Barley is the fourth most important crop in the world. Development of a regeneration
system using immature embryos is both time consuming and laborious. The present study was initiated
with a view to develop a regeneration system in six genotypes of Indian barley (Hordeum vulgare)
cultivars as a prerequisite to transformation. The mature embryos were excised from seeds and
cultured on MS medium supplemented with high and low concentrations of cytokinins and auxins
respectively. The MS medium containing 3 mg/L N6-benzylaminopurine (BA) and 0.5 mg/L 2,4-
dichlorophenoxyacetic acid (2,4-D) was found to be the most effective for multiple shoot formation
in HOR7231 cultivar that could produce 12 shoots per explant. The other cultivars HOR4409 and
HOR3844 produced a minimum number of adventitious shoots (1.33 and 1.67 respectively) on MS
medium supplemented with 1 mg/L BA and 0.3 mg/L 2,4-D. The elongated shoots were separated
and successfully rooted on MS medium containing 1 mg/L indole-3-acetic acid (IAA). The response
of different barley cultivars was found to be varying with respect to multiple shoot production. This
is the first report of multiple shoot induction and plantlet regeneration in Indian cultivar of barley
which would be useful for genetic transformation.