Optimization of in vitro germination conditions for seeds of the rare species Ligularia sibirica cass. L. and Senecio besserianus minder. (Asteraceae)

DOI: https://doi.org/10.7124/visnyk.utgis.23.1-2.1778
Keywords: seed germination, in vitro culture, sterilization, growth regulators, stratification, pre-sowing treatment, bubbling

Abstract

Aim. To select an effective sterilization protocol and optimize seed germination conditions for the rare and vulnerable species Ligularia sibirica (L.) Cass. and Senecio besserianus Minder., listed in the Red Data Book of Ukraine, for their subsequent introduction into in vitro culture and microclonal propagation. Methods. Four seed sterilization schemes were compared, and the effects of five variants of seed pre-sowing treatment (distilled water, indole-3-butyric acid, succinic acid, gibberellic acid, and bubbling) were tested, followed by inoculation on half-strength Murashige and Skoog medium (MS/2). The following germination parameters were evaluated: final germination percentage (FGP), mean daily germination (MDG), mean germination time (MGT), and germination index (GI). Data were expressed as mean and standard error (M ± SE). The significance of differences between treatments was assessed at p < 0.05. Results. For both species, an effective sterilization scheme involved the use of 96 % ethanol, 15 % hydrogen peroxide, and a benomyl-based fungicide (systemic fungicide Fundazol), ensuring 100 % sterility. In L. sibirica, the highest germination parameters were recorded after pre-sowing treatment with succinic acid and indole-3-butyric acid (FGP 86.67 %). For S. besserianus, soaking in distilled water proved optimal (FGP 40 %). Gibberellic acid at the tested concentration (0.1 mg mL⁻¹) did not have a positive effect on seed germination in either species. Bubbling generally increased MGT and reduced GI. Conclusions. The proposed sterilization scheme using a benomyl-based fungicide provides reliable production of sterile and viable seed material of L. sibirica and S. besserianus. For S. besserianus, comparable results were obtained using a sterilization scheme based on 15 % hydrogen peroxide without benomyl. Based on four germination indexes succinic acid was the most effective stimulant of L. sibirica seed germination, whereas soaking in distilled water was optimal for S. besserianus.

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