The diversity of fruit color in sweet cherry (Prunus avium L.) has been attributed to the presence of either anthocyanin or carotenoid. We profiled the anthocyanin and carotenoid metabolites to investigate the different pigments and the underlying regulatory mechanisms of differential expression genes (DEGs) between red and yellow fruits of sweet cherry. We profiled two cultivars, ‘Jiangnanhong’(JNH, red fruits) and ‘Chaoyang’(CY, yellow fruits) to establish their anthocyanin and carotenoid metabolites by LC-MS/MS and transcriptome analysis by RNA-seq to test the difference in gene expression and metabolic substances between the two varieties. Cyanidin-3-O-rutinoside was the most different pigment between two cultivars, the content of which in red fruit was significantly higher than in the yellow one during the whole ripening stage (stage 3 and stage 4). The total carotenoid content in the two color types of fruits was close, but the content in yellow fruit was shown to be more stable after harvest. Based on the transcriptome data, the heatmap of selected structural DEGs showed that all of the anthocyanin genes expressed significantly higher levels in red fruits than that in yellow fruits. Two unigenes encoding chalcone synthase (CHS) and UDP glucose-flavonoid 3-O-glucosyltransferase (UFGT) were expressed 1134.58 and 1151.24 times higher in red than in yellow fruits at stage 4, respectively. Correlation analysis showed that anthocyanin genes in JNH were negatively correlated with those in CY; by contrast, there were some strong correlations observed between the two cultivars in carotenoid genes. Thus, the coloration of sweet cherry was mainly attributed to anthocyanin-related genes.

Gibberellin (GA) is an important plant hormone that is involved in various physiological processes during plant development. Sweet cherries planted in southern China have always encountered difficulty in bearing fruit. In recent years, gibberellin has successfully solved this problem, but there has also been an increase in malformed fruits. This study mainly explores the mechanism of malformed fruit formation in sweet cherries. By analyzing the synthesis pathway of gibberellin using metabolomics and transcriptomics, the relationship between gibberellin and the formation mechanism of deformed fruit was preliminarily determined. The results showed that the content of GA₃ in malformed fruits was significantly higher than in normal fruits. The differentially expressed genes in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were mainly enriched in pathways such as “plant hormone signal transduction”, “diterpenoid biosynthesis”, and “carotenoid biosynthesis”. Using Quantitative Real-Time Reverse Transcription PCR (qRT-PCR) analysis, the gibberellin hydrolase gene GA2ox and gibberellin synthase genes GA20ox and GA3ox were found to be significantly up-regulated. Therefore, we speculate that the formation of malformed fruits in sweet cherries may be related to the accumulation of GA₃. This lays the foundation for further research on the mechanism of malformed sweet cherry fruits.

This study aimed to characterize carotenoid profiles and unravel the genetic mechanisms underlying flesh color variation in white and yellow-fleshed peaches, with a focus on the hybrid cultivar ‘ZY29’ derived from two white-fleshed parents (‘Yulu’ and ‘Hujing Honey Dew’). Using UPLC-APCI-MS/MS, we quantified carotenoids in the pericarp (exocarp) and flesh (mesocarp) of parental and hybrid fruits. Results showed that ‘ZY29’ accumulated significantly higher levels of β-carotene and lutein compared to its white-fleshed parents. Transcriptome analysis revealed upregulation of carotenoid biosynthesis genes (PSY, LCYB, and ZDS) and downregulation of the carotenoid cleavage gene CCD4 in ‘ZY29’, explaining enhanced carotenoid accumulation. Integrative metabolome-transcriptome analysis identified core regulatory networks associated with metabolic shifts, including transcription factors (MYB and WRKY). These findings provide novel insights into the molecular basis of yellow flesh formation in peaches, offering potential targets (PSY and LCYB) and metabolic markers (β-carotene and lutein) for breeding nutritionally enriched cultivars. These findings contribute to a better understanding of the genetic factors and parental regulatory mechanisms involved in the formation of yellow flesh color in peaches. Our results have important implications for breeding new peach varieties with desirable color and nutritional qualities and may provide valuable insights for future research in this area.

The flower color of Rhododendron is primarily determined by anthocyanin biosynthesis, with cytochrome P450 CYP75 family members, particularly flavonoid 3′,5′-hydroxylase (F3′5′H), playing a central role. However, the composition and functional characterization of CYP75 genes in Rhododendron remain insufficiently explored. This study performed genome-wide identification of the CYP75 gene family using the Rhododendron simsii reference genome and functionally characterized the corresponding F3′5′H homolog cloned from Rhododendron × hybridum petals (red cultivar and pink cultivar). Seven RsCYP75 genes were identified, categorized into two subfamilies: RsCYP75A (A1–A5) and RsCYP75B (B1–B2), with a prominent cluster on chromosome 13. All encoded proteins contained a conserved cytochrome P450 domain and typical heme-binding motifs. Among these, RhCYP75A2 showed the highest expression level in red petals at full blooming period and was designated as RhF3′5′H. RhF3′5′H encodes a basic membrane protein with the characteristic F3′5′H motif, with its transcript most abundant in flowers. Transient overexpression of RhF3′5′H in red R. × hybridum petals resulted in a 9.74-fold increase in its transcript levels and a 1.25-fold increase in anthocyanin content compared to that in the control accompanied by the up-regulation of CHS, F3H, DFR and ANS. Conversely, RhF3′5′H silencing reduced anthocyanin accumulation but increased CHS and F3H transcript levels, suggesting a compensatory transcriptional response in the upstream anthocyanin pathway. Moreover, RhF3′5′H was heterologously expressed in E. coli Rosetta as an MBP fusion protein, purified, and identified by LC-MS/MS and ELISA. The protein showed the ability to promote anthocyanin accumulation. Molecular docking analysis demonstrated that RhF3′5′H can bind to naringenin and dihydrokaempferol. These results confirm that RhF3′5′H is a functional F3′5′H-type CYP75A enzyme and a positive regulator of anthocyanin accumulation in Rhododendron petals. This work enriches the CYP75 gene catalog in Rhododendron and provides candidate genes for future studies on flower color regulation and molecular breeding.