Μicropropagation and cultivation of Salvia sclarea for essential oil and sclareol production in northern Greece

Abstract

Salvia sclarea L. is valuable for its essential oils and other perfumery products used in the food and pharmaceutical industries. The aim of this study was to develop a rapid micropropagation protocol and cultivate S. sclarea in northern Greece for essential oil and sclareol production. The initial plant material used was S. sclarea seeds (biotype SC1) collected from a local population in Bulgaria. Apical meristems from seedlings germinated in the greenhouse were disinfected and established in vitro. For shoot proliferation, six different modified basal culture media supplemented with plant growth regulators (benzyladenine, auxins, and gibberellic acid) at different concentrations and combinations were tested. After 30 d, the highest proliferation rate (1.78) was achieved with an MS medium supplemented with 1.5 μM benzyladenine (BA) and 0.15 μM IBA. The MS medium with 5 μM IBA was the most effective treatment for rooting (40% rooting percentage, one root/microshoot, and 1.3-cm root length after 12 d of culture). Within 2 mo, rooted shoots (93–95%) were successfully acclimatized and survived ex vitro, whereas the remaining shoots rooted and survived in vivo after treatment with 5 μM IBA or α-naphthaleneacetic acid (NAA) (51–53%). During the 2-yr pilot cultivation, the yield of inflorescences in dry weight within the first year was 7 g/plant (129.50 kg/ha) and 33.3 g/plant (616.05 kg/ha) in the second year. The essential oil was found to be rich in secondary metabolites (linalyl acetate, trans-caryophyllene and its oxide, a-copaene and germacrene D). The extraction yield of sclareol from the blossoms was 1.5–2 g/100 g of dry mass (10–15 kg/ha). This micropropagation protocol results in plant propagation in 330 d and field cultivation with the first harvest in 450 d.