Dynamic changes in DNA methylation occur in TE regions and affect cell proliferation during leaf-to-callus transition in <i>Arabidopsis</i>

Plant somatic cells can be reprogrammed into pluripotent cell mass, called callus, through a two-step <i>in vitro</i> tissue culture method. Incubation on callus-inducing medium triggers active cell proliferation to form pluripotent callus. Notably, DNA methylation is implicated during callus formation, but a detailed molecular process regulated by DNA methylation remains to be fully elucidated. Here, we compared genome-wide DNA methylation profiles between leaf and callus tissues in <i>Arabidopsis</i> using whole genome bisulfite-sequencing. Global distribution of DNA methylation showed that CHG methylation was increased, whereas CHH methylation was reduced especially around transposable element (TE) regions during leaf-to-callus transition. We further analyzed differentially expressed genes around differentially methylated TEs (DMTEs) during leaf-to-callus transition and found that genes involved in cell cycle regulation were enriched and also constituted a coexpression gene network along with pluripotency regulators. In addition, a conserved DNA sequence analysis for upstream <i>cis</i>-elements led us to find a putative transcription factor associated with cell fate transition. CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1) was newly identified as a regulator of plant regeneration, and consistently, the <i>cca1lhy</i> mutant displayed altered phenotypes in callus proliferation. Overall, these results suggest that DNA methylation coordinates cell cycle regulation during callus formation, and CCA1 may act as a key upstream coordinator at least in part in the processes.