miR319-targeted LsTCP4 and non-target LsTCP17 act in parallel to promote leaf senescence in lettuce

Leaf senescence directly affects lettuce quality and postharvest shelf life, but the regulatory roles of miR319-targeted and non-target CIN-TCP transcription factors remain unclear. Here, we examined whether the miR319-TCP module controls lettuce leaf senescence through separable genetic branches. MIR319 overexpression delayed dark-induced senescence, whereas STTM-mediated miR319 suppression accelerated chlorophyll loss, photosynthetic decline, and senescence-marker activation. Disruption of the miR319-targeted gene LsTCP4 phenocopied MIR319 overexpression, supporting LsTCP4 as a pro-senescence factor downstream of miR319. We further found that the miR319 non-target CIN gene LsTCP17 also promoted senescence, as tcp17 leaves retained more chlorophyll than wild type during dark treatment. Genetic combinations showed that tcp17 enhanced chlorophyll retention in the OX319 background and partially rescued the accelerated senescence phenotype of S319, indicating that LsTCP17 acts through a route separable from the miR319-targeted branch. Together, these results reveal a split CIN-TCP architecture in which miR319-targeted LsTCP4 and non-target LsTCP17 provide parallel pro-senescence inputs, offering a genetic framework for targeted improvement of lettuce quality.