The m⁶A reader SlYTH2 negatively regulates tomato fruit aroma by impeding the translation process.

Abstract

<i>N</i>⁶-methyladenosine (m⁶A) is a fundamentally important RNA modification for gene regulation, whose function is achieved through m⁶A readers. However, whether and how m⁶A readers play regulatory roles during fruit ripening and quality formation remains unclear. Here, we characterized SlYTH2 as a tomato m⁶A reader protein and profiled the binding sites of SlYTH2 at the transcriptome-wide level. SlYTH2 undergoes liquid-liquid phase separation and promotes RNA-protein condensate formation. The target mRNAs of SlYTH2, namely m⁶A-modified <i>SlHPL</i> and <i>SlCCD1B</i> associated with volatile synthesis, are enriched in SlYTH2-induced condensates. Through polysome profiling assays and proteomic analysis, we demonstrate that knockout of <i>SlYTH2</i> expedites the translation process of <i>SlHPL</i> and <i>SlCCD1B</i>, resulting in augmented production of aroma-associated volatiles. This aroma enrichment significantly increased consumer preferences for CRISPR-edited fruit over wild type. These findings shed light on the underlying mechanisms of m⁶A in plant RNA metabolism and provided a promising strategy to generate fruits that are more attractive to consumers.