To explore the mechanisms of the color formation of different colored persimmon fruits, we used two different colored persimmon cultivars (yellow-peeled persimmon fruit ‘Zhongshi No.6’ and red-peeled persimmon fruit ‘Hongdenglong’) as materials to study the synthesis and accumulation of carotenoids in three stages (full green, color transition, and full yellow or red) using targeted metabolomic and transcriptomic methods. A total of 14 carotenoids and 30 carotenoid lipids were identified in the peel of the two persimmon cultivars. After analysis, it was found that the total carotenoid content of the red persimmon cultivar was higher than that of the yellow persimmon cultivar. The contents of lycopene, α-carotenoid, β-carotenoid, (E/Z)-phytoene, and β-cryptoxanthin are the main reasons for the difference in total carotenoid content between the two persimmon cultivars, especially lycopene. Twelve structural genes involved in the metabolism of carotenoids were also found in this study. In comprehensive metabolome and transcriptome analysis, we found that, between the two persimmon cultivars, lycopene was the key metabolite responsible for the color difference, and PSY, LCYE, and ZDS were the key genes that regulated the differences in lycopene accumulation. The results of this study provide us with new information regarding persimmon fruit synthesis and accumulation. In addition, they also provide a theoretical foundation for improving persimmon fruit germplasm.

Persimmon fruit has a high nutritional value and significantly varies between pollination-constant astringent (PCA) and pollination-constant non-astringent (PCNA) persimmons. The astringency type affects sugar, flavonoids, and tannin accumulation and is well known in persimmon fruit. However, the impact of the fruit astringency type on ascorbic acid (AsA) accumulation is limited. In this study, typical PCA varieties (‘Huojing’ and ‘Zhongshi5’) and PCNA varieties (‘Yohou’ and ‘Jiro’) of persimmon fruit were sampled at four developing stages (S1–S4) to provide valuable information on AsA content variation in PCA and PCNA persimmon. Persimmon fruit is rich in ascorbic acid; the AsA content of the four varieties ‘Zhongshi5’, ‘Huojing’, ‘Jiro’, and ‘Youhou’ mature fruit reached 104.49, 48.69, 69.69, and 47.48 mg/100 g. Fruit of the same astringency type persimmon showed a similar AsA accumulation pattern. AsA content was significantly higher in PCA than PCNA fruit at S1–S3. The initial KEGG analysis of metabolites showed that galactose metabolism is the major biosynthetic pathway of AsA in persimmon fruit. There were significant differences in galactose pathway-related metabolite content in developing PCA and PCNA fruit, such as Lactose, D-Tagatose, and D-Sorbitol content in PCA being higher than that of PCNA. Combined gene expression and WGCNA analyses showed that the expression of the GME (evm.TU.contig4144.37) gene was higher in PCA-type than in PCNA-type fruit in S1–S3 and exhibited the highest correlation with AsA content (r = 690 **, p < 0.01). Four hub genes, including the DNA methylation gene, methyltransferase gene, F-box, and Actin-like Protein, were identified as potential regulators of the GME gene. These results provide basic information on how astringency types affect AsA accumulation and will provide valuable information for further investigation on AsA content variation in persimmon fruit.

Transcription factors are crucial for plant growth and development. MADS-box, WRKY, NAC, and SBP-box, some of the most vital transcription factors, are not well identified in Diospyros spp., and their floral development and sex differentiation related functions remain unknown. We identified and analysed 53 MADS-box, 66 WRKY, 83 NAC, and 17 SBP-box transcription factors using the chromosomal D. oleifera genome. There were six DolSBPs identified as miR156 and miR157 targets. According to the multiple sequence alignments of Arabidopsis and D. oleifera proteins and their conserved domains and motifs, DolMADSs were divided into 23 type I and 30 type II. The DolWRKYs, DolNACs, and DolSBPs were divided into 7, 16, and 6 subgroups, respectively. It was found that one DolMADS, five DolWRKYs, one DolNAC, and four DolSBPs may promote unisexual female flowers development, while three DolMADSs, four DolWRKYs, and one DolNAC may enhance unisexual male flowers development. The functions among the different members of the same family may, thus, vary widely. The results will help to elucidate the evolution of the MADS-box, WRKY, NAC, and SBP-box genes families in D. oleifera and to determine their functions in flower development for Diospyros spp.