ALMT (aluminum-activated malate transporter) proteins play crucial roles in the transport of organic acids and have significant implications for plant stress responses and development. While extensively studied in some plants, the characteristics and functional divergence of the ALMT gene family have not yet been thoroughly explored in Prunus mume and some other Rosaceae species. In this study, we systematically analyzed the ALMT gene family across nine Rosaceae species to explore their evolutionary relationships, structural characteristics, and functional roles. A total of 138 ALMT genes were identified and categorized into four groups based on a phylogenetic analysis. The motif analysis confirmed the accuracy of the phylogenetic grouping. The collinearity analysis indicated that whole-genome duplication events were the primary drivers of ALMT gene expansion in these species. Furthermore, the cis-acting element analysis revealed diverse regulatory elements associated with environmental responses, including abscisic acid, light, and jasmonic acid. The gene expression correlation analysis showed that PmALMT1 is primarily associated with malic acid accumulation, whereas PmALMT8 is related to citric acid accumulation. Further transient expression in Nicotiana benthamiana validated the above conclusion. This comprehensive analysis provides valuable insights into the evolution, function, and regulation of the ALMT gene family in Rosaceae species.

KANADI (KAN) transcription factors are pivotal regulators of lateral organ polarity establishment in plants. Although extensively studied in herbaceous plants, the role of KAN genes in woody plant development remains unclear. This study conducts the first comprehensive analysis of 26 PmKAN genes in Prunus mume, elucidating their evolutionary trajectories, structural configurations, tissue-specific expression patterns and potential roles in root and fruit development. Phylogenetic analysis of four Rosaceae species and Arabidopsis thaliana clustered these PmKANs into five subfamilies, with conserved motif patterns supporting this classification. Chromosomal localization revealed that all PmKAN members are distributed across eight chromosomes, with tandem duplications events and syntenic relationships indicating functional diversification driven by gene family expansion. Cis-regulatory element analysis identified light-responsive, hormone-associated, stress-related, and developmental motifs, suggesting PmKAN genes are involved in regulating plant physiological processes and development. The qRT-PCR analysis revealed tissue-specific expression heterogeneity among PmKAN genes, with markedly elevated expression particularly observed in roots and fruits. Further expression profiling across fruit developmental stages suggests potential stage-specific functional divergence of PmKAN genes during fruit development. This study provides a theoretical foundation for further investigating the evolutionary relationships and molecular regulatory mechanisms of the PmKAN gene family.

Organic acid accumulation is a key determinant of fruit flavor and processing quality in Prunus mume, yet its underlying molecular mechanisms remain poorly understood. P-type ATPases are membrane proteins that drive the active transport of ions and molecules via ATP hydrolysis and play important roles in organic acid accumulation However, their genome-wide features and functions in P. mume remain poorly understood. Here, 49 P-type ATPase genes were identified from the P. mume pan-genome, including 29 core, 6 near-core, 11 dispensable, and 3 private genes, and classified into six subfamilies. Dispersed and whole-genome duplication were the major force driving gene family expansion, and most genes experienced strong negative selection. Expression profiling identified the P3A-type gene PmAHA3 as highly expressed in fruit, with vacuolar membrane localization. Both transient RNAi silencing in fruit and CRISPR/Cas9 knockout in callus significantly reduced the contents of malic and citric acid, demonstrating their roles in organic acid accumulation. These results provide insight into the evolution and function of P-type ATPases, offering potential targets for improving fruit quality.

Potassium (K⁺) is a vital macronutrient for plant growth and stress resilience, with KT/HAK/KUP transporters playing a central role in its homeostasis. Although these transporters are known to influence photosynthesis, the molecular mechanisms by which fertilization promotes assimilate accumulation in peach crops remain poorly understood. In this study, 17 PpHAK genes were identified based on the peach genome and classified into four distinct clades through phylogenetic analysis, a classification further supported by conserved gene structures and motifs. Interspecific collinearity analysis revealed that transporters are highly conserved among Rosaceae species. Physiological measurements demonstrated that foliar application significantly enhanced photosynthetic capacity, as evidenced by a 33% increase in net photosynthetic rate (Pn) and improved photoelectron yield (Y(II)). At the same time, the transcript levels of the transporters PpHAK1, PpHAK5, and PpHAK9 were significantly upregulated, as confirmed by quantitative real-time RT-PCR (qRT-PCR) analysis. Furthermore, the expression of genes involved in sugar metabolism and transport, particularly PpPLT5-1, was significantly induced. Collectively, these results indicate that foliar K⁺ application enhances photosynthesis and promotes assimilate accumulation by modulating the expression of both K⁺ and sugar transporters. These findings offer a theoretical basis for optimizing nutrient management to improve fruit quality in stone fruit production.