The objective of this study was to design a protocol of the successful establishment of plants of Begonia fischeri var. palustris obtained in the in vitro seeds culture, to evaluate flavonoid content and antimicrobial properties of in vitro plantlets. The significant increase of percentage of seed germination (92.5%) was recorded in the half strength MS medium and 1% agar medium (83.3%) in comparison with 50-60% in greenhouse conditions. Flavonoid composition of the leaves of in vitro plantlets

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... and greenhouse stock plants had no substantial differences. Significant differences (p<0.05) were not found between flavonoid contents of the leaves (13.6 and 15.5 mg·g −1 of Dry Weight (DW), respectively). Aqueous ethanol extracts of plants showed antimicrobial effects against reference strains of Bacillus subtilis, Streptococcus pyogenes and Staphylococcus aureus. Concentration of flavonoids in acetone and ethanol extracts of exudative compounds of the leaves of in vitro plantlets was 0.02 and 2.0 mg·g −1 DW, respectively. Our results indicated that this protocol can be used for the production of a large number of plants of B. fischeri var. palustris with rich flavonoid composition and high quantities of flavonoids, including exudative compounds, with healing and air cleaning properties.