Investigatıon of callus induction from different explant of endemic ajuga vestita boiss and biological activite of callus

Ajuga vestita BOISS. is a species of genus Ajuga which is medical importance, endemic and in the category of ‘’EN-dengered’’. In this study which have not been studied before in literature, it was aimed to determine the potential of shoots regeneration from callus by establishment and optimization of callus cultures from different explants of in vitro shoots of Ajuga vestita BOISS. For this purpose; firstly surface- sterilized seeds of Ajuga vestita BOISS. were germinated in a hormone-free 1/4 MS medium. Axenic shoots tips were propagated in MS medium supplemented with 0,125 mg/L-1 Kin and obtained in vitro shoots were used as a starting materials for callus initiation studies. The leaves, stem and roots of in vitro shoots, and broken seeds were cultured in 1/1 MS medium containing differents concentrations of cytokinin (Kin, BAP) and oxin (2,4-D). As a result; callus formation was observed in the hormone combinations tested for all of explants types. However; it was determined that the best callus formation was MS medium supplemented with 0,5 mg/L-1 Kin+2.0 mg/L-1 2,4-D in all explants. However, leaf and broken seeds cultured in 0.5 mg/L-1 Kin+2.0 mg/L-1 2,4-D medium were found to have the best response to callus culture. The callus obtained from all explants were cultured in the hormone-free MS medium (4 week) to development of callus and shoot regeneration from callus. It was observed that the callus formed in root and internod explants completely darkened, where as leaf and seed explants were create yellow and light green callus. For this reason, the callus obtained from leaf and seed explants were cultured in MS medium containing different concentrations of BAP and Kin (0.125, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 10.0 mg/L-1) for shoot regeneration studies. Shoot formations was occured in MS medium supplemented with 0.125 ve 0.5 mg/L-1 BAP 5 weeks after culturing. In 4.0-10.0 mg/L-1 BAP containing medium and in most of the Kin treatments, root formation was detected in the callus. In addition, the total phenolic and flavonid contents of the callus obtained from different concentrations of BAP and Kin were determined as equivalent to gallic acid and quercetin respectisely The total phenolic and flavonoid content of acetone extract was higher then methanol extract. However, total antioxidant activity of acetone and methanol extracts prepared from callus was determined by using DPPH free radical activity method. The highest antioxidant activity was detected in the methanol extract at a concentration of 500 μg/ L-1with 88.91% inhibition. Acetone and methanol extracts at a concentration of 250 and 500 μg/ L-1 were found to have a higher effect than BHT which is used as positive control at the same concentrations.