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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 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.
A two-factorial experiment was conducted at Agricultural University in Szczecin. The first factor of the experiment was the level of magnesium fertilization: MgO - Og of Mg per flowerpot (Mitcherlich type), Mgl - 0.30g of Mg per flowerpot and Mg2 - 0.90g of Mg per flowerpot. The second factor of experiment was the level of soil moisture: 30% and 60% of maximum water capacity. The content and chemical composition of essential oil were determined in the leaves of S. officinalis L. The leaves of tested plant were harvested twice every year. The results of the present study showed that the content of essential oil in the leaves of medical sage depended on the doses of magnesium fertilization. Chemical composition of the oil was similar for all experimental combinations, except for combination Mg2 30% maximum water capatity.
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.
Badania prowadzono w celu określenia, czy inokulacja grzybami tworzącymi mikoryzę arbuskularną (AMF) i stosowanie kompostu poprawi tolerancyjność szałwii omszonej na NaCl, CaCl2 i Na2CO3. Rośliny nieinokulowane i inokulowane AMF uprawiano w podłożu bez kompostu i w podłożu wzbogaconym w kompost i traktowano NaCl, CaCl2 i Na2CO3 w stężeniu 0, 11,68 i 23,36 g∙dm-3. Traktowanie NaCl nieznacznie obniżało suchą masę i znacznie hamowało wzrost elongacyjny pędów, opóźniało kwitnienie szałwii oraz obniżało procent zasiedlenia korzeni przez grzyby mikoryzowe. Traktowanie CaCl2 wpływało niekorzystnie na suchą masę roślin inokulowanych i uprawianych w podłożu z dodatkiem kompostu, ale nie wpływało na wysokość roślin, termin kwitnienia i liczbę kwiatostanów. Traktowanie Na2CO3 obniżało suchą masę szałwii inokulowanej AMF i uprawianej w podłożu z dodatkiem kompostu. Wpływ Na2CO3 na wysokość roślin i liczbę kwiatostanów był niewielki. Mikoryzacja nie wpływała korzystnie na wzrost i kwitnienie szałwii. Dodatek kompostu poprawiał wszystkie parametry wzrostu roślin i zwiększał liczbę kwiatostanów, niezależnie od traktowania roślin solami i inokulacji AMF. Najbardziej szkodliwe dla wartości dekoracyjnej szałwii okazały się traktowania NaCl i Na2CO3 w stężeniu 23,36 g∙dm-3, powodujące nekrozę liści. Traktowanie Na2CO3 w stężeniu 23,36 g∙dm-3 powodowało pojawianie się brązowych plam na liściach dolnych. Zarówno mikoryzacja, jak i dodatek kompostu, stosowane oddzielnie, nie zapobiegały ujemnemu oddziaływaniu testowanych soli na jakość liści szałwi omączonej. Liście szałwii inokulowanej AMF i uprawianej w podłożu z dodatkiem kompostu i podlewanej roztworami CaCl2 lub Na2CO3 w stężeniu 23,36 g∙dm-3 nie miały nekroz i brązowych plam.
Salvia officinalis L. (Dalmatian sage) is used traditionally in medicine as herbal tea and as a tonic, antiseptic and for inflammations and infections in the mouth. Also, sage is widely used as food flavouring, either as dried leaves or as the essential oil or oleoresin. Thus, any heavy metal accumulation by this plant would increase the possibility of heavy metal toxicity to consumers. In Montenegro, sage and some other aromatic herbs grow wild in close proximity to major roads. This study aims to establish the impact of pollution on the composition of sage leaves and this first investigation acts as a platform from which further work will follow. The total contents of Pb, Cd, Cu and Zn in Salvia officinalis leaves and in essential oil, solvent extract, tea brew and tea infusion at selected distances from a heavy metal pollution source was examined. Generally, it was found that the concentration of heavy metals decreases at 50 m, i.e. 100 m from the edge of a major road. Therefore the impact of road traffic through the pollution of aromatic herbs was noted. The concentration of hazardous heavy metals Pb and Cd in all investigated sage extracts was bellow detection limits of the analytical technique used while the solubility of heavy metals Zn and Cu was the highest in tea infusion extract.
Background. Natural antioxidants become increasingly important in fish processing and particularly in the preservation phase of fish raw materials. Some of them can protect lipids, containing essential unsaturated fatty acids. The aim of the present study was to determine antioxidant properties of aqueous extracts of rosemary and sage, compared with the activity of BHA (butylated hydroxyanisole) and Endox in lipids of fresh Baltic fishes (herring, sprat, flounder) and those subjected to frozen storage at -25°C for 6 months. Materials and Methods. Antioxidant activity of the additives was determined based on β-carotene changes in fish lipids after 10, 20, 40, and 60 min of heating at 50°C. Results. The plant-origin extracts and the synthetic antioxidants used, inhibited most extensively β-carotene changes in the lipids of non-frozen herring. They were the least effective in non-frozen sprat lipids. The frozen storage lowered the antioxidant action of the additives in relation to β-carotene in lipids of herring and flounder, whereas in sprat lipids the antioxidants used, continued their protective action for β-carotene, during heating. Elongation of the heating time from 10 to 60 min increased the β-carotene losses and their pace followed the logarithmic scale. The activity of the antioxidants in fish lipids decreased along with the increase of polyunsaturated fatty acids (C20 : 5, C22 : 5, and C22 : 6) contents Conclusion. The antioxidant properties of plant-origin additives and synthetic antioxidants decreased along with the increased content on unsaturated fatty acids of fish lipids. Six-month-long frozen storage of the fish raw materials resulted in a decrease of the protective properties of the additives used, in relation to the lipids of Baltic herring and flounder. The best protective properties for lipids of flounder and sprat exhibited Endox, while aqueous extract of rosemary was the most effective in relation to sprat lipids.
The objective of the study was to perform micromorphological analyses of the secretory structures of leaves and stems of oil-bearing industrial plants from the Lamiaceae family such as lemon balm (Melissa officinalis L.), peppermint (Mentha × piperita L.), sage (Salvia officinalis L.), marjoram (Origanum majorana L., syn. Origanum dubium Boiss.), rosemary (Rosmarinus officinalis L.) and common thyme (Thymus vulgaris L.) using light microscope and scanning electron microscope. In addition, an estimation of the content of volatile substances in the plant species under study was performed using GC-MS, as well as the qualitative and quantitative analysis of essential oil, that is an important component in terms of the estimation of raw material applicability for use in the industry. In the epidermal cells of studied plants, 2 types of Lamiaceae-type glandular trichomes were identified: short- and long-stalked capitate glandular trichomes with single- and bicellular secretory capitulum, and peltate glandular trichomes with eight- and over a dozen-cell secretory capitulum. Capitate trichomes were densely distributed on the surface of the epidermis, while peltate trichomes were sparse, though regular, and were situated in depressions. Glandular trichomes were found more frequently on leaves than on stems. The cuticle of the abaxial of leaf was characterized in most cases by the occurrence of larger average diameter peltate trichomes compared to the cuticle of the adaxial side of leaf. Peppermint produced the largest structures accumulating essential oil on the leaves (average diameter of peltate trichomes – 78.48 µm on the adaxial side of leaf, up to 96.43 µm), while on the stem, the highest average diameter of the peltate trichomes was observed in sage (an average of 75.53 µm, up to 85.99 µm). The lemon balm was characterized by the presence of capitate and peltate trichomes with the smallest diameter (an average of 44.26 µm). Lemon balm was characterized by the greatest density of glandular trichomes compared to other plant species. Among the plants studied, the highest content of oil was noted in the case of thyme and peppermint (2.22% and 2.20% v/w, respectively), and the lowest in green parts of lemon balm (0.17% v/w). The isolated essential oils contained predominantly components from the groups of monoterpenes and sesquiterpenes, and it is the presence of those substances that determines the possibility of utilizing the plants studied for a variety of purposes.
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