Transcriptome-Wide Analysis of RNA m⁶A Methylation and Gene Expression Changes Among Two Arabidopsis Ecotypes and...

The remodeling of transcriptome, epigenome, proteome, and metabolome in hybrids plays an important role in heterosis. N(6)-methyladenosine (m⁶A) methylation is the most abundant type of post-transcriptional modification for mRNAs, but the pattern of inheritance from parents to hybrids and potential impact on heterosis are largely unknown. We constructed transcriptome-wide mRNA m⁶A methylation maps of Arabidopsis thaliana Col-0 and Landsberg erecta (Ler) and their reciprocal F₁ hybrids. Generally, the transcriptome-wide pattern of m⁶A methylation tends to be conserved between accessions. Approximately 74% of m⁶A methylation peaks are consistent between the parents and hybrids, indicating that a majority of the m⁶A methylation is maintained after hybridization. We found a significant association between differential expression and differential m⁶A modification, and between non-additive expression and non-additive methylation on the same gene. The overall RNA m⁶A level between Col-0 and Ler is clearly different but tended to disappear at the allelic sites in the hybrids. Interestingly, many enriched biological functions of genes with differential m⁶A modification between parents and hybrids are also conserved, including many heterosis-related genes involved in biosynthetic processes of starch. Collectively, our study revealed the overall pattern of inheritance of mRNA m⁶A modifications from parents to hybrids and a potential new layer of regulatory mechanisms related to heterosis formation.