysis of Aquilegia flavescens Across the Central and Northern Rocky Mountains

Billman, Peter; Lavin, Matt

Study of yellow columbine genetic diversity across the Rocky Mountains. During the summer months in 2014 and 2015, I hiked to subalpine-‐‑alpine regions that were historically documented for containing Aquilegia flavescens populations on the Consortium of Pacific Northwest Herbaria’s website (http://www.pnwherbaria.org). Several sites were selected that appeared to have suitable climate, terrain, and elevation (but weren’t yet noted in the websites database for having Aq. fl.) and these areas were also sampled to complement the previously documented sites. I collected healthy tissue from one basal leaf per plant. These leaves were then immediately placed in envelopes and pressed to preserve the DNA. 46 populations were sampled across the U.S. and Canadian Rockies and one sample of Aquilegia coerulea was used as an outgroup. The A. coerulea sample was collected from the Wind River Mountains of Wyoming. Glacier National Park in Montana was generous enough to allow me to collect three leaves from plants that were grown at their greenhouse location. These plants were grown from seeds collected near the Big Bend area of the park. The University of British Columbia (UBC) in Vancouver allowed me to obtain leaf tissue from seven herbarium-‐‑pressed specimens that spanned across the Canadian Rockies. Without their assistance, we would not have been able to analyze this expansive region of the continent that we weren’t able to access in only two summer seasons. Back in the lab, we isolated the DNA utilizing a DNEasy Plant Mini Kit (Qiagen, Valencia, CA, USA). We used the instructions included in the kit to isolate and amplify the DNA. trnL primers and ITS 6 and 9 primers were then added and the polymerase chain reaction (PCR) was ran. The samples were then sent off to Washington state to be sequenced. We analyzed the sequenced DNA using Sequencer and Se-‐‑Al software after we received the DNA back.