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Crop: Groundnut is one of the proteins and oil-producing legume crop of arid and semi-arid regions of the world.The Objective of the study: To estimate the genotypic coefficient of variation, heritability and genetic advance for physio-biochemical and root related traits among groundnut germplasm under the irrigated condition and To put guideline to establish a breeding program for developing genotype (s) with higher yield using suitable selection criteria through genetic variability studies. Research location and season:The College of Horticulture, Hiriyur during rabi 2016-17. Experimental design: Randomized complete block design with two replications. Methodology: forty groundnut genotypes including ICRISAT based advanced breeding lines and existed cultivars were evaluated under field condition. The crop was taken care of as per the standard recommended package of practices. Data was recorded on physio-biochemical and root related parameters at different stages of crop growth. The resulted mean data was subjected to statistical analysis using Genstat 14.1 software. Results: Results revealed highly significant differences for all the traits under study indicating the presence of genetic variation among the genotypes. The characters like MSI (99.89 & 112.99%), leaf phenols (99.63 & 74.26%), seed phenols (99.06 & 73.05%) and root to shoot ratio (86.19 & 21.04%) exhibited high heritability coupled with high genetic advance over mean. Conclusion:It can be concluded that the presence of additive gene action in governing these traits. Thus, there is ample scope for improving these characters based on the direct selection from the genetic stock studied which means if these characters are subjected to any selection scheme for exploiting fixable genetic variance; a widely adopted genotype can be developed.