Characterization of CBL-CIPK signaling networks and their response to abiotic stress in sugarcane

Calcineurin B-like proteins (CBLs) perceive calcium signals triggered by abiotic stress and interact with CBLinteracting protein kinases (CIPKs) to form a complex signal network. This study identified 21 SsCBL and 89 SsCIPK genes in Saccharum spontaneum, and 90 ScCBL and 367 ScCIPK genes in the sugarcane cultivar ZZ1. Phylogenetic analysis classified CBL genes into three groups and CIPK genes into twenty-five groups, with wholegenome duplication events promoting their expansion in sugarcane. RNA-seq analysis revealed their involvement in abiotic stress responses through ABA, JA, and SA pathways. Four ScCBLs and eight ScCIPKs were cloned from ZZ1. Three CBL-CIPK interactions were detected using a yeast two-hybrid system and Firefly luciferase complementation imaging, showing CBLs as membrane proteins and CIPKs as nuclear proteins. Spatial expression profiles indicate these genes are expressed in various tissues, with the highest expression in roots. Gene expression analyses suggested that CBL-CIPK signaling networks are involved in responses to drought, salt, and reactive oxygen species, possibly through Ca2+-induced hormone pathways. These findings establish three CBLCIPK signaling networks responding to abiotic stress, providing a molecular basis for improving sugarcane stress resistance.