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The shoots of Salvia officinalis growing in MS liquid medium supplemented with IAA 0.1 mg l-1) and BAP (0.45 mg l-1) were treated with methyl jasmonate (MeJA) to increase production of compounds with antioxidant activity (carnosic acid, carnosol and rosmarinic acid). The increase in metabolite production depended on MeJA concentration, the period of exposure to elicitor and type of compound. The MeJA action was observed 24 h after elicitation. It was found that the maximum level of diterpenoids, calculated as the sum of CA and Car (about 8 mg g-1 dry wt) was achieved at 3 days after elicitation with 20 µM methyl jasmonate. The highest amount of rosmarinic acid (about 41 mg g-1 dry wt) was achieved with 50 or 100 μM methyl jasmonate on the 5th day after elicitation. It was almost 2-fold higher compared to the control (cultures treated with only ethanol).
Shoot tips excised from shoot culture of Salvia officinalis were encapsulated in 2% or 3% (w/v) sodium alginate and exposed to 50 mM calcium chloride for complexation. Immediately or after 6, 12 or 24 weeks of storage at 4°C, the synthetic seeds were cultured for 6 weeks on half-strength MS medium supplemented with indole-3- acetic acid (IAA) (0.1 mg/l) and solidified with 0.7% agar. The frequency of shoot and root emergence from encapsulated shoot tips was affected by the concentrations of sodium alginate and additives in the gel matrix (sucrose, gibberellic acid, MS nutrient medium) as well as duration of storage. The frequency of shoot and root induction of non-stored synthetic seeds was highest with shoot tips encapsulated with 2% sodium alginate containing 1.5% sucrose and 0.5 mg/l gibberellic acid (GA3). Shoot tips maintained their viability and ability to develop shoots even after 24 weeks of storage when they were encapsulated in 3% alginate with 1/3 MS medium, sucrose (1.5%) and GA3 (0.25 mg/l). Root formation tended to decrease with storage time. Overall, 90% of the plantlets derived from stored and non-stored synthetic seeds survived in the greenhouse and grew to phenotypically normal plants. This procedure can enable the use of synthetic seed technology for germplasm conservation of S. officinalis, a plant species of high medical and commercial value.
The study focused on the production of compounds with antioxidant activity in hairy root and shoot cultures of Salvia officinalis grown in laboratory-scale sprinkle nutrient bioreactors. HPLC analysis showed that production of rosmarinic acid in transformed roots (34.65±1.07 mg l-1) was higher that in shoot culture (26.24±0.48 mg l-1). In the latter diterpenoids: carnosic acid (1.74±0.02 mg l-1) and carnosol (1.34±0.01 mg l-1) were also found. Biomass accumulation after a growth period in the bioreactor was also studied. An 18-fold increase in hairy root biomass was recorded after 40 days of culture. In sage shoot culture, biomass increased 43 times after 21 days of bioreactor run. The current operating conditions of the bioreactor were not suitable for the propagation of Salvia officinalis mainly due to the hyperhydricity problem of leaves and stems.
Liquid shoot culture of Salvia officinalis L. in MS medium containing IAA (0.1 mg l-1) and BAP (0.45 mg l-1) was developed and evaluated in relation to shoot multiplication and antioxidant compound (carnosic acid, carnosol and rosmarinic acid) accumulation. In the liquid medium, on average, 3 new shoots per explant (shoot tip) were obtained within 3 weeks. The shoots produced 8.2±0.02 mg of diterpenoids and 31.2±0.29 mg of rosmarinic acid per gram of dry weight. Shoot proliferation and diterpenoid content increased when triacontanol (5, 10 or 20 pg l-1) was added to the liquid medium. In optimum conditions (at 20 pg l-1 TRIA) almost 7 shoots were formed per explant after 3 weeks. An increase in diterpenoid production (expressed as the sum of carnosol and carnosic acid) ranged from 30% to 50% and dependended on triacontanol concentration tested. The level of diterpenoids in triacontanol-treated shoots was similar to the content of compounds in commercial herbal product (dried leaves of S. officinalis) (10-12 mg g-1 dry wt). Triacontanol did not increase rosmarinic acid production, but the content of the phenolic as compound in shoots grown in liquid culture (31 mg g-1 dry wt) was even 24 times higher compared to samples of dried leaves of S. officinalis plants. We also demonstrated that the highest amounts of CA, Car and RA were accumulated in young, top parts of sage shoots. This observation could be useful for improving the selection of material for the extraction of natural antioxidants from S. officinalis.
Celem autorów była diagnoza stanu obiektów przemysłowych i poprzemysłowych warszawskiej dzielnicy Praga-Południe oraz opracowanie strategii dalszych działań. Przygotowano studia analityczne badanego terenu pod kątem uwarunkowań planistycznych, komunikacyjnych, funkcjonalnych, przyrodniczych i społecznych, a także analizę SWOT, będącą podsumowaniem pozytywnych i negatywnych cech badanego terenu. Efektem badań są proponowane kierunki praktycznych działań i wytyczne do projektu zagospodarowania przestrzennego dla wybranego fragmentu analizowanej dzielnicy. Opracowano również rekomendacje dla kształtowania procesu rewitalizacji na tym obszarze. Głównymi metodami wykorzystanymi podczas opracowania były: kwerenda dokumentów prawnych (w tym zwłaszcza planistycznych) i literatury, badania terenowe oraz proste analizy geoprzestrzenne z kategorii algebry map, których wyniki zostały zaprezentowane w formie opracowań kartograficznych. Na podstawie badań wyodrębniono do szczegółowego opracowania os. Kamionek, położone w północno-zachodniej części Pragi-Południe i zawierające w swych granicach większość dziedzictwa poprzemysłowego tej dzielnicy. Ze względu na wielość problemów zidentyfikowanych na opisywanym obszarze (zły stan techniczny budynków i infrastruktury, brak zieleni urządzonej, skomplikowane uwarunkowania prawne zabytkowych obiektów poprzemysłowych oraz niski poziom przedsiębiorczości mieszkańców) wymaga on rewitalizacji w jej kompleksowym wymiarze. Postulując rozwiązanie tych problemów, sporządzono wytyczne dla dalszego rozwoju tego obszaru, obejmujące m.in. uporządkowanie zagospodarowania przestrzennego, wykorzystanie miejscowych walorów kulturowych oraz wprowadzenie nowych funkcji celem jego ożywienia gospodarczego.
This report describes the effect of triacontanol on shoot multiplication and production of antioxidant compounds (carnosic acid, carnosol and rosmarinic acid) in S. officinalis cultures grown on MS basal medium (agar solidified medium supplemented with 0.1 mg l-1 IAA, 0.45 mg l-1 BAP). It was found that shoot proliferation significantly increased when triacontanol at concentrations of 5, 10 or 20 µg l-1 was added to the medium. HPLC analysis of acetone and methanolic extracts of sage shoots showed that the production of diterpenoids, carnosic acid/carnosol ratio, as well as, contents of rosmarinic acid were also affected by the treatment with triacontanol. The highest stimulation effect of triacontanol was observed on the production of carnosol, where the treatment with 20 µg l l-1 increased the content of this diterpenoid 4.5-fold compared to that in the control (sage shoots growing on MS basal medium, only).
The concentrations of carnosic acid, carnosol and rosmarinic acid in different materials from differentiated (multiple shoot cultures and regenerated plants) and undifferentiated (callus and cell suspension) in vitro cultures of Salvia officinalis were determined by HPLC. The results suggested that diterpenoid (carnosic acid and carnosol) production is closely related to shoot differentiation. The highest diterpenoid yield (11.4 mg g-1 for carnosic acid and 1.1 mg g-1 for carnosol) was achieved in shoots of 10-week-old micropropagated plants. The levels were comparable to those found in shoots of naturally growing plants. Undifferentiated callus and cell suspension cultures produced only very low amounts of carnosol (ca. 0.05 mg g-1 of dry weight). In contrast, content of rosmarinic acid in callus and suspension cultures as well as shoots growing in vitro and in vivo was similar and ranged between 11.2 and 18.6 mg g-1 of dry weight.
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