Establishing an efficient explant superficial sterilization protocol for in vitro micropropagation of bear’s garlic ( Allium ursinum L.)

Abstract

Introduction: Allium ursinum L. has a commercial value due to its high contents of bio-active compounds and mild, garlic-like taste. In vitro culture played an important role in obtaining Allium species with the desired characteristics and in the production of healthy reproductive material. Objective: Developing an effective method of sterilization bear’s garlic bulbs. Methods: To obtain sterile shoots of garlic several methods of sterilization involving such factors as ACE, H 2 O 2 , HgCl 2 , and UV-C were tested. Results: In order to obtain sterile shoots of bear’s garlic, several sterilization procedures were tested. The best procedure was based on a two-step disinfection, where the whole onions were treated with ethanol and H 2 O 2 for 20 min. Thereafter, the isolated apical buds were sterilized in ACE for 10 min, rinsed in double-distilled water and transferred onto MS medium for growing. Up to 95% of the inoculated explants formed shoots, which were sub-cultured on MS with 4 mg dm −3 BAP in order to enable further propagation. Conclusion: After optimization and stabilization, this procedure may become the basic concept of a proper and reliable propagation method of bear’s garlic on commercial scale.