The Gibberellic Acid Stimulated Arabidopsis/Gibberellin Stimulated Transcript (GASA/GAST) gene family is a group of plant-specific genes encoding cysteine-rich peptides essential to plant growth, development, and stress responses. Although GASA family genes have been identified in various plant species, their functional roles in Prunus mume are still unknown. In this study, a total of 16 PmGASA genes were identified via a genome-wide scan in Prunus mume and were grouped into three major gene clades based on the phylogenetic tree. All PmGASA proteins possessed the conserved GASA domain, consisting of 12-cysteine residues, but varied slightly in protein physiochemical properties and motif composition. With evolutionary analysis, we observed that duplications and purifying selection are major forces driving PmGASA family gene evolution. By analyzing PmGASA promoters, we detected a number of hormonal-response related cis-elements and constructed a putative transcriptional regulatory network for PmGASAs. To further understand the functional role of PmGASA genes, we analyzed the expression patterns of PmGASAs across different organs and during various biological processes. The expression analysis revealed the functional implication of PmGASA gene members in gibberellic acid-, abscisic acid-, and auxin-signaling, and during the progression of floral bud break in P. mume. To summarize, these findings provide a comprehensive understanding of GASA family genes in P. mume and offer a theoretical basis for future research on the functional characterization of GASA genes in other woody perennials.

The CCD gene family plays a crucial role in the cleavage of carotenoids, converting them into apocarotenoids. This process not only impacts the physiology and development of plants but also enhances their tolerance toward different stresses. However, the character of the PmCCD gene family and its role in ornamental woody Prunus mume remain unclear. Here, ten non-redundant PmCCD genes were identified from the P. mume genome, and their physicochemical characteristics were predicted. According to the phylogenetic tree, PmCCD proteins were classified into six subfamilies: CCD1, CCD4, CCD7, CCD8, NCED and CCD-like. The same subfamily possessed similar gene structural patterns and numbers of conserved motifs. Ten PmCCD genes were concentrated on three chromosomes. PmCCD genes exhibited interspecific collinearity with P. armeniaca and P. persica. Additionally, PmCCD genes had obvious specificity in different tissues and varieties. Compared with white-flowered ‘ZLE’, PmCCD1 and PmCCD4 genes were low-expressed in ‘HJH’ with yellow petals, which suggested PmCCD1 and PmCCD4 might be related to the formation of yellow flowers in P. mume. Nine PmCCD genes could respond to NaCl or PEG treatments. These genes might play a crucial role in salt and drought resistance in P. mume. Moreover, PmVAR3 and PmSAT3/5 interacted with PmCCD4 protein in yeast and tobacco leaf cells. This study laid a foundation for exploring the role of the PmCCD gene family in flower coloration and stress response in P. mume.

Callose is an important polysaccharide composed of beta-1,3-glucans and is widely implicated in plant development and defense responses. Callose synthesis is mainly catalyzed by a family of callose synthases, also known as glucan synthase-like (GSL) enzymes. Despite the fact that GSL family genes were studied in a few plant species, their functional roles have not been fully understood in woody perennials. In this study, we identified total of 84 GSL genes in seven plant species and classified them into six phylogenetic clades. An evolutionary analysis revealed different modes of duplication driving the expansion of GSL family genes in monocot and dicot species, with strong purifying selection constraining the protein evolution. We further examined the gene structure, protein sequences, and physiochemical properties of 11 GSL enzymes in Prunus mume and observed strong sequence conservation within the functional domain of PmGSL proteins. However, the exon–intron distribution and protein motif composition are less conservative among PmGSL genes. With a promoter analysis, we detected abundant hormonal responsive cis-acting elements and we inferred the putative transcription factors regulating PmGSLs. To further understand the function of GSL family genes, we analyzed their expression patterns across different tissues, and during the process of floral bud development, pathogen infection, and hormonal responses in Prunus species and identified multiple GSL gene members possibly implicated in the callose deposition associated with bud dormancy cycling, pathogen infection, and hormone signaling. In summary, our study provides a comprehensive understanding of GSL family genes in Prunus species and has laid the foundation for future functional research of callose synthase genes in perennial trees.

Sugars play a pivotal regulatory role in floral bud dormancy release in Prunus mume, a process that critically determines subsequent flowering time. However, the precise molecular mechanisms linking sugar metabolism to this developmental transition remain poorly understood. To address this gap, we integrated physiological profiling and transcriptomic sequencing using two cultivars with contrasting flowering phenologies: the early-flowering ‘Chaotang Gongfen’ (CTGF) and the late-flowering ‘Shichu Jin’ (SCJ). Exogenous sugar treatments were applied separately to floral buds of the cultivar ‘Yilian’ to evaluate the effect of sugars on dormancy release. During dormancy release, glucose and sucrose contents increased progressively and showed significant positive correlations with bud break rates in both CTGF and SCJ (r > 0.75). Consistently, exogenous application of glucose and sucrose significantly accelerated bud break in ‘Yilian’, whereas mannose exhibited an inhibitory effect. Transcriptome analysis of CTGF and SCJ revealed significant enrichment of starch and sucrose metabolism, hormone signal transduction, and stress-responsive pathways. Key metabolic genes, notably the α-amylase gene PmAMY1-2 and the cell wall invertase genes PmCWINV1/4, were upregulated during this transition. Weighted gene co-expression network analysis (WGCNA) further identified PmFRK4, PmSUS6, and the aforementioned invertases as candidate genes within a sugar-associated regulatory module. Collectively, these findings support a model in which glucose and sucrose accumulation promotes endodormancy release via the transcriptional activation of starch and sucrose catabolic pathways. This study provides a theoretical framework for deciphering dormancy regulation in woody perennials and offers potential targets for the precise manipulation of flowering time.