The effects of polysaccharide elicitors such as chitin, pectin, and dextran on the production of phenylpropanoids (phenolics and flavonoids) and naphtodianthrones (hypericin and pseudohypericin) inHypericum perforatumshoot cultures were studied. Nonenzymatic antioxidant properties (NEAOP) and peroxidase (POD) activity were also observed in shoot extracts. The activities of phenylalanine ammonia lyase (PAL) and chalcone-flavanone isomerase (CHFI) were monitored to estimate channeling in phenylpropanoid/flavonoid pathways of elicited shoot cultures. A significant suppression of the production of total phenolics and flavonoids was observed in elicited shoots from day 14 to day 21 of postelicitation. This inhibition of phenylpropanoid production was probably due to the decrease in CHFI activity in elicited shoots. Pectin and dextran promoted accumulation of naphtodianthrones, particularly pseudohypericin, within 21 days of postelicitation. The enhanced accumulation of naphtodianthrones was positively correlated with an increase of PAL activity in elicited shoots. All tested elicitors induced NEAOP at day 7, while chitin and pectin showed increase in POD activity within the entire period of postelicitation. The POD activity was in significantly positive correlation with flavonoid and hypericin contents, suggesting a strong perturbation of the cell redox system and activation of defense responses in polysaccharide-elicitedH. perforatumshoot cultures.

Hypericum perforatum L. is a medicinal plant considered as an important natural source of secondary metabolites with a wide range of pharmacological attributes. Hairy roots (HR) were induced from root segments of in vitro grown seedlings from H. perforatum after cocultivation with Agrobacterium rhizogenes A4. Investigations have been made to study the production of phenolic compounds in dark‐grown (HR1) and photoperiod‐exposed (HR2) cultures. The chromatographic analysis of phenolic acids, flavonols, flavan‐3‐ols, and xanthones revealed marked differences between HR1 and HR2 cultures. The production of quinic acid, kaempferol, and seven identified xanthones was increased in HR2. Moreover, HR2 showed a capability for de novo biosynthesis of two phenolic acids (3‐p‐coumaroylquinic acid and 3‐feruloylquinic acid), three flavonol glycosides (kaempferol hexoside, hyperoside, and quercetin acetylglycoside), and five xanthones (tetrahydroxy‐one‐methoxyxanthone, 1,3,5‐trihydroxy‐6‐methoxyxanthone, 1,3,5,6‐tetrahydroxy‐2‐prenylxanthone, paxanthone, and banaxanthone E). On the other side, HR1 cultures were better producers of flavan‐3‐ols (catechin, epicatechin, and proanthocyanidin dimers) than HR2. This is the first comparative study on phenolic profile of H. perforatum HR cultures grown under dark and photoperiod conditions.

Three Hypericum perforatum hairy root lines (HR B, HR F and HR H) along with non-transformed roots were analyzed for phenolic compounds composition and in vitro enzyme inhibitory properties. In silico molecular modeling was performed to predict the interactions of the most representative phenolic compounds in HR clones with enzymes related to depression, neurodegeneration and diabetes. Chromatographic analyses revealed that HR clones represent an efficient source of quinic acid and hydroxybenzoic acids, epicatechin and procyanidin derivatives, quercetin and kaempferol glycosides, as well numerous xanthones. In vitro antidepressant activity of HR extracts through monoamine oxidase A (MAO-A) inhibition was attributed to the production of oxygenated and prenylated xanthones. The neuroprotective potential of HR extracts was related to the accumulation of quercetin 6-C-glucoside, epicatechin, procyanidins and γ-mangostin isomers as potential inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Vanillic acid and prenylated xanthones in HR clones as promising inhibitors of tyrosinase additionally contributed to the neuroprotective activity. Five preeminent xanthones in HR (γ-mangostin, mangiferin, garcinone C, garcinone E and 1,3,7-trihydroxy-6-metoxy-8-prenyl xanthone) along with the flavonol quercetin 6-C-glucoside effectively inhibited α-amylase and α-glucosidase indicating the antidiabetic properties of HR extracts. Transgenic roots of H. perforatum can be exploited for the preparation of novel phytoproducts with multi-biological activities.

The aim of this study was to evaluate the non-enzymatic and enzymatic antioxidant response of fifteen Hypericum perforatum L. dark-grown (HR1 A-HR1 O) and photoperiod-exposed (HR2 A-HR2 O) hairy root clones. Dark-grown HR1 clones were characterized with high biomass accumulation and up-regulated phenylpropanoid metabolism through phenylalanine ammonia lyase (PAL)-mediated production of flavonoids, flavan-3-ols and anthocyanins. These groups of phenolics showed superior hydrogen-donating capability and significantly contributed to the antioxidant capacity of dark-grown HR1 clones. Photoperiod-exposed HR2 clones showed green coloration with shoot regenerative potential and reduced biomass accumulation. Photoperiod exposition improved the production of hypericins, as well as ferrous chelating properties and lipid peroxidation inhibition activity in HR2 cultures. Furthermore, HR2 clones were represented with enhanced activity of antioxidant enzymes (guaiacol peroxidase, catalase and superoxide dismutase) that resulted in suppression of oxidative stress markers (hydrogen peroxide, superoxide anion and malondialdehyde). These observations revealed the involvement of an efficient antioxidant defense system in the adaptive response of HR to photooxidative stress. Altogether, photoperiod-exposed H. perforatum HR2 clones were considered as a promising alternative for further scale-up production of naphthodianthrones that could be used in the pharmaceutical industry.