Cloning and Functional Analysis of NoMYB60 Gene Involved in Flavonoid Biosynthesis in Watercress (Nasturtium officinale R. Br.).

The MYB60 gene belongs to the R2R3-MYB subfamily, which includes the MYB31/30/96/94 genes. Although these genes have been shown to respond to heat and drought stresses, their role in flavonoid synthesis remains unclear. In this study, NoMYB60 was cloned from watercress and its structure and function were analyzed. Sequence structure analysis showed that NoMYB60 had a highly conserved R2R3 DNA-binding region at the N-terminus. Under the treatment of ABA, SA or MeJA, the expression level of NoMYB60 first significantly increased and then decreased, indicating that ABA, SA and MeJA positively regulated NoMYB60. The subcellular localization of NoMYB60-GFP indicated that NoMYB60 was localized in the nuclear region, which is consistent with the molecular characterization of the transcription factor. Gene silencing experiments were also performed to further test the function of NoMYB60. The result showed that virus-induced silencing of NoMYB60 affected the expression of enzyme genes in flavonoid synthesis pathways and promoted the synthesis of flavonoids. Moreover, we discovered that NoMYB60 interacts with NoBEH1/2. In this study, provides a reference for research on the regulation mechanism of flavonoid synthesis in Cruciferae and other crops.

Formation and stability of isothiocyanate protein conjugates at different pH values and bread types enriched with nasturtium (Tropaeolum majus L.).

Brassicaceae vegetables are rich in glucosinolates (GLS), which degrade into various breakdown products, including isothiocyanates (ITC), during food processing. ITC are associated with health-promoting properties; therefore, producing food products enriched with a high content of these compounds is of interest for improving and maintaining human health. The present study aimed at evaluating a potential increase in ITC formation in Brassicaceae-enriched bread while minimizing ITC-protein conjugates. The influence of pH on GLS degradation was evaluated in different bread types and pH-adjusted model breads. In all samples, ITC, nitriles, and ITC-amino acid conjugates were analyzed by GC-MS and LC-ESI-MS/MS, respectively. The highest ITC-Lys levels with 33.9 µmol BITC-Lys/g bread could be found in a (more alkaline soda) bread with a pH of 7.2, while "free" BITC content was the lowest. However, this finding could not be directly correlated to pH value, because in model breads no significant relation between the pH and the BITC content could be identified. Especially the baking process impacted the ITC content as it was reduced from dough to the bread by 98%. Therefore, a heated food product is not suitable for an enrichment with GLS-rich vegetables to achieve a high content on BITC. Nevertheless, in the bread matrix itself the degradation products and BITC-Lys conjugates were stable during storage.

Analysis of Flavonoid Metabolites in Watercress (Nasturtium officinale R. Br.) and the Non-Heading Chinese Cabbage (Brassica rapa ssp. chinensis cv. Aijiaohuang) Using UHPLC-ESI-MS/MS.

Flavonoids from plants play an important role in our diet. Watercress is a special plant that is rich in flavonoids. In this study, four important watercress varieties were compared with non-heading Chinese cabbage by ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS). A total of 132 flavonoid metabolites (including 8 anthocyanins, 2 dihydroflavone, 3 dihydroflavonol, 1 flavanols, 22 flavones, 11 flavonoid carbonosides, 82 flavonols, and 3 isoflavones) were detected. Flavonoid metabolites varied widely in different samples. Both the non-heading Chinese cabbage and the variety of watercress from Guangdong, China, had their own unique metabolites. This work is helpful to better understand flavonoid metabolites between the non-heading Chinese cabbage and the other four watercress varieties, and to provide a reliable reference value for further research.

Precursor-Boosted Production of Metabolites in Nasturtium officinale Microshoots Grown in Plantform Bioreactors, and Antioxidant and Antimicrobial Activities of Biomass Extracts.

The study demonstrated the effects of precursor feeding on the production of glucosinolates (GSLs), flavonoids, polyphenols, saccharides, and photosynthetic pigments in Nasturtium officinale microshoot cultures grown in Plantform bioreactors. It also evaluated the antioxidant and antimicrobial activities of extracts. L-phenylalanine (Phe) and L-tryptophan (Trp) as precursors were tested at 0.05, 0.1, 0.5, 1.0, and 3.0 mM. They were added at the beginning (day 0) or on day 10 of the culture. Microshoots were harvested after 20 days. Microshoots treated with 3.0 mM Phe (day 0) had the highest total GSL content (269.20 mg/100 g DW). The qualitative and quantitative profiles of the GSLs (UHPLC-DAD-MS/MS) were influenced by precursor feeding. Phe at 3.0 mM stimulated the best production of 4-methoxyglucobrassicin (149.99 mg/100 g DW) and gluconasturtiin (36.17 mg/100 g DW). Total flavonoids increased to a maximum of 1364.38 mg/100 g DW with 3.0 mM Phe (day 0), and polyphenols to a maximum of 1062.76 mg/100 g DW with 3.0 mM Trp (day 0). The precursors also increased the amounts of p-coumaric and ferulic acids, and rutoside, and generally increased the production of active photosynthetic pigments. Antioxidant potential increased the most with 0.1 mM Phe (day 0) (CUPRAC, FRAP), and with 0.5 mM Trp (day 10) (DPPH). The extracts of microshoots treated with 3.0 mM Phe (day 0) showed the most promising bacteriostatic activity against microaerobic Gram-positive acne strains (MIC 250-500 µg/mL, 20-21 mm inhibition zones). No extract was cytotoxic to normal human fibroblasts over the tested concentration range (up to 250 µg/mL).

Bioaccumulation of selected macro- and microelements and their impact on antioxidant properties and accumulation of glucosinolates and phenolic acids in in vitro cultures of Nasturtium officinale (watercress) microshoots.

The study evaluated bioaccumulation capacity of macro- and microelements, their impact on the production of glucosinolates and phenolic acids and antioxidant properties in a microshoot culture model of Nasturtium officinale. Elements: calcium, chromium, copper, iron, lithium, magnesium, selenium and zinc were supplemented in different salt concentrations to culture media. Bioaccumulation of elements [mg/100 gDW] varied from 1.24 (Li,1 mg/l) to 498.62 (Cr,50 mg/l) and was dependent on the type of element and its concentration. The bioconcentration factor (BCF) ranged from 11.37 (Li,25 mg/l) to 4467.00 (Ca,1 mg/l). The total glucosinolate contents [mg/100gDW] varied from 108.11 (Cr,1 mg/l) to 172.90 (Ca,1 mg/l). The presence of four phenolic acids was confirmed in the microshoots. Their total contents [mg/100gDW] ranged from 19.35 (Mg,10 mg/l) to 139.21 (Fe,50 mg/l). The highest antioxidant activity [nM trolox/mgDW], as evaluated by CUPRAC and QUENCHER-CUPRAC methods, was equal to 55.50 (Cu,1 mg/l) and 161.10 (Li,5 mg/l), respectively. The results proved good correlations between all studied parameters.

Enhancement of Glucosinolate Production in Watercress ( Nasturtium officinale) Hairy Roots by Overexpressing Cabbage Transcription Factors.

Glucosinolates are secondary metabolites that play important roles in plant defense and human health, as their production in plants is enhanced by overexpressing transcription factors. Here, four cabbage transcription factors (IQD1-1, IQD1-2, MYB29-1, and MYB29-2) affecting genes in both aliphatic and indolic glucosinolates biosynthetic pathways and increasing glucosinolates accumulation were overexpressed in watercress. Five IQD1-1, six IQD1-2, five MYB29-1, six MYB29-2, and one GUS hairy root lines were created. The expression of all genes involved in glucosinolates biosynthesis was higher in transgenic lines than in the GUS hairy root line, in agreement with total glucosinolates contents, determined by high-performance liquid chromatography. In transgenic IQD1-1 (1), IQD1-2 (4), MYB29-1 (2), and MYB29-2 (1) hairy root lines, total glucosinolates were 3.39-, 3.04-, 2.58-, and 4.69-fold higher than those in the GUS hairy root lines, respectively. These results suggest a central regulatory function for IQD1-1, IQD1-2, MYB29-1, and MYB29-2 transcription factors in glucosinolates biosynthesis in watercress hairy roots.

The hydroalcoholic extract of watercress attenuates protein oxidation, oxidative stress, and liver damage after bile duct ligation in rats.

Cholestatic liver disease is recognized by extreme collagen formation and deposition, which is mediated by free radicals. The aim of the current study was to investigate the probable hepatoprotective effects of hydroalcoholic extract of watercress (WC) against oxidative stress and liver injury in bile duct ligation (BDL)- induced cholestatic rats. A total of 32 male Wistar rats were divided into four groups; sham control (SC), BDL, SC + hydroalcoholic extract of WC and BDL + hydroalcoholic extract of WC. WC-treated rats received daily WC 500 mg/kg/day for 10 days. Biochemical tests, hepatic oxidative stress markers, and antioxidant enzymes activity were estimated. Further, liver hydroxyproline content was assayed and histological analysis was made. The BDL model markedly elevated the protein carbonyl (PCO) and hydroxyproline contents and decreased the glutathione peroxidase (GPx) activity. Hydroalcoholic extract of WC significantly decreased the surge in liver PCO and hydroxyproline levels and increased the reduced GPx enzyme activity contents in the hepatic tissue. As determined by hematoxylin and eosin staining, BDL considerably induced hepatocyte necrosis. Moreover, these changes were significantly attenuated by the hydroalcoholic extract of WC treatment. Our data indicate that the hydroalcoholic extract of WC extract attenuated liver damage in BDL rats by decreasing the hydroxyproline content and histopathological indexes. Also, it reduced oxidative stress by preventing the hepatic protein oxidation and enhancing the activity of the GPx enzyme via antioxidative effect and free-radical scavenging. Our findings suggest that hydroalcoholic extract of WC could be a beneficial new curative agent for cholestatic liver damage.

Chloroplast-selective gene delivery and expression in planta using chitosan-complexed single-walled carbon nanotube carriers.

Plant genetic engineering is an important tool used in current efforts in crop improvement, pharmaceutical product biosynthesis and sustainable agriculture. However, conventional genetic engineering techniques target the nuclear genome, prompting concerns about the proliferation of foreign genes to weedy relatives. Chloroplast transformation does not have this limitation, since the plastid genome is maternally inherited in most plants, motivating the need for organelle-specific and selective nanocarriers. Here, we rationally designed chitosan-complexed single-walled carbon nanotubes, utilizing the lipid exchange envelope penetration mechanism. The single-walled carbon nanotubes selectively deliver plasmid DNA to chloroplasts of different plant species without external biolistic or chemical aid. We demonstrate chloroplast-targeted transgene delivery and transient expression in mature Eruca sativa, Nasturtium officinale, Nicotiana tabacum and Spinacia oleracea plants and in isolated Arabidopsis thaliana mesophyll protoplasts. This nanoparticle-mediated chloroplast transgene delivery tool provides practical advantages over current delivery techniques as a potential transformation method for mature plants to benefit plant bioengineering and biological studies.

Metabolic targets of watercress and PEITC in MCF-7 and MCF-10A cells explain differential sensitisation responses to ionising radiation.

PURPOSE: Watercress is a rich source of phytochemicals with anticancer potential, including phenethyl isothiocyanate (PEITC). We examined the potential for watercress extracts and PEITC to increase the DNA damage caused by ionising radiation (IR) in breast cancer cells and to be protective against radiation-induced collateral damage in healthy breast cells. The metabolic events that mediate such responses were explored using metabolic profiling. METHODS: 1H nuclear magnetic resonance spectroscopy-based metabolic profiling was coupled with DNA damage-related assays (cell cycle, Comet assay, viability assays) to profile the comparative effects of watercress and PEITC in MCF-7 breast cancer cells and MCF-10A non-tumorigenic breast cells with and without exposure to IR. RESULTS: Both the watercress extract and PEITC-modulated biosynthetic pathways of lipid and protein synthesis and resulted in changes in cellular bioenergetics. Disruptions to the redox balance occurred with both treatments in the two cell lines, characterised by shifts in the abundance of glutathione. PEITC enhanced the sensitivity of the breast cancer cells to IR increasing the effectiveness of the cancer-killing process. In contrast, watercress-protected non-tumorigenic breast cells from radiation-induced damage. These effects were driven by changes in the cellular content of the antioxidant glutathione following exposure to PEITC and other phytochemicals in watercress. CONCLUSION: These findings support the potential prophylactic impact of watercress during radiotherapy. Extracted compounds from watercress and PEITC differentially modulate cellular metabolism collectively enhancing the therapeutic outcomes of radiotherapy.

A Nanobionic Light-Emitting Plant.

The engineering of living plants for visible light emission and sustainable illumination is compelling because plants possess independent energy generation and storage mechanisms and autonomous self-repair. Herein, we demonstrate a plant nanobionic approach that enables exceptional luminosity and lifetime utilizing four chemically interacting nanoparticles, including firefly luciferase conjugated silica (SNP-Luc), d-luciferin releasing poly(lactic-co-glycolic acid) (PLGA-LH2), coenzyme A functionalized chitosan (CS-CoA) and semiconductor nanocrystal phosphors for longer wavelength modulation. An in vitro kinetic model incorporating the release rates of the nanoparticles is developed to maximize the chemiluminescent lifetimes to exceed 21.5 h. In watercress (Nasturtium officinale) and other species, the nanoparticles circumvent limitations such as luciferin toxicity above 400 µM and colocalization of enzymatic reactions near high adenosine triphosphate (ATP) production. Pressurized bath infusion of nanoparticles (PBIN) is introduced to deliver a mixture of nanoparticles to the entire living plant, well described using a nanofluidic mathematical model. We rationally design nanoparticle size and charge to control localization within distinct tissues compartments with 10 nm nanoparticles localizing within the leaf mesophyll and stomata guard cells, and those larger than 100 nm segregated in the leaf mesophyll. The results are mature watercress plants that emit greater than 1.44 × 1012 photons/sec or 50% of 1 µW commercial luminescent diodes and modulate "off" and "on" states by chemical addition of dehydroluciferin and coenzyme A, respectively. We show that CdSe nanocrystals can shift the chemiluminescent emission to 760 nm enabling near-infrared (nIR) signaling. These results advance the viability of nanobionic plants as self-powered photonics, direct and indirect light sources.

Effects of live Myriophyllum aquaticum and its straw on cadmium accumulation in Nasturtium officinale.

The purpose of this study is to determine whether the allelopathy of living Myriophyllum aquaticum and its straw has the same effects; two pot experiments were conducted to study the effects of intercropping using M. aquaticum and its straw on the growth and cadmium (Cd) accumulation of Nasturtium officinale. Different planting ratios (1:3, 2:2 and 3:1) of N. officinale and M. aquaticum led to an increase in the biomass of both plant species and increased the Cd content in roots and shoots of N. officinale, but led to a reduction in the Cd content in roots and shoots of M. aquaticum. When the intercropping ratio of N. officinale and M. aquaticum was 3:1, the Cd amount in whole plants reached the maximum of 293.96 µg pot-1. Mulching the straw of M. aquaticum roots on the soil surface increased the biomass of N. officinale, but mulching the straw of M. aquaticum stems and leaves led to a decrease. Mulching the straw of roots, stems and leaves of M. aquaticum reduced Cd content and amounts in roots and shoots of N. officinale. Intercropping with M. aquaticum can improve the Cd uptake ability of N. officinale, but mulching M. aquaticum straw can reduce its Cd uptake ability.

De novo transcriptome analysis and glucosinolate profiling in watercress (Nasturtium officinale R. Br.).

BACKGROUND: Watercress (Nasturtium officinale R. Br.) is an aquatic herb species that is a rich source of secondary metabolites such as glucosinolates. Among these glucosinolates, watercress contains high amounts of gluconasturtiin (2-phenethyl glucosinolate) and its hydrolysis product, 2-phennethyl isothiocyanate, which plays a role in suppressing tumor growth. However, the use of N. officinale as a source of herbal medicines is currently limited due to insufficient genomic and physiological information. RESULTS: To acquire precise information on glucosinolate biosynthesis in N. officinale, we performed a comprehensive analysis of the transcriptome and metabolome of different organs of N. officinale. Transcriptome analysis of N. officinale seedlings yielded 69,570,892 raw reads. These reads were assembled into 69,635 transcripts, 64,876 of which were annotated to transcripts in public databases. On the basis of the functional annotation of N. officinale, we identified 33 candidate genes encoding enzymes related to glucosinolate biosynthetic pathways and analyzed the expression of these genes in the leaves, stems, roots, flowers, and seeds of N. officinale. The expression of NoMYB28 and NoMYB29, the main regulators of aliphatic glucosinolate biosynthesis, was highest in the stems, whereas the key regulators of indolic glucosinolate biosynthesis, such as NoDof1.1, NoMYB34, NoMYB51, and NoMYB122, were strongly expressed in the roots. Most glucosinolate biosynthetic genes were highly expressed in the flowers. HPLC analysis enabled us to detect eight glucosinolates in the different organs of N. officinale. Among these glucosinolates, the level of gluconasturtiin was considerably higher than any other glucosinolate in individual organs, and the amount of total glucosinolates was highest in the flower. CONCLUSIONS: This study has enhanced our understanding of functional genomics of N. officinale, including the glucosinolate biosynthetic pathways of this plant. Ultimately, our data will be helpful for further research on watercress bio-engineering and better strategies for exploiting its anti-carcinogenic properties.

Uranium accumulation in aquatic macrophytes in an uraniferous region: Relevance to natural attenuation.

Phytoremediation potential of uranium (U) was investigated by submerged, free-floating and rooted emergent native aquatic macrophytes inhabiting along the streams of Horta da Vilariça, a uraniferous geochemical region of NE Portugal. The work has been undertaken with the following objectives: (i) to relate the U concentrations in water-sediment-plant system; and (ii) to identify the potentialities of aquatic plants to remediate U-contaminated waters based on accumulation pattern. A total of 25 plant species culminating 233 samples was collected from 15 study points along with surface water and contiguous sediments. Concentrations of U showed wide range of variations both in waters (0.61-5.56 µg L(-1), mean value 1.98 µg L(-1)) and sediments (124-23,910 µg kg(-1), mean value 3929 µg kg(-1)) and this is also reflected in plant species examined. The plant species exhibited the ability to accumulate U several orders of magnitude higher than the surrounding water. Maximum U concentrations was recorded in the bryophyte Scorpiurium deflexifolium (49,639 µg kg(-1)) followed by Fontinalis antipyretica (35,771 µg kg(-1)), shoots of Rorippa sylvestris (33,837 µg kg(-1)), roots of Oenanthe crocata (17,807 µg kg(-1)) as well as in Nasturtium officinale (10,995 µg kg(-1)). Scorpiurium deflexifolium displayed a high bioconcentration factor (BF) of ∼2.5 × 10(4) (mean value). The species Fontinalis antipyretica, Nasturtium officinale (roots) and Rorippa sylvestris (shoots) exhibited the mean BFs of 1.7 × 10(4), 5 × 10(3) and 4.8 × 10(3) respectively. Maximum translocation factor (TF) was very much pronounced in the rooted perennial herb Rorippa sylvestris showing extreme ability to transport U for the shoots and seems to be promising candidate to be used as bioindicator species.

Unveiling the first indole-fused thiazepine: structure, synthesis and biosynthesis of cyclonasturlexin, a remarkable cruciferous phytoalexin.

As a mechanism of defense against pathogens and other types of stress, watercress plants produce a variety of elicited chemical defenses generally known as phytoalexins. Herein the chemical structure, synthesis, biosynthesis and antifungal activity of cyclonasturlexin, the most intriguing indolyl phytoalexin isolated to date, are reported.

Impact of spiked concentrations of Cd, Pb, As and Zn in growth medium on elemental uptake of Nasturtium officinale (Watercress).

This study is aimed at investigating the impact of water quality on the uptake and distribution of three non-essential and toxic elements, namely, As, Cd and Pb in the watercress plant to assess for metal toxicity. The plant was hydroponically cultivated under greenhouse conditions, with the growth medium being spiked with varying concentrations of As, Cd and Pb. Plants that were harvested weekly for elemental analysis showed physiological and morphological symptoms of toxicity on exposure to high concentrations of Cd and Pb. Plants exposed to high concentrations of As did not survive and the threshold for As uptake in watercress was established at 5 ppm. Translocation factors were low in all cases as the toxic elements accumulated more in the roots of the plant than the edible leaves. The impact of Zn on the uptake of toxic elements was also evaluated and Zn was found to have an antagonistic effect on uptake of both Cd and Pb with no notable effect on uptake of As. The findings indicate that phytotoxicity or death of the watercress plant would prevent it from being a route of human exposure to high concentrations of As, Cd and Pb in the environment.

Elemental distribution and uptake by watercress (Nasturtium aquaticum) as a function of water quality.

Watercress (Nasturtium aquaticum), is an edible plant commonly found in Southern Africa, which grows both in terrestrial and aquatic environments. The elemental concentrations in the plant and surrounding water (growth solution) were investigated to determine the plants nutritional value and to establish impact of water quality on elemental uptake by the plant. The concentrations in the leaves, roots and water were analyzed at eight different sites along the east coast of KwaZulu-Natal, South Africa (30.0000° S, 25.0000° E). Elemental concentrations in the leaves/roots (µg g(-1), dry mass) and water exchangeable/total (µg L(-1)) at Verulam (situated in the northern part of KwaZulu-Natal) were: As (1.2/7.5 and 0.053/0.09), Ca (18272/26091 and 0.336/7.15), Cd (0.9/0.8 and 0.004/0.01), Co (37.2/34 and 0/0), Cr (7/35 and 0.003/0.03), Cu (3/19 and 0.016/0.16), Fe (528/4308 and 0.375/0.6), Mg (3444/1141 and 6.4/7), Mn (110/667 and 0/0), Ni (20/63 and 0/0.01), Pb (16/17 and 0.01/0.02), Se (28/11 and 0.003/0.01) and Zn (102/116 and 0.099/0.36). Elemental uptake was controlled by the plant but water quality did have an impact on uptake. Concentrations of metals in the leaves were in decreasing order of Ca > Mg > Fe > Mn > Zn > Cu > Cr > Ni > Pb > Se > Cd > As > Co. The watercress plant was found to be a rich source of essential elements especially Fe and Cr and contained low concentrations of the toxic metals investigated thereby making it safe for human consumption.

Biodegradation of C.I. Acid Blue 92 by Nasturtium officinale: Study of Some Physiological Responses and Metabolic Fate of Dye.

The present study was conducted to evaluate the potential of aquatic vascular plant, Nasturtium officinale, for degradation of C.I. Acid Blue 92 (AB92). The effect of operational parameters such as initial dye concentration, plant biomass, pH, and temperature on the efficiency of biological decolorization process was determined. The reusability of the plant in long term repetitive operations confirmed the biological degradation process. The by-products formed during biodegradation process were identified by GC-MS technique. The effects of the dye on several plant physiological responses such as photosynthetic pigments content and antioxidant enzymes activity were investigated. The content of chlorophyll and carotenoids was significantly reduced at 20 mg/L of the dye. The activities of superoxide dismutase and peroxidase were remarkably increased in the plant root verifying their importance in plant tolerance to the dye contamination.

Arsenic accumulation by edible aquatic macrophytes.

Edible aquatic macrophytes grown in arsenic (As)-contaminated soil and sediment were investigated to determine the extent of As accumulation and potential risk to humans when consumed. Nasturtium officinale (watercress) and Diplazium esculentum (warabi) are two aquatic macrophytes grown and consumed in Hawaii. Neither has been assessed for potential to accumulate As when grown in As-contaminated soil. Some former sugarcane plantation soils in eastern Hawaii have been shown to have concentrations of total As over 500 mg kg(-1). It was hypothesized that both species will accumulate more As in contaminated soils than in non-contaminated soils. N. officinale and D. esculentum were collected in areas with and without As-contaminated soil and sediment. High soil As concentrations averaged 356 mg kg(-1), while low soil As concentrations were 0.75 mg kg(-1). Average N. officinale and D. esculentum total As concentrations were 0.572 mg kg(-1) and 0.075 mg kg(-1), respectively, corresponding to hazard indices of 0.12 and 0.03 for adults. Unlike previous studies where watercress was grown in As-contaminated water, N. officinale did not show properties of a hyperaccumulator, yet plant concentrations in high As areas were more than double those in low As areas. There was a slight correlation between high total As in sediment and soil and total As concentrations in watercress leaves and stems, resulting in a plant uptake factor of 0.010, an order of magnitude higher than previous studies. D. esculentum did not show signs of accumulating As in the edible fiddleheads. Hawaii is unique in having volcanic ash soils with extremely high sorption characteristics of As and P that limit release into groundwater. This study presents a case where soils and sediments were significantly enriched in total As concentration, but the water As concentration was below detection limits.

Chemiluminescent examination of abiotic oxidative stress of watercress.

Watercress (Nasturtium officinale) is an aquatic plant that readily bioaccumulates heavy metals that may be found in contaminated aquatic systems. Toxic effects of contaminants on the physiological processes cause changes in oxidase enzymatic activity in watercress, which can be measured with a luminometer. The luminometer uses the reaction produced when peroxidases break down hydrogen peroxide into water and an oxygen radical. The resulting oxyradical binds to and oxidizes phenolic groups, producing a measureable luminescent reaction. Nasturtium officinale plants were exposed to 3 different concentrations of heavy metals, including lead, nickel, copper, and manganese for 24 h, 48 h, and 72 h. Aquatic exposure to the 4 heavy metals caused a significant increase in oxidative enzyme production. Fluorometric and morphometric measurements were also conducted to compare plant stress with the oxidative enzyme analyses. Fluorometric measurements performed on plants stressed by exposure to heavy metals revealed no significant decreases in photosystem II efficiency. Morphometric measurements of root length showed decreased root growth resulting from exposures to Ni, Cu, and Mn.

Turning the 'mustard oil bomb' into a 'cyanide bomb': aromatic glucosinolate metabolism in a specialist insect herbivore.

Plants have evolved a variety of mechanisms for dealing with insect herbivory among which chemical defense through secondary metabolites plays a prominent role. Physiological, behavioural and sensorical adaptations to these chemicals provide herbivores with selective advantages allowing them to diversify within the newly occupied ecological niche. In turn, this may influence the evolution of plant metabolism giving rise to e.g. new chemical defenses. The association of Pierid butterflies and plants of the Brassicales has been cited as an illustrative example of this adaptive process known as 'coevolutionary armsrace'. All plants of the Brassicales are defended by the glucosinolate-myrosinase system to which larvae of cabbage white butterflies and related species are biochemically adapted through a gut nitrile-specifier protein. Here, we provide evidence by metabolite profiling and enzyme assays that metabolism of benzylglucosinolate in Pieris rapae results in release of equimolar amounts of cyanide, a potent inhibitor of cellular respiration. We further demonstrate that P. rapae larvae develop on transgenic Arabidopsis plants with ectopic production of the cyanogenic glucoside dhurrin without ill effects. Metabolite analyses and fumigation experiments indicate that cyanide is detoxified by ß-cyanoalanine synthase and rhodanese in the larvae. Based on these results as well as on the facts that benzylglucosinolate was one of the predominant glucosinolates in ancient Brassicales and that ancient Brassicales lack nitrilases involved in alternative pathways, we propose that the ability of Pierid species to safely handle cyanide contributed to the primary host shift from Fabales to Brassicales that occured about 75 million years ago and was followed by Pierid species diversification.