Flavonoid, Nitrate and Glucosinolate Concentrations in Brassica Species Are Differentially Affected by Photosynthetically Active Radiation, Phosphate and Phosphite.

We evaluated the effects of phosphate (Pi-deficiency: 0.1 mM; Pi-sufficiency: 0.5 mM), phosphite (low-Phi: 0.1 mM; medium-Phi: 0.5 mM; and high-Phi: 2.5 mM), and two mean daily photosynthetically active radiations (lower PAR: 22.2 mol ⋅ m-2 ⋅ d-1; higher PAR: 29.7 mol ⋅ m-2 ⋅ d-1), as well as their interactions, on flavonoid, nitrate and glucosinolate (GL) concentrations and growth characteristics in hydroponically grown Brassica campestris cv. Mibuna Early and Brassica juncea cv. Red Giant. As expected, higher PAR increased dry matter and contrariwise decreased number of leaves but only in B. campestris. Total flavonoid and individual flavonoid compounds increased with the higher PAR value in B. campestris. Pi-sufficiency resulted in a lower quercetin concentration in both species, the isorhamnetin and total flavonoid concentrations in B. campestris, and the cyanidin concentration in B. juncea, in comparison to Pi-deficiency. Similarly, Pi-sufficient plants exhibited lower GL concentration, especially alkyl-GLs in B. campestris and alkenyl-GLs and an aryl-GL in B. juncea. Pi did not affect the nitrate concentration in either species, and nor did Phi influence the flavonoid concentrations in either species. In B. campestris, medium Phi (0.5 mM) increased the 1-methoxyindol-3-ylmethyl GL concentration by 28.3%, as compared to that observed at low Phi. In B. juncea, high Phi level increased the but-3-enyl-GL concentration by 18.9%, in comparison to values recorded at medium Phi. B. campestris plants exposed to higher PAR increased total flavonoids concentration. In both Brassica species, higher PAR stimulated the alkyl-, alkenyl-, and indole-GLs. The interaction of lower PAR and increasing Phi significantly decreased flavonoid concentration in B. juncea, whereas increasing Phi at higher PAR increased such concentration in this species. The same combination reduced the concentration of 2-phenylethyl- and indol-3-ylmethyl-GL in B. juncea. The highest indol-3-ylmethyl-GL concentration was observed when Pi was deficient combined with medium Phi in B. juncea. Thus, PAR, Pi and Phi may modulate flavonoid, GL and nitrate concentrations in Brassica species, which may be a useful tool to improve the nutraceutical quality of these leafy vegetables if properly managed.

Influence of Light and Temperature on Gene Expression Leading to Accumulation of Specific Flavonol Glycosides and Hydroxycinnamic Acid Derivatives in Kale (Brassica oleracea var. sabellica).

Light intensity and temperature are very important signals for the regulation of plant growth and development. Plants subjected to less favorable light or temperature conditions often respond with accumulation of secondary metabolites. Some of these metabolites have been identified as bioactive compounds, considered to exert positive effects on human health when consumed regularly. In order to test a typical range of growth parameters for the winter crop Brassica oleracea var. sabellica, plants were grown either at 400 µmol m(-2) s(-1) or 100 µmol m(-2) s(-1) at 10°C, or at 400 µmol m(-2) s(-1) with 5 or 15°C. The higher light intensity overall increased flavonol content of leaves, favoring the main quercetin glycosides, a caffeic acid monoacylated kaempferol triglycoside, and disinapoyl-gentiobiose. The higher temperature mainly increased the hydroxycinnamic acid derivative disinapoyl-gentiobiose, while at lower temperature synthesis is in favor of very complex sinapic acid acylated flavonol tetraglycosides such as kaempferol-3-O-sinapoyl-sophoroside-7-O-diglucoside. A global analysis of light and temperature dependent alterations of gene expression in B. oleracea var. sabellica leaves was performed with the most comprehensive Brassica microarray. When compared to the light experiment much less genes were differentially expressed in kale leaves grown at 5 or 15°C. A structured evaluation of differentially expressed genes revealed the expected enrichment in the functional categories of e.g. protein degradation at different light intensities or phytohormone metabolism at different temperature. Genes of the secondary metabolism namely phenylpropanoids are significantly enriched with both treatments. Thus, the genome of B. oleracea was screened for predicted genes putatively involved in the biosynthesis of flavonoids and hydroxycinnamic acid derivatives. All identified B. oleracea genes were analyzed for their most specific 60-mer oligonucleotides present on the 2 × 105 K format Brassica microarray. Expression differences were correlated to the structure-dependent response of flavonoid glycosides and hydroxycinnamic acid derivatives to alterations in either light or temperature. The altered metabolite accumulation was mainly reflected on gene expression level of core biosynthetic pathway genes and gave further hints to an isoform specific functional specialization.

Nitrogen Limited Red and Green Leaf Lettuce Accumulate Flavonoid Glycosides, Caffeic Acid Derivatives, and Sucrose while Losing Chlorophylls, Β-Carotene and Xanthophylls.

Reduction of nitrogen application in crop production is desirable for ecological and health-related reasons. Interestingly, nitrogen deficiency can lead to enhanced concentrations of polyphenols in plants. The reason for this is still under discussion. The plants' response to low nitrogen concentration can interact with other factors, for example radiation intensity. We cultivated red and green leaf lettuce hydroponically in a Mediterranean greenhouse, supplying three different levels of nitrogen (12 mM, 3 mM, 0.75 mM), either in full or reduced (-50%) radiation intensity. In both red and green lettuce, we found clear effects of the nitrogen treatments on growth characteristics, phenolic and photosynthetic compounds, nitrogen, nitrate and carbon concentration of the plants. Interestingly, the concentrations of all main flavonoid glycosides, caffeic acid derivatives, and sucrose increased with decreasing nitrogen concentration, whereas those of chlorophylls, ß-carotene, neoxanthin, lactucaxanthin, all trans- and cis-violaxanthin decreased. The constitutive concentrations of polyphenols were lower in the green cultivar, but their relative increase was more pronounced than in the red cultivar. The constitutive concentrations of chlorophylls, ß-carotene, neoxanthin, all trans- and cis-violaxanthin were similar in red and green lettuce and with decreasing nitrogen concentration they declined to a similar extent in both cultivars. We only detected little influence of the radiation treatments, e.g. on anthocyanin concentration, and hardly any interaction between radiation and nitrogen concentration. Our results imply a greater physiological plasticity of green compared to the red lettuce regarding its phenolic compounds. They support the photoprotection theory regarding anthocyanins as well as the theory that the deamination activity of phenylalanine ammonia-lyase drives phenylpropanoid synthesis.

Inoculation with arbuscular mycorrhizal fungi improves the nutritional value of tomatoes.

Arbuscular mycorrhizal (AM) fungi can affect many different micronutrients and macronutrients in plants and also influence host volatile compound synthesis. Their effect on the edible portions of plants is less clear. Two separate studies were performed to investigate whether inoculation by AM fungi (Rhizophagus irregularis, Funneliformis mosseae, or both) can affect the food quality of tomato fruits, in particular common minerals, antioxidants, carotenoids, a suite of vitamins, and flavor compounds (sugars, titratable acids, volatile compounds). It was found that AM fungal inoculation increased the nutrient quality of tomato fruits for most nutrients except vitamins. Fruit mineral concentration increased with inoculation (particularly N, P, and Cu). Similarly, inoculated plants had fruit with higher antioxidant capacity and more carotenoids. Furthermore, five volatile compounds were significantly higher in AM plants compared with non-AM controls. Taken together, these results show that AM fungi represent a promising resource for improving both sustainable food production and human nutritional needs.

Interaction of moderate UV-B exposure and temperature on the formation of structurally different flavonol glycosides and hydroxycinnamic acid derivatives in kale (Brassica oleracea var. sabellica).

Kale has a high number of structurally different flavonol glycosides and hydroxycinnamic acid derivatives. In this study we investigated the interaction of moderate UV-B radiation and temperature on these compounds. Kale plants were grown at daily mean temperatures of 5 or 15 °C and were exposed to five subsequent daily doses (each 0.25 kJ m(-2) d(-1)) of moderate UV-B radiation at 1 d intervals. Of 20 phenolic compounds, 11 were influenced by an interaction of UV-B radiation and temperature, e.g., monoacylated quercetin glycosides. Concomitantly, enhanced mRNA expression of flavonol 3'- hydroxylase showed an interaction of UV-B and temperature, highest at 0.75 kJ m(-2) and 15 °C. Kaempferol glycosides responded diversely and dependent on, e.g., the hydroxycinnamic acid residue. Compounds containing a catechol structure seem to be favored in the response to UV-B. Taken together, subsequent exposure to moderate UV-B radiation is a successful tool for enhancing the flavonoid profile of plants, and temperature should be considered.

Unlike quercetin glycosides, cyanidin glycoside in red leaf lettuce responds more sensitively to increasing low radiation intensity before than after head formation has started.

This study investigated the effect of low-level photosynthetic photon flux density (PPFD; 43-230 µmol m(-2) s(-1)) on the major phenolic compounds of red leaf lettuce in three growth stages, before, during, and after head formation, using HPLC-DAD-ESI-MS(2) and evaluating via multiple regression analysis. Generally, the light-related increase of flavonoid glycosides was structure and growth stage-dependent. In detail, an interaction was detected between plant age and PPFD regarding cyanidin-3-O-(6"-O-malonyl)-glucoside concentration: the increase was strongest before head formation. The relationship between PPFD and quercetin-3-O-(6"-O-malonyl)-glucoside concentration was linear, whereas the increase of quercetin-3-O-glucoside and -3-O-glucuronide concentrations abated with increasing PPFD. Independent of growth stage, the caffeic acid derivatives concentration was not related to PPFD. All major phenolic compounds decreased with plant age. These results show the differential regulation of cyanidin, quercetin, and caffeic acid derivatives in lettuce, although closely connected biosynthetically, and emphasize the importance of ontogeny in the study of plant physiology.

Topsoil drying combined with increased sulfur supply leads to enhanced aliphatic glucosinolates in Brassica juncea leaves and roots.

The decrease of water availability is leading to an urgent demand to reduce the plants' water supply. This study evaluates the effect of topsoil drying, combined with varying sulfur (S) supply on glucosinolates in Brassica juncea in order to reveal whether a partial root drying may already lead to a drought-induced glucosinolate increase promoted by an enhanced S supply. Without decreasing biomass, topsoil drying initiated an increase in aliphatic glucosinolates in leaves and in topsoil dried roots supported by increased S supply. Simultaneously, abscisic acid was determined, particularly in dehydrated roots, associated with an increased abscisic acid concentration in leaves under topsoil drying. This indicates that the dehydrated roots were the direct interface for the plants' stress response and that the drought-induced accumulation of aliphatic glucosinolates is related to abscisic acid formation. Indole and aromatic glucosinolates decreased, suggesting that these glucosinolates are less involved in the plants' response to drought.

Cool-cultivated red leaf lettuce accumulates cyanidin-3-O-(6″-O-malonyl)-glucoside and caffeoylmalic acid.

Cultivating lettuce in greenhouses at low temperatures improves its CO2-balance and may increase its content of flavonoid glycosides and phenolic acids. We cultivated 5weeks old red leaf lettuce seedlings at 20/15°C (day/night) or 12/7°C until plants reached comparable growth stages: small heads were harvested after 13 (warm) and 26 (cool)days, while mature heads were harvested after 26 (warm) or 52 (cool)days. Additionally, some plants were cultivated first cool then warm and vice versa (39days). Cool-cultivated small heads had higher concentrations of cyanidin-3-O-(6″-O-malonyl)-glucoside and caffeoylmalic acid than warm-cultivated ones but we detected no differences concerning quercetin and luteolin glycosides or di-O-caffeoyltartaric and 5-O-caffeoylquinic acid. Regarding mature heads, there were only differences concerning cyanidin-3-O-(6″-O-malonyl)-glucoside. We therefore suggest that only cyanidin-3-O-(6″-O-malonyl)-glucoside was truly responsive to temperatures alone. Previously reported contrasting effects may rather be due to comparison of different growth stages or interactive effects with radiation.

Verticillium suppression is associated with the glucosinolate composition of Arabidopsis thaliana leaves.

The soil-borne fungal pathogen Verticillium longisporum is able to penetrate the root of a number of plant species and spread systemically via the xylem. Fumigation of Verticillium contaminated soil with Brassica green manure is used as an environmentally friendly method for crop protection. Here we present a study focused on the potential role of glucosinolates and their breakdown products of the model plant Arabidopsis thaliana in suppressing growth of V. longisporum. For this purpose we analysed the glucosinolate composition of the leaves and roots of a set of 19 key accessions of A. thaliana. The effect of volatile glucosinolate hydrolysis products on the in vitro growth of the pathogen was tested by exposing the fungus to hydrated lyophilized plant tissue. Volatiles released from leaf tissue were more effective than from root tissue in suppressing mycelial growth of V. longisporum. The accessions varied in their efficacy, with the most effective suppressing mycelial growth by 90%. An analysis of glucosinolate profiles and their enzymatic degradation products revealed a correlation between fungal growth inhibition and the concentration of alkenyl glucosinolates, particularly 2-propenyl (2Prop) glucosinolate, respectively its hydrolysis products. Exposure of the fungus to purified 2Prop glucosinolate revealed that its suppressive activity was correlated with its concentration. Spiking of 2Prop glucosinolate to leaf material of one of the least effective A. thaliana accessions led to fungal growth suppression. It is suggested that much of the inhibitory effect observed for the tested accessions can be explained by the accumulation of 2Prop glucosinolate.

The hydroxycinnamic acid content of barley and brewers' spent grain (BSG) and the potential to incorporate phenolic extracts of BSG as antioxidants into fruit beverages.

The hydroxycinnamic acid (HA) content of starting barley for brewers' spent grains (BSG), whole BSG and phenolic extracts from BSG was measured using high performance liquid chromatography (HPLC) and correlated with antioxidant potential. The effect of BSG phenolic extracts on antioxidant activity of fruit beverages was also assessed (using the total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays). The concentration of HA present in barley extract and BSG was in the order of ferulic acid (FA), p-coumaric acid (p-CA) derivatives, FA derivatives, p-CA, caffeic acid (CA) and CA derivatives. Results suggested that brewing and roasting decreased the HA content. Antioxidant activity was significantly (P<0.05) correlated with caffeic acid (R(2)=0.8309) and total HA (R(2)=0.3942) concentrations. Addition of extracts to fruit beverages resulted in a significant (P<0.05) increase in antioxidant activity of cranberry juice, measured by the FRAP assay. In vitro digestion significantly (P<0.05) reduced TPC, DPPH and FRAP activity of the fruit beverages.

Temporary reduction of radiation does not permanently reduce flavonoid glycosides and phenolic acids in red lettuce.

Applying transparent daytime screens in greenhouses in cool seasons reduces the amount of energy needed for heating, but also the solar radiation available for crops. This can reduce yield and product quality of leafy vegetables because of constrained photosynthesis and altered biosynthesis. To study this, we cultivated five-week old red leaf lettuce (Lactuca sativa L.) for four weeks in growth chambers under a photosynthetic photon flux density (PPFD) of 225 and 410 µmol m(-2) s(-1), respectively. Some plants were exchanged between radiation intensities after two weeks. We investigated the concentration of five flavonoid glycosides, three caffeic acid derivatives, reducing sugars as well as plant growth. Remarkably, no significant influence of radiation intensity on the concentration of phenolic acids or anthocyanin glycosides was observed. In contrast, quercetin and luteolin glycoside concentration was between 14 and 34% lower in plants growing under lower compared to higher PPFD. Already after two weeks of cultivation, plants grown under lower PPFD contained less quercetin and luteolin glycosides but they completely compensated if subsequently transferred to higher PPFD until harvest. Hence, marketable lettuce heads which experienced temporary shading followed by an unshaded phase did not contain lower concentrations of flavonoid glycosides or phenolic acids. Also, there was no reduction of head mass in this variant. Our results suggest that saving energy in early growth stages is feasible without losses in yield or health promoting phenolic substances. In addition, there was a close correlation between the concentration of reducing sugars and some flavonoid glycosides, indicating a close metabolic connection between their biosynthesis and the availability of carbohydrates.

Low and moderate photosynthetically active radiation affects the flavonol glycosides and hydroxycinnamic acid derivatives in kale (Brassica oleracea var. sabellica) dependent on two low temperatures.

Kale (Brassica oleracea var. sabellica) contains a large number of naturally occurring structurally different non-acylated and acylated flavonol glycosides as well as hydroxycinnamic acid derivatives. The objective of this study was to determine the effect of low and moderate photosynthetic active radiation (PAR) and how these levels interact with low temperature in these phenolic compounds. Juvenile kale plants were treated with PAR levels from 200 to 800 µmol m(-2) s(-1) at 5 and 10 °C under defined conditions in climate chambers. Of the investigated 20 compounds, 11 and 17 compounds were influenced by PAR and temperature, respectively. In addition, an interaction between PAR and temperature was found for eight compounds. The response of the phenolic compounds to PAR was structure-dependent. While quercetin triglycosides increased with higher PAR at 5 and 10 °C, the kaempferol triglycosides exhibited the highest concentrations at 400 µmol m(-2) s(-1). In contrast, kaempferol diglycosides exhibited the highest concentrations at increased PAR levels of 600 and 800 µmol m(-2) s(-1) at 10 °C. However, key genes of flavonol biosynthesis were influenced by temperature but remained unaffected by PAR. Furthermore, there was no interaction between the PAR level and the low temperature in the response of hydroxycinnamic acid derivatives in kale with the exception of caffeoylquinic acid, which decreased with higher PAR levels of 600 and 800 µmol m(-2) s(-1) and at a lower temperature. In conclusion, PAR and its interaction with temperature could be a suitable tool for modifying the profile of phenolic compounds.

Thermal-induced changes of kale's antioxidant activity analyzed by HPLC-UV/Vis-online-TEAC detection.

Generally, boiling of vegetables is assumed leading to lower nutritional values because of leaching effects and activity loss of bioactive compounds. Kale (Brassica oleracea var. sabellica) reveals a great diversity of flavonoids, which have been shown to be good antioxidants. As Brassica vegetables are mainly consumed cooked, the influence of boiling on kale's flavonoids and their antioxidant activity was investigated. Therefore, three kale cultivars were cooked at 100°C for 2 and 4h prior to analysis. The total phenolic content (TPC) and the total antioxidant activity (TEAC assay and EPR spectrometry) of each cultivar were determined and revealed no change, independent of cooking time, although kale samples visually altered. Using the HPLC-UV/Vis-online-TEAC approach, distinct changes in composition and antioxidant activity of the flavonoids were detectable. Thus, it was observable, that the antioxidant activities of the reaction products compensated the "loss" of the antioxidant activity of the original compounds of the uncooked material.

Genotypic variation of the glucosinolate profile in pak choi (Brassica rapa ssp. chinensis).

Thirteen different pak choi (Brassica rapa ssp. chinensis) cultivars were characterized regarding their glucosinolate profile analyzed by HPLC-DAD-MS. The identified glucosinolates were subjected to principal component analysis, and three distinct groups of pak choi sprouts were identified. Group differences were marked mainly by variations in the aliphatic glucosinolate profile such as differing levels of 3-butenyl glucosinolate and 2-hydroxy-3-butenyl glucosinolate as well as by their varying proportional ratios. In addition, the three groups of pak choi sprouts varied by the presence or absence of 2-hydroxy-4-pentenyl glucosinolate and in level and composition of butyl glucosinolates. This classification is reflected by relative mRNA expression level of 2-oxoacid-dependent dioxygenase. As in sprouts, the major glucosinolates in mature leaves were found to be the aliphatic glucosinolates. However, unlike in sprouts, an additional aliphatic glucosinolate, 5-methylsulfinylpentyl glucosinolate, was detected as characteristic ontogenetic variation in mature leaves in 12 of the 13 pak choi cultivars analyzed.

Determination of bioactive, free isothiocyanates from a glucosinolate-containing phytotherapeutic agent: a pilot study with in vitro models and human intervention.

Isothiocyanates (ITCs) derived from plants of the order Brassicales are known for their antibacterial, anti-inflammatory or anticarcinogenic potential. Although only the free ITCs exert bioactivity, quantification in vivo is almost exclusively performed on total ITC/metabolite content. We therefore investigated in a pilot study the amount of free ITC at different steps critical for therapeutic efficacy. A sensitive and specific GC-MS/MS method for the simultaneous quantification of individual free ITC after solid-phase extraction (SPE) was developed. We show here that release of biologically active ITC from plants occurs at not only alkaline but also acidic pH. Furthermore, in human urine conversion of the ultimate, inactive mercapturic acid conjugate back into its corresponding bioactive form is increased at alkaline as compared to neutral pH. This was also observed in the urine of human volunteers, where - in correlation with the pH value - a mean of 0.16 to 1.03 µmol ITC was detected after oral application of a phytotherapeutic agent containing 30.4 µmol of the initial pro-drugs. The amounts of free ITC being necessary for bioactivity in vitro were found to be indeed achieved in vivo. These data might be helpful to better understand the beneficial effects of ITC observed in vivo.

UV-B irradiation changes specifically the secondary metabolite profile in broccoli sprouts: induced signaling overlaps with defense response to biotic stressors.

Only a few environmental factors have such a pronounced effect on plant growth and development as ultraviolet light (UV). Concerns have arisen due to increased UV-B radiation reaching the Earth's surface as a result of stratospheric ozone depletion. Ecologically relevant low to moderate UV-B doses (0.3-1 kJ m(-2) d(-1)) were applied to sprouts of the important vegetable crop Brassica oleracea var. italica (broccoli), and eco-physiological responses such as accumulation of non-volatile secondary metabolites were related to transcriptional responses with Agilent One-Color Gene Expression Microarray analysis using the 2×204 k format Brassica microarray. UV-B radiation effects have usually been linked to increases in phenolic compounds. As expected, the flavonoids kaempferol and quercetin accumulated in broccoli sprouts (the aerial part of the seedlings) 24 h after UV-B treatment. A new finding is the specific UV-B-mediated induction of glucosinolates (GS), especially of 4-methylsulfinylbutyl GS and 4-methoxy-indol-3-ylmethyl GS, while carotenoids and Chl levels remained unaffected. Accumulation of defensive GS metabolites was accompanied by increased expression of genes associated with salicylate and jasmonic acid signaling defense pathways and up-regulation of genes responsive to fungal and bacterial pathogens. Concomitantly, plant pre-exposure to moderate UV-B doses had negative effects on the performance of the caterpillar Pieris brassicae (L.) and on the population growth of the aphid Myzus persicae (Sulzer). Moreover, insect-specific induction of GS in broccoli sprouts was affected by UV-B pre-treatment.

Effect of differential N and S competition in inter- and sole cropping of Brassica species and lettuce on glucosinolate concentration.

Field and greenhouse pot experiments were conducted to evaluate the potential to use intercropping as an alternative method to increase glucosinolates in Brassicas by manipulating nitrogen (N) and sulfur (S) balance by intercropping with lettuce (Lactuca sativa L. var. capitata). In both experiments, four combinations of N and S fertilization were used. In the field experiment no effect of intercropping on the total glucosinolate concentration was found as the growing lettuce was strongly inhibited by the presence of broccoli (Brassica oleracea L. var. italic). In contrast to this, in the pot experiment both total and individual glucosinolate concentrations in red leaf mustard (Brassica juncea L.) increased by intercropping. Fertilization treatments influenced glucosinolate concentrations in both experiments, and an interaction between N and S fertilization was noticed.

Structurally different flavonol glycosides and hydroxycinnamic acid derivatives respond differently to moderate UV-B radiation exposure.

The aim of this study was to investigate the modifying influence of moderate ultraviolet-B (UV-B) radiation exposure on structurally different flavonol glycosides and hydroxycinnamic acid derivatives during pre-harvest using kale, a leafy Brassica species with a wide spectrum of different non-acylated and acylated flavonol glycosides. Juvenile kale plants were treated with short-term (1 day), moderate UV-B radiation [0.22-0.88 kJ m⁻² day⁻¹ biologically effective UV-B (UV-B(BE))]. Twenty compounds were quantified, revealing a structure-specific response of flavonol glycosides and hydroxycinnamic acid derivatives to UV-B radiation. A dose- and structure-dependent response of the investigated phenolic compounds to additional UV-B radiation was found. The investigated quercetin glycosides decreased under UV-B; for kaempferol glycosides, however, the amount of sugar moieties and the flavonol glycoside hydoxycinnamic acid residue influenced the response to UV-B. Monoacylated kaempferol tetraglucosides decreased in the investigated UV-B range, whereas the monoacylated kaempferol diglucosides increased strongly with doses of 0.88 kJ m⁻² day⁻¹ UV-B(BE) . The UV-B-induced increase in monoacylated kaempferol triglucosides was dependent on the acylation pattern. Furthermore, the hydroxycinnamic acid glycosides disinapoyl-gentiobiose and sinapoyl-feruloyl-gentiobiose were enhanced in a dose-dependent manner under UV-B. While UV-B radiation treatments often focus on flavonol aglycones or total flavonols, our investigations were extended to structurally different non-acylated and acylated glycosides of quercetin and kaempferol.

Isothiocyanate-containing mustard protects human cells against genotoxins in vitro and in vivo.

BACKGROUND: Human intervention trials in which cytogenetic biomarkers are used as intermediate endpoints in carcinogenesis are implicitly required to support the assumption of chemo-preventive efficacy. METHODS: To evaluate the genotoxic and anti-genotoxic properties of defined isothiocyanate-containing mustard, we first used a human liver cell-line and then conducted a controlled pilot human intervention trial. Blood from volunteers served as surrogate tissue for time-kinetic analysis of the chemo-preventive effect of mustard consumption. RESULTS: Mustard extracts displayed significant anti-genotoxicity against benzo(a)pyrene in human HepG2 hepatoma cells. At high concentrations, the extracts induced genotoxicity by themselves without compromising cell viability. The protective effect of mustard supplementation against DNA damage induced ex vivo was detected in blood of volunteers within 12h after the start of the intervention, and increased over time. No genotoxicity was induced in human peripheral mononuclear blood cells by mustard intake over the whole period of the study. Also, liver parameters remained within the normal range at all times. Although no change in total plasma GST activity was detected, plasma alpha-GST levels increased over time, peaking at 48 h. CONCLUSIONS: The results suggest the capacity of small amounts of isothiocyanate-containing food to protect cells from DNA damage, even with short-term application.

Phytochemical changes induced by different nitrogen supply forms and radiation levels in two leafy Brassica species.

The effect of three different nitrogen (N) supply forms differing in their ammonium-to-nitrate (NH(4):NO(3)) ratio (100% NH(4), 50% NH(4) + 50% NO(3), 100% NO(3)) under three different levels of daily photosynthetic active radiation (PAR) (low, 5.0; medium, 6.8; high, 9.0 mol m(-2) day(-1)) on a range of desirable health-promoting phytochemicals in Brassica rapa subsp. nipposinica var. chinoleifera and Brassica juncea was determined. The 100% NH(4) supply under medium PAR levels led to the highest concentration of glucosinolates based on a low nitrogen/sulfur ratio as well as high levels of carotenoids in the leaves of both Brassica species. However, the 100% NH(4) supply under low and medium PAR levels resulted in low concentrations of flavonoids based on high N concentration in the leaves. Thus, the data provided here have strong implications for crop management strategies aimed at optimizing both the concentration and composition of a range of phytochemicals.