The ERF subfamily, a significant part of the APETALA2/ethylene-responsive element-binding factor (AP2/ERF) transcription family, plays a crucial role in plant growth, development, and stress responses. Despite its importance, research on this gene family in sweet cherry (Prunus avium L.) is limited. This study identified and analyzed the sweet cherry ERF subfamily in terms of classification, physicochemical properties, structural characteristics, chromosome distribution, gene replication and collinearity, Cis-acting elements, and potential protein interactions. Preliminary investigations of transcription during fruit cracking and normal development were also conducted. Fifty ERFs (PatiERF1~50) were identified, distributed unevenly across eight chromosomes and classified into ten groups with nineteen conserved motifs. Collinearity analysis with other plant species revealed homology, with the highest number of ERF orthologous genes found in apple (Malus domestica L.). Cis-acting elements, particularly abscisic acid response factor, were abundant in PatiERF promoters. Weighted gene co-expression network analysis (WGCNA) and quantitative real-time PCR (RT-qPCR) analysis indicated the involvement of PatiERFs in sweet cherry fruit development and cracking, and nine and four significant candidates related to these processes were speculated, respectively. Furthermore, four other classes of transcription factors (TFs), namely MYB, GRAS, BHLH, and BZIP, as well as 23 structure genes, were predicted to have co-expression and interaction relationships with PatiERFs during fruit development. This suggests their potential synergistic regulation with ERFs in the cherry fruit development process. Our study represents the first comprehensive genome-wide analysis of the ERF subfamily in sweet cherry, laying a crucial foundation for a deeper understanding of the molecular mechanisms correlated with fruit growth, development, and cracking mediated by ERF genes.

Nitrogen (N) is the most important nutrient for plant growth and development. However, the mechanisms by which the form and supply of N regulate the growth and N utilization of cherry rootstock are unclear at present. We investigated the effects of different N supply levels and N forms on the growth, N uptake, assimilation and distribution, and photosynthetic N use efficiency (PNUE) of Gisela 6 cherry rootstock seedlings. The results showed that a high N level and a single supply of either nitrate N or ammonium N hindered N uptake and assimilation, increased photosynthetic limitation, reduced PNUE and ¹⁵ N use efficiency, and inhibited cherry rootstock growth. Further experiments showed that a mixed supply of nitrate N and ammonium N maintained high transcription levels of nitrate and ammonium transporters as well as N metabolism enzyme activities, thereby increasing the net inflow rates of NO ₃ ⁻ and NH ₄ ⁺ into roots and the soluble protein content of leaves. In addition, a mixed N supply reduced oxidative damage to leaves by maintaining an appropriate nitrate/ammonium ratio, increased the proportion of leaf N allocated to photosynthetic N, decreased leaf cell wall thickness, and enhanced stomatal conductance, mesophyll conductance, and the maximum carboxylation efficiency. This resulted in reduced leaf photosynthetic limitation, increased leaf net photosynthetic rate and PNUE, and ultimately enhanced the growth of Gisela 6 cherry rootstock seedlings. Our results provide a basis for optimizing N management strategies in cherry cultivation.

Exogenous glucose functions not only as a carbon source but also as a key signaling molecule involved in regulating root development and metabolism in plants. To elucidate the molecular mechanisms underlying this response in cherry rootstock (Prunus cerasus), we performed RNA-seq on lateral roots collected at 0, 6, 12, 24, 48, and 72 h after glucose treatment. Transcriptome profiling revealed a dynamic and sustained transcriptional reprogramming, with a total of 461 differentially expressed genes (DEGs) consistently altered across all post-treatment time points relative to the control (T0). Weighted gene co-expression network analysis identified five modules strongly correlated with glucose exposure, notably enriched for genes involved in nitrogen, carbon, and sulfur metabolism. Functional enrichment analyses further revealed a pronounced overrepresentation of pathways associated with nutrient utilization, as well as carbon fixation, glycolysis, amino acid biosynthesis, and stress-responsive processes such as glutathione metabolism and MAPK signaling. Intriguingly, key transcription factors and signaling components were consistently co-enriched across multiple functional categories, suggesting the presence of a tightly coordinated regulatory network that links sugar sensing to metabolic reprogramming, redox homeostasis, and developmental plasticity. Notably, glucose treatment induced both activation and repression of nitrogen-related genes in distinct co-expression modules, indicating fine-tuned modulation of nutrient uptake in response to carbon availability. Together, these findings suggest that exogenous glucose triggers a systems-level shift in root physiology, coordinating primary metabolism with stress adaptation and growth regulation through tightly interconnected carbon–nitrogen–sulfur metabolic circuits.