In vitro culture of Zingeria biebersteiniana (Claus) P. Smirn. as a method for conservation and restoration of genetic diversity

Abstract

Aim. The aim of the work was to determine the conditions for development of in vitro culture, induction of callus formation, and long-term tissue culture of Zingeria biebersteiniana (Claus) P. Smirn. Methods. In vitro clonal propagation, tissue culture techniques. Results. The seed germination rate was found to increase significantly after long-term cold stratification. The protocol for seed sterilization was developed, which yielded 57.3% of aseptic plants. Collections of in vitro and pot cultured plants were created. Experiments on the adaptation of in vitro propagated plants to pot culture conditions revealed a high level of their survival. The optimal medium for in vitro clonal propagation was MS, supplemented with 0.1 mg/L NAA; while the most effective media for induction of callus formation and for long-term tissue culture was B5 supplemented with 2 mg/L 2,4-D and 0.1 mg/L BAP. Conclusions. The protocols and conditions for seed germination, in vitro clonal propagation, induction of callus formation, as well as long-term tissue culture of Z. biebersteiniana have been developed. The developed techniques of in vitro culture can be used for conservation and restoration of genetic diversity of the species, as well as to obtain sufficient plant material for further studies.