Abstract
C methylation of DNA, which occurs at CG, CHG, and CHH (H = A, C, or T) sequences in plants, is a hallmark for the epigenetic repression of repetitive sequences. The chromatin-remodeling factor DECREASE IN DNA METHYLATION1 (DDM1) is essential for DNA methylation, especially at CG and CHG sequences. However, its potential roles in RNA-directed DNA methylation (RdDM) and in chromatin function are not completely understood in rice (Oryza sativa). In this work, we used high-throughput approaches to study the function of rice DDM1 (OsDDM1) in RdDM and the expression of noncoding RNA. We show that loss of function of OsDDM1 results in ectopic CHH methylation of transposable elements and repeats. The ectopic CHH methylation was dependent on rice DOMAINS REARRANGED METHYLTRANSFERASE2, a DNA methyltransferase involved in RdDM. Mutations in OsDDM1 lead to decreases of histone H3K9me2 and increases in the levels of heterochromatic small RNA and long noncoding RNA. In particular, OsDDM1 was found to be essential to repress the transcription of the two repetitive sequences, Centromeric Retrotransposons of Rice1 and the dominant centromeric CentO repeats. These results suggest that OsDDM1 antagonizes RdDM at heterochromatin and represses the tissue-specific expression of noncoding RNA from repetitive sequences in the rice genome.