Abstract: One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.
Abstract: This experiment was performed to optimize the medium for tissue culture of Taraxacum kok-saghyz Rodin. Different tissue culture approaches such as shoot regeneration from seed, callus formation from leaf explants and plant regeneration from callus were investigated in this study. All the explants were cultured on MS basal medium supplemented with 20g/l sucrose, 7g/l agar and different plant growth regulators. Seeds of Taraxacum kok-saghyzwere cultured on media containing different levels of BA and 2,4-D (0.5, 1.0 and 3.0mg/L) to direct shoot regeneration study. Leaf explants were cultured in different combination of BA (at three levels: 0.5, 1.0 and 3.0mg/L) and zeatin (at two levels: 0.5 and 1.0mg/L) to examine callus formation. After the callus formation the formed calli were cultured on different combinations of BA and NAA for shoot regeneration. BA at three levels (0.5 and 1.0 and 3.0mg/L) and NAA at two levels (0.5 and 1.0mg/L) in all possible combinations were used for shoot regeneration from callus. The results showed that the treatment containing 1.0mg/L 2,4-D in combination with 1.0mg/L BA was found to be the best one for shoot regeneration from seeds. The treatment with 1.0mg/L BA in combination with 1.0mg/L zeatin were found to be suitable treatments for callus production from leaf explants, as well. Moreover, 0.5mg/L BA alone or in combination with 1.0mg/L NAA were found to be the best treatments for shoot regeneration from callus.
Abstract: Quercetin and (+)-catechin are metabolites present in Phyllanthus niruri plant, have potential in medicinal uses as anticancer and antioxidant agents. Studies on production of quercetin and (+)-catechin from P. niruri callus culture via in vitro technique were carried out and the results were compared to the intact plant. P. niruri explants were cultured on Murashige and Skoog (MS) solidified media supplemented with several phytohormone combinations for one month. The metabolites were extracted from P. niruri callus and intact plant by using carbon dioxide supercritical fluid extraction (SFE) with ethanol as modifier and solvent extraction techniques. The extracts were analyzed by means of HPLC method. Results showed that P. niruri callus culture was successfully established. The highest content of quercetin (1.72%) was found from P. niruri callus grown in media supplemented with 0.8mg/L kinetin and 0.2mg/L 2,4-dicholophenoxyacetic acid (2,4-D), which was 1.2 fold higher than intact plant. Meanwhile, the highest amounts of (+)-catechin (0.63%) was found from P. niruri callus grown in media with addition of 0.2mg/L 1-naphthalene acetic acid (NAA) and 0.8mg/L 2,4-D. The SFE condition in this study showed better extraction efficiency when higher contents of selected metabolites were found in all SFE extracts compared to the common solvent extracts.
Abstract: The extract of milk thistle contains a mix of flavonolignans termed silymarine.. In order to analysis influence of growth regulators, genotype, explant and subculture on the accumulation of flavonolignans, a study was carried out by using two genotype (Budakalszi and Noor abad moghan cultivars), cotyledon and hypocotyle explants, solid media of MS supplemented by different combinations of two growth regulators; Kinetin (0.1, 1 mg/l) and 2,4-D (1, 2 mg/l). Seeds of the plant were germinated in MS media whitout growth regulators in growth chamber at 26°C and darkness condition. In order to callus induction, the culture media was supplemented whit different concentrations of 2,4-D and kinetin. Calli obtained from explants were sub-cultured four times into the fresh media of the first experiment. flavonoides was extracted from calli in four subcultures. The flavonoid components were determined by high- performance liquid choromatography (HPLC) and separated into Taxifolin, Silydianin+Silychristin, Silybin A+B and Isosilybin A+B. Results showed that with increasing callus age, increased accumulation of silybin A+B, but reduced Isosilybin A+B content. Highest accumulation of Taxifolin was observed at first calli. Calli produced from cotyledon explant of Budakalszi cultivar were superior for Silybin A+B, where calli from hypocotyl explant produced higher amount of Taxifolin and Silydianin+Silychristin. The best cultivar for Silymarin production in this study was Budakalszi cultivar. High amount of SBN A+B and TXF were obtained from hypocotil explant.