*2.1. Isolation and Analysis of PheDof12-1*

Based on the moso bamboo genome database, *PheDof12-1* was isolated from moso bamboo. The full-length CDS of *PheDof12-1* is 1299 bp, encoding 432-amino acids, with predicted molecular weight (MW) and isoelectric point (pI) of 46.37 kDa and 8.32, respectively. Structure analysis showed that PheDof12-1 contains one intron and two exons (Figure 1A). The deduced proteins contain the conserved zf-Dof domain. Furthermore, phylogenetic analysis of PheDof12-1 and homologous proteins from other plants shows that PheDof12-1 and other Dofs from monocotyledons belong to the same clade (Figure 1B). The amino acid sequence of PheDof12-1 shows 83% and 84% identity with *Brachypodium* (XP\_003558722) and rice (XP\_015690912), respectively. This result was consistent with the findings in the stated phylogeny and classification of plants. All these proteins contain the conserved zf-Dof domain (Supplementary Figure S1).

*Int. J. Mol. Sci.* **2019**, *20*, x FOR PEER REVIEW 3 of 13

**Figure 1.** Characterization and preliminary expression analysis of *PheDof12-1*. (**A**) Gene structure of *PheDof12-1.* (**B**) Phylogenetic analysis of PheDof12-1 with other DNA binding with one finger (Dof) proteins. (**C**) qRT-PCR analysis of *PheDof12-1* in different tissues of moso bamboo. (**D**) Expression profile of *PheDof12-1* in different flower developmental stages: F1: floral bud formation stage; F2: inflorescence growing stage; F3: blooming stage; F4: flowers are withered. (**E**) Glucuronidase (GUS) **Figure 1.** Characterization and preliminary expression analysis of *PheDof12-1*. (**A**) Gene structure of *PheDof12-1*. (**B**) Phylogenetic analysis of PheDof12-1 with other DNA binding with one finger (Dof) proteins. (**C**) qRT-PCR analysis of *PheDof12-1* in different tissues of moso bamboo. (**D**) Expression profile of *PheDof12-1* in different flower developmental stages: F1: floral bud formation stage; F2: inflorescence growing stage; F3: blooming stage; F4: flowers are withered. (**E**) Glucuronidase (GUS) staining of *ProPheDof12-1*-*GUS* in transgenic Arabidopsis seedling. (**F**) GUS staining of *ProPheDof12-1*-*GUS* plants showing PheDof12-1 localization in flower and pollen.
