Using appropriate isolation and fractionation procedure, and UHPLC-MS/MS as powerful analytical tool, comprehensive phenolic acid profile of leaves and flowers of Hypericum perforatum L., growing wild in Bosnia and Herzegovina, has been analyzed for the first time. The phenolic acids were isolated from plant tissue, liberated from esters and glycosidic bonds, and then analyzed as free acids. Results show that free forms of caffeic and chlorogenic acid were the most abundant. Generally, the content of acids is higher in flowers than in leaves, but environmental factors do not significantly influence chemical composition of this medicinal plant.

Knautia arvensis (family Dipsacaceae) is usually found in grasslands of Europe, but can also be found in some parts of Africa and Asia, usually in dry meadows, pastures, dry hills, and open woods. The chemical composition of this species is relatively unknown. The aim of this study was to give a phenolic profile of this plant, and to show how its phenolic composition varies depending upon plant organ and geographic origin. The chemical analysis included quantification of free phenolics soluble in methanol, esters and glycosides, and phenolics that are insoluble in methanol. Fourteen different phenolic acids and eight flavonoids were quantified in total. According to this study, the chemical composition of individual plant parts can differ tremendously within one population, which does not have to be in correspondence with chemical variability between populations. As shown in this study, the variation in chemical composition between plant parts can exceed that between different populations (from different climates, altitudes, with different environmental factors), which implies that microhabitat conditions can greatly affect the composition of some plant parts, which was shown also after PCA and HCA analysis.

The effects of plant growth regulators on Mentha piperita explants cultured in vitro were studied for the purpose of analyse growth, secondary metabolite production, antioxidant and antimicrobial activities in micropropagated plants. The basal medium was experimentally supplemented with the auxin, indole-3-butyric acid (IBA) and the cytokinin, N6- benzyladenine (BAP) individually and in combination. Treatment with BAP and IBA resulted in an increased shoot and root number. The production of phenolic compounds was affected by the addition of the highest concentration of BAP, while antioxidant and antimicrobial activities were affected by several BAP and IBA treatments. Our results demonstrate that the application of growth regulators increases growth and secondary metabolite productions in the medicinal herb M. piperita.

The effect of aluminium and cadmium on growth and physiological parameters in 60 days old basil seedlings (Ocimum basilicum L.) grown in polluted soil were investigated. Control plants, grown in unpolluted soil, were used for evaluation of the heavy metal effect. Growth parameters were analysed immediately after sample collection, leaf area and plant height were recorded. Analysis of secondary metabolite, proline, total carbohydrates content and antioxidant capacity in 60 days old seedlings was performed. The antioxidant activity of basil extracts was determined by the DPPH method, and results showed an increase of antioxidative capacity in basil seedlings exposed to heavy metal stress due to the activation of antioxidative system in plant cells. Maximum production of secondary metabolites was recorded for seedlings exposed to 100 mg/kg soil Cd and Al. Results showed that cadmium stress increased concentration of proline, while aluminium did not had that effect. The content of total carbohydrates remained almost unchanged, with exception of basil seedlings grown on soil containing 50mg/kg AlCl3.

Knautia sarajevensis, Dipsacaceae, is an endemic species found at wood margins and meadows only on mountains of Dinaric Alps. Members of this family are widely used in traditional medicine as rich sources of pharmacologically important substances. Since it is well known that flavonoid compounds are one of the main carriers of biological activities of plant extracts, the aim of this study was to investigate cytokinin effects in concentration changes of flavonoid constituents. Presence of four different flavonoid constituents was noticed: quercetin, naringenin, hesperetin and rutin in extracts of K. sarajevenis shoots cultivated on three in vitro treatments (control, 1.0 mg L -1 kinetin and 10.0 mg L -1 kinetin), but only rutin content was quantified. All extracts were prepared using dried material and 80% methanol HPLC grade. Analysis of rutin indicated that high cytokinin concentrations did induce improvement of rutin content, but these concentrations are still lower than those recorded for control treatment. Further analysis using different types and concentrations of cytokinins are necessary to establish a pattern of cytokinin induced concentration changes in content of these four investigated flavonoids in Knautia sarajevensis.

Karalija, E., Paric, A.: The effect of BA and IBA on the secondary metabolite production by shoot cultureof Thymus vulgaris L.. Biologica Nyssana, 2 (1), September 2011: 29-35. In vitro shoots of the common thyme ( Thymus vulgaris L.) were established, and the effects of differentconcentrations of the cytokinin BA and the auxin IBA on secondary metabolite production were investigated.The highest number of shoots was obtained through cultivation on the MS medium containing 2 mg/L BAand 0.1 mg/L IBA and 4 mg/L BA and 0.1 mg/L IBA (14.3 and 13.3 respectively). Quantitative changes inchlorophyll a, chlorophyll b and carotenoid content were recorded in response to the effect of varyingconcentrations of growth regulators in the medium. Furthermore, after addition of plant growth regulators(0.5 mg/L BA + 0.1 mg/L IBA) to the medium, elicitation of phenolic compounds was recorded in plantlets.The concentration range of BA from 2 to 4 mg/L improved the production of flavonoids (0.61 and 0.64 mg/gFW). In contrast, plantlets cultivated on the same treatment, showed a decrease in monomeric anthocyanins. Key words : common thyme, in vitro, micropropagation, secondary metabolite production

Organogenic capacity of leaves and bulb explants of Lilum bosniacum was tested. For direct shoot formation MS medium with combinations of 0,5 mg/LBA and 0,2 mg/LIBA as well as 0,5 mg/LTDZ and 0,2 mg/L IBA were used. Both combinations supported shoot regeneration from bulb explants, but only the BA + IBA combination encouraged sho ot regeneration from leaf explants. Rhizogenesis was induced on MS basal medium with 0,2 mg/LIB A. Plantlets were successfully acclimated under greenhouse conditions.

Karalija, E., Trbojevic, S., Paric, A.: Somatic embryogenesis and in vitro plantlet regeneration of Lilium martagon L. var. cattaniae Vis.. Biologica Nyssana, 1 (1-2), December 2010: 57-60. In this study organogenic capacity of two different explants type (leaves and whole bulbs) of Lilium martagon L. var. cattaniae Vis. was examined. For induction of in vitro somatic embryogenesis and adventitive regeneration different concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzilaminopurine (from 0,25 mg/l to 8,00 mg/l) added to MS basal medium were used. Our results indicate that concentration of 0,5 mg/l 2,4-dichlorophenoxyacetic acid and 4 mg/l 6- benzilaminopurine promoted somatic embryogenesis from leaves of Lilium martagon var. cattaniae, while all other concentrations promoted direct shoot regeneration from bulb explants. Root formation was induced on MS basal medium with 0,2 mg/l indole butyric acid. These plantlets were acclimatized well in a greenhouse conditions.

Karalija, E., Trbojević, S., Parić, A.: Somatic embryogenesis and in vitro plantlet regeneration of Lilium martagon L. var. cattaniae Vis.. Biologica Nyssana, 1 (1-2), December 2010: 57-60. In this study organogenic capacity of two different explants type (leaves and whole bulbs) of Lilium martagon L. var. cattaniae Vis. was examined. For induction of in vitro somatic embryogenesis and adventitive regeneration different concentrations of 2,4-dichlorophenoxyacetic acid and 6-benzilaminopurine (from 0,25 mg/l to 8,00 mg/l) added to MS basal medium were used. Our results indicate that concentration of 0,5 mg/l 2,4-dichlorophenoxyacetic acid and 4 mg/l 6benzilaminopurine promoted somatic embryogenesis from leaves of Lilium martagon var. cattaniae, while all other concentrations promoted direct shoot regeneration from bulb explants. Root formation was induced on MS basal medium with 0,2 mg/l indole butyric acid. These plantlets were acclimatized well in a greenhouse conditions.