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Castilleja tenuiflora Benth. (Orobanchaceae) in vitro cultures are alternative sources of phenylethanoid glycosides (PhGs), promising natural products for chronic diseases treatment because of their extensive range of biological activities. To increase the yields of these bioactive compounds, the identification of factors affecting their biosynthesis is required. Here, we show that N deficiency stimulates PhGs biosynthesis by increasing the activity of PAL. The contents of two PhGs, verbascoside and isoverbascoside, were enhanced under N deficiency compared with control conditions. The maximum PAL activity in shoots cultured under N deficiency was also increased. Furthermore, we found de novo anthocyanin synthesis under N deficiency suggesting an additional impact of this stress factor on flavonoid metabolism. N deficiency negatively affected the shoot growth (length, biomass, and total chlorophyll content) and multiplication rate, and inhibited the root formation of C. tenuiflora shoots. Our results demonstrate that N deficiency increases PhGs biosynthesis in C. tenuiflora.
The aim of this study was to determine if the increase of the initial sucrose concentration (ISC) improves cell growth and arabinogalactan protein (AGP) secretion of Beta vulgaris L. cultures. ISC tested were 43.8, 87.6 and 131.4 mM. Cell growth and specific growth rate were improved increasing the ISC. Cell cultures grown with ISC 43.8 mM were fed with sucrose, and cellular growth was enhanced twofold, revealing the stimulatory effect of sucrose on cell growth. The AGP secretion was stimulated, increasing the ISC. This event was partially associated with the exponential growth phase of the culture. AGP precipitation with Yariv reagent of cell cultures inhibited cell growth without changes in viability. The assay of sucrose feeding confirmed the relationship between cell growth and AGP secretion. These observations suggest that AGPs may be required for cell division. The increase of AGP secretion by ISC coincided with a higher cellular aggregation, suggesting a possible role of AGP as cellular adhesion molecules. To determine whether AGP secretion is also stimulated by an osmotic effect, mannitol was fed to raise the osmotic potential from 23.78 to 95.97 mOsm kg⁻¹. Mannitol was not used for cell growth, but AGP secretion was stimulated sixfold in relation to the control. These results are important for understanding the possible factors involved in the AGP secretion of plant cell culture and that may be considered to improve the AGP production.
The content of total phenolic compounds and flavonoids was determined in methanol extracts of root, stem, leaves, and inflorescences from wild growing and greenhouse cultivated plants of Castilleja tenuiflora. The antioxidant activity in each extract was evaluated using three in vitro models: scavenging of free radicals with 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), and reducing power by the phosphomolybdenum assay. Both, antioxidant activity and phytochemicals content were influenced significantly (P<0.05) by the source of the plant material and the organ. Cultivated plants had the highest content of phenolic compounds (37.95 mg gallic acid equiv. g⁻¹ dry weight, P<0.05) and the strongest antioxidant activity. Total phenolic compounds content correlated significantly with the antioxidant activity for all studied plant material and organs (P<0.05). TLC profile using DPPH as a detection reagent indicated that the phenylethanoids verbascoside and isoverbascoside are the main contributors to the free-radical scavenging of C. tenuiflora. Cultivated plants of C. tenuiflora are an alternative source of natural antioxidants to wild growing plants. The antioxidant properties of C. tenuiflora may be associated with its traditional use to treat conditions consistent with radical-related diseases (e.g. Inflammation, tumors).
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