Regeneration is one of the most important methods in the massive proliferation of ornamental plants through micropropagation and genetic modification. Thin Cell Layering (TCL) technique, is a new introduction of micropropagation efficiency in producing optimized mass scale of crops. The horizontally placed TCL explants (tTCLs) was examined in Van der Salm (VS) basal medium in combination with different plant Adjuvants (Proline, Glutamic Acid, Casein Hydrolysate) to investigate their effect on callus induction and shoot regeneration of the explants. In order to produce healthy propagules of Rosa damascena Mill. on a large scale, a new cost-effective method of tissue culture has been established. The results indicated the positive effect of Proline (0.5 gr/l) and Glutamic Acid (300 mg/l) in both callus induction and regeneration stage. Also, in rooting phase, different concentrations of IBA and NAA (0, 0.3, 0.6, 0.9 mg/l) in basal ½ Van der Salm (VS) medium were tested. It was observed that the best results were related to 0.3 mg/l IBA+0.3 mg/l NAA in ½ Van der Salm (VS) medium.

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