Browsing by Subject "Carrot suspension cultured cells"

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    Effect of diflufenican on total carotenoid and phytoene production in carrot suspension‑cultured cells
    (Springer, 2018-08-06) Fraser, Paul D.; Almagro Romero, Lorena; Miras Moreno, Begoña; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
    This work analyzes the efect of difufenican, an inhibitor of phytoene desaturase, on the gene expression profles of the biosynthetic pathway of carotenoids related with the production of these compounds in carrot cell cultures. The results showed that the presence of 10 µM difufenican in the culture medium increased phytoene levels, which was 493-fold higher than in control cells after 7 days of treatment but did not alter cell growth in carrot cell cultures. The maximal production of phytoene was reached with 10 µM difufenican after 7 days of incubation in the presence of light and with 30 g/L sucrose in the culture medium. Moreover, difufenican decreased the expression of phytoene synthase and phytoene desaturase genes at all the times studied. This difufenican-induced inhibition of phytoene desaturase gene expression in carrot cell cultures resulted in an increased production of phytoene. Our results provide new insights into the action of difufenican in carrot cell cultures, which could represent an alternative more sustainable and environmentally friendly system to produce phytoene than those currently used.
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    Effect of terbinafine on the biosynthetic pathway of isoprenoid compounds in carrot suspension cultured cells
    (Springer, 2018-04-21) Almagro Romero, Lorena; Miras Moreno, Begoña; Pedreño García, María Ángeles; Sabater Jara, Ana Belén; Biología Vegetal
    Plant sterols are essential components of membrane lipids, which contributing to their fluidity and permeability. Besides their cholesterol-lowering properties, they also have anti-inflammatory, antidiabetic and anticancer activities. Squalene, which is phytosterol precursor, is widely used in medicine, foods and cosmetics due to its anti-tumor, antioxidant and anti-aging activities. Nowadays, vegetable oils constitute the main sources of phytosterols and squalene, but their isolation and purification involve complex extraction protocols and high costs. In this work, Daucus carota cell cultures were used to evaluate the effect of cyclodextrins and terbinafine on the production and accumulation of squalene and phytosterols as well as the expression levels of squalene synthase and cycloartenol synthase genes. D. carota cell cultures were able to produce high levels of extracellular being phytosterols in the presence of cyclodextrins (12 mg/L), these compounds able to increase both the secretion and accumulation of phytosterols in the culture medium. Moreover, terbinafine induced a significant increase in intracellular squalene production, as seen after 168 h of treatment (497.0 ± 23.5 µg g dry weight−1) while its extracellular production only increased in the presence of cyclodextrins.The analysis of sqs and cas gene expression revealed that cyclodextrins did not induce genes encoding enzymes involved in the phytosterol biosynthetic pathway since the expression levels of sqs and cas genes in cyclodextrin-treated cells were lower than in control cells. The results, therefore, suggest that cyclodextrins were only able to release phytosterols from the cells to the extracellular medium, thus contributing to their acumulation. To sum up, D. carota cell cultures treated with cyclodextrins or terbinafine were able to produce high levels of phytosterols and squalene, respectively, and, therefore, these suspension-cultured cells of carrot constitute an alternative biotechnological system, which is at the same time more sustainable, economic and ecological for the production of these bioactive compounds.