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Heat shock transcription factors (HSFs) contribute significantly to thermotolerance acclimation. Here, we identified and cloned a putative HSF gene (<i>HSFA2h</i>) of 1218 nucleotide (acc. no. KP257297.1) from wheat <i>cv.</i> HD2985 using a de novo transcriptomic approach and predicted sHSP as its potential target. The expression of <i>HSFA2h</i> and its target gene (<i>HSP17</i>) was observed at the maximum level in leaf tissue under heat stress (HS), as compared to the control. The <i>HSFA2h</i>-pRI101 binary construct was mobilized in Arabidopsis, and further screening of T3 transgenic lines showed improved tolerance at an HS of 38 °C compared with wild type (WT). The expression of <i>HSFA2h</i> was observed to be 2.9- to 3.7-fold higher in different Arabidopsis transgenic lines under HS. <i>HSFA2h</i> and its target gene transcripts (<i>HSP18.2</i> in the case of Arabidopsis) were observed to be abundant in transgenic Arabidopsis plants under HS. We observed a positive correlation between the expression of <i>HSFA2h</i> and <i>HSP18.2</i> under HS. Evaluation of transgenic lines using different physio-biochemical traits linked with thermotolerance showed better performance of HS-treated transgenic Arabidopsis plants compared with WT. There is a need to further characterize the gene regulatory network (GRN) of <i>HSFA2h</i> and sHSP in order to modulate the HS tolerance of wheat and other agriculturally important crops.