Cloning and Phylogenetic Analysis of OoNAC72

The sequence of OoNAC72 was obtained by our research group from the O. ochrocephala’s transcriptome sequencing data (He et al., 2015 (link)). Using the specific primers, we amplified the ORF of OoNAC72 (Supplementary Table S1). The PCR condition was as follows: 3 min at 95°C; 34 cycles of 30 s at 95°C, 30 s at 55°C, and 30 s at 72°C; and then 10 min at 72°C. The resulting PCR products were cloned to the pGEM-T Easy Vector (TaKaRa, Beijing, China) and sequenced by Sangon Biotech Co., Ltd., (Shanghai, China). Multiple sequence alignment of OoNAC72 with NAC TFs in other species was performed with DNAMAN 8.0. A phylogenetic tree was constructed using a neighbor-joining (NJ) method with 1000 bootstrap replicates in MEGA 5.0.