Genetic improvement along with widened crop base necessitates for the detailed understanding of the genetic diversity and population structure in wheat. The present investigation reports the discovery of a total of 182 alleles by assaying 52 simple sequence repeats (SSRs) on 40 genotypes of bread wheat. Unweighted neighbour-joining method grouped these genotypes into two main clusters. Highly heat tolerant and intermediate tolerant cultivars were grouped in the same cluster, whereas remaining genotypes, particularly sensitive ones, were assigned different cluster. Similarly, the entire population was structured into two sub-populations (K = 2), closely corresponding with the other distance-based clustering patterns. The marker-trait association was discovered for four important physiological parameters, viz. canopy temperature depression, membrane thermostability index (MSI), normalized difference vegetation index and heat susceptibility index, indicating for heat stress (HS) tolerance in wheat. Both general and mixed linear models of association studies during 2017 and 2018, revealed the association of SSR markers, wmc222 (17.60%, PV) and gwm34 (20.70%, PV) with the mean phenotypic value of MSI. Likewise, SSR markers barc183, gwm75, gwm11 and cfd7 revealed a unique relationship with four selected physiological traits. Candidate genes discovered using in silico tools had nine SSR markers within the genic regions reported to play a role in heat and drought stress responses in plants. The information generated about these genic regions may be explored further in expression studies in-vivo to impart HS tolerance in bread wheat.