Watermelon (Citrullus lanatus) is a globally important Cucurbitaceae crop in which grafting is commonly used to improve stress tolerance and enhance nutrient utilization. However, the mechanism underlying grafting-enhanced nutrient assimilation remains unclear. Here, we demonstrate the possible involvement of a novel Cucurbitaceae miRNA, ClmiR86, in grafting-enhanced phosphate-starvation tolerance via CALCINEURIN B-LIKE INTERACTING PROTEIN KINASE 5 (ClCIPK5) suppression in watermelon. Transcript analyses revealed that the induction of ClmiR86 expression was correlated with the downregulation of ClCIPK5 in squash-grafted watermelon under phosphate starvation. In addition, the differential expression of ClmiR86 in various watermelon genotypes was consistent with their phosphate utilization efficiency. Furthermore, ClmiR86 overexpression in Arabidopsis enhanced root growth and phosphate uptake under phosphate starvation and promoted inflorescence elongation under normal conditions. These results suggest that the ClmiR86–ClCIPK5 axis is involved in phosphate starvation response as well as grafting-enhanced growth vigor and phosphate assimilation. The present study provides valuable insights for investigating long-distance signaling and nutrient utilization in plants.

Strigolactone (SL) plays essential roles in regulating plant growth, development, and stress response. This study was conducted to evaluate the effect of exogenous SL on watermelon resistance against herbicides penoxsulam (PXL) and bensulfuron-methyl (BSM). These herbicides were found to negatively impact watermelon root growth and photosynthetic pigments, probably due to the ultrastructural damage and cell death in leaf and root tissues under herbicide stresses. The activation of SL-related gene expression suggested that the SL pathway may mitigate herbicide toxicity in watermelon. The exogenous SL dose-dependently reversed the PXL- or BSM-induced antioxidant activity, suggesting that SL may participate in maintaining antioxidant enzyme homeostasis under herbicide stresses. The up-regulation of herbicide metabolization and detoxification-related genes (cytochrome P450 and acetolactate synthase) by exogenous SL also in part explained how this phytohormone alleviates herbicide toxicity in watermelon. Our findings will provide valuable information underlying the regulatory effects of SL on herbicide tolerance in Cucurbitaceae crops.