A genome-wide survey of DNA methylation reveals hyper-methylation regulates after-ripening and dormancy of recalcitrant Panax notoginseng seeds.

DNA methylation plays a crucial role in regulating fruit ripening and seed development. It remains unknown about the dynamic characteristics of DNA methylation and its regulation mechanisms in morpho-physiological dormancy (MPD)-typed seeds with recalcitrant characteristics. The Panax notoginseng seeds are defined by the MPD and are characterized by a strong sensitivity to dehydration during the after-ripening process. We performed DNA methylomes, siRNA profiles, and transcriptomes of embryo and endosperm in P. notoginseng seeds at different after-ripening stages. Herein, we find that the hyper-methylation contributes to the increase in DNA methylation during the after-ripening process. The endosperm genome is hyper-methylated compared to the embryo genome. The hyper-methylation is caused by the high expression level of DNA methyltransferase PnCMT2 in the seeds. The hyper-methylation alters gene transcription levels to regulate the after-ripening and dormancy of recalcitrant seeds. For example, it inhibits the expression of genes in embryo development to make seeds maintain a dormant status. Together, our findings reveal an increase in DNA methylation and its vital driver in gene expression, and thus elucidate how hyper-methylation regulates the after-ripening in recalcitrant MPD-typed seeds. This work establishes a key role for epigenetics in regulating the dormancy of MPD-typed seeds with recalcitrant characteristics.