Abiotic stress plays a substantial role in morphological and physiological changes of plants leading to yield loss. Drought and salinity tolerance (DST) is a zinc finger protein serving an indispensable function in stress resilience through modulation of stomatal closure. Yet, no known report of their roles in sugarcane exists. This research presents a comprehensive study on isolation, characterization and identification of ShDST genes by employing a combination of in silico and experimental methods. The isolated full-length coding region of the ShDST gene transcribed a single open reading frame of 810 base pairs encoding 269 amino acids possessing a molecular weight of 27.7 kilo Daltons and an isoelectric point (Pi) of 7.70. Subcellular localization studies highlighted the presence of ShDST within the nucleus. A molecular docking study revealed a significant affinity between ShDST and Hydrogen peroxide suggesting a potential interaction and implication for oxidative stress response. In the course of this investigation, a total of 21 genes were identified utilizing Hidden Markov Models, for comprehensive genome-wide identification, contributing to a deeper understanding of genetic diversity and variation within the Saccharum species. Comparative evolutionary analysis, including phylogeny, collinearity, and synteny analysis was employed to compare the Saccharum DST gene families with various crops under Poaceae family. A heat map representation of Cis-element occurrence further elucidated the distribution and abundance of the regulatory elements, providing a valuable tool for assessing potential transcriptional activity and regulatory significance. This first systematic analysis of DST families in sugarcane provides a basis for further functional study and provides valuable insights into the potential to enhance crop breeding practices for improved sustainability and adaptability to challenging environmental conditions in sugarcane.