Cultivated roses have the largest global market share among ornamental crops. Postharvest release of ethylene is the main cause of accelerated senescence and decline in rose flower quality. To understand the molecular mechanism of ethylene-induced rose petal senescence, we analyzed the transcriptome of rose petals during natural senescence as well as with ethylene treatment. A large number of differentially expressed genes (DEGs) were observed between developmental senescence and the ethylene-induced process. We identified 1207 upregulated genes in the ethylene-induced senescence process, including 82 transcription factors and 48 protein kinases. Gene Ontology enrichment analysis showed that ethylene-induced senescence was closely related to stress, dehydration, and redox reactions. We identified a calcineurin B-like protein (CBL) interacting protein kinase (CIPK) family gene in Rosa hybrida, RhCIPK6, that was regulated by age and ethylene induction. Reducing RhCIPK6 expression through virus-induced gene silencing significantly delayed petal senescence, indicating that RhCIPK6 mediates petal senescence. In the RhCIPK6-silenced petals, several senescence associated genes (SAGs) and transcription factor genes were downregulated compared with controls. We also determined that RhCIPK6 directly binds calcineurin B-like protein 3 (RhCBL3). Our work thus offers new insights into the function of CIPKs in petal senescence and provides a genetic resource for extending rose vase life.

In cut rose (Rosa hybrida), the flower-opening process is closely associated with vase life. Auxin induces the expression of transcription factor genes that function in petal growth via cell expansion. However, the molecular mechanisms underlying auxin effect during flower opening are not well understood. Here, we identified the auxin-inducible transcription factor gene RhMYB6, whose expression level is high during the early stages of flower opening. Silencing of RhMYB6 delayed flower opening by controlling petal cell expansion through downregulation of cell expansion-related genes. Furthermore, we demonstrated that the auxin response factor RhARF2 directly interacts with the promoter of RhMYB6 and represses its transcription. Silencing of RhARF2 resulted in larger petal size and delayed petal movement. We also showed that the expression of genes related to ethylene and petal movement showed substantial differences in RhARF2-silenced petals. Our results indicate that auxin-regulated RhARF2 is a critical player that controls flower opening by governing RhMYB6 expression and mediating the crosstalk between auxin and ethylene signaling.

Flower senescence is genetically regulated and developmentally controlled. The phytohormone ethylene induces flower senescence in rose (Rosa hybrida), but the underlying signaling network is not well understood. Given that calcium regulates senescence in animals and plants, we explored the role of calcium in petal senescence. Here, we report that the expression of calcineurin B-like protein 4 (RhCBL4), which encodes a calcium receptor, is induced by senescence and ethylene signaling in rose petals. RhCBL4 interacts with CBL-interacting protein kinase 3 (RhCIPK3), and both positively regulate petal senescence. Furthermore, we determined that RhCIPK3 interacts with the jasmonic acid response repressor jasmonate ZIM-domain 5 (RhJAZ5). RhCIPK3 phosphorylates RhJAZ5 and promotes its degradation in the presence of ethylene. Our results reveal that the RhCBL4-RhCIPK3-RhJAZ5 module mediates ethylene-regulated petal senescence. These findings provide insights into flower senescence, which may facilitate innovations in postharvest technology for extending rose flower longevity.