Tissue-specific expression and genome-wide analysis of RALF gene family in pea (Pisum sativum L.) under different stresses

• Genes from RALF family are promising targets for improving abiotic stress tolerance in crops • This work analysed structural features and expression patterns of RALF genes under a range of stresses • Reported data lays the groundwork for developing climate-resilient pea varieties. The rapid alkalinization factor (RALF) gene family plays a crucial role in plant growth, development, and stress responses, yet its characterization in pea ( Pisum sativum L.), an agronomically important legume, remains limited. This study identified eight PsRALF genes in the pea genome, phylogenetically clustered into 8 distinct groups, and analyzed their structural features, evolutionary relationships, and expression patterns under various abiotic stresses. Chromosomal localization and collinearity analyses revealed both conserved and lineage-specific duplication events, while promoter analysis identified stress- and hormone-responsive cis -elements, suggesting their roles in stress adaptation. Under abiotic stress conditions such as drought, salinity, and aluminum toxicity, PsRALF2 exhibits significant upregulation, highlighting the potential of this gene as a stress-responsive candidate. Subcellular localization predictions indicate that members of this family are predominantly localized extracellularly, with one member targeted to the chloroplast, suggesting that they may possess diverse functional roles. Additionally, All analyzed PsRALFs possesed prion-like domain (PrD) features with varying prominence, suggesting their potential to drive protein aggregation or liquid-liquid phase separation (LLPS) that may mechanistically regulate signaling functions. These findings provide foundational insights into the PsRALF family, offering valuable targets for breeding climate-resilient pea varieties to enhance sustainable agriculture and meet the growing demand for plant-based proteins. Abiotic stress tolerance has been present in wild crop relatives but then lost during their domestication. As a result, all crops are extremely sensitive to climate-driven environmental constraints such as drought, salinity, flooding, and various nutritional disorders. This is specifically true for pea – an important legume crop. This work aims to offer crop breeders new valuable targets for breeding climate-resilient pea varieties to enhance sustainable agriculture and meet the growing demand for plant-based proteins, focusing on genes from the RALF (rapid alkalinization factor) family. By detailed analysis of the phylogenetic relationships, structural features, and tissue- specific and stress-responsive expression patterns undebiotci stresses of RALF genes, our work provides the first comprehensive resource on PsRALF genes, bridging a critical knowledge gap in legume stress biology and lays the groundwork for future genetic engineering or marker-assisted selection to develop climate-resilient pea varieties.