Vitrification during development of clonal micropropagation technology for rare plant species of Salvia scabiosifolia Lam.

Abstract

Aim. Anatomical structure comparison of normal and vitrified Salvia scabiosifolia Lam plants in vitro; nutrient medium growth regulator selection for vitrification minimization. Methods. In vitro culture methods were used. Plants were grown on MS nutrient medium, enriched by peroxyacetic and indoleacetic acids and BAP with different combinations and concentrations. Anatomical leaf slices 10 µm thick were dyed with aceto-orcein and safranin. Results. Anatomical differences between thew structures of normal and vitrified S. scabiosifolia leaves were detected. Minimal vitrification was achieved by using nutrient mediums with 0.2 mg/l IAA + 1 mg/l BAP; 0.2 mg/l IAA + 2.5 mg/l BAP; 0.2 mg/l IAA + 0.1 mg/l NAA + 2 mg/l BAP. Conclusions. Anatomical structural features of vitrified leaves indicate abnormal transpiration, division and growth speed of epidermis and mesophyll cells. Vitrification can be lowered by variation of growth regulation content in the nutrient medium. Major role in causing vitrification in S. scabiosifolia is played by auxins, but combining them with cytokinins lowers vitrification significantly.
 Keywords: Salvia scabiosifolia Lam., in vitro, plant anatomy, vitrification, auxins, BAP.