Background: The Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) subfamily, a specialized group within receptor-like kinases (RLKs), was initially identified in C. roseus cell cultures. CrRLK1L plays an important role in the growth, development and stress response of plants. Although CrRLK1L genes have been characterized across multiple plant species, their biological and genetic functions in potatoes (Solanum tuberosum) remains poorly elucidated. Methods: a genome-wide investigation, phylogenetic analysis, chromosome localization, exon–intron structure, conserved motifs, stress-responsive cis-elements, tissue-specific expression patterns, and their effects on pathogen associated molecular patterns (PAMPs) induced reactive oxygen species (ROS) production were analyzed. Results: A total of 29 CrRLK1L genes were identified in the S. tuberosum genome, unevenly distributed across 10 chromosomes and divided into three groups. Tissue-specific expression analysis revealed seven genes highly expressed in all tissues, while CrRLK1L13 was specific to stamens and flowers. Under stress conditions (mannitol, salt, hormone, and heat), StCrRLK1L genes exhibited diverse expression patterns. Functional characterization in Nicotiana benthamiana identified seven ROS suppressors and four ROS enhancers, implicating their roles in PAMP-triggered immunity. Conclusions: This study provides valuable insights into the StCrRLK1L gene family, enhancing our understanding of its functions, particularly in plant innate immunity.