Genome wide identification of Dof transcription factors in Carmine radish reveals RsDof33 role in cadmium stress and anthocyanin biosynthesis.

Carmine radish (Raphanus sativus L.) is cultivated in Fuling, Chongqing, for its red color. Dof-TFs are critical in regulating plant growth, development, stress responses, and signal transduction.This work comprehensively examined the structure, evolution, and expression of the carmine radish Dof gene and its behavior under cadmium (Cd) stress. The radish genome has 59 RsDofs, which are divided into nine clusters (A: 8, B1: 10, B2: 10, C1: 3, C2.1: 5, C2.2: 4, C3: 11, D1: 4, and D2: 4). Phylogenetic tree analysis revealed significant Dof gene family resemblance between Arabidopsis thaliana and Brassica napus. Perhaps segment duplication resulted in RsDof gene family expansion. Cd stress-induced RsDof expression patterns were studied using an RNA-seq atlas and qRT-PCR. The majority of RsDofs were tissue-specific and Cd-sensitive. The involvement of RsDof genes in Cd stress response and anthocyanin synthesis was verified using qRT-PCR. RsDof33 is involved in Cd stress response and anthocyanin synthesis. A. thaliana overexpressed the recombinant fusion protein RsDof33-GFP, which was localized to the nucleus, resulting in fewer rosette leaves, delayed flowering, and higher anthocyanin concentration. RsDof33-expressing plants had significantly higher transcript levels of the auxin biosynthetic genes YUCCA (AtYUC2), auxin efflux carrier (AtPIN4), and AtKNAT2, which are involved in leaf shape development, as well as AtPAL, AtCHS, AtCHI, AtDFR, AtLDOX, and AtUF3GT. These findings indicate that RsDofs are critical to plant development and stress responses.