Sampling
Seeds of Aquilegia viridiflora were collected from 20 locations
covering its current distribution range, and all voucher specimens were
identified by Dr Hongxing Xiao and deposited in the Northeast Normal
University Herbaria (Figure 1; Table S1). Each of the maternal plants
was separated from the others by > 50 m. Furthermore, to
determine whether the A. viridiflora materials collected shared
the most recent common ancestor (MRCA), we also collected A.
amurensis , A. ecalcarata , A. japonica , A.
oxysepala var. kansuensis , A. oxysepala var.oxysepala , and A. yabeana seeds, as reported in our
previous study (W. Zhang, Wang, Dong, Zhang, & Xiao, 2021).
Additionally, seeds of Paraquilegia microphylla were collected as
an outgroup. All the seeds were grown in the experimental field of
Northeast Normal University. Fresh leaves and buds were used to extract
DNA and RNA, respectively. These materials were quickly frozen in liquid
nitrogen and stored at -80 ℃.