ENDOGLUCANASE SlCEL2 and EXPANSIN SlEXP1 synergistically affect cellulose degrading and tomato fruit softening.

Delayed fruit softening in tomato (Solanum lycopersicum) is highly desirable for extending shelf life, facilitating long-distance transportation, and reducing post-harvest losses caused by mechanical damage. Fruit softening is a natural ripening process characterized by the increased expression of genes involved in cell wall modification, leading to the breakdown of cell wall polysaccharides and the gradual disintegration of cellular structure. The yft1 mutant (yellow-fruited tomato 1, originally designated n3122) exhibits inhibited ethylene production, preventing normal ripening and resulting in firmer fruit. Concurrently, yft1 shows significant downregulation of several genes associated with cell wall degradation, including endoglucanase SlCEL2 and EXPANSIN SlEXP1. Both genes exhibit similar expression patterns, peaking during ripening, suggesting their importance in fruit softening. To investigate this further, RNAi silencing lines targeting SlCEL2 and SlEXP1 were generated. The double mutant, slcel2 slexp1, displayed increased firmness at the red ripe stage (54 days post-anthesis, dpa), whereas the single mutants showed similar softening to the wild-type M82. Anatomical analysis at 54 dpa revealed enhanced cell wall structure, slightly increased cuticle thickness, and significantly higher pericarp cellulose content in slcel2 slexp1 compared to M82, slcel2, and slexp1. Furthermore, this study found that SlEXP1 expression was significantly upregulated in slcel2 fruit, compared to M82 (wild type), at 54 dpa. This suggests a compensatory transcriptional regulation between these two genes in tomato fruit, potentially aimed at maintaining normal softening during ripening. These findings demonstrate that SlCEL2 and SlEXP1 act synergistically in cellulose degradation during tomato ripening, and promoting fruit softening.