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Ca²⁺ serves as a ubiquitous second messenger regulating several aspects of plant growth and development. A group of unique calcium sensor proteins, calcineurin B-like (CBL), interact with CBL-interacting protein kinases (CIPKs) to decode the Ca²⁺ signature inside the cell. Although CBL-CIPK signaling toolkit has been shown to play significant roles in the responses to numerous stresses in different plants, the information about pineapple CBL-CIPK remains obscure. In the present study, a total of eight <i>Ac</i>CBL and 21 <i>Ac</i>CIPK genes were identified genome-wide in pineapple. The identified genes were renamed on the basis of gene ID in ascending order and phylogenetic analysis divided into five groups. Transcriptomic data analysis showed that <i>Ac</i>CBL and <i>Ac</i>CIPK genes were expressed differentially in different tissues. Further, the expression analysis of <i>Ac</i>CBL1 in different tissues showed significant changes under various abiotic stimuli. Additionally, the ectopic expression of <i>Ac</i>CBL1 in <i>Arabidopsis</i> resulted in enhanced tolerance to salinity, osmotic, and fungal stress. The present study revealed the crucial contribution of the CBL-CIPK gene in various biological and physiological processes in pineapple.