Establishment and optimization of cell suspension cultures of juvenile mastic tree (Pistacia lentiscus L.)

To develop an effective protocol for the initiation and optimization of callus cultures in Pistacia lentiscus L., the surface-sterilized seeds were germinated in an MS medium supplemented with 1 mg/l IBA and then the axenic apical shoots obtained were propagated in MS medium containing 1 mg/l BAP + 0.5 mg/l GA3. The roots obtained by in vitro germination of the seeds and the axenic leaves from in vitro propagated cultures were used as explant source for callus induction. The effect of the combination of different auxin (IAA, IBA, NAA and 2,4-D) and cytokinin (BAP, Kin, TDZ and 2IP) each one at 0.25, 0.5, 1.0 and 2 mg /l concentrations, respectively were tested. for callus formation. The effects of different nutrient media types (MS, WPM, SH, B5) prepared in 1/1 strength were also tested. In addition, different types of sugar (glucose and sucrose) and their different concentrations (15, 30, 50 mg / l), different pHs (4.5, 5, 5.8, 6.5, 7), different light intensities (20,40,80 μmol/s), and different temperature (10,20,25,30,35 ° C) effects were tested for callus development. Besides these experiments, different types of nutrient media (MS, WPM, SH, B5) prepared in (0.25, 0.5, 1 and 2 mg/l) strength were also tested. In the study of callus cultures initiation, it was found that the MS medium for both root (80%) and leaf (84%) explants gave the best results as nutrient medium type. In terms of different types of PGR, again an MS medium containing 1mg/l Kin ve 1 mg/l 2,4-D for both root (80%) and leaf (80%) explants gave the best results. In the optimization studies on callus cultures development, the highest root and leaf explants in application results of different sugar types and concentrations from 1/1 MS medium in root (80%) and leaf (100%) explants in which different food types were searched. The optimal value for root (96%) and leaf (100%) explants was different from pH 5.8 medium between different pH applications and 20 μmol (100%) for root explants among different light intensity applications from MS medium containing 15 mg/l sucrose 80% (100%) light applications for explants and the best result for different temperature applications was obtained from both roots (76%) and leaves (100%) at 25°C temperature. In the optimization studies on callus cultures development, in experiments investigating the different media types, the root (80%) and leaf (100%) of explants that 1/1 MS medium, different sugar types and their different concentrations the root and leaf of explants at 15 mg / l sucrose, different pH applications . root (96%) and leaf (100%) explants at pH 5.8, different light intensity applications the root (100%) at 20 μmol and leaf (100%) at 80 μmol, different temperature applications, both root (76%) and leaf (100%) explants at 25 ° C temperature gave the best results was determined. In summary, this study allows to develop a protocol that can routinely be used for the initiation and optimization of callus cultures, which at the same time will shed light on the production of valuable secondary metabolites by P. lentiscus L. callus cultures.