2.2. Classification and Phylogeny of Aquilegia coerulea Aquaporins

Multiple sequence alignment of AQPs was performed using the CLUSTALW tool implemented in the MEGA7 software suite [37]. A phylogenetic tree of AQPs was generated using the maximum likelihood (ML) method, and 1000-bootstraps was performed to measure the stability of branch nodes in the ML tree. The classification of AQP subgroups was performed in accordance with the nomenclature of known AQPs from rice, Arabidopsis, and poplar, which were used as a query. A combined phylogenetic tree of AQPs from Aquilegia, Arabidopsis, rice, and poplar was also constructed. Phylogenetic tree of plant orders was generated based on National Center for Biotechnology Information (NCBI) taxonomy using the online server phyloT (http://phylot.biobyte.de/).