In vitro propagation of onobrychis germanicopolitana HUB.-MOR. ET SIMON

In this study, in vitro propagation of Onobrychis germanicopolitana HUB.-MOR. ET SIMON was carried out, a species identified as an endangered plant according to the Red List of the International Union for Conservation of Nature. O. germanicopolitana is a local endemic plant that displays a natural growth on Çankırı gypsum hills located in the A4 square according to the Grid system created by Davis. Shoot tips and internodes were used as an explant source. For in vitro propagation purpose, to 1) Murashige and Skoog (MS) and 2) Gamborg's B5 basal media were added 1) 6-benzylaminopurine (BAP) (0 mg/L, 0.5 mg/L, 1 mg/L) and 2) Kinetin (KIN) (0 mg/L, 0.5 mg/L, 1 mg/L) as a source of cytokinin and, 3) Indole-3-butyric acid (IBA) (0 mg/L, 0.25 mg/L, 0.5 mg/L) and 4) 2,4-Dichlorophenoxy acetic acid (2,4-D) (0 mg/L, 0.25 mg/L, 0.5 mg/L) as well as an auxin source. A total of 36 different plant growth regulator combinations were obtained and added to the basal media. Gelrite gelling agent (2.1g/L) was added to the media and 30 g/L sucrose was also as a source of carbon. Due to the manifestation of infections occurring in the initial or advanced stages of the study, the experiment was repeated twice and a more appropriate sterilization protocol for the plant was developed. Plant regeneration could not be achieved, and as a result, non-organogenic calli were obtained. The best callus formation belonged to MS medium containing 1 mg/L BAP + 0.25 mg/L IBA, 0.5 mg/L BAP + 0.5 mg/L 2,4-D, 1 mg/L BAP + 0.5 mg/L 2,4-D, 0.5 mg/ L KIN + 0.5 mg/L 2,4-D plant growth regulator combinations. The fact that no in vitro propagation studies of this specific endemic species have been carried out has a unique value in terms of offering the groundwork for future studies.