Root rot disease of sesame caused by Macrophomina phaseolina (Tassi.) Gold, a screening programme was done in artificial (in vitro) and sick plot (in vivo) condition to identify the resistant genotype. In artificial screening method, the seedlings grown in autoclaved roll towel were dipped in a suspension of Macrophomina phaseolina culture for one minute with an up-down movement and again placed in roll towel at 35±10C for ten days before observing for the reaction. In sick plot condition, the genotypes were grown along the susceptible cultivars after every 4 rows of test entries. The genotypes namely ORM 7, ORM 14 and ORM 17 were identified as resistant to root rot which could be used as cultivars or used in hybridization programme. The 36 F1’s obtained by crossing the three resistant genotypes (as testers) with twelve susceptible genotypes (as lines) found to be susceptible which indicated that the resistance trait may be governed by the recessive gene.