The tissue lines derived from the Arieeni, Retezat and Valea Sebeului bilberry genotypes grown on a WPM medium supplemented with 40, 60 and 80 mg/l AS, were subject to molecular analysis through amplification of DNA samples with ISSR and RAPD primers. The results of agarose gel electrophoresis of the amplification products show the existence of significant differences in the model of polymorphic bands obtained from tissue lines, compared to the mother plant. Based on the obtained results, the presence of somatic variability, induced in the tissue lines under the influence of AS, is emphasized. It consists in the absence of binding sites of the HB-12 ISSR marker compared to the mother plant. Considering the statements in the specialty literature, concerning the mutagenic effect of growth, we can state that AS caused changes within the DNA level in the bilberry calluses selected by us under the experimental conditions.

In vitro cells culture is a good method to induce somatic variability in plants. The selection of plant tissues with a good capacity of cellular biosynthesis is necessary to obtain the important secondary metabolites by in vitro culture system. In our experiments we used biochemical methods to evaluate the variability induced by in vitro callus culture at three blueberry populations native in Romania. Callus obtained from Arieseni, Retezat and Valea Sebesului populations, on WPM medium was sub-cultured in presence of ANA, BAP and AS. Antioxidant capacity using the FRAP method, glutathion (GSH), total anthocyanin, macroelements, microelements and heavy metals contents from callus lines selected were investigated. The AS concentration positively affects the activity of glutathione, with a significantly positive correlation coefficient (r = 0.54, p <0.05). The use of this hormone facilitates the obtaining of blueberry callus biomass with an increased antioxidant activity. There was also found a strong positive correlation between the total content of anthocyanin and FRAP values (r = 0.60, p <0.05) and a positive correlation with the level of GSH, respectively (r = 0.47, p <0.05).

Our research aimed the obtaining a proliferative tissue at 7 bilberry (Vaccinium mytillus L.) local population, to select some anthocyanins producing tissue lines by in vitro culture. The tissue lines selected from Arie�i eni, Semenic, Retezat, Valea Sebe�i ului, Buces-Vulcan, Vadul Dobrii and Cornereva local population, with good results of callus growth capacity in subculture, produced anthocyanins (646 mgC3GE/100g). These results demonstrated the possibility to elaborate a production system of these metabolites in controlled conditions.

The biological and medical effects of bilberry fruit (Vaccinium myrtillus L.) are mainly due to high anthocyanin content of tissues. Calli containing anthocyanins, derived from bilberry plants, may represent a potential source of natural colouring matter, pharmaceutical and natural antioxidants. In the present study we investigated the occurrence of differences in elemental composition and antioxidant capacity of the three local populations of mountain bilberry collected in the western region of Romania (Arieseni, Retezat and Sebes Valley) in order to compare the anthocyanin production of plant and callus tissues originated from various plant populations. It was found that K, Fe and Zn content was higher in calli than in intact plant leaves. The excess of latter two microelements, Fe and Zn can induce oxidative stress, and, as a result of the accumulation of reactive oxygen species, various antioxidant mechanisms. The total antioxidant capacity of callus cultures determined by FRAP method (ferric reducing antioxidant power) could be enhanced as a function of increasing adenine sulphate (AS) concentration in the culture medium and it depended on the origin of mother plants. The leaves of intact plants contained higher amount of total non-protein sulfhydryl groups than calli, and the decrease was especially significant in tissue cultures originated from the Retezat region. In contrast, depending on the AS concentration, the anthocyanin content could increase in callus cultures. The tissues originated from various populations exhibited different AS concentration optimum. This suggests that bilberry callus cultures can be a suitable source of the anthocyanins.

The bilberry fruit (Vaccinium myrtillus L.) has biological and medical effects, mainly due to high anthocyanin content of tissues. Calli containing anthocyanins, derived from bilberry plants, may represent a potential source of natural colouring matter, pharmaceutical and natural antioxidants. In the present study we investigated the occurrence of differences in elemental composition and antioxidant capacity of the three local populations of mountain bilberry collected in the western region of Romania (Arieseni, Retezat and Sebes Valley) in order to compare the anthocyanin production of plant and callus tissues originated from various plant populations. The total antioxidant capacity of callus cultures determined by FRAP method (ferric reducing antioxidant power) could be enhanced as a function of increasing adenine sulphate (AS) concentration in the culture medium and it depended on the origin of mother plants. The leaves of intact plants contained higher amount of total gluthatione than calli, and the decrease was especially significant in tissues cultures originated from the Retezat region. In contrast, depending on the AS concentration, the anthocyanin content could increase in callus cultures. The tissues originated from various populations exhibited different AS concentration optimum. This suggests that bilberry callus cultures can be a suitable source of the anthocyanins.

In the present research, proteins were determined in a relatively large and diverse number of gemotherapeutic preparations from different medicinal plants with anti-diabetes properties such as: bitter melon (Momordica charantia), white mulberry (Morus alba), and bilberry (Vaccinium myrtillus), as well as from certain edible and medicinal mushrooms (Agaricus bisporus, Pleurotus ostreatus, Lentinula edodes-shiitake) used in the treatment of a wide range of conditions, some of which very serious, such as different forms of cancer (particularly mammary and prostates), kidney failure, atherosclerosis, hypercholesterolemia, cardiovascular diseases, Heliobacter pylon infections, etc. Gemoderivatives were presented under the form of hydroglycerin alcoholic solutions in the first decimal dilution (DHI) and were prepared from plant parts during growth: sprouts, buds and young roots, or buttons (primordia) in the case of the mushrooms. To determine protein content, we used a rapid colorimetric method based on a Cu2+ ion reagent and the Folin-Ciocalteu reagent.