Explants of apical buds of sandy everlasting Helichrysum arenarium L. Moench sterilized with calcium hypochlorite – Ca(OCl2) were placed onto Murashige-Skoog (MS) growth medium enriched with 1 mg∙dm^-3 KIN (kinetin) in two consecutive passages. To optimize the procedure of in vitro micropropagation, the axillary shoots of Helichrysum arenarium L. Moench were transferred during the third passage onto 9 different combinations of MS medium without plant growth regulators (PGRs-free medium), with different concentrations of KIN (1.0, 3.0, 4.0, 5.0 mg∙dm^-3) and KIN (1.0, 3.0, 4.0, 5.0 mg∙dm^-3) with IAA (indole-3-acetic acid) (0.5 mg dm^-3). The highest mean number of shoots (24.7) was observed on MS medium with 5 mg∙dm^-3 KIN and 0.5 mg∙dm^-3 IAA. The achieved branched shoots were rooted and acclimatized. Rhizogenesis was intensified by the presence of growth hormones: 0.5 mg∙dm^-3 IAA or 0.5 mg∙dm^-3 IBA (indole-3-butyric acid). At the stage of acclimation of plantlets the application of a water solution of MS salts (25%) for watering the plants increased the efficacy of plant acclimation from 56 to 75%.

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