Awareness of genetic diversity in wheat is crucial for the selection of parental genotypes that could yield heterotic combinations. So, the purpose of this study is to assess the genetic diversity of forty wheat advance lines chosen from diverse geographic regions. Field experimentation was carried out at Wheat and barley Section, Department of Genetics and Plant Breeding, CCS HAU, Hisar during Rabi, 2019-20 with three replications. The data were recorded for 15 different morpho-physiological parameters. The results revealed that first seven PCs with Eigen values >1 contributed to 75.54 % of the variance among genotypes. Among these PC1 contributes to the greatest variation (23.39%) followed by PC2 (12.10%) and PC3 (10.83%). Maximum positive component loading towards PC1 was contributed by NDVI, SL/S, ET/m, G/S, DH and DM. Maximum number of positive component loading for PC2 was contributed by HI, TGW and GY/P. Cluster analysis divides the experimental material into five major clusters with cluster Ⅱ, Ⅲ and Ⅳ possessed nine genotypes in each. Higher inter-cluster distance was observed than intra-cluster distance. The maximum intra-cluster distance was observed in cluster Ⅲ (49.83) and the maximum inter cluster distance was observed between cluster Ⅰ and cluster V (88.22). Based on current study, it can be suggested that the important morpho-physiological traits could serve as effective phenotypic markers for the selection of high-yielding wheat varieties, while the genotypes from clusters I and V with the most diverse parents could be used in the hybridization programme to develop the most promising hybrids.