In vitro regeneration protocol for endemic Campanula leblebicii Yıldırım

Campanula leblebicii, discovered by Yıldırım, is found only in the village of Ovacık in the Kemalpaşa district of İzmir province, and it is one of the valuable endemic species for Turkey flora. Petiole and leaf explants were cultured on full-strength Murashige and Skoog (MS) medium containing 30.0 g L⁻¹ sucrose; 4.0 g L⁻¹ Gelrite; 0.3 mg L⁻¹ 1-naphthaleneacetic acid (NAA); 0, 0.5, 1.0, 2.0, or 3.0 mg L⁻¹ zeatin (ZEA); and 0, 0.5, 1.0, 2.0, or 3.0 mg L⁻¹ 6-benzyladenine (BA) for callus initiation. To obtain shoot formation, callus was transferred to full-strength MS medium containing 0.3 mg L⁻¹ NAA; 0, 0.5, 1.0, 2.0, or 3.0 mg L⁻¹ gibberellic acid (GA₃); and 0, 0.5, 1.0, 2.0, 3.0 mg L⁻¹ BA. Efficient callus (93.33%) and shoot formation (73.33) were observed from petiole explants cultured on MS medium containing 0.3 mg L⁻¹ NAA and 1.0 mg L⁻¹ BA. Rooting was obtained spontaneously in all shoot initiation medium, but efficient rooting occurred on MS medium supplemented with 0.3 mg L⁻¹ NAA and 1.0 mg L⁻¹ BA. Plants were acclimatized to the pots including sterile peat and perlite (1:1, v/v), and, in total, 12 plants were acclimatized from one genotype.