Abstract
The Dof gene family represents a class of plant-specific transcription factors that play crucial regulatory roles in various biological processes, including plant growth, development, and responses to abiotic stress. However, genome-wide identification and functional characterization of the Dof gene family remain unexplored in Dendrobium officinale. In this study, we performed a genome-wide identification and functional analysis of the DoDof gene family. A total of 28 Dof family members were identified and named DoDof1-28 based on genome annotation data. Phylogenetic analysis classified these genes into four major groups (A-D) and further subdivided them into nine subfamilies. Gene structure analysis revealed that most DoDofs lack introns, with no distinct specificity observed among different subfamilies and considerable diversity within the same subfamily. Sequence alignment analysis demonstrated that all DoDof proteins contain a conserved Dof domain consisting of 52 amino acids, which includes a C2-C2 zinc finger motif and a DNA-binding domain. MEME analysis revealed that the conserved motif composition exhibits a certain degree of conservation among DoDof proteins, but significant differences exist across subfamilies. Expression pattern analysis demonstrated that DoDofs have exhibited diverse expression profiles across different developmental stages, tissues, and under abiotic stresses (such as low temperature, salinity, and drought) in D. officinale, suggesting their potential roles in plant development and stress responses. Subcellular localization analysis indicated that DoDof15, DoDof22, and DoDof24 are localized exclusively in the nucleus. Yeast one-hybrid assays revealed that DoDof22 binds to the promoter of the ABA receptor DoPYL9, while DoDof15 and DoDof24 bind to the promoter of the bHLH transcription factor DobHLH68. These results suggest that DoDof proteins may regulate the growth, development, and stress response processes of D. officinale by binding to the promoters of target genes. This study provides critical insights into the functional roles of Dof transcription factors in Orchidaceae family and establishes a theoretical foundation for molecular breeding and stress resistance improvement in D. officinale.