Dağ çileğinde (Fragaria vesca L.) in vitro rejenerasyon,biyolojik aktivite değerlendirilmesi ve fenolik bileşiklerin belirlenmesi

Fragaria vesca L. is a perennial herb in the family Rosaceae that has been used in the treatment of liver and kidney complaints, as well as in the treatment of rheumatism in folk medicine. A highly efficient in vitro culture protocol of F. vesca was developed via multiple shoot formation. Leaf and petiole explants obtained from field-grown plants and sterile seedlings were cultured on MS medium containing different concentrations and combinations of various plant growth regulators and regeneration enhancers (silver nitrate, tween 20, tween 80 or pluronic F-68). Shoot regeneration was observed with explants obtained from sterile seedlings. However, field-grown plant materials were not responsive for shoot regeneration. The regeneration efficiency was found to be higher in leaf explants than petiole explants. Supplementing media with pluronic F-68 significantly increased shoot number and frequency of shoot formation. Among leaf explants, the best shoot proliferation was obtained with 1.0 mg/l TDZ in combination with 0.5 mg/l IBA and 0.1 % pluronic. Among petiole explants, best shoot proliferation was obtained with 1.5 mg/l TDZ in combination with 0.5 mg/l IBA and 0.1% pluronic. Regenerated shootwere rooted on MS medium containing different concentration of IAA, IBA, 2,4-D or NAA. Best root formation was observed with MS basal medium (control). Comparing various auxins for rooting, IBA and IAA were more effective. After rooting, plantlets were transferred to soil for hardening.The phenolic contents in the methanolic extracts of field-grown leaves, fruits and in vitro-grown leaves and callus of F. vesca were analysed using LC-ESI-MS/MS. According to the results, concentrations of phenolic compounds in field-grown leaf extracts (NFM and NM) were at least five times higher than in other extracts. Five different biological assays (antibacterial, anti-tumor, toxicity, anticancer and antioxidant activity) were used for assessing the biological activities of F.vesca extracts. Extracts obtained from field-grown leaves and fruits indicated better antibacterial activity than extracts of in vitro grown leaves and callus against E. cloacae. The best antitumor activity was observed with aqueous (NW) and methanolic (NM) extracts of field-grown leaves and hot ethanolic extract (FFHE) of fresh fruit via potato disc diffusion assay. In brine shrimp toxicity assay, extracts of field-grown leaves and fruits were more toxic than in the extracts of in vitro-grown leaves and callus.The best anticancer activity on MCF-7 cell lines was observed with extracts of field-grown leaves and fruits. Furthermore, the free radical scavenging potency of methanolic extracts of F. vesca was determined with DPPH method. The total phenolic and flavonoid contents were also investigated.Keywords: Fragaria vesca L., In vitro regeneration, LC-ESI-MS/MS, Biological activities, Antibacterial, Antioxidant, Toxicity, Antitumor, Anticancer.