Interspecific hybridization in Brassica species leads to changes in agronomic traits through the regulation of gene expression by chromatin accessibility and DNA methylation.

Interspecific hybridization is a common method in plant breeding to combine traits from different species, resulting in allopolyploidization and significant genetic and epigenetic changes. However, our understanding of genome-wide chromatin and gene expression dynamics during allopolyploidization remains limited. This study generated two Brassica allotriploid hybrids via interspecific hybridization. We observed that accessible chromatin regions (ACRs) and DNA methylation interact to regulates gene expression after interspecific hybridization, ultimately influencing the agronomic traits of the hybrids. In total, 234,649 ACRs were identified in the parental lines and hybrids; the hybridization process induces changes in the distribution and abundance of their accessible chromatin regions, particularly in gene regions and their proximity. Genes associated with proximal ACRs were more highly expressed than those associated with distal and genic ACRs. More than half of novel ACRs drove transgressive gene expression in the hybrids, and the transgressive upregulated genes showed significant enrichment in metal ion binding, especially magnesium ion, calcium ion, and potassium ion binding. We also identified Bna.bZIP11 in the single-parent activation ACR, which binds to BnaA06.UF3GT to promote anthocyanin accumulation in F1 hybrids. DNA methylation plays a role in repressing gene expression, and unmethylated ACRs are more transcriptionally active. Additionally, the A-subgenome ACRs were associated with genome dosage rather than DNA methylation. The interplay among DNA methylation, transposable elements, and sRNA contributes to the dynamic landscape of ACRs during interspecific hybridization, resulting in distinct gene expression patterns on the genome.