Special Issue-"Bioactive Compounds from Natural Sources II".

In recent decades, there has been a great interest in bioactive compounds from natural sources [...].

Distinguishing features of Lycium L. species (family Solanaceae) distributed in Egypt based on their anatomical, metabolic, molecular, and ecological characteristics.

Among the 70-80 species of the genus Lycium (family Solanaceae) disjunctly distributed around the world, only three are frequently distributed in different locations in Egypt. Due to the morphological similarities between these three species, there is a need for alternative tools to distinguish them. Thus, the objective of this study was to revise the taxonomic features of Lycium europaeum L., Lycium shawii Roem. & Schult., and Lycium schweinfurthii var. aschersonii (Dammer) Feinbrun in consideration of their anatomical, metabolic, molecular, and ecological characteristics. In addition to analysis of their anatomical and ecological features, DNA barcoding was performed for molecular characterization through internal transcribed spacer (ITS) sequencing and start codon targeted (SCoT) markers. Furthermore, metabolic profiling of the studied species was conducted based on gas chromatography-mass spectrometry (GC-MS). The observed anatomical features of the adaxial and abaxial epidermal layers, type of mesophyll, crystals, number of palisade and spongy layers, and the vascular system showed variations between the studied species. Beyond this, the anatomy of the leaves showed an isobilateral structure in the studied species, without distinct differences. Species were molecularly identified in terms of ITS sequences and SCoT markers. The ITS sequences were deposited in GenBank with accession numbers ON149839.1, OP597546.1, and ON521125.1 for L. europaeum L., L. shawii, and L. schweinfurthii var. aschersonii, respectively. The sequences showed variations in GC content between the studied species; this was 63.6% in L. europaeum, 61.53% in L. shawii, and 63.55% in L. schweinfurthii var. aschersonii. A total of 62 amplified fragments, including 44 polymorphic fragments with a ratio of 70.97%, were obtained in the SCoT analysis, as well as unique amplicons in L. europaeum L., shawii, and L. schweinfurthii var. aschersonii of 5, 11, and 4 fragments, respectively. Through GC-MS profiling, 38 compounds were identified with clear fluctuations in the extracts of each species. Of these, 23 were distinguishing chemicals that could help in chemical identification of the extracts of the studied species. The present study succeeds in identifying alternative clear and diverse characteristics that can be used to distinguish between L. europaeum, L. shawii, and L. schweinfurthii var. aschersonii.

Potential of Low Cost Agro-Industrial Wastes as a Natural Antioxidant on Carcinogenic Acrylamide Formation in Potato Fried Chips.

Acrylamide is classified as a toxic and a prospective carcinogen to humans, and it is formed during thermal process via Maillard reaction. In order to find innovative ways to diminish acrylamide formation in potato chips, several extracts of agricultural wastes including potato peels, olive leaves, lemon peels and pomegranate peels extracts were examined as a soaking pre-treatment before frying step. Total phenolic, total flavonoids, antioxidant activity, and the reduction in sugar and asparagine contents were additionally performed. Proximate composition of these wastes was found to be markedly higher in fat, carbohydrate and ash contents. Lemon peels and potato peels showed almost similar phenolic content (162 ± 0.93 and 157 ± 0.88 mg GAE /g, respectively) and exhibited strong ABTS and DPPH radical scavenging activities than the other wastes. The reduction percentage of reducing sugars and asparagine after soaking treatment ranged from 28.70 to 39.57% and from 22.71 to 29.55%, respectively. HPLC results showed higher level of acrylamide formation in control sample (104.94 mg/kg) and by using the wastes extracts of lemon peels, potato peels, olive leaves, and pomegranate peels succeeded to mitigate acrylamide level by 86.11%, 69.66%, 34.03%, and 11.08%, respectively. Thus, it can be concluded that the soaking of potato slices in the tested wastes extracts as antioxidant as pre-treatment before frying reduces the formation of acrylamide and in this way, the risks connected to acrylamide consumption could be regulated and managed.

Special Issue "Bioactive Compounds from Natural Sources (2020, 2021)".

In recent decades, there has been a huge level of interest in bioactive compounds from natural sources [...].

Genetic Stability, Phenolic, Flavonoid, Ferulic Acid Contents, and Antioxidant Activity of Micropropagated Lycium schweinfurthii Plants.

Lycium schweinfurthii is a Mediterranean wild shrub rich in plant secondary metabolites. In vitro propagation of this plant may support the production of valuable dietary supplements for humanity, introduction of it to the world market, and opportunities for further studies. The presented study aimed to introduce an efficient and reproducible protocol for in vitro micropropagation of L. schweinfurthii and assess the genetic stability of micropropagated plants (MiPs) as well as to estimate phenolic, flavonoid, ferulic acid contents, and the antioxidant activity in leaves of micropropagated plants. Two DNA-based techniques, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR), and one biochemical technique, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), were used to assess the genetic stability in MiPs. Spectrophotometric analysis was performed to estimate total phenolic and flavonoid contents and antioxidant activity of MiPs leaves, while ferulic acid content was estimated using high-performance thin-layer chromatography (HPTLC). Sufficient shoot proliferation was achieved at MS (Murashige and Skoog) medium supplemented with 0.4 mg L-1 kinetin and rooted successfully on half-strength MS medium fortified with 0.4 mg L-1 Indole-3-butyric acid (IBA). The Jaccard's similarity coefficients detected in MiPs reached 52%, 55%, and 82% in the RAPD, ISSR, and SDS-PAGE analyses, respectively. In the dried leaves of MiPs, the phenolic, flavonoid, and ferulic acid contents of 11.53 mg gallic acid equivalent, 12.99 mg catechin equivalent, and 45.52 mg were estimated per gram, respectively. However, an IC50 of 0.43, and 1.99 mg mL-1 of MiP dried leaves' methanolic extract was required to scavenge half of the DPPH, and ABTS free radicals, respectively. The study presented a successful protocol for in vitro propagation of a valued promising plant source of phenolic compounds.

Biological Evaluation of Golden Delicious Apples Exposure to UV Lights in Rats.

BACKGROUND AND OBJECTIVE: Anthocyanin is responsible for the red color of apple. Ultraviolet light plays a key role in activating the genes responsible for anthocyanin biosynthesis. However, the most important concern is using UV light irradiation on fruit to increase anthocyanins level and its nutritional quality. In this study, the accumulation of anthocyanin in green apple using UV-B and UV-C was investigated and its biological influence was evaluated in rats. MATERIAL AND METHOD: Green Golden delicious apples were irradiated with doses of UV-C and UV-B light for a period of 3 h/day each for 3 days. Two Groups of rats were fed on balanced diet or balanced diet supplemented with 10% apple exposure to UV (AP-UV) for a month. RESULTS: The HPTLC and spectrophotometric determination of anthocyanin revealed that color development was significantly increased by 90% in treated apple compared to the control apples. Histological difference was observed between the 2 groups. Plasma levels of uric acid, the activity of transaminases (ALT and AST) as well as malondialdehyde (MDA) were significantly elevated in AP-UV rats. Plasma total cholesterol, triglycerides and creatinine level did not differ among the 2 groups. Liver MDA and catalase levels were eminent in AP-UV rats compared to control. Gene expression of selected inflammatory cytokines (TNF-α, IL-6 and IL-1ß) was significantly up-regulated in liver of AP-UV rats in comparison to control rats. CONCLUSION: The result revealed that there is a health-hazard linked to feeding rats on diet containing irradiated-apple with UV-B and UV-C, which represented by body weight reduction, inflammation development, liver function and oxidative stress elevation.

Bioactive Phytochemicals and Antioxidant Properties of the Grains and Sprouts of Colored Wheat Genotypes.

The grains and sprouts of colored wheat genotypes (having blue, purple and yellow colored grains) contain specific anthocyanidins, such as pelargonidin and cyanidin derivatives, that produce beneficial health effects. The objective of the presented study is to compare the antioxidant capacity and contents of bioactive phytochemicals in grains and sprouts of wheat genotypes that differ in grain color. The methods α, α-diphenyl-ß-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) scavenging activities, together with spectrophotometrical and high-performance thin-layer chromatography (HPTLC) methods, were used to study the presence of total phenolics, flavonoids, anthocyanins and anthocyanidins (pelargonidin, peonidin, cyanidin, delphinidin) content. It was predicted that the sprouts of all colored wheat genotypes would have significantly higher total flavonoids, total phenolics, anthocyanidin levels and antioxidant activity than the grains. The correlation results between antioxidant activity and contents of bioactive phytochemicals in grains and sprouts of colored wheat genotypes have shown a high correlation for cyanidin and pelargonidin, especially in grains, as well as quercetin in sprouts. It was found that total anthocyanin, quercetin and pelargonidin contents were significantly higher in the sprouts of the purple wheat genotypes than in the blue or yellow wheat genotypes. Delphinidin was detected at a higher level in the grains than in the sprouts of the blue wheat genotypes. Peonidin was present at very low quantities in the grains of all colored wheat genotypes. The sprouts of the purple wheat genotypes, among the colored wheat genotypes, had the highest pelargonidin, cyanidin and quercetin contents and, therefore, can be a promising source for functional food use.

Determination of lignans, phenolic acids and antioxidant capacity in transformed hairy root culture of Linum usitatissimum.

Hairy root culture is a promising alternative method for the production of secondary metabolites. In this study, transformed root of Linum usitatissimum was established using Agrobacterium rhizogenes A4 strain from root cultures for lignans, phenolic acids and antioxidant capacity determination. Total lignin content (secoisolariciresinol diglucoside, secoisolariciresinol and matairesinol) was 55.5% higher in transformed root cultures than in the non-transformed root culture. Secoisolariciresinol was detected in higher concentration (2.107 µmol/g DM) in the transformed root culture than non-transformed culture (1.099 µmol/g DM). Secoisolariciresinol diglucoside and matairesinol were exclusively detected in the transformed root culture, but were not found in the non-transformed root culture. The overall production of phenolic acids in transformed roots was approximately 3.5 times higher than that of the corresponding non-transformed culture. Free radical scavenging DPPH˙ and ABTS˙+ assays showed 2.9-fold and 1.76-fold higher anti-oxidant activity in transformed root culture as compared to non-transformed.

Enhancement of silymarin and phenolic compound accumulation in tissue culture of Milk thistle using elicitor feeding and hairy root cultures.

In the present study, the effects of the metabolite elicitors chitosan, methyl jasmonate (MeJA) and salicylic acid (SA) as well as the hairy root transformation were tested for silymarin and phenolic compound accumulation in in vitro cultures of Milk thistle. For callus induction, leaf explants were cultured on MS medium supplemented with 5 mg/l NAA + 2 mg/l Kin + 0.1 mg/l GA3. Chitosan, SA and MeJA were added separately in three concentrations 200, 400 and 800 mg/l; 10, 20 and 40 mg/l; 20, 40 and 80 mg/l, respectively, to hormone free B5 medium. Alternatively, cotyledons of 12 day old seedlings were transformed with Agrobacterium rhizogenes A4 strain. Overall, increasing the concentrations of the three elicitors dramatically increased the total silymarin content. Remarkably, the elicitors mainly enhanced the accumulation of silybine A&B that were not detected in un-treated callus culture (control). In addition, the hairy root culture triggered the accumulation of silybine A&B, and silydianin, which was not detected in the non-transgenic roots. The hairy root culture was superior in production of the phenolic compounds in comparison to the control and elicitor treatments. The hairy root cultures showed also higher antioxidant capacities than non-transformed cultures and/or chemically elicited-callus cultures. Thus hairy root provide instrumental in enhancing the production of economically valuable metabolite.

Effects of different encapsulation agents and drying process on stability of betalains extract.

Red beet plants are rich in betalains that can be used as food natural colorants. Betalains were extracted from red beet and encapsulated with different carrier agents and freeze or spray dried. Effect of different encapsulating agents as maltodextrin, guar gum, gum Arabic, pectin and xanthan gum with different concentration (as encapsulating agents) were studied on the betalain stability. Encapsulated betalains with xanthan gum with maltodextrin showed about 65 % more recovery than the control. Encapsulation showed a higher recovery of betalains during freeze drying by 1.3 times than during spray drying. Spray dried samples has L* (lightness) higher than the freeze dried samples. The variations of maltodextrin with xanthan and guar gum freeze dried have highest chroma value of 21. The stabilization of pure betalain pigments may boost the use of these colouring molecules in the food industry and promote their application.

Effect of chlorocholine chlorid on phenolic acids accumulation and polyphenols formation of buckwheat plants.

BACKGROUND: Effect of chlorocholine chloride (CCC) on phenolic acids composition and polyphenols accumulation in various anatomical parts (stems, leaves and inflorescences) of common buckwheat (Fagopyrum esculentum Moench) in the early stages of vegetation period were surveyed. RESULTS: Treatment of buckwheat seeds with 2% of CCC has been increased content of total phenolics in the stems, leaves and inflorescences. On analyzing the different parts of buckwheat plants, 9 different phenolic acids - vanilic acid, ferulic acid, trans-ferulic acid, chlorogenic acid, salycilic acid, cinamic acid, p-coumaric acid, p-anisic acid, methoxycinamic acid and catechins were identified. The levels of identified phenolic acids varied not only significantly among the plant organs but also between early stages of vegetation period. Same changes as in contents of chlorogenic acid, ferulic acid, trans-ferulic acid were found for content of salycilic acid. The content of these phenolic acids has been significant increased under effect of 2% CCC treatment at the phase I (formation of buds) in the stems and at the phase II (beginning of flowering) in the leaves and then inflorescences respectively. The content of catechins as potential buckwheat antioxidants has been increased at the early stages of vegetation period after treatment with 2% CCC. CONCLUSIONS: The obtained results suggest that influence of CCC on the phenolics composition can be a result of various mechanisms of CCC uptake, transforming and/or its translocation in the buckwheat seedlings.

Effect of chlorocholine chlorid on phenolic acids accumulation and polyphenols formation of buckwheat plants

BACKGROUND: Effect of chlorocholine chloride (CCC) on phenolic acids composition and polyphenols accumulation in various anatomical parts (stems, leaves and inflorescences) of common buckwheat (Fagopyrum esculentum Moench) in the early stages of vegetation period were surveyed. RESULTS: Treatment of buckwheat seeds with 2% of CCC has been increased content of total phenolics in the stems, leaves and inflorescences. On analyzing the different parts of buckwheat plants, 9 different phenolic acids - vanilic acid, ferulic acid, trans-ferulic acid, chlorogenic acid, salycilic acid, cinamic acid, p-coumaric acid, p-anisic acid, methoxycinamic acid and catechins were identified. The levels of identified phenolic acids varied not only significantly among the plant organs but also between early stages of vegetation period. Same changes as in contents of chlorogenic acid, ferulic acid, trans-ferulic acid were found for content of salycilic acid. The content of these phenolic acids has been significant increased under effect of 2% CCC treatment at the phase I (formation of buds) in the stems and at the phase II (beginning of flowering) in the leaves and then inflorescences respectively. The content of catechins as potential buckwheat antioxidants has been increased at the early stages of vegetation period after treatment with 2% CCC. CONCLUSIONS: The obtained results suggest that influence of CCC on the phenolics composition can be a result of various mechanisms of CCC uptake, transforming and/or its translocation in the buckwheat seedlings.

Enhanced resveratrol production in Vitis vinifera cell suspension cultures by heavy metals without loss of cell viability.

The effects of heavy metal ions (Co(2+), Ag(+), Cd(2+)) on cell viability and secondary metabolite production, particularly anthocyanins and phenolic acids in Vitis vinifera cell suspension cultures, were investigated. Of these, Co at all three used concentrations (5.0, 25, and 50 µM), Ag, and Cd at low concentration (5.0 µM) were most effective to stimulate the phenolic acid production, increasing the 3-O-glucosyl-resveratrol up to 1.6-fold of the control level (250.5 versus 152.4 µmol/g), 4 h after the treatments. Meanwhile, the elicitors at effective concentrations did not suppress cell growth, while the cell viability maintained. In contrast, Ag and Cd at high concentrations (25 and 50 µM) remarkably reduced the cell viability, decreasing the cell viability up to about 15 % of the control level, 24 h after the treatments. The heavy metal ions did not affect the anthocyanin production. These observations show how, in a single system, different groups of secondary products can show distinct differences in their responses to potential elicitors. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, peroxidase activity, medium pH value, and conductivity were only slightly elevated by the heavy metal ions. The results suggest that some of the secondary metabolites production was stimulated by the used elicitors, but there was not a stress response of the cells.

The effect of tea fermentation on rosmarinic acid and antioxidant properties using selected in vitro sprout culture of Orthosiphon aristatus as a model study.

Orthosiphon aristatus, an Indonesian medicinal plant, is normally used as a traditional herbal tea. Recently, this plant has begun to attract attention due to its antioxidant properties. However, little is known about tea fermentation effect on antioxidant properties of this plant. Thus, to extend the tea fermentation study, in vitro sprout culture of this plant was established as a new feature model. This model plant was selected based on three reasons. Firstly, as a native tropical plant, to grow this plant in sub-tropic area is considered difficult. Secondly, the in vitro sprout culture is more genetically stable compared to other types of in vitro cultures. Thirdly, results showed that this in vitro sprout culture grew faster and produced higher biomass than in vitro tissue culture. Both characteristics are important in producing tea leaves. Accordingly, the aim of the current study was twofold. First was to establish high rosmarinic acid line of in vitro sprout culture of Orthosiphon aristatus by elicitation. Second was to evaluate the effect of tea fermentation on antioxidant properties of this plant. Results showed that yeast extract (5 g/L) elicitation resulted in the highest production of rosmarinic acid. This elicited plant was subjected to partial and full tea fermentation. Results revealed that both tea fermentations decreased the level of rosmarinic acid, total phenolic compounds, flavonoids, and flavonols. These decreases were concomitant with reduced antioxidant activities as measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, Trolox equivalent antioxidant capacity (TEAC), and Superoxide dismutase (SOD)-like activity assays. HPLC results showed that the longer the tea fermentation was, the greater reduction rosmarinic acid was found. High correlation value of 0.922 between rosmarinic acid and antioxidant activities was also observed. These results indicated that rosmarinic acid is the major contributor to the antioxidant activities of Orthosiphon aristatus. These results may provide useful information, in particular, for the food and pharmaceutical industries in the development of functional foods desiring maximum potential health benefits from Orthosiphon aristatus.

Effects of phytohormones and jasmonic acid on glucosinolate content in hairy root cultures of Sinapis alba and Brassica rapa.

Although some study have established hairy root cultures from brassicaceous plants with glucosinolates (GS) as characteristic secondary metabolite, studies are missing which compare hairy roots with the corresponding mother plants. Therefore, two different plant species-Sinapis alba and Brassica rapa subsp. rapa pygmeae teltoviensis-were transformed with the Agrobacterium rhizogenes strain A4. Aliphatic and indolyl GS were present in B. rapa, exhibiting larger quantities in leaves than in roots. Aromatic p-hydroxybenzyl GS were found particularly in the leaves of S. alba. However, the proportion of indolyl GS increased suddenly in transformed hairy roots of S. alba and B. rapa. Cultivation with the phytohormone kinetin (0.5 mg L(-1)) enhanced GS accumulation in B. rapa hairy roots, however not in S. alba, but 2,4-D (0.4 mg L(-1)) induced de-differentiation of roots in both species and reduced GS levels. GS levels especially of 1-methoxyindol-3ylmethyl GS increased in hairy roots in response to JA, but root growth was inhibited. While 2 weeks of cultivation in 100 to 200 µM JA were determined at optimum for maximum GS yield in S. alba hairy root cultures, 4 weeks of cultivation in 50 to 100 µM JA was the optimum for B. rapa.

Investigation of Phenolic Acids in Suspension Cultures of Vitis vinifera Stimulated with Indanoyl-Isoleucine, N-Linolenoyl-L-Glutamine, Malonyl Coenzyme A and Insect Saliva.

Vitis vinifera c.v. Muscat de Frontignan (grape) contains various high valuable bioactive phenolic compounds with pharmaceutical properties and industrial interest which are not fully exploited. The focus of this investigation consists in testing the effects of various biological elicitors on a non-morphogenic callus suspension culture of V. vinifera. The investigated elicitors: Indanoyl-isoleucine (IN), N-linolenoyl-L-glutamine (LG), insect saliva (IS) and malonyl coenzyme A (MCoA) were aimed at mimicking the influence of environmental pathogens on plants in their natural habitats and at provoking exogenous induction of the phenylpropanoid pathway. The elicitors' indanoyl-isoleucine (IN), N-linolenoyl-L-glutamine (LG) and insect saliva (IS), as well as malonyl coenzyme A (MCoA), were independently inoculated to stimulate the synthesis of phenylpropanoids. All of the enhancers positively increased the concentration of phenolic compounds in grape cells. The highest concentration of phenolic acids was detected after 2 h for MCoA, after 48 h for IN and after 24 h for LG and IS respectively. At the maximum production time, treated grape cells had a 3.5-fold (MCoA), 1.6-fold (IN) and 1.5-fold (IS) higher phenolic acid content compared to the corresponding control samples. The HPLC results of grape cells showed two major resveratrol derivatives: 3-O-Glucosyl-resveratrol and 4-(3,5-dihydroxyphenyl)-phenol. Their influences of the different elicitors, time of harvest and biomass concentration (p < 0.0001) were statistically significant on the synthesis of phenolic compounds. The induction with MCoA was found to demonstrate the highest statistical effect corresponding to the strongest stress response within the phenylpropanoid pathway in grape cells.

The Effect of Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM Fermentation on Antioxidant Properties of Selected in Vitro Sprout Culture of Orthosiphon aristatus (Java Tea) as a Model Study.

High rosmarinic acid (RA) productivity has been achieved by applying jasmonic acid and yeast extract elicitors to the in vitro sprout culture of Orthosiphon aritatus (IOSC). The highest RA accumulation from three solvents was detected in IOSC after treatment with yeast extract (5 g/L). HPLC analysis clearly confirmed a drastic increase in RA subjected to yeast extract elicitation. Therefore, this yeast extract elicited IOSC was chosen for a lactic acid bacteria (LAB) fermentation study as a model system. This selected IOSC was subjected to different types of LAB fermentations (Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM) for different periods of time 24, 48 and 72 h. The LAB fermentations consisted of solid state fermentations (SSF) and liquid state fermentations (LSF) in a Digital Control Unit (DCU) fermenter system. The aim was to determine the effect of fermentation on the antioxidant properties of the plant extract. Results indicated that all types of LAB fermentation decreased the level of RA and total phenolics, however, a slight increase in total flavonoids and flavonols was observed in SSF samples. HPLC results confirmed that the longer the fermentation, the greater the reduction in RA content. The highest reduction was obtained in the sample of LSF inoculated with L. plantarum for a period of 72 h. The temperature of fermentation (37 °C) was predicted as contributing to the declining level in RA content. The loss in RA was concomitant with a loss of total antioxidant activity (1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, Trolox Equivalent Antioxidant Capacity (TEAC), and Superoxide Dismutase (SOD)-like activity). These results indicate that RA is the major contributor to the antioxidant activity of this plant.

Exudation: an expanding technique for continuous production and release of secondary metabolites from plant cell suspension and hairy root cultures.

This review addresses methods of obtaining secondary metabolites from plant cell suspension and hairy root cultures and their exudates, particularly the physiological mechanisms of secondary metabolites release and trafficking. The efficiency for product recovery of metabolites can be increased by various methods, based on the principle of continuous product release into the cultivation medium. The most common methods for metabolite recovery are elicitation, influencing membrane permeability, and in situ product removal. The biosynthetic pathways can be influenced by cultivation conditions, transformation, or application of elicitors. The membrane permeability can be altered through the application of chemical or physical treatments. Product removal can be greatly increased through a two-phase system and the introduction of absorbents into the cultivation medium. In this review, we describe some improved approaches that have proven useful in these efforts.

Enhanced anthocyanins and resveratrol production in Vitis vinifera cell suspension culture by indanoyl-isoleucine, N-linolenoyl-L-glutamine and insect saliva.

The effects of two synthetic elicitor indanoyl-isoleucine (In-Ile), N-linolenoyl-L-glutamine (Lin-Gln) and one biotic elicitor insect saliva (from Manduca sexta larvae) on plant cell cultures with respect to the induction of secondary metabolite production were investigated. Stimulated production of secondary metabolites, particularly anthocyanins in plant cells and phenolic acids in culture medium, was studied by using suspension culture of Vitis vinifera L. cv. Gamay Fréaux as a model system. In the treatments with In-Ile, the production of anthocyanins was enhanced 2.6-fold. In-Ile, Lin-Gln and saliva significantly elevated the accumulation of phenolic acids, particularly 3-O-glucosyl-resveratrol. The used elicitors did not suppress cell growth. Secondary metabolites were differently responsive to elicitation. 3-O-glucosyl-resveratrol was the predominant phenolic acid in V. vinifera cell culture, and its production was significantly stimulated by saliva, with 7.0-fold of the control level 24 h after treatment. The production of 4-(3,5-dihydroxy-phenyl)-phenol was significantly stimulated by In-Ile with 6.4-fold of the control level 24 h after treatment.

Enhanced glucosinolates in root exudates of Brassica rapa ssp. rapa mediated by salicylic acid and methyl jasmonate.

Elicitation studies with salicylic acid (SA) and methyl jasmonate (MJ) inducing a targeted rhizosecretion of high levels of anticarcinogenic glucosinolates in Brassica rapa ssp. rapa plants were conducted. Elicitor applications not only led to an accumulation of individual indole glucosinolates and the aromatic 2-phenylethyl glucosinolate in the turnip organs but also in turnip root exudates. This indicates an extended systemic response, which comprises the phyllosphere with all aboveground plant organs and the rhizosphere including the belowground root system and also root exudates. Both elicitor applications induced a doubling in 2-phenylethyl glucosinolate in root exudates, whereas application of MJ enhanced rhizosecreted indole glucosinolates up to 4-fold. In addition, the time course study revealed that maximal elicitation was observed on the 10th day of SA and MJ treatment. This study may provide an essential contribution using these glucosinolates as bioactive additives in functional foods and nutraceuticals.