Innate mobility of plants restrained their potential to circumvent heat stress and required them to withstand stress through inherent defence abilities in which heat shock proteins (HSPs) played essential roles by acting as chaperones. HSPs mediate the disaggregation of denatured proteins leading to protein homeostasis and their expression is regulated by heat shock transcription factors (HSFs). Prosopis cineraria is a phreatophyte distributed across arid and semi-arid regions of India and can tolerate high temperatures due to its adaptive physiological and biochemical mechanisms. Therefore, P. cineraria represents a repository of genes for abiotic stress tolerance. Two months old P. cineraria plants were subjected to heat stress at two different high temperatures and their genome-wide transcriptome sequencing was conducted to identify and validate the expression of HSPs with real-time qPCR. It was noted that small HSPs (sHSPs) including HSP15.7, HSP17.9, HSP18.5, HSP22.7 and HSP26.5 are major chaperone proteins upregulated immediately after heat stress. A few HSFs including HSFA3a, HSFA6b and HSFA7a were upregulated which control the expression of HSPs. These results provide new insight into good candidate genes for crop improvement. Identified genes were cloned from P. cineraria and mobilized in pCAMBIA1300-UbiP-NosT for plant transformation. To elucidate the function of sHSPs, we constructed recombinant pRSETA vectors and transformed into E. coli BL21 (DE3) pLysS. These sHSPs will be induced in E. coli and purified His-tagged sHSPs will be harvested. The interaction studies of purified sHSPs with their interactome will be carried out by mass spectrometry.