To overcome the problems in liverwort collecting such as small size and easily mixed with other species in the wild, we have successfully cultivated Marchantia polymorpha L. under in vitro conditions in the previous study. The aim of this study is to evaluate the biological activities of this in vitro biomass as a confirmation of the sufficient protocol in cultivation this species. Cultured biomass was dried at a temperature of 45-50 oC to constant weight and ground into a fine powder. The coarse powder was extracted with organic solvents of increasing polarization including n-hexane, chloroform, ethyl acetate, and ethanol using the maceration technique. Four extracts were investigated antioxidant (iron reduction power, DPPH), antibacterial (agar diffusion), tyrosinase inhibitory activity, anti-proliferation on MCF-7 cells. Additionally, the presence of natural metabolite groups of the extracts was detected by using specific reagents. For antioxidant activity, ethyl acetate fraction extract had the highest iron reducing power and DPPH free radical scavenging ability with IC50 = 439.31 µg ml-1. All three n-hexane, chloroform, and ethyl acetate extracts possessed resistance to the bacterial strain tested. At a concentration of 2 mg ml-1, n-hexane and chloroform extracts had the highest percentage of tyrosinase inhibition (69.54 and 69.10%, respectively). The n-hexane extract is a potent extract that inhibits the proliferation of MCF-7 cells with the lowest IC50 of 38.15 µg ml-1. A preliminary chemical composition survey showed that the cultured biomass liverwort contains many bioactive compounds, particularly the compounds of range of non- and less-polarized fractions.

Catharanthus roseus L. is a medicinal plant that produces numerous indole terpenoid alkaloids, including vincristine and vinblastine, which are used for cancer treatment. The effect of specified precursors (L-phenylalanine, L-tyrosine) and elicitors (chitosan, methyl jasmonate) on C. roseus hairy roots (CHR) growth has been examined in order to increase the content of vincristine and vinblastine. Our results showed that CHR generated by an Agrobacterium rhizogenes strain isolated in Vietnam was capable of producing both vincristine and vinblastine when subjected to precursors, but only vinblastine when exposed to elicitors. However, both precursors and elicitors were evaluated to have an effect on increasing the accumulation of TIAs in CHR. In particular, the use of elicitors required more time to find the appropriate induction conditions, while the use of precursors gave outstanding efficiency in the treatment with 1 µM phenylalanine. The greatest yields of vincristine (51.99 µg g-1 DW) and vinblastine (699.92 µg g-1 DW) were obtained in the 7th week (with 0.306 g DW biomass). This result is the first time we might boost the levels of vincristine and vinblastine in our CHR clone generated by the Vietnam strain of A. rhizogenes.

Aeroponics is considered as a potential method for the culture of herbal plants due to the high growth rate, quantity and quality enhancement of secondary metabolites, and substantial environmental progress associated with this method. The aim of this study was to develop a sufficient protocol for successful Urena lobata hairy root induction by Agrobacterium rhizogenes ATCC 15834, using a precursor and elicitor to enhance α-glucosidase inhibitory activity (GIA) of aeroponic hairy roots (AHRs) in greenhouse conditions. In this study, we found that the optimized procedure (10 min, Woody plant medium (WPM), 1/25 salt strength) had an outstanding effect with a reduction in the rooting time (RT), promotion of the rooting rate (RR), and increase in the fresh weight (FW) and dry weight (DW) compared with the original procedure (30 min, Murashige and Skoog (MS) medium, 1/25 salt strength) after 30 days of culture. The highest DW, GIA, flavonoid (FLA) and phenolic (PHEL) contents were observed for individual addition of 10 mM phenylalanine (PA) or 50 mM chitosan (CS) in the late exponential phase (eighth week) with 15 days of elicitation compared to the control AHRs. However, individual treatment was less effective than the combination of the two. Positive correlations among the GIA, FLA and PHEL indicate that AHRs accumulated phenolic compounds, leading to an increase in the GIA by a synergistic effect. In conclusion, the culture of Urena lobata AHRs with PA and CS is an efficient procedure to produce GIA material in greenhouse conditions. The treatment of type II diabetes is sophisticated in terms of the variety of risk factors, one of which is the considerably rapid development of postprandial hyperglycemia [5]. Postprandial hyperglycemia occurs through the reaction of α-glucosidase and α-amylase in order to produce glucose. Therefore, by restricting or reducing the activity of these enzymes, the occurrence of postprandial hyperglycemia should decrease. The inhibition of α-glucosidase activity leads to a deterioration in disaccharide hydrolysis which has substantial long-term effects on glycemic index control in diabetic patients [6]. In recent years, natural medicines from plants have been used as useful treatments without any side effects.Hairy roots (HRs) are established from the interaction between Agrobacterium rhizogenes and the host plant (gene transformation from bacteria to the genome of the plant) [7]. There are numerous merits that accrue from the collaboration; in particular, a high growth rate without the dependence of any exogenous hormones, genetic stability, and a high branching rate. In addition, the ability to produce secondary metabolites remains stable. Hence, the HR cultural technique is considered to be an attractive substitute for the manufacture of tremendously valuable natural secondary metabolites instead of bud or callus cultures [8].Accompanied by the significant progress of modern techniques, aeroponics is a new method that has been adapted from classical hydroponics. Compared with hydropon...

White mulberry (Morus alba L.) root was reported to contain potential tyrosinase inhibitors such as norartocarpetin, streppogenin, oxyresveratrol (5, 7, 2′, 4′-tetrahydroxy-flavavone) and 2-arylbenzofuran. This study aims at investigating Morus alba L. hairy root culture to create a source of tyrosinase inhibitors for skin-whitening application. Morus alba L. leaf (including mesophyll and petiole) and stem explants were randomly wounded and inoculated with Agrobacterium rhizogenes ATCC 15834 in 10-40 minutes, and coculturing for 4 days. After inoculation, Agrobacterium rhizogenes were removed by using MS medium adding 200mg/L cefotaxime as antibiotics. Then, fresh inducted hairy roots were grown in nonphytohormone liquid medium (MS, White, Gamborg B5 or WPM) on a shaker (80rpm) at 25 ±1°C. Inducted hairy roots were tested for the present of rolB by PCR analysis and its effect on tyrosinase activity using a 96-well plate reader. The suitable explants for hairy root induction are leaves, especially petioles in 10 minutes (root induction frequency: 83%). After induction, hairy roots on MS was grown more efficiently (increased 3.933 times in FW) than on WPM, B5 and White medium. Inducted hairy roots were successfully transgenes by the present of rolB in their genome and had the same anti-tyrosinase activity as the natural roots without significance differences (IC50: 3.860 and 3.597 μg/mL, respectively). Cultured Morus alba L. hairy roots are potential source of tyrosinase inhibitors for future skin-whitening ingredient.

Paramignya trimera (Oliv.) Guill., a woody climber commonly known as "Xao tam phan", has been used in Vietnamese folk for the treatment of numerous cancers. Due to word of mouth about the anticancer properties of this plant, its stems and roots have been overexploited leading to the serious decline of this species in Phu Yen, Khanh Hoa and Ninh Thuan provinces. The aim of the study was to establish an in vitro propagation protocol for the conservation of P. trimera. In this research shoot clusters (5–8 shoots/cluster) were regenerated from axillary bud explants of 1–3 year-old trees after 3 months of cultures on the WPM (woody plant medium) supplemented with STS 3 and BA 5–7 mg/L. STS (silver thiosulfate) was used to prevent the leaf abscission. These shoot clusters grew slowly and reached 1–3 cm in heights after 4 months of the cultures. These shoot clusters did not form any roots after 2 months of culture on the rooting media with IBA and/or NAA 1–5 mg/L. However, there was 51 % of the treated shoot clusters acclimatized and produced new stem and leaves after 2 months growing in greenhouse. WPM supplemented with STS 3, BA 5 and IBA 5 mg/L showed the best response for callus induction in leaf explants after 3 months of cultures. Among the callus types, the milky white compact calli were induced at the cut surface of leaf explants after 3 months of the cultures and became the compact and nodulated calli within 4 weeks later.