Receptor-like cytoplasmic kinases (RLCKs) play important roles in various developmental processes and stress responses in plants. Whereas, the detailed information of this family in cassava has not clear yet. In this study, A total of 322 MeRLCK genes were identified in the cassava genome, and they could be divided into twelve clades (Clades I-XII) according to their phylogenetic relationships. Most RLCK members in the same clade have similar characteristics and motif compositions. Over half of the RLCK s possess cis-elements in their promoters that respond to ABA, MeJA, defense reactions, and stress. Under Xpm 11 infection, the expression levels of four genes show significant changes, suggesting their involvement in Xpm 11 resistance. Two RLCK (MeRLCK 11 and MeRLCK 84) genes potentially involved in resistance to cassava bacterial blight were identified through VIGS experiments. This work laid the foundation for studying the function of the cassava RLCK genes, especially the genes related to pathogen resistance. • We have identified a total of 322 MeRLCK genes in the cassava genome, then we performed bioinformatics analysis and expression pattern analysis on these genes. • We have found Two RLCK (MeRLCK 11 and MeRLCK 84) genes potentially involved in resistance to cassava bacterial blight were identified through VIGS experiments. • This work laid the foundation for studying the function of the cassava RLCK gene, especially the genes related to pathogen resistance. [ABSTRACT FROM AUTHOR]