Micropropagation of Baskil Reverse Tulip ( Fritillaria baskilensis Behçet) via tissue culture

The aim of this study is to provide in vitro micropropagation of Fritilla baskilensis Behçet which is an endemic plant of our country and has a risk of becoming extinct. Parts of bulb scales of Fritillaria baskilensis petals of not open flowers and flower stalks were used as the explant source. In experiments, MS (with vitamins) basic nutrition medium, %3 and %6 sucrose and plant growth regulators as NAA, IAA and BA were used. Ethyl alcohol and sodium hypochlorite (NaOCl) were used for surface sterilization. Explants were allowed to grow at 20 ±1 oC. A Photoperiod of 8 hours dark+16 hours light were applied. At the end of first 30 days of incubation period, the formation of callus was observed on %38.4 and formation of bulblets was observed on %8.6 of survived bulb explants (survived bulb explants: %85.94). At the end of 120 days of incubation period, the best result was obtained from the medium (1 BA + 0.6 NAA + 0.4 IAA) suplemented with high cytokinin and %3 sucrose. In this medium, direct and indirect organogenesis was observed on %96.42 of bulb explants and callus formation was observed on the % 3.57. When PGR free basic medium does not contain sucrose, ratio of callus formation was be %48.13 but the organogenesis was not observed. As a comparison, medium containing %3 sucrose to medium containing %6 sucrose; at medium containing %3 sucrose is more effective on formation of callus + shoot (relatively %147.2 more effective) and on formation of bulblet (relatively %153.3 more effective). When %3 sucrose and high cytokinin (1 mg/BA + 0.6 mg/L NAA + 0.4 mg/L lAA) are used together, the probability of organogenesis is increased. Medium supplemented with high cytokinin and %3 sucrose is more effective on rate of formation of callus and shoot; medium supplemented with low cytokinin and %3 sucrose (0.1 BA + 0.6 NAA + 0.4 IAA) is more effective on rate of formation of callus and bulblet. Quantity of shoot formed by indirect organogenesis was increased %50 by medium supplemented with high cytokinin and %3 sucrose. Average 13.7 units of shoot formation per explant were obtained in medium supplemented with high cytokinin and %3 sucrose; average 13.9 units of bulblet formation per explant were obtained in medium supplemented with low-cytokinin and %3 sucrose. Explants which observed formation shoot and bulblet were allowed to take root in medium supplemented with ½ MS+ % 0,6 agar+ %3 sucrose + 5 mg/L NAA+0.5 mg/L BA. At the end of first 30 days of incubation period, rootings were observed on %48.32 of 329 units of explants and at the end of 60 days of incubation period, rootings were observed on %77.50 of 329 units of explants. Significant result regarding adaptation of little plants obtained from bulblet explants in vitro conditions to open field could not be obtained. %98 of little plants were died after transportation to external environment (flowerpots). In addition, callus formation and direct + indirect organogenesis was not observed in the experiments where cleistogamus and pedicles were used as explants.