Investigation of potential targets of TaSPL6-D in wheat

Here, we performed the cleavage under targets and tagmentation (CUT&Tag) assay, a recently developed technology for protein-DNA interaction, in protoplasts extracted from wheat root tissues to genome-widely identity the potential targets of TaSPL6-D. Using CUT-Tag-seq, we obtained 7,403 confidential peaks from two biological replicates, with which 2688 genes was associated. Zooming in the promoter region, GTAC motif was significantly enriched in the peaks. Gene Ontology analysis of genes with binding peaks in the promoter region revealed that, the most enriched gene term is ‘response to temperature/heat stimuli’, which is consistent with previous studies on the functions of TaSPL6-D orthologs in rice and Arabidopsis. The following enriched terms are genes of ‘regulation of response to salt stress’, ‘regulation of response to osmotic stress’, and particularly ‘positive regulation of response to salt stress’, implying TaSPL6-D is an upstream regulator of salt stress response. DNA library established by Cleavage under targets and tagmentation (CUT&Tag) assay in wheat protoplasts was used for sequencing to genome-widely identity the potential targets of TaSPL6-D.