Propagation of fritillaria minima via plant tissue culture

In this study, the in vitro propagation of Fritillaria minima Rix which is endemic for our country and endangered was aimed. In experiments where the bulb, unopened flower hills and flower stalks were used as explant source, the MS (with vitamin) medium was used as the nutrient medium. Callus and organogenesis formation rates of different explants of Fritillaria minima were determined by using 3% and 6% sucrose and plant growth regulators (PGR) such as NAA, IAA and BA. Surface sterilization procedures were performed by using ethyl alcohol and sodium hypochlorite (NaOCl) at different concentrations. Explants were allowed to develop at 20 ± 1 °C. 8 hours dark and 16 hours light photoperiod are applied. At the end of the first 30-day incubation, the infection rate in all bulb explants was kept at 6 %. Then, explants with no development and infections were evaluated in the category of explants that did not develop. As a result, the survival rate of the explants was determined as 92.06 %. Callus formation was observed in 25.43% of the bulb explants of the surviving explants, and bulblet formation was observed in 9.05%. At the end of the 120-day incubation period, the best results were obtained in high cytokinin medium (1 BA + 0.6 NAA + 0.4 IAA) containing 3% sucrose. Direct or indirect organogenesis was observed in 94.29% of the bulb explants whereas only callus formation was observed in the remaining 5.71% of the bulb explants. Callus formation was obtained in 37.93% when sucrose was not present in the PGR-free primary fattening medium. In the high cytokinin medium containing 3% sucrose, 40% shoot formation was observed by indirect organogenesis. The highest bulblet formation by indirect organogenesis was obtained in the low cytokinin medium containing 3% sucrose (35.29%). High cytokinin medium containing 6% sucrose was identified as the most effective medium for shoot formation by direct organogenesis (14.28%). On the other hand, it was observed that the most effective medium for the formation of the bulblet by direct organogenesis was a low cytokinin medium containing 3% sucrose (23.52%). Indirect shoot formation per explant was highest in the high cytokinin medium containing 3% sucrose (13.1), and indirect bulblet formation per explant was highest in the low cytokinin medium containing 3% sucrose (14.1). Explants with shoot and bulblet formation were rooted in medium containing ½ MS + 0.6% agar + 3% sucrose + 5 mg/L NAA + 0.5 mg/L BA. At the end of the first 30-day incubation in rooting medium, 45% of all explants were rooted while 76.25% of all explants were rooted after 60 days of incubation. No significant results were obtained from the adaptation of the whole plantlets to outdoor environment obtained from the bulb explants in vitro. In addition, no formation was observed in the experiments where unopened flower petals and flower stalks were used as explants.