Genome-wide re-identification of the CrRLK1L family in soybean and functional characterization of GmCrRLK1L2 in salt stress response

The Catharanthus roseus receptor-like kinase 1-like (CrRLK1L), a member of the plant receptor-like kinase family, plays crucial roles in plant development and stress response. Despite previous screenings of soybean CrRLK1Ls, their specific function in stress response remains unclear. Utilizing recent soybean whole genome sequencing data, a comprehensive genome-wide analysis was performed to identify CrRLK1Ls in soybean. This study identified 45 CrRLK1L members in soybean, with CrRLK1L2 singled out for further functional investigation. Phylogenetic analysis categorized GmCrRLK1L genes into six classes, some of which exhibited homologs in Arabidopsis. Gene structure and motif analysis revealed a predominance of intron-lacking and relatively conserved GmCrRLK1L genes, with conserved motif patterns across members. Collinearity analysis indicated that both gene tandem and segmental duplication events drove the expansion of the GmCrRLK1L gene family, with segmental duplications being the primary driver. Expression profiling under salt treatment revealed significant up or downregulation of several GmCrRLK1L genes. Overexpression of GmCrRLK1L2 in soybean resulted in reduced plant height and salt tolerance, while knockout did not affect salt tolerance, suggesting potential genetic compensations by other genes. RNA-seq data showed that GmCrRLK1L2 overexpression influenced the expression of metabolic pathways and salt stress-responsive genes, including ACS3, ACO1, PER64, PER53, SAT3, and BCAT2. These findings lay the groundwork for elucidating the biological roles of GmCrRLK1Ls in soybean growth, development, and response to salinity stress.