Wheat is an important cereal crop that often suffers from osmotic stress under various growing conditions. The objective of this study was to investigate the effects of PEG-induced osmotic stress at the phytomer level on root growth and root hair morphology of 22 hydroponically grown wheat varieties. Two treatments, 0% and 10% PEG, were imposed for 15 days duration at 20 days old wheat seedlings. PEG stress significantly reduced plant height, number of live leaves per tiller, chlorophyll content, shoot dry weights, number of root bearing phytomers and roots per tiller. By contrast, PEG stress significantly increased leaf injury scores, root dry weight, main axis length and diameter of developed roots, length and diameter and density of both first and second order lateral roots, density and length of root hairs. Principal component analysis was conducted taking three less-affected, high yielding varieties and three highly-affected, low yielding varieties under 10% PEG stress compared to control. PC1 separated PEG-treated wheat genotypes from control treated genotypes for high and positive co-efficient of density of lateral roots and root hairs, length and diameter of main axis and first order later roots and leaf injury scores indicating that these traits are associated with osmotic stress responsive root traits. PC2 separated high-yielding and tolerant wheat genotypes from low-yielding and susceptible genotypes for high co-efficient of root dry weight, density of root hairs and second order lateral roots, length of main axis and first order lateral roots indicating that these traits are associated with osmotic stress tolerance of high-yielding wheat genotypes. Evolved information of this research could be exploited for identifying osmotic stress tolerant QTL and in developing abiotic tolerant cultivars of wheat.