Drought is one of the important abiotic stresses affecting the all growth stages of wheat (Triticum aestivum L.) crop; it becomes more severe during the flowering and grain-filling phases (terminal drought) and can cause up to 78% yield loss. Across the world various breeding work is underway to develop drought tolerant varieties. Being a complex trait and governed by many genes it is highly affected by the environment hence precise phenotyping is prerequisite. A total 500 genotypes were evaluated for post anthesis (Terminal) drought at MACS- ARI, Hol using non-destructive image-based shoot phenotyping. The field experiment was conducted in two different environments to study genotype-by-environment (G×E) interaction. One is under well-irrigated (control) and another under terminal drought by stopping irrigation after the anthesis. The population was screened by using Infrared (IR) imaging and Relative Water Content (RWC) and other genetic traits to study significant physiological responses. Additionally, Seedling stage RSA (Root system architecture) was investigated to study its relation to drought tolerance. Based on the data obtained from this study, it was found that relative water content (RWC) during grain filling was found to be positively and significantly correlated with the spike characters of genotype under both moisture stress and control conditions. The grain appearance trait like grain area found to be positively and significantly correlated with grain yield and stress tolerance index (STI) under drought conditions. The spikelet’s per spike was negatively correlated with grain area. One of the important outcomes of this study are image-based techniques that are standardized and can be successfully used for seminal root trait phenotyping.