Background: Soybean, Glycine max has gained paramount importance in the agriculture of this country. Contribution of soybean in the oilseed basket of the world has been maximum. In India, of late, the cultivation and utilization of soybean has picked up rapidly. Insect infestation is considered to be the single largest impediment among several abiotic and biotic factors. Among them, stem fly, Melanagromyza sojae (Zehnter) is the number one pest of soybean in southern India causing infestation up to a level of 75 to 90 per cent. Hence, the study was carried out to identify the potential donar of resistance in soybean to stem fly. Methods: The trial was carried out at Main Agricultural Research Station, University of Agricultural Sciences, Dharwad, Karnataka in randomized block design (RBD) with two replications during kharif 2022 and 2023. Twenty-one soybean genotypes were screened against stem fly, M. sojae with one check variety JS 335 under in-vivo condition. Observation on seedling mortality was recorded at weekly intervals up to 30 days after germination. Stem fly incidence and stem tunneling due to stem fly at flowering and harvesting stage were recorded and percentage was worked out. Result: The pooled data of two years concluded that, the soybean genotypes DSb 34, DLSb 1, KDS 1096 and NRC 197 could be found under resistant genotype, while DLSb 3, JS 23-09, MAUS 795, NRC 195, NRC 196 and MAUS 791 as moderately resistant. JS 23-03, JS 335, VLS 102, DSb 21, RSC 11-42 and RVS 13-20 were categorized as low resistant genotypes as against SL 1282, RSC 11-35 and PS 1682 as susceptible and CAUMS-2 and AS 24 genotypes as highly susceptible irrespective of the crop growth stages.