RNA N6-Methyladenosine Responds to Low-Temperature Stress in Tomato Anthers

Cold stress is a serious threat to subtropical crop pollen development and induces yield decline. N6-methyladenosine (m⁶A) is the most frequent mRNA modification and plays multiple physiological functions in plant development. However, whether m⁶A regulates pollen development is unclear, and its putative role in cold stress response remains unknown. Here, we observed that moderate low-temperature (MLT) stress induced pollen abortion in tomato. This phenotype was caused by disruption of tapetum development and pollen exine formation, accompanied by reduced m⁶A levels in tomato anther. Analysis of m⁶A-seq data revealed 1,805 transcripts displayed reduced m⁶A levels and 978 transcripts showed elevated m⁶A levels in MLT-stressed anthers compared with those in anthers under normal temperature. These differentially m⁶A enriched transcripts under MLT stress were mainly related to lipid metabolism, adenosine triphosphatase (ATPase) activity, and ATP-binding pathways. An ATP-binding transcript, SlABCG31, had significantly upregulated m⁶A modification levels, which was inversely correlated to the dramatically downregulated expression level. These changes correlated with higher abscisic acid (ABA) levels in anthers and disrupted pollen wall formation under low-temperature stress. Our findings characterized m⁶A as a novel layer of complexity in gene expression regulation and established a molecular link between m⁶A methylation and tomato anther development under low-temperature conditions.