Investigation on i̇n vitro clonal propagation of the medicinal plant Calendula officinalis L.

INVESTIGATION ON IN VITRO CLONAL PROPAGATION OF THE MEDICINAL PLANT CALENDULA OFFICINALIS L. Betül KURTULUŞ Biology Department, M. S. Thesis, 2015 Thesis Supervisor: Assist. Prof. Burcu ÇETIN SUMMARY The present study was conducted to investigate the effects of different concentrations and combinations of plant growth regulators (PGRs) on callus induction and micropropagation of Calendula officinalis L., a medicinal and aromatic herb. Leaf, hypocotyl, cotyledon and cotyledon node explants, excised from the plantlets obtained by in vitro germinated seeds. The seeds were surface sterilized with 70 % ethanol for 3 mins, 10 % commercial bleach for 5 mins. subsequently, 3 times rinsed with sterile water for 3 mins. The seeds were germinated in Murashige ve Skoog (MS) medium. In the first experiment; the effects of plant growth regulators were researched on callus induction in leaf and cotyledon explants. The explants were cultured on MS medium supplemented with different concentrations and combinations of cytokinins (0.5, 1.0 mg/L 2,4 D; 0.5, 1.0 mg/L kin) and auxin (0.5 mg/L IAA). The best callus induction from cotyledon explants was observed at MS media containing 1 mg/L 2,4 D + 0,5 mg/L kin + 0,5 mg/L IAA and from the leaf explants at MS media containing 1 mg/L 2,4 D + 1 mg/L kin + 0,5 mg/L IAA. In the second experiment; hypocotyl, cotyledon and cotyledon node explants were cultured on MS media supplemented with various concentrations of cytokinins (BAP; 1.0, 2.0 mg/L) and auxin (IBA; 0.1, 0,5 mg/L) for callus induction and regeneration. The best results were achieved with cotyledon nodes. The highest number of shoots (13.14 ± 2,72) was observed in a treatment with 2 mg/L BAP + 0,5 mg/L IBA and the longest shoots (1,62 ± 0,13) was observed in a treatment with 1 mg/L BAP, after eight weeks of culture. The shoots were treatment with various IBA (0.3, 0.5 and 1 mg/L) concentration for rooting. The best rooting was observed in the MS medium supplemented with 0.3 mg/L IBA (77 %). Keywords: Calendula officinalis L., Plant growth regulators (PGRs), Leaf, Hypoctyl, Cotyledon, Cotyledon node, In vitro, Murashige ve Skoog (MS), Cytokinin, Auxin.