Leaves of Begonia semperflorens accumulate anthocyanins and turn red under low temperature (LT). In the present work, LT increased H ₂ O ₂ content and superoxide anions production rate, causing significant increases in the activities of enzymes and contents of reduced components involved in the ascorbate-glutathione cycle (AsA-GSH cycle). As a result, LT-exposed seedlings increased the expression of genes involved in anthocyanin biosynthesis, and accumulated anthocyanin. Based on LT condition, application of N,N'-dimethylthiourea (DMTU) decreased reactive oxygen species (ROS) content, and unbalanced the AsA-GSH-controlled redox homeostasis. As a result, seedlings in the LT + DMTU group did not accumulate anthocyanin. Our results suggest that ROS may act as an important inducer in LT-induced anthocyanin biosynthesis.

Low temperature (LT) is an important elicitor that triggers anthocyanin biosynthesis. To investigate whether the reactive oxygen species (ROS) produced via RBOH are involved in this process, we analysed the function and mechanism of ROS produced via RBOH during LT-induced anthocyanin biosynthesis in Begonia semperflorens Link & Otto. The results showed that BsRBOHD transcription was upregulated in LT-grown seedlings at the 3rd hour, which was followed by the upregulation of anthocyanin-biosynthesis genes at the 5–9th hour, leading to anthocyanin accumulation on the 2nd day. The LT-induced increases in ROS production, BsRBOHD and anthocyanin-biosynthesis gene transcription, and anthocyanin content were abolished by the pre-treatment of seedlings with DPI [an inhibitor of nicotinamide adenine nucleoside phosphorylase (NADPH) oxidase or DMTU (a H₂O₂ scavenger)], but were promoted by pre-treatment with NADPH (a substrate of NADPH oxidase). Changes in the chlorophyll fluorescence parameters showed that pre-treatment of DPI or DMTU alleviated the LT-induced decrease in the seedling chlorophyll content and a/b ratio, which subsequently alleviated the LT-induced decreases in the ABS/CSm, TRo/CSm, RC/CSm, ETo/CSm and REo/CSm values. In contrast, NAPDH pre-treatment intensified these changes. Therefore, we suggest that ROS produced via BsRBOHD may be involved in the LT-induced anthocyanin biosynthesis by strengthening the overaccumulation of ROS produced by the overexcitation of PSII reaction centres and overflux from \({\text{Q}}_{{\text{A}}}^{ - }\) to NADP⁺ in B. semperflorens.

The phosphatidylethanolamine-binding protein (PEBP) gene family, known for its pivotal role in controlling floral transition, regulates flowering time, and, thus, shapes the continuous-flowering trait in ornamental plants. In this study, we conducted the first genome-wide identification and bioinformatics analysis of the PEBP gene family in Begonia semperflorens ‘Super Olympia’, a variety that exhibits year-round flowering. Via phylogenetic analysis, a total of 10 BsPEBP genes were identified and categorized into four subfamilies: the FT-like (two members), TFL1-like (three members), PEBP-like (three members), and MFT-like (two members) subfamilies. Gene structure analysis revealed highly conserved motif compositions among family members, and protein tertiary structure prediction indicated the dominance of random coils in their structures. Promoter cis-acting element analysis revealed light-responsive, hormone-responsive (ABA, GA, and JA), and abiotic stress-responsive elements in the BsPEBP genes, suggesting their potential integration into multiple regulatory pathways. The tissue-specific expression profiles revealed that BsPEBP6 was significantly upregulated in floral organs, whereas TFL1-like subfamily members were predominantly expressed in vegetative tissues. These findings imply that the FT-like and TFL1-like genes antagonistically regulate the continuous-flowering trait of B. semperflorens ‘Super Olympia’ through their respective roles in promoting and repressing flowering. Our findings provide a preliminary theoretical foundation for elucidating the molecular mechanisms by which the PEBP gene family regulates flowering time in ornamental plants and offer valuable insights for developing breeding strategies aimed at flowering time modulation.