RhLHY-RhTPPF-Tre6P Inhibits Flowering in Rosa hybrida Under Insufficient Light by Regulating Sugar Distribution

Light is crucial for flower bud development in plants, serving as both signal and energy source. However, the mechanisms by which daylength and light intensity regulate flowering in modern roses remain unclear. In Rosa hybrida 'Carola', insufficient light delays flowering and reduces the sugar content in terminal buds. RNA sequencing identified the Trehalose-6-phosphate phosphatase F (RhTPPF) gene as a key responder to insufficient light, modulating Tre6P metabolism. Overexpression of RhTPPF in rose calli enhanced sugar accumulation and suppressed the synthesis of RhCO/FT. In tobacco, overexpression of RhTPPF delayed the transition from vegetative growth to flowering, while silencing RhTPPF in roses accelerated flowering. Silencing RhTPPF in roses elevated trehalose-6-phosphate (Tre6P) levels and decreased trehalose. Transcriptome data showed that the expression level of RhTPPF was highly correlated with the circadian rhythm gene LATE ELONGATED HYPOCOTYL (RhLHY). Yeast one-hybrid assays, dual luciferase assays and EMSA revealed that RhLHY directly binds to the RhTPPF promoters. Overexpression of RhLHY suppressed flowering, while silencing RhLHY promoted flowering. Furthermore, altering the expression of RhLHY influenced Tre6P synthesis and the expression of sucrose-related transport genes. These findings suggest a RhLHY-RhTPPF-Tre6P regulatory module that maintains sugar balance and inhibits flower formation under reduced light conditions by modulating sugar distribution.