Investigation of in vitro mikcropropagation methods of endemic Hypericum spectabile grown in Southeast Anatolia region

In this study,the main aim was to develop a method for Hypericum spectabile?s in vitro micropropagation of its seed that an endemic sepecies grown in East and Southeast Anatolia Region.In the first phase of study; seeds of Hypericum spectabile, used as a starting material, a sterilization process was done waiting respectively in 70% ethyl alcohol for 30 seconds, 5% NaOCI solution for 10 minutes. Sterile seeds were germinated based culture on MS medium free plant growth regulators (PGRs) .In the second phase of study; different concentrations (0.25, 0.5, 1.0, 1.5, 2.0, 2.5 mg/l) of benzylaminopurine (BAP) and Kinetin (Kn) that are used as a cytokinin tested separately. Among the concentrations 0.25 mg/l BAP (49.80 shoots per explant) was found ideal for shoot proliferation.Compared to Kin concentrations, the most shoots were obtained on MS medium supplemented with 0.5 mg/l Kin (27.33 shoots per explant). Although more shoots were obtained in this medium,growth of shoots were not good.In addition , in order to improve and increase the number of shoots nine different types of medium was prepared to investigate combined effects of BAP and Kin. The data obtained in accordance with previous experiments, BAP (0.25 mg/l ) and Kin (1.5 mg/l ) together with different concentrations of (0.25, 0.5,1.0, 2.0 mg/l) naphthalene acetic acid (NAA) and indole acetic acid (IAA) were tested seperately. Compared hormone combinations tested, were not a meaning ful increase in the number of shoots.In the third phase of study; leaf and root explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of benzilaminopurin (BAP; 0.5, 1.0, 1.5 mg/l) and 2,4-dichlorophenoxyacetic acid (2,4-D; 0.5, 1.0, 2.0 mg/l). The best callus induction for leaf explants was observed in the treatment containing 1.0 mg/l BAP + 2.0 mg/l 2.4-D on the other hand for root explants 1.5 mg/l BAP + 0.5 mg/l 2,4-D culture medium was optimal rates.For shoot formation from calli, calli were transferred into different BAP concentrations (0.25, 0.5, 1.0, 1.5 mg/l ) separately. Although the highest number of shoots produced on medium supplemented with 1.5 mg/l BAP (25.15 shoots per explant), however, morfology of shoots were not good. The best result for shoot morfology was observed in the treatment containing 0.5 mg/l BAP (22.75 per explant), in this medium shoots were more vibrant and more healthy.In the fourth phase of study; for the shoot rooting four different concentrations of NAA and IAA (0.25, 0.5, 1.0, 2.0 mg/l) together with the force of the MS medium (1 / 1 MS, ½ MS) was investigated. Optimum result were taken on MS medium supplemented with 0.25 mg/l for rooted of seedling.Adaptation to soil of Obtained rooted seedling were succesfully provided with acclimatization works.Keywords : Hypericum spectabile, micropropagation, callus, PGRs