Green synthesized Ag nanoparticles stimulate gene expression and paclitaxel production in Corylus avellana cells.

Synthesis of nanoparticles (NPs) through plant extracts has been suggested as an effective and nature-friendly method. Paclitaxel is one of the most valuable secondary metabolites with therapeutic uses, and hazelnut has been suggested as one of the sustainable resources for producing this metabolite. In the present study, we synthesized Ag NPs using the ethanolic extract of C. avellana leaves and were characterized using UV-visible, FTIR, XRD, EDX, DLS, SEM, and TEM analyses. In addition, we investigated the effect of green synthesized Ag (GS Ag) NPs (5 and 10 mg/L), para-aminobenzoic acid (PABA) (20 mg/L), and AgNO3 (10 mg/L) on cell viability, physiological characteristics, gene expression, and biosynthesis of secondary metabolites in hazelnut cell cultures. The results showed that 10 mg/L Ag NPs and AgNO3 significantly affected the cell viability, the content of ROS, peroxidation of lipids, antioxidant capacity, secondary metabolite production, and expression pattern of the genes involved in the taxanes biosynthesis pathway in the hazelnut cells. The cytotoxicity increased by increasing the GS Ag NPs concentration from 5 to 10 mg/L, which was associated with reduced membrane integrity and cell viability. Elicitation of the cells with 10 mg/L Ag NPs combined with 20 mg/L PABA (as a precursor) remarkably excited the expression of TAT and GGPPS genes and the production of secondary metabolites as well as paclitaxel. So that the highest expression of TAT and GGPPS genes (3.71 and 3.69) and the highest amount of taxol (230.21 µg g-1 FW) and baccatin (1025.8 µg g-1 FW) were observed in this treatment. KEY POINTS: • For the first time, we assessed and reported the molecular and physiological responses of C. avellana cells to GS Ag NPs, AgNO3, and PABA. • In hazel cells, GS Ag NPs stimulate several physiological and molecular responses. • In addition to increasing antioxidant activity, GS Ag NPs significantly increased the expression of genes involved in the paclitaxel biosynthesis pathway and the production of secondary metabolites.

Biologically synthesized CuO nanoparticles induce physiological, metabolic, and molecular changes in the hazel cell cultures.

The utilization of plant extracts in nanoparticle (NP) synthesis has been suggested as a nature-friendly method and an efficient alternative to the conventional approaches such as physical and chemical methods. Taxol is a valuable medicinal compound, and hazelnut has been suggested as one of the sustainable resources for producing this metabolite. In the present research, copper oxide (CuO) nanoparticles (NPs) were biologically synthesized by utilizing hazelnut leaf extracts. FTIR, XRD, EDAX, DLS, and SEM analyses were used for characterizing and confirming the synthesized NPs. The effect of biosynthesized CuO NPs (10 and 90 ppm), para-aminobenzoic acid (PABA) (20 ppm), and CuSO4 (10 ppm) on the cell viability, biochemical properties, expression of TAT and GGPPS genes, and accumulation of taxol and baccatin III in hazelnut cell cultures was investigated. The results indicated that biosynthesized CuO NPs significantly influenced the cell viability, amount of ROS, antioxidant capacity, lipid peroxidation, secondary metabolite production, and expression pattern of the genes engaged in the biosynthesis pathway of taxanes in the C. avellana L. cells. The cytotoxicity of CuO NPs to cells was dose dependent and increased with increasing its concentration, as evidenced by a decline in the survival rate and cell membrane integrity. Furthermore, the utilization of 10 ppm CuSO4 caused more toxicity in the cells than the same concentration of CuO NPs. This result could be attributed to the fact that plant extracts components act as a coating for the NPs and reduce their toxicity. Treatment of the cell cultures with CuO (10 ppm) + PABA (20 ppm) and CuO (10 ppm) induced the highest radical scavenging activity. The activity of antioxidant enzymes was increased with increasing the copper oxide NPs level from 10 to 90 ppm. Contrariwise, the cell's survival rate, radical scavenging activity, and amount of secondary metabolites were significantly reduced in the higher levels of copper oxide NPs (90 ppm) compared to the 10 ppm. The combined utilization of 10 ppm copper oxide NPs and 20 ppm PABA considerably stimulated the TAT and GGPPS genes expression and produced the highest amount of taxol and baccatin III. KEY POINTS: • CuO NPs were biologically synthesized using the hazel leaf extracts and confirmed by FTIR, XRD, EDAX, DLS, and SEM analyses. • CuO NPs significantly affected the amount of ROS, antioxidant capacity, and lipid peroxidation in C. avellana L. cells. • Treatment of the hazel cells with CuO NPs increased the production of secondary metabolites including taxol and baccatin III and expression of the genes involved in taxol and baccatin III biosynthesis (TAT and GGPPS).

Factors affecting the growth, antioxidant potential, and secondary metabolites production in hazel callus cultures.

Hazelnut is one of the most important nut plants recently suggested as a sustainable source for paclitaxel. In the present study, the effect of the concentration and combination of PGRs, different basal medium and ultrasonic waves on callus induction and growth, physiological characteristics, and taxol and baccatin III production in hazelnut callus cultures were investigated. The results indicated that combining 2,4-D (2 mg/L) and Kin (0.2 mg/L) with the sonication of explants for 1 min provides an optimized condition for callus induction and growth. Hazelnut explants exhibited different callus production and biochemical and metabolic characteristics depending on the basal medium type, ultrasound treatment, and inclusion of ascorbic acid in the medium. So that, the highest percentage of callogenesis (100%) observed in ½ MS + 1 min US, ½ MS + 150 mg/L AA, B5 + 1 min US and B5 + 150 mg/L AA, and also ½ MS salt + Nitsch vitamins + 150 mg/L AA. Furthermore, the highest callus growth (7.86 g FW) was obtained from ½ MS + 1 min US. The highest amount of baccatin III production (147.98 and 147.85 mg/L) was obtained from the WPM and MS basal media; the highest taxol production (44.89 mg/L) was observed in the WPM basal medium. The cultures in the MS, WPM, and MS salts + Nitsch vitamins media, had the highest H2O2 and MDA content, antioxidant enzymes activity, and phenolic compounds. In conclusion, culture media nutrient composition and concentration not only affect the cell growth and physiological status of the cultures but also improve secondary metabolites production and accumulation.