The present study was conducted for karyomorphological investigation in 15 lines of two species of Hordeum. All the lines showed a diploid chromosome number of 14. Differences were observed regarding chromosome length, total chromatin length (TCL), type etc. among the 15 entries. The longest (11.06 m) chromosomes were observed in BTON-10 while the shortest (4.23 m) in BEL-4 and BEL-36 of Hordeum vulgare. Extra large chromosomes were found in BEL-4 (1 pair) and BTON-10 (5 pairs) of H. vulgare. Large type chromosomes were found in BHV-105, BTON-10 and conquest of H. vulgare. Medium type and relatively short type chromosomes were absent in BEL-36, BHV-1 and BTON-10 of H. vulgare, respectively. More metacentric chromosomes (7 pairs) were found in BHV-1 of H. vulgare. Maximum chromatin length (70.21 m) was also found in BTON-10 and minimum (40.96 m) in BEL-36 of H. vulgare. Maximum (47.76%) and minimum (39.88%) total fractional percentage (TF %) were found in BHV-1 and API-19 of H. vulgare, respectively. Each line has a specific chromosomal morphology which might be responsible for the variations of Hordeum species.