Response of Saponaria officinalis L. hairy roots to the application of TiO2 nanoparticles in terms of production of valuable polyphenolic compounds and SO6 protein.

Saponaria officinalis L. is a perennial plant from the Caryophyllaceae family whose various parts are used in traditional medicine as the treatment agent of skin diseases, blood purifier, diuretic, sudorific, and bile purifier. The cultivation system of hairy roots is a proper alternative for improving the valuable pharmaceutical compounds production compared to other in-vitro methods. The extensive nanotechnology applications in hairy roots cultivation is a sustainable production foundation to produce such active elements. In this study, the effect of various concentrations of titanium dioxide nanoparticles (TiO2 NPs) (0, 10, 20, 30, 50 mg L-1) with two treatments (24 and 48 h) was examined on the growth level, antioxidant capacity, total phenol and flavonoid contents, antioxidant enzyme activities, certain polyphenol compounds and SO6 protein in hairy roots of S. officinalis. According to the results, the maximum (3.09 g) and minimum (0.96 g) fresh weight (FW) of hairy roots were observed in treated culture media with 10 and 20 mg L-1 of TiO2 NPs after 24 and 48 h of exposure times, respectively. The highest rate of total phenol (9.79 mg GLA g-1 FW) and total flavonoid contents (1.06 mg QE g-1 FW) were obtained in the treated hairy roots with 50 and 30 mg L-1 of nano elicitor in 24 and 48 h of treatments, respectively. The maximum level of most polyphenols, such as rosmarinic acid, cinnamic acid, and rutin, was produced in 24 h of treatment. The use of TiO2 NP for 48 h with 50 mg L-1 concentration showed the highest production level of SO6 protein.

Inductive effect of titanium dioxide nanoparticles on the anticancer compounds production and expression of rosmarinic acid biosynthesis genes in Dracocephalum kotschyi transformed roots.

Methoxylated flavonoids, mainly xanthomicrol and cirsimaritin that can be extracted from Dracocephalum kotschyi Boiss, have anticancer, antispasmodic and antiplatelet effects. The production of these valuable pharmaceutical compounds is one of the major goals of biotechnology studies. In this work, induced transformed roots were influenced by various concentrations of titanium dioxide nanoparticles (TiO2 NPs) at 24 or 48 h exposure time. The effects of TiO2 NPs were assessed on growth rate, activity of antioxidant enzymes, total phenol and flavonoid content (TPC and TFC) and rosmarinic acid (RA) and some flavonoids accumulation. The gene expression level of phenylalanine ammonia-lyase (pal) and rosmarinic acid synthase (ras) genes were assessed by real time PCR analysis. The transformed roots biomass was substantially increased in elicited roots in comparison with the control. The TPC, TFC and antioxidant enzymes activitywere affected by TiO2 NPs concentration and exposure time. Valuable flavonoids with anticancer characteristics along with xanthomicrol, cirsimaritin and isokaempferide exhibited an increase (70, 34.28 and 7.81-fold, respectively) versus the control. The maximum content of RA (530.5 µg g-1 FW), which was 4.30 times as great as that of control was detected in samples treated with TiO2 NPs (50 mg L-1) 24 h after elicitation. Real-time PCR analysis revealed a considerable increase in pal and ras expression rate engaged by TiO2 NPs levels and exposure time. Overall D. kotschyi transformed roots elicitation by TiO2 NPs led to a massive increment in the production of valuable anticancer flavonoids such as xanthomicrol, cirsimaritin and RA as polyphenol.

Iron oxide nanoparticles: a novel elicitor to enhance anticancer flavonoid production and gene expression in Dracocephalum kotschyi hairy-root cultures.

BACKGROUND: Dracocephalum kotschyi Boiss. is a valuable source of rosmarinic acid (RA) and methoxylated hydroxyflavones (such as xanthomicrol and cirsimaritin) with antioxidative and antiplatelet effects and with antiproliferative potential against various cancer cells. The extensive application of nanotechnology in hairy root cultures is a new sustainable production platform for producing these active constituents. In the present study, hairy roots derived from 4-week-old leaves and Agrobacterium rhizogenes strain ATCC15834 were used to investigate the impact of various concentrations of iron oxide nanoparticles (Fe NPs) in two elicitation time exposures (24 and 48 h) on growth, antioxidant enzyme activity, total phenolic and flavonoid content (TPC and TFC), and some polyphenols. Gene expression levels of phenylalanine ammonia-lyase (pal) and rosmarinic acid synthase (ras) were also analyzed. RESULTS: Iron nanoparticles enhanced biomass accumulation in hairy roots. The treatment time and Fe NP dosage largely improved the activity of antioxidant enzymes, TPC and TFC. The highest RA (1194 µg g-1 FW) content (9.7-fold), compared to controls, was detected with 24 h of exposure to 75 mg L-1 Fe NP, which was consistent with the expression of pal and ras genes under the influence of elicitation. The xanthomicrol, cirsimaritin, and isokaempferide content was increased 11.87, 3.85, and 2.27-fold, respectively. CONCLUSION: Stimulation of D. kotschyi hairy roots by Fe NPs led to a significant increase in the induction and production of important pharmaceutical compounds such as rosmarinic acid and xanthomicrol. © 2019 Society of Chemical Industry.