A novel in vitro transformation of Lepidium draba L. using rapid direct shoot regeneration

The present research is carried out to study Lepidium draba gene transformation for the first time, using direct shoot explants. As a prerequisite for gene transformation, the regeneration conditions in L. draba were optimized. We achieved an efficient and reproducible protocol for successful direct shoot regeneration without intervening callus formation. The results indicate that L. draba is the insistent species of Brassicaceae in direct shoot regeneration. Various explants of L. draba were genetically transformed with different strains of Agrobacterium tumefaciens, viz., LBA4404, GV3850, GV3101, and EHA105, using the vector pBI121. Expression of GUS reporter protein was assayed by histochemical staining. In addition, using the PCR method with specific primers proved the integration of GUS gene into the plants. The highest transformation efficiency was achieved with Agrobacterium strain GV3850. Moreover, we found that infected hypocotyl and root explants of L. draba interestingly yielded higher transformation efficiency, so that in hypocotyls on average exceeded 70% of the explants. This study showed that L. draba, in addition to the numerous desirable traits, has a high potential for gene transfer.