Fig. 7

Identification of DNA and histone demethylation of OsZIP1 in rice exposed to excess Zn, Cu, Mn, Fe and Cd. a Diagram of the OsZIP1 genomic region with CG, CHG and CHH demethylation pattern in R2 with Cd (+Cd) and without Cd (−Cd) exposure. b DNA methylation status At R1-R3 of OsZIP1 with -Cd and + Cd was determined by McrBC-qPCR assays (Note: a lower value of DNA amplification of (+McrBC/−McrBC) indicates the higher methylation). c: Histone methylation state was detected using an anti-H3K9me2 ChIP-qPCR assay at R1-R3 of the genomic regions of OsZIP1 under –Cd and + Cd conditions. d DNA methylation status at R1-R3 of OsZIP1 with the normal and excessive Zn, Cu, Mn and Fe supply. Two weeks-old young rice plants were grown in the nutrient solution supplemented with 0.4 (normal) and 1000 (high) μM Zn, 0.2 (normal) and 20 (high) μM Cu, 0.5 (normal) and 500 (high) μM Mn, and 10 (normal) and 1000 (high) μM Fe, or with 0 and 80 μM Cd for 4 d. The rice Ubi2 and Actin1 were tested in parallel as negative controls. Anti-H3 was used as an internal reference in the ChIP-qPCR assay. Vertical bars represent standard deviation. Asterisks indicate that the mean values of three replicates are significantly different between the metal treatments and control (p < 0.05)