The fruit skin pubescence of Prunus persica is an economically important characteristic and comprises the classification criteria. The mapping and identification of a complete linkage marker to the fruit skin trichome trait locus of peach fruit are critical for the molecular marker-assisted selection for peach/nectarine. In this study, the BC₁ population was constructed from the parents “Zhongyou No. 4”, the recurrent parent, and “Baihuashanbitao”, the non-recurrent parent. Based on the 38 BC₁ individuals’ phenotypes and their genotyping using next-generation sequencing, the G (Glabrous skin) locus of the gene was first identified between 14.099 and 16.721 Mb on chromosome 5. Using other individuals of this population, the gene was fine-mapped in the range of 481 kb with SNP markers. Based on the resequencing data of other cultivars (lines), the candidate SNP in the gene Prupe.5G196400 was obtained. Subsequently, the SNP marker was designed and applied to natural and hybrid peach populations. Via genotyping analysis, we confirmed co-segregation between the peach/nectarine phenotype, which was used in the identification of peach or nectarine with 100% accuracy.

Malic acid is the major organic acid in loquat fruit, contributing to the sourness of fruit and affecting fruit flavor. However, the transcriptional regulation of malic acid in loquat is not well understood. Here, we discovered a MADS-box transcription factor (TF), EjAGL18, that regulated malic acid accumulation in loquat. EjAGL18 is a nucleus-localized TF without transcriptional activity. The expression of EjAGL18 increased during fruit ripening, opposite to the accumulation pattern of malic acid in loquat. The transient overexpression of EjAGL18 in loquat fruit downregulated malic acid accumulation and the transcriptional level of the tonoplast dicarboxylate transporter EjtDT1. Conversely, silencing EjAGL18 in loquat fruit upregulated the malic acid content and EjtDT1 expression level. Dual-luciferase assays and yeast one-hybrid experiments further confirmed that EjAGL18 could bind to the promoter of EjtDT1 and repress its transcriptions. Furthermore, the transient overexpression of EjtDT1 in loquat fruit increased the malic acid content. These results revealed that EjAGL18 negatively regulates malic acid content by repressing EjtDT1 in loquat. This study broadens the understanding of the MADS-box TF’s regulatory mechanisms in malic acid and provides new insights into fruit flavor improvement in loquat.