Lepidium campestre (L.) R. Br. of the Brassicaceae family is being developed as a novel oil crop as well as ornamental garden plant. The goal of this study was to develop an efficient shoot regeneration protocol for this species. Inducing callus formation with 2,4-D was an absolute requirement for shoot regeneration to occur. A temporary desiccation treatment was also shown to further enhance the shoot regeneration frequency. However, the composition of plant growth regulators in the shoot induction medium seems to have little effect. The optimized shoot regeneration protocol includes culturing of 5-day old cotyledons for 3 days on 1.0 mg l^-1 2,4-D, shoot regeneration on MS medium containing 0.5 mg l^-1 zeatin and a temporary desiccation treatment for 2 days, applied after 2 weeks on shoot induction medium. Using this protocol indirect adventitious shoot regeneration could be obtained at a frequency of 68.3% with 2.4 shoots per explant. The five studied accessions of L. campestre showed varied shoot regeneration frequencies, and accessions Öland and Budapest demonstrated the highest shoot regeneration responses.