Microclonal propagation and callus induction of some species of Carlina L. genus

Abstract

Aim. The aim of the research was to choose the conditions for microclonal propagation and obtain callus cultures from Carlina аcaulis L., Carlina cirsioides Klok and Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl plants in vitro. Methods. For microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia we used rosettes of 2–3-month specimens and planted them on semi-solid Murashige and Skoog (MS) medium with decreased macro- and microsalts concentrations (MS/2) supplemented with kinetin (Кin) (from 1–3 mg/l) and 0.1 mg/l of 1-naphthaleneacetic acid (NAA). For induction of callus formation, we used root, stem explants from С. acaulis, C. cirsioides and C. onopordіfolia, and planted them on nutrient media MS, MS/2, and Gamborg and Eveleigh (В5) supplemented with different concentrations of cytokinins – 6-benzylaminopurine (BAP) or Кin and auxins – 2.4-dichlorophenoxyacetic acid (2.4-D) or NAA and indole-3-acetic acid (IAA). Results. MS/2 medium supplemented with growth regulators of NAA and Кin were the most efficient to provide the formation of microclones. For C. сirsioides plants, this indicator was 6.6–6.8 rosettes per graft after 6 months of cultivation and for С. acaulis and C. onopordіfolia – 4.2–5.0 and 4.8–5.2 respectively. To raise the percentage of rooting for microclones of Carlina species, it was expedient to steep them preliminarily in the solution of indole-3-butyric acid (IBA) with 1000 mg/l concentration for a minute. Optimal for obtaining callus tissue from Carlina plants was nutrient medium MS supplemented with 3 mg/l IAA, 0.5 mg/l NAA and 0.5 mg/l Kin and MS/2 with 0.1 mg/l BAP and 0.5 mg/l 2.4-D; under such conditions the percentage of callus induction exceeded 90 % for all types of explants. Conclusions. There were chosen the conditions for microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia and worked out the schemes for enrooting obtained microclones in vitro. Capable of growing rapidly callus cultures from root and stem explants of the investigated plant species were obtained.
 Keywords: Carlina аcaulis L., Carlina cirsioides Klok, Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl, in vitro, microclonal propagation, callus induction.