Providing organ regenerations under in vitro conditions in rown imperial (Fritillaria imperialis L.)

Crown imperial (Fritillaria imperialis L.) which belongs to Fritillaria genus of Liliaceae family, is grown as semi-endemic in our country. Since crown imperial (F. imperialis L.) is used as ornamental plant for a long time because of its splendid flowers, it is exported to Europe especially Holland and it is forbidden to remove from nature. Aim of this study is to propagate using various in vitro nutrient mediums and different tissue culture techniques in F. imperialis species, which includes lots of restrictions in both vegetative and generative production. For this purpose, organ regeneration under in vitro conditions was performed by using various explants (bulb, leave, shoot, root) of F. imperialis L. For the green leaf explants, the maximum callus formation ratio (59.26%) was obtained on B5 medium containing 1.0 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA and 60 g/l sucrose, while for the petal explants, the maximum callus formation ratio (33.3%) was detected on MS medium with the same content. In the studies was made by using the green leaf explants, it is recorded that regenerated callus in MS based nutrient media had more tendency to form both somatic embryo (the maximum embryogenic callus ratios 92.31% and 56.25%, respectively) and shoot regeneration (the maximum shoot forming callus ratios 88.89% and 37.50%, respectively) than regenerated callus in B5 based nutrient media. Among these media, MS and B5 media supplemented with higher concentration of BAP (1.0 mg/l) combined with auxins (0.5 mg/l NAA + 0.4 mg/l IAA) were more successful (88.89% and 37.50%, respectively) than low concentration BAP (0.1 mg/l) combined with same auxins (53.85% and 16.67%, respectively) regarding callus ratios forming shoot. In the studies which used green leaf explants as starting material, the medium supplemented with BAP cytokinin and auxins was more efficient in shoot regeneration, while the medium containing Kin or TDZ cytokinins and auxins was more effective in callus stimulation. At the F. imperialis species that using MS medium include 1 mg/l NAA + 2 mg/l 2,4-D and roots as initial explants the first time which was grown by themselves on bulbs during storage and, photoperiod conditions of 16 hours light/8 hours dark were more suitable than fully dark conditions in terms of callus regeneration. The green leaf explants were subculture after six weeks of culture, 1.35 shoots per green leaf explant and total 146 shoots regenerated. Total 62 bulblets were obtained from callus that developed on green leaf explants which maintained 36 weeks in culture without subculture, while the petal leaves formed no bulblets. At the end of the 36 weeks culture periods, 1.89 bulblets were obtained per green leaf explant (at the FS7 coded medium which was included B5 + 1 mg/l BAP + 0.5 mg/l NAA + 0.4 mg/l IAA).