Microclonal propagation and rooting in vitro of rare Gentiana acaulis L. species

Abstract

Aim. The alternative methods enabling to restore natural habitats of rare plant species, including Gentiana acaulis L., encompass repatriation of grown in vitro plants into disturbed populations. Therefore, the aim of the work was to choose conditions for microclonal propagation and in vitro rooting the shoots of G. acaulis, a rare alpine Carpathian species. Methods. Cultivation of plant objects in vitro. Results. There were chosen conditions for microclonal propagation of G. acaulis plants: agarized nutrient medium MS/2 supplemented with 0.2 mg/l of 6-benzylaminopurine (BAP) and 0.2 kinetin (Kin; for plants from Rebra mountain, the Ukrainian Carpathians) and medium MS/2 with double concentration of СaCl₂, supplemented with 0.5 mg/l BAP and 0.1 mg/l Kin (for plants from Turkul mountain, the Ukrainian Carpathians). Optimal for rooting G. acaulis plants in vitro was nutrient medium MS/2 with half concentration of NH₄NO₃without vitamins and sucrose supplemented with 3 g/l of mannite and 0.1 mg/l of 1-naphthaleneacetic acid, the maintaining substrate was agar (4 g/l) combined with perlite (16 g/l). Conclusions. The efficient method of microclonal propagation and rooting G. acaulis plants in vitro was found and tested. The results of the research will be used for obtaining a sufficient amount of viable rooted plants able to efficiently adapt to conditions ex vitro and in situ.

Keywords: Gentiana acaulis L., microclonal propagation, rooting in vitro.