This study describes the influence of bio-synthesized silver nanoparticles (AgNPs) on phytochemicals and their pharmacological activities in the cell suspension cultures (CSC) of bitter gourd. To standardize the effect of sucrose, plant growth regulators, medium, AgNPs and growth kinetics for the biomass and bioactive compounds accumulation in CSC of bitter gourd. The medium comprising MS salts, sucrose (30 g/L) with 2,4-D (1.0 mg/L) and TDZ (0.1 mg/L) at 28 days of CSC was appropriate for biomass and bioactive compound accumulation. The contents of silver, malondialdehyde and hydrogen peroxide were highly elevated in AgNPs (10 mg/L)-elicited CSC when compared with non-elicited CSC. AgNPs (5 mg/L) elicited CSC extracts had significantly enhanced the production of total phenolic (3.5 ± 0.2 mg/g), and flavonoid (2.5 ± 0.06 mg/g) contents than in the control CSC extracts (2.5 ± 0.1 and 1.6 ± 0.05 mg/g). AgNPs (5 mg/L) elicited CSC showed a higher amount of flavonols (1822.37 µg/g), hydroxybenzoic (1713.40 µg/g) and hydroxycinnamic (1080.10 µg/g) acids than the control CSC (1199, 1394.42 and 944.52 µg/g, respectively). Because of these metabolic changes, the pharmacological activities (antioxidant, antidiabetic, antibacterial, antifungal and anticancer) were high in the AgNPs (5 mg/L)-elicited CSC extracts in bitter gourd. The study suggested the effectiveness of elicitation process in enhancing the accumulation of phenolic compounds and pharmacological activities. AgNPs-elicited CSC offered an effective and favorable in vitro method to improve the production of bioactive compounds for potential uses in pharmaceutical industries.

Turnip (Brassica rapa ssp. rapa) is an important vegetable crop producing glucosinolates (GSLs) and phenolic compounds. The GSLs, phenolic compound contents and transcript levels in hairy root cultures, as well as their antioxidant, antimicrobial and anticancer activity were studied in turnip. Transgenic hairy root lines were confirmed by polymerase chain reaction (PCR) and reverse transcription-PCR. GSLs levels (glucoallysin, glucobrassicanapin, gluconasturtiin, glucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin and 4-hydroxyglucobrassicin) and their gene expression levels (BrMYB28, BrMYB29, BrMYB34, BrMYB51, BrMYB122, CYP79 and CYP83) significantly increased in hairy roots compared with that in non-transformed roots. Furthermore, hairy roots efficiently produced several important individual phenolic compounds (flavonols, hydroxybenzoic and hydroxycinnamic acids). Colorimetric analysis revealed that the highest levels of total phenol, flavonoid contents, and their gene expression levels (PAL, CHI and FLS) in hairy roots than non-transformed roots. Our study provides beneficial information on the molecular and physiological active processes that are associated with the phytochemical content and biosynthetic gene expression in turnip. Moreover, antioxidant activity, as measured by DPPH scavenging activity, reducing potential, phosphomolybdenum and ferrous ion chelating ability assays was significantly higher in hairy roots. Hairy root extracts exhibited higher antimicrobial activity against bacterial and fungal species. The extract of hairy roots showed inhibition of human breast and colon cancer cell lines.

a b s t r a c tAn efficient protocol for hairy root induction of spine gourd (Momordica dioica) was established using Agrobacterium rhizogenes (KCTC 2703). This study evaluates the phenolic compound production, antioxidant and antimicrobial (antibacterial, antifungal and antiviral) activities of transgenic hairy root cultures in M. dioica . Hairy roots were induced from leaves, petiole, and internodal explants. Molecular analysis of PCR and gene sequencing using specific primers of rolC and aux1 revealed T-DNA integration in the hairy root clones and RT-PCR analysis confirmed the expression of hairy root inducible genes (rolC and aux1). The greatest biomass accumulation of hairy roots on MS liquid medium supplemented with 3% sucrose was observed at 22 days. Ultra-HPLC was used to compare the individual phenolic compound contents of transgenic and non-transgenic roots. Moreover, transgenic hairy roots efficiently produced several phenolic compounds, such as flavonols, hydroxycinnamic acid and hydroxybenzoic acid derivatives. The total phenolic, flavonoid contents and biological (antioxidant, antibacterial, antifungal and antiviral) activities were higher in hairy roots compared to non-transformed roots. These results demonstrate the greater potentiality of M. dioica hairy root cultures for the production of valuable phenolic compounds and for studies of their biological activity.

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Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry. To determine a suitable cell line for IBDV infection, replication, and growth kinetics of the virus, DF-1 cells and chicken embryo fibroblasts (CEF) were used. The population doubling per day (Pd/D) was found to be higher in DF-1 as compared to CEF cells. A suitable time of infection (TOI) was established for increased production of virus and greater infectivity titers. The DF-1 and CEF cells were found to be susceptible to infection by producing marked cytopathic effects (CPEs), and the growth curves of IBDV in DF-1 and CEF cells were evaluated by infectivity assay using tissue culture infectious dose (TCID50). The cytopathic effects of the virus in DF-1 and CEF cells were found to be similar, but higher viral titers were detected in the DF-1 cells as compared to CEF. Thus the DF-1 cell line had a higher growth potential and infectivity, which will be of advantage in vaccine production.