The Laboratory experiment carried out in the Tissue Culture Laboratory, Department of Agronomy, Faculty of Agriculture, the University of Brawijaya, Malang, Indonesia in January-March 2019. The objective of the experiment was germination performance and response to thirty genotypes of Bambara groundnut (Vigna subterranea (L.) Verdc.) for osmotic stress induced by polyethylene glycol (PEG6000) by using three concentrations (0%, 5%, and 10%). A randomized complete block design was used for this experiment with three replicates. The 10% PEG was the most suitable concentration to select Bambara groundnut lines for drought tolerance. The result showed that osmotic stress caused by a concentration of 10% PEG inhibits germination for ten genotypes and inhibits the formation of leaflets for three genotypes, these genotypes of Bambara were regarded as drought sensitive. As well as, it lessens the number of leaflets (NL), hypocotyl length stress tolerance index (HLSI), root length stress tolerance index (RLSI) and dry matter stress tolerance index (DMSI) for all genotypes which have the ability of germination in this concentration compared with control 0%. It was noted there was a difference between genotypes in water imbibition rate because of osmotic stress caused on concentrations PEG and the genotypes showed differences in drought tolerance. The genotypes BBL 1.1, BBL 5.1.1, BBL 5.2, CKB 1, PWBG 4.1.1, PWBG 5.2.1, PWBG 5.3.1, SS 1.4.1, SS 2.4.2, SS 3.4.2 and TKB 1 performed better and were classified as drought tolerant.