Transcriptome-wide analysis of gene expression using detached first-pair rosette leaves before culture (time 0), at 10min to 12h after detachment Col-0, coi1-2 and sdg8-2 seedlings.

To reveal the early molecular events upon wounding during DNRR from Arabidopsis leaf explants cultured on B5 medium without exogenous hormones, we carried out an RNA-seq experiment using leaf explants before culture (i.e. time 0), at 10 and 30 min after detachment, and at 1 to 12 h after detachment. The results showed that the upregulation of JA-related genes occurred as an early response to wounding (mainly within 1 h), prior to the upregulation of auxin-related genes (from 2 to 12 h). These results indicated that JA responds rapidly to wounding and earlier than does auxin. To analysis whether SDG8-mediated H3K36me3 is generally involved in JA-mediated gene upregulation in response to wounding. We evaluated changes in the transcriptome in sdg8-2 and coi1-2 leaf explants (2 h vs. time 0) by analyzing RNA-seq data, and compared them with transcriptomic changes in wild-type. Overall design: Arabidopsis seeds were germinated on ½ MS medium at 22°C under a 16-h light/8-h dark photoperiod. Detached leaf explants from 12-day-old seedlings were cultured on B5 medium without sucrose at 22°C under 24-h light conditions. Biological duplicates were performed for each timepoint analyzed.

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