Glucosinolates

Glucosinolates (GLS) are secondary plant metabolites occurring in cruciferous vegetables. Their biologically active breakdown products show cancer preventive properties in animal and cell studies. So far, epidemiologic studies, using consumption of cruciferous vegetables as proxy for GLS intake, yielded inconsistent results. Here, we evaluated the association between dietary intake of GLS in comparison with consumption data of GLS-containing foods and the risk of prostate cancer. The study population comprised 11,405 male participants of the prospective EPIC-Heidelberg cohort study. During a mean follow-up time of 9.4 years, 328 incident cases of prostate cancer occurred. At recruitment, habitual food consumption was assessed by a validated food frequency questionnaire, and intake of individual GLS was estimated by means of a newly compiled database on food content of GLS. Adjusted hazard ratios (HR) for prostate cancer were calculated using the Cox proportional hazard model. Median daily intake of total GLS was 7.9 mg/day (interquartile range 5.1-11.9 mg/day). The risk of prostate cancer decreased significantly over quartiles of total GLS intake (multivariate HR [4th vs. 1st quartile] 0.68, 95% CI 0.48-0.97, p trend 0.03). Associations with GLS-containing food intake were weaker. Among GLS subgroups, aliphatic GLS showed the strongest inverse association with cancer risk. Analyses stratified by tumor stage and grade gave hint to inverse associations for localized and low-grade cancers. This study shows an inverse association between dietary intake of GLS and the risk of prostate cancer. Because this is the first prospective study using individual GLS intake data, confirmation in other studies is warranted.

This study was conducted to identify and quantify the sensory characteristics and chemical profile of organically and conventionally grown pac choi (Brassica rapa var. Mei Qing Choi), also called bok choy, at 3 stages of growth (2.5, 4.5, and 6.5 wk). Sensory and instrumental data were correlated using partial least squares regression. Pac choi was grown in late spring. Descriptive sensory analysis was conducted by a highly trained panel and compounds were identified and quantified using a gas chromatograph/mass spectrometer. The findings of the study indicate that the differences in sensory characteristics and chemical profiles among stages of growth are more substantial than the differences between organic and conventional production. Green-unripe, musty/earthy, lettuce, and sweet flavors are representative in pac choi at early stages of growth. When older, pac choi has higher intensities of green-grassy/leafy, bitter, cabbage, and sulfur flavors that are associated with the increase of (Z)-3hexen-1-ol, octyl acetate, 1-nonanol, 2-decanone, 1-penten-3-ol, linalool, camphor, menthol, isobornyl acetate, geranylacetone, and cedrol compounds. Conventional pac choi was higher than organic pac choi in green overall, bitter, and soapy flavors only at 2.5 wk of age. This may be associated with the presence of (Z)-3-hexenal, 2-hexyn-1-ol, and (E)-2-hexenal compounds. Practical Application: The increased popularity of organic production has amplified the need for research that will help in understanding how this production system affects the final quality of food products. This study suggests that the stage of development has a much larger impact on sensory quality than organic or conventional growing of pac choi. Findings from this study promote consumer choice by showing that comparable sensory quality can be obtained using either production system, making the ultimate choice not only based on sensory quality but consumer choice related to environmental beliefs or economics.

Myrosinase is a cytosolic plant enzyme present in daikon (Raphanus sativus, Japanese white radish) roots that hydrolyzes 4-methylthio-3-butenyl glucosinolate (MTBGLS) into the natural pungent agent 4-methylthio-3-butenyl isothiocyanate (MTBITC), which possesses antimicrobial, antimutagenic, and anticarcinogenic properties. The concentration of MTBGLS, myrosinase activity, and production of MTBITC in seven daikon varieties (one conventional and six heirlooms) were determined to rank the activity of the glucosinolate-myrosinase system and identify critical factors influencing the production of MTBITC. The six heirloom varieties produced 2.0-11.5 times higher levels of MTBITC as compared to the conventional variety, Aokubi, which is consumed by the present Japanese population. The myrosinase was located exclusively in the outer epidermal layer in Aokubi, and MTBGLS was widely distributed throughout the root tissue. Although the skin is a potentially rich source of myrosinase in Aokubi, the skin is usually peeled off in the current practice of preparing daikon for cooking. New practices are therefore proposed for the preparation of daikon tubers that eliminate the peeling of the skin to avoid removing the enzyme needed to convert MTBGLS to the health-beneficial MTBITC. It is also concluded that the consumption of heirloom daikon varieties in addition to changes in food preparation will optimize the health benefits of daikon.

El presente articulo cubre una revision bibliografica realizada durante un seminario sobre las curtiembres y su impacto ambiental. El texto va desde la descripcion del proceso y las caracteristicas de los residuos de cadaetapa del proceso de curtido, las consecuencias ambientales que tiene el vertido de estos residuos al ambiente, algunos aspectos toxicologicos y un breve analisis de la realidad peruana de esta industria.

Glucosinolate levels in leaves were determined in a collection of 113 varieties of turnip greens (Brassica rapa L.) from northwestern Spain grown at two sites. Sensorial attributes were also assessed by a consumer panel. The objectives were to determine the diversity among varieties in total glucosinolate content and glucosinolate profile and to evaluate their sensory attributes in relation to glucosinolate content for breeding purposes. Sixteen glucosinolates were identified, being the aliphatic glucosinolates, gluconapin and glucobrassicanapin the most abundant. Other aliphatic glucosinolates, such as progoitrin, glucoalyssin, and gluconapoleiferin were relatively abundant in varieties with a different glucosinolate profile. Indolic and aromatic glucosinolate concentrations were low and showed few differences among varieties. Differences in total glucosinolate content, glucosinolate profile and bitterness were found among varieties, with a total glucosinolate content ranging from 11.8 to 74.0 lmol g À1 dw at one site and from 7.5 to 56.9 lmol g À1 dw at the other site. Sensory analysis comparing bitterness with variation in glucosinolate, gluconapin and glucobrassicanapin concentrations suggested that these compounds and their breakdown products are not the only determinants of the characteristic flavour of this vegetable. Other phytochemicals are probably involved on the characteristic bitter flavour. The varieties MBG-BRS0132, MBG-BRS0082, MBG-BRS0173, and MBG-BRS0184 could be good candidates for future breeding programs since they had high total glucosinolate content and good agronomic performance. The presence of glucoraphanin in some varieties should be studied more extensively, because this aliphatic glucosinolate is the precursor of sulforaphane, a potent anti-cancer isothiocyanate.

Glucosinolate content of leaves and roots, diversity in leaf pubescence, and resistance to two near-isogenic lines of the flea beetle Phyllotreta nemorum with or without an R-gene, were determined for 27 accessions of 7 Barbarea taxa, i.e. B. stricta, B. orthoceras, B. intermedia, B. verna, B. vulgaris var. vulgaris, the G-type of B. vulgaris var. arcuata and the P-type of B. vulgaris var. arcuata. Four variable glucosinolate biosynthetic characters were deduced. For (formally) homophenylalanine-derived glucosinolates: (1) Presence or absence of 2-hydroxylation, and if present, R-or S-configuration of 2-hydroxylation; (2) presence or absence of p-hydroxylation; and for tryptophan-derived glucosinolates: (3) presence or absence of N-methoxyglucobrassicin; and (4) presence or absence of 1,4-dimethoxyglucobrassicin. Three phenotypes of leaf-pubescence were observed; (1) glabrous to glabrate leaves;

The glucosinolate-myrosinase system found in plants of the order Brassicales is one of the best studied plant defense systems. Hydrolysis of the physiologically inert glucosinolates by hydrolytic enzymes called myrosinases, which only occurs upon tissue disruption, leads to the formation of biologically active compounds. The chemical nature of the hydrolysis products depends on the presence or absence of supplementary proteins, such as epithiospecifier proteins (ESPs). ESPs promote the formation of epithionitriles and simple nitriles at the expense of the corresponding isothiocyanates which are formed through spontaneous rearrangement of the aglucone core structure. While isothiocyanates are toxic to a wide range of organisms, including insects, the ecological significance of nitrile formation and thus the role of ESP in plant-insect interactions is unclear. Here, we identified ESP-expressing cells in various organs and several developmental stages of different Arabidopsis thaliana ecotypes by immunolocalization. In the ecotype Landsberg erecta, ESP was found to be consistently present in the epidermal cells of all aerial parts except the anthers and in S-cells of the stem below the inflorescence. Analyses of ESP expression by quantitative real-time PCR, Western blotting, and ESP activity assays suggest that plants control the outcome of glucosinolate hydrolysis by regulation of ESP at both the transcriptional and the post-transcriptional levels. The localization of ESP in the epidermal cell layers of leaves, stems and reproductive organs supports the hypothesis that this protein has a specific function in defense against herbivores and pathogens.

Broccoli inflorescences have been recognized as components of healthy diets on the basis of their high content of fiber, vitamin C, carotenoids, and glucosinolates/isothiocyanates. Broccoli sprouts have been recently shown to have high levels of glucoraphanin (4-methylsulfinylbutyl glucosinolate), the precursor of the chemoprotective isothiocyanate, sulforaphane. This study evaluated the effects of temperature and developmental stage on the glucosinolate content of broccoli sprouts. Seedlings cultivated using a 30/15°C (day/night) temperature regime had significantly higher glucosinolate levels (measured at six consecutive days postemergence) than did sprouts cultivated at lower temperatures (22/15 and 18/12°C; p < 0.001). Both higher (33.1°C) and lower (11.3°C) constant temperatures induced higher glucosinolate levels in sprouts grown to a uniform size. Glucosinolate levels were highest in cotyledons and lowest in roots of sprouts dissected both early and late in the 11 day developmental span investigated. Nongerminated seeds have the highest glucosinolate levels and concordantly greater induction of mammalian phase 2 detoxication enzymes. Levels decline as sprouts germinate and develop, with consistently higher glucosinolate content in younger developmental stages, independent of the temperature regime. Temperature stress or its associated developmental anomalies induce higher glucosinolate levels, specific elevations in glucoraphanin content, and parallel induction of phase 2 chemoprotective enzymes.

Sulphur (S) fertilization is essential for primary and secondary metabolism in cruciferous foods. Deficient, suboptimal, or excessive S affects the growth and biosynthesis of secondary metabolites in adult plants. Nevertheless, there is little information regarding the influence of S fertilization on sprouts and seedlings. An experiment was set up to evaluate the effect of S fertilization, supplied as K 2 SO 4 at 0, 15, 30, and 60 mg/L, on the glucosinolate content of broccoli sprouts during the germination course of 3, 6, 9, and 12 d after sowing. Glucosinolate concentration was strongly influenced by germination, causing a rapid increase during the first 3 d after sowing, and decreasing afterwards. The S supply increased aliphatic and total glucosinolate content at the end of the monitored sprouting period. S-treated sprouts, with S 15 , S 30 , and S 60 at 9 and 12 d after sowing presented enhanced glucosinolate content. Overall, both germination time and S fertilization were key factors in maximizing the bioactive health-promoting phytochemicals of broccoli.

Glucosinolates from the genus Brassica can be converted into bioactive compounds known to induce phase II enzymes, which may decrease the risk of cancers. Conversion via hydrolysis is usually by the brassica enzyme myrosinase, which can be inactivated by cooking or storage. We examined the potential of three beneficial bacteria, Lactobacillus plantarum KW30, Lactococcus lactis subsp. lactis KF147, and Escherichia coli Nissle 1917, and known myrosinase-producer Enterobacter cloacae to catalyze the conversion of glucosinolates in broccoli extract. Enterobacteriaceae consumed on average 65% glucoiberin and 78% glucoraphanin, transforming them into glucoiberverin and glucoerucin, respectively, and small amounts of iberverin nitrile and erucin nitrile. The lactic acid bacteria did not accumulate reduced glucosinolates, consuming all at 30−33% and transforming these into iberverin nitrile, erucin nitrile, sulforaphane nitrile, and further unidentified metabolites. Adding beneficial bacteria to a glucosinolate-rich diet may increase glucosinolate transformation, thereby increasing host exposure to bioactives.

Glucosinolates have attracted significant interest due to the chemopreventive properties of some of their transformation products. Numerous protocols for the extraction and analysis of glucosinolates have been published, but limited effort has been devoted to optimize and validate crucial extraction parameters and sample preparation steps. We carried out a systematic optimization and validation of a quantitative assay for the direct analysis of intact glucosinolates in Isatis tinctoria leaves (woad, Brassicaceae). Various parameters such as solvent composition, particle size, temperature, and number of required extraction steps were optimized using pressurized liquid extraction (PLE). We observed thermal degradation of glucosinolates at temperatures above 50°C, and loss of &gt;60% within 10min at 100°C, but no enzymatic degradation in the leaf samples at ambient temperature. Excellent peak shape and resolution was obtained by reversed-phase chromatography on a Phenomenex Aqua column...

Moringa oleifera is a medicinal plant and an excellent dietary source of micronutrients (vitamins and minerals) and healthpromoting phytochemicals (phenolic compounds, glucosinolates and isothiocyanates). Glucosinolates and isothiocyanates are known to possess anti-carcinogenic and antioxidant effects and have attracted great interest from both toxicological and pharmacological points of view, as they are able to induce phase 2 detoxification enzymes and to inhibit phase 1 activation enzymes. Phenolic compounds possess antioxidant properties and may exert a preventative effect in regards to the development of chronic degenerative diseases.

In this review, we provide an overview of the role of glucosinolates and other phytochemical compounds present in the Brassicaceae in relation to plant protection and human health. Current knowledge of the factors that influence phytochemical content and profile in the Brassicaceae is also summarized and multi-factorial approaches are briefly discussed. Variation in agronomic conditions (plant species, cultivar, developmental stage, plant organ, plant competition, fertilization, pH), season, climatic factors, water availability, light (intensity, quality, duration) and CO(2) are known to significantly affect content and profile of phytochemicals. Phytochemicals such as the glucosinolates and leaf surface waxes play an important role in interactions with pests and pathogens. Factors that affect production of phytochemicals are important when designing plant protection strategies that exploit these compounds to minimize crop damage caused by plant pests and pathogens. Brassicaceous pl...

Cooking induces many chemical and physical modifications in foods; among these the phytochemical content can change. Many authors have studied variations in vegetable nutrients after cooking, and great variability in the data has been reported. In this review more than 100 articles from indexed scientific journals were considered in order to assess the effect of cooking on different phytochemical classes. Changes in phytochemicals upon cooking may result from two opposite phenomena: (1) thermal degradation, which reduces their concentration, and (2) a matrix softening effect, which increases the extractability of phytochemicals, resulting in a higher concentration with respect to the raw material. The final effect of cooking on phytochemical concentration depends on the processing parameters, the structure of food matrix, and the chemical nature of the specific compound. Looking at the different cooking procedures it can be concluded that steaming will ensure better preservation/extraction yield of phenols and glucosinolates than do other cooking methods: steamed tissues are not in direct contact with the cooking material (water or oil) so leaching of soluble compounds into water is minimised and, at the same time, thermal degradation is limited. Carotenoids showed a different behaviour; a positive effect on extraction and the solubilisation of carotenes were reported after severe processing.

T his chapter is a comprehensive review of the health promoting phytochemicals commonly found in our daily food. These include carotenoids, phenolics, phytoestrogenes, polyunsaturated fatty acids, conjugated linoleic acids, tocols, allicin, glucosinolates, limonene and capsaicinoids. The review encompasses the main food sources of these chemicals in the diet, the possible mechanisms of their activity, evidence for potential health promoting activity and possible harmful effects. The newly emerged interest in these phytochemicals in animal nutrition as substitutes for synthetic antibiotic growth promoters has also been addressed.

The glucosinolate content of various organs of the model plant Arabidopsis thaliana (L.) Heynh., Columbia (Col-0) ecotype, was analyzed at different stages during its life cycle. Significant differences were noted among organs in both glucosinolate concentration and composition. Dormant and germinating seeds had the highest concentration (2.5-3.3% by dry weight), followed by inflorescences, siliques (fruits), leaves and roots. While aliphatic glucosinolates predominated in most organs, indole glucosinolates made up nearly half of the total composition in roots and late-stage rosette leaves. Seeds had a very distinctive glucosinolate composition. They possessed much higher concentrations of several types of aliphatic glucosinolates than other organs, including methylthioalkyl and, hydroxyalkyl glucosinolates and compounds with benzoate esters than other organs. From a developmental perspective, older leaves had lower glucosinolate concentrations than younger leaves, but this was not due to decreasing concentrations in individual leaves with age (glucosinolate concentration was stable during leaf expansion). Rather, leaves initiated earlier in development simply had much lower rates of glucosinolate accumulation per dry weight gain throughout their lifetimes. During seed germination and leaf senescence, there were significant declines in glucosinolate concentration. The physiological and ecological significance of these findings is briefly discussed. #

This paper reviews the properties of the most cultivated species of the Moringaceae family, Moringa oleifera Lam. The paper takes a critical look at the positive and the associated negative properties of the plant, with particular emphasis on its chemistry, selected medicinal and nutritional properties, as well as some ecological implications of the plant. The review highlights the importance of glucosinolates (GS) compounds which are relatively unique to the Moringa species family, with glucomoriginin and its acylated derivative being the most abundant. We highlight some new research findings revealing that not all M. oleifera cultivars contain an important flavonoid, rutin. The review also focuses on phenolic acids, tannin, minerals and vitamins, which are in high amounts when compared to most vegetables and fruits. Although there are numerous benefits of using M. oleifera for medicinal purposes, there are reports of contraindications. Nonetheless, we note that there are no major harmful effects of M. oleifera that have been reported by the scientific community. M. oleifera is suspected to be potentially invasive and moderately invasive in some regions of the world because of its ability to grow in a wide range of environmental conditions. However, the plant is currently classified as a low potential invasive species and thus there is a need to constantly monitor the species. Despite the numerous benefits associated with the plant, there is still a paucity of data on clinical trials proving both the positive and negative effects of the plant. We recommend further clinical trials to ascertain the properties associated with the plant, especially regarding long term use.

El presente artículo cubre una revisión bibliográfica realizada durante un seminario sobre las curtiembres y su impacto ambiental. El texto va desde la descripción del proceso y las características de los residuos de cada etapa del proceso de curtido, las consecuencias ambientales que tiene el vertido de estos residuos al ambiente, algunos aspectos toxicológicos y un breve análisis de la realidad peruana de esta industria.

Glucosinolates are sulfur-rich secondary metabolites characteristic of the Brassicales order with important biological and economic roles in plant defense and human nutrition. Application of systems biology tools continues to identify genes involved in the biosynthesis of glucosinolates. Recent progress includes genes in all three phases of the pathway, i.e. side-chain elongation of precursor amino acids, formation of the core glucosinolate structure and side-chain decoration. Major breakthroughs include the ability to produce glucosinolates in Nicotiana benthamiana, the finding that specific glucosinolates play a key role in Arabidopsis innate immune response, and a better understanding of the link between primary sulfur metabolism and glucosinolate biosynthesis.

Glucosinolates present classical examples of plant compounds affecting insect-plant interactions. They are found mainly in the family Brassicaceae, which includes several important crops. More than 120 different glucosinolates are known. The enzyme myrosinase, which is stored in specialized plant cells, converts glucosinolates to the toxic isothiocyanates. Insect herbivores may reduce the toxicity of glucosinolates and their products by excretion, detoxification, or behavioral adaptations. Glucosinolates also affect higher trophic levels, via reduced host or prey quality or because specialist herbivores may sequester glucosinolates for their own defense. There is substantial quantitative and qualitative variation between plant genotypes, tissues, and ontogenetic stages, which poses specific challenges to insect herbivores. Even though glucosinolates are constitutive defenses, their levels are influenced by abiotic and biotic factors including insect damage. Plant breeders may use knowledge on glucosinolates to increase insect resistance in Brassica crops. State-of-the-art techniques, such as mutant analysis and metabolomics, are necessary to identify the exact role of glucosinolates. 57 Annu. Rev. Entomol. 2009.54:57-83. Downloaded from www.annualreviews.org by Swedish University of Agricultural Sciences on 01/18/11. For personal use only.

The glucosinolate composition and content in various tissues of Arabidopsis thaliana (L.) Heynh. ecotype Columbia during development from seeds to bolting plants were determined in detail by highperformance liquid chromatography. Comparison of the glucosinolate profiles of leaves, roots and stems from mature plants with those of green siliques and mature seeds indicated that a majority of the seed glucosinolates were synthesized de novo in the silique. A comparison of the glucosinolate profile of mature seeds with that of cotyledons indicated that a major part of seed glucosinolates was retained in the cotyledons. Turnover of glucosinolates was studied by germination of seeds containing radiolabelled p-hydroxybenzylglucosinolate (p-OHBG). Approximately 70% of the content of [ 14 C]p-OHBG in the seeds was detected in seedlings at the cotyledon stage and [ 14 C]p-OHBG was barely detectable in young plants with rosettes of six to eight leaves. The turnover of p-OHBG was found to coincide with the expression of the glucosinolate-degrading enzyme myrosinase, which was detectable at very low levels in seedlings at the cotyledon stage, but which dramatically increased in leaves from plants at later developmental stages. This indicates that there is a continuous turnover of glucosinolates during development and not only upon tissue disruption.

Glucosinolates are an important class of secondary plant metabolites, possessing health-promoting properties. Young broccoli plants are a very good source of glucosinolates with concentrations several times greater than in mature plants. The aim of our study was to develop a liquid chromatography-mass spectrometry and liquid chromatography/tandem mass spectrometry qualitative and quantitative method for the measure of glucosinolates in broccoli sprouts. The described method provides high sensitivity and specificity, allowing a rapid and simultaneous determination of 14 glucosinolates. The proposed method has been validated for eight glucosinolates: glucobrassicin, glucoraphanin, glucoiberin, glucoerucin, progoitrin, gluconapin, sinigrin and glucocheirolin. The linear range was 1-150 mg ml À1 , the intra-day and inter-day precision values are within 6% and 8% at the lower limit of quantification, while the overall recovery of the eight glucosinolates was 99 AE 9%. This validated method was used successfully for analysis of glucosinolates content of broccoli sprouts grown in different conditions. Precision and accuracy were evaluated at three concentration levels for each compound through triplicate intra-day assays and inter-day assays over 3 days LC-MS determination of glucosinoloma

The formation of plant-derived biomolecules during sauerkraut fermentation was studied. Cabbage was fermented with a starter culture, and the results were compared to the results of spontaneous fermentation. The concentration of flavonoids and glucosinolates was analyzed by HPLC, and that of the glucosinolate breakdown products, by GC-MS. Of the 20 different flavonoids tested, only kaempferol was found (0.9 mg/ kg FW, fresh weight). The content of kaempferol remained constant in the cabbage fiber matrix over the fermentation process. The nitrite concentration was below the detection limit in both fermentations. The total glucosinolate content in the raw material was 3.71 µmol/g DW, dry weight. Glucosinolates were totally decomposed in both fermentations during two weeks, and different types of breakdown products were formed. Isothiocyanates, indole-3-carbinol, goitrin, allyl cyanide, and nitriles were determined in the fermented cabbage. Isothiocyanates and allyl cyanide were the predominant breakdown products in both fermentations. Sulforaphane nitrile and goitrin were found only in small quantities in the end products.

Sulforaphane, a naturally occurring cancer chemopreventive, is the hydrolysis product of glucoraphanin, the main glucosinolate in broccoli. The hydrolysis requires myrosinase isoenzyme to be present in sufficient activity; however, processing leads to its denaturation and hence reduced hydrolysis. In this study, the effect of adding mustard seeds, which contain a more resilient isoform of myrosinase, to processed broccoli was investigated with a view to intensify the formation of sulforaphane. Thermal inactivation of myrosinase from both broccoli and mustard seeds was studied. Thermal degradation of broccoli glucoraphanin was investigated in addition to the effects of thermal processing on the formation of sulforaphane and sulforaphane nitrile. Limited thermal degradation of glucoraphanin (less than 12%) was observed when broccoli was placed in vacuum sealed bag (sous vide) and cooked in a water bath at 100°C for 8 and 12 min. Boiling broccoli in water prevented the formation of any significant levels of sulforaphane due to inactivated myrosinase. However, addition of powdered mustard seeds to the heat processed broccoli significantly increased the formation of sulforaphane.

As part of our analysis of branched-chain amino acid metabolism in plants, we analyzed the function of Arabidopsis thaliana BRANCHED-CHAIN AMINOTRANSFERASE4 (BCAT4). Recombinant BCAT4 showed high efficiency with Met and its derivatives and the corresponding 2-oxo acids, suggesting its participation in the chain elongation pathway of Met-derived glucosinolate biosynthesis. This was substantiated by in vivo analysis of two BCAT4 T-DNA knockout mutants, in which Met-derived aliphatic glucosinolate accumulation is reduced by ;50%. The increase in free Met and S-methylmethionine levels in these mutants, together with in vitro substrate specificity, strongly implicate BCAT4 in catalysis of the initial deamination of Met to 4-methylthio-2-oxobutyrate. BCAT4 transcription is induced by wounding and is predominantly observed in the phloem. BCAT4 transcript accumulation also follows a diurnal rhythm, and green fluorescent protein tagging experiments and subcellular protein fractions show that BCAT4 is located in the cytosol. The assignment of BCAT4 to the Met chain elongation pathway documents the close evolutionary relationship of this pathway to Leu biosynthesis. In addition to BCAT4, the enzyme methylthioalkylmalate synthase 1 has been recruited for the Met chain elongation pathway from a gene family involved in Leu formation. This suggests that the two pathways have a common evolutionary origin.

Barbarea vulgaris R. Br. has been proposed as a dead-end trap crop for diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), because its larvae do not survive on this plant species despite being highly preferred for oviposition. We compared plants of several species, varieties, and types in the genus Barbarea (Brassicaceae) to study their potential as trap crops for P. xylostella. In terms of insect behavior, Barbarea plants were assessed based on the criteria of high oviposition preference by P. xylostella moths (compared to other Barbarea plants and to three Brassica oleracea L. crop varieties) and low survival of P. xylostella larvae. Barbarea plants were also assessed based on the criteria of high content of glucosinolates, which stimulate adult oviposition and larval feeding in P. xylostella, and high content of saponins, which are detrimental to survival of P. xylostella larvae. All Barbarea plants tested were preferred over cabbage by ovipositing P. xylostella. Among Barbarea plants, few significant differences in oviposition preference by P. xylostella were found. Ovipositing P. xylostella preferred B. vulgaris plants containing mainly 2-phenylethylglucosinolate over B. vulgaris plants containing mainly (S)-2-hydroxy-2-phenylethylglucosinolate, and P-type B. vulgaris var. arcuata plants over Barbarea rupicola and B. vulgaris var. variegata plants. Despite containing a lower content of saponins than other Barbarea plants tested, Barbarea verna did not allow survival of P. xylostella larvae. Our studies show that, except for B. rupicola and P-type B. vulgaris var. arcuata, which allowed survival of P. xylostella larvae, all Barbarea plants tested have potential as dead-end trap crops for P. xylostella.

In this review, we provide an overview of the role of glucosinolates and other phytochemical compounds present in the Brassicaceae in relation to plant protection and human health. Current knowledge of the factors that influence phytochemical content and profile in the Brassicaceae is also summarized and multi-factorial approaches are briefly discussed. Variation in agronomic conditions (plant species, cultivar, developmental stage, plant organ, plant competition, fertilization, pH), season, climatic factors, water availability, light (intensity, quality, duration) and CO(2) are known to significantly affect content and profile of phytochemicals. Phytochemicals such as the glucosinolates and leaf surface waxes play an important role in interactions with pests and pathogens. Factors that affect production of phytochemicals are important when designing plant protection strategies that exploit these compounds to minimize crop damage caused by plant pests and pathogens. Brassicaceous pl...

In the present study analytical and preparative supercritical fluid chromatography (SFC) were used for investigation of myrosinase catalysed degradation of 4-hydroxybenzylglucosinolate (sinalbin). Sinalbin occurs as a major glucosinolate in seeds of Sinapis alba L., in various mustards and other food products. The degradation products were identified and quantified by analysis based on a developed SFC method using a bare silica column. Determinations comprised transformation products of sinalbin, produced both during degradation of isolated sinalbin, and during autolysis of meal from S. alba seeds. The conditions in the developed SFC method were used as basis for the preparative SFC procedure applied for isolation of the components prior to their identification by nuclear magnetic resonance (NMR) spectroscopy. Myrosinase catalysed sinalbin hydrolysis resulted in the reactive 4-hydroxybenzyl isothiocyanate as an initial product at pH values from 3.5 to 7.5 whereas 4-hydroxybenzyl cyanide was one of the major products at low pH values. 4-Hydroxybenzyl isothiocyanate was found to disappear from the aqueous reaction mixtures in a few hours, as it reacted easily with available nucleophilic reagents. 4-Hydroxybenzyl alcohol was found as the product from reaction with water, and with ascorbic acid, 4-hydroxybenzylascorbigen was produced.

This article reviews the mechanisms by which glucosinolate breakdown products are thought to inhibit carcinogenesis. It describes how isothiocyanates, thiocyanates, nitriles, cyano-epithioalkanes and indoles are produced from glucosinolates through the actions of myrosinase, epithiospecifier protein and epithiospecifier modifier protein released from cruciferous vegetables during injury to the plant. The various biological activities displayed by these phytochemicals are described. In particular, their abilities to induce cytoprotective genes, mediated by the Nrf2 (NF-E2 related factor 2) and AhR (arylhydrocarbon receptor) transcription factors, and their abilities to repress NF-κB (nuclear factor-κB) activity, inhibit histone deacetylase, and inhibit cytochrome P450 are outlined. Isothiocyanates appear to alter gene expression through modification of critical thiols in regulatory proteins such as Keap1 (Kelch-like ECH-associated protein 1) or IKK (IκB kinase), causing activation of Nrf2 and inactivation of NF-κB, respectively. Certain indoles act as ligands for AhR. Isothiocyanates and indoles are also capable of affecting cell cycle arrest and stimulating apoptosis. The mechanisms responsible for these anti-proliferative responses are discussed.

Dietary glucosinolates are under intensive investigation as precursors of cancer-preventive isothiocyanates. Quantitation of the dose and bioavailability of glucosinolates and isothiocyanates requires a comprehensive analysis of the major dietary glucosinolates, isothiocyanates, and related metabolites. We report a liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) analytical method for the comprehensive analysis of the seven major dietary glucosinolates, related isothiocyanates, and putative amine degradation products. The parent glucosinolates were sinigrin, gluconapin, progoitrin, glucoiberin, glucoraphanin, glucoalyssin, and gluconasturtiin. The LC-MS/ MS analysis method for these compounds was developed and validated; a standard addition analysis protocol was used generally to avoid the requirement for stable isotopic standards. Where stable isotopic standards were available, internal standardization with these gave estimates in agreement with those obtained by the standard addition analysis protocol. For glucosinolates, negative ion electrospray LC-MS/ MS analysis was performed. Isothiocyanates and amines were prederivatized to the corresponding thiourea and N-acetamides, respectively, and were quantiWed by positive ion electrospray LC-MS/MS. The limits of detection were 0.5-2 pmol; the recoveries for glucosinolates, isothiocyanates, and amines were 85-90%, 50-85%, and 60-70%, respectively; and the intra-and interbatch coeYcients of variation were 1-4% and 3-10%, respectively. These methods provide facile access to comprehensive analytical data on the major dietary glucosinolates and related metabolites to quantify inputs and metabolic formation of these compounds in cancer prevention and related studies.  Glucosinolates are -thioglycosides and plant secondary metabolites that are implicated in decreasing the risk of cancer with a vegetable-rich diet. They are found in all members of the Cruciferous plants, including the Brassica crops-Brussels sprouts, broccoli, cauliXower, cabbage, watercress, oilseed rape, and mustard . Glucosinolates are converted to the corresponding aglycone by enzymatic hydrolysis with myrosinase (thioglucoside glucohydrolase, EC 3.2.3.1), which then decomposes to isothiocyanates, thiocyanates, or nitriles, depending on the substrate, pH, and availability of ferrous ions . At physiological pH, isothiocyanates are the major product. Glucosinolates and myrosinase are segre-* Corresponding

Numerous epidemiological studies indicate that Brassica vegetables in general and broccoli in particular protect humans against cancer; they are rich sources of glucosinolates and posses a high content on flavonoids, vitamins, and mineral nutrients. The contents of total intact glucosinolates, total phenolics, vitamin C, and minerals (potassium, sodium, calcium, magnesium, iron, manganese, zinc, and copper) in the edible portions of freshly harvested broccoli (florets), which was subjected to stir-frying treatments, were evaluated. In the present work, the stir-fry cooking experiments were carried out using different edible oils from plant origin (refined olive oil, extra virgin olive oil, sunflower oil, peanut oil, soyabean oil, and safflower oil) known and used worldwide. Results showed that during stir-frying, phenolics and vitamin C were more affected than glucosinolates and minerals. Stir-fry cooking with extra virgin olive, soybean, peanut, or safflower oil did not reduce the total glucosinolate content of the cooked broccoli compared with that of the uncooked sample. The vitamin C content of broccoli stir-fried with extra virgin olive or sunflower oil was similar to that of the uncooked sample, but greater than those samples stir-fried with other oils.

The seeds of Brassicaceae or Cruciferae family contain secondary plant products with various biological effects, including beneficial and harmful ones. These products contain glucosinolates (Gls). They have many types, but the bitter taste type and the anti-nutritive effect are thioglucosides, which contain a sulfate group on the aromatic ring. The seeds of the watercress plant, which were used in the current study in order to try to get rid of the impact of those substances, by cracking by the enzyme myrosinase and is able to crack and decompose Glycoside bond, a water enzyme, and Using six fungal strains: Sclerotinia sclerotiorum, Macrophomina phaseolina, Trichoderma longibrachiatum, Penicillium digitatum, and Fusarium oxysporum in addition to one chemical treatment using hydrothermal copper sulphate (CuSO 4), and comparing the previous seven fungal and chemical biological parameters with a control treatment in which the parasite was treated The seeds of watercress are the same as the pre-treatment in the preparation of the solid fermentation environment but without fertilization of the environment in any fungal chain and the control treatment was incubated in the same conditions as the six fungal treatments. Because the seeds of watercress seeds are rich in antioxidants and anti-cancer, These materials are in all experimental and knowledgeable transactions The following tests were performed: Total polyphenols, Total flavonoids, Reducing power, ABTS + action radical, DPPH radical, and Hydrogen peroxide scavenging activity, in addition to analysis The results of the various chemical treatments from the normal chemical analysis of protein, fiber, ash, fat and carbohydrates as well as the major mineral elements, to determine the effect of each treatment on the seedling of watercress seeds (Eruca sativa meal). The biological treatment included natural thermal treatment by sterilization of the plant material and biological fungal treatment by pollination and incubation and then the secretion of the water enzyme Capable of cracking glucosinolate for other substances. The results of this study showed that there are three biological factors which are T2 (Macrophomina phaseolina), T4 (Trichoderma longibrachiatum), and T6 (Penicillium digitatum) achieved the highest ratios that exceed the control treatment (Non-biologically and chemically treated), in the measurements of total polyphenols and total flavonoids content, Reducing power, (ABTS +) action radical, (DPPH) radical, and Hydrogen peroxide (H 2 O 2) scavenging activity of rocket meal seed (RMS) in different treatments.

The aim of this study was to investigate the effect of cold storage of Chinese cabbage on the
content of glucosinolates (GSLs). High performance liquid chromatography (HPLC) separation
method guided to identify and quantify eleven GSLs including six aliphatic (progoitrin,
sinigrin, gluconapoleiferin, gluconapin, glucocochlearin and glucobrassicanapin), four indolyl
(4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin and neoglucobrasscin)
and one aromatic (gluconasturtiin) respectively. Storage at 4 °C for 72 h, the content of total
GSLs were gradually decreased from 13.1 to 7.41 μmol/g dry weight (DW), whereas, approximately
63 and 55% content of aliphatic and indolyl GSLs were reduced. This finding suggests
that the nutritional contents of Chinese cabbage, especially GLSs could not be preserved for a
long time in the cold storage condition.

The large white butterfly, Pieris brassicae (L.),
is an important pest of Indian mustard, Brassica juncea (L.)
Czern., and inflicts heavy damage to all the above ground
plant parts with strong yield reducing impacts. Farmers
have few practical options other than to spray insecticides
to protect their crop. In this study, an attempt was made
during 2012–2013 and 2013–2014 crop seasons at Ludhiana,
India, to evaluate Ethiopian mustard, Brassica carinata
A. Braun as a trap crop to manage this pest as an
alternate pest management strategy. B. carinata borders
surrounding B. juncea were compared to chemically protected
B. juncea and control (without B. carinata borders)
plots for their effects on P. brassicae infestation and grain
yield. Oviposition preference of adult butterflies was
studied in a two-choice test in field cages, while the effect
of two host plants on larval performance was studied under
laboratory conditions under no choice conditions. B. juncea
plots bordered with B. carinata harbored significantly
lower larval population compared to control B. juncea plots
during both the seasons. The grain yield in the bordered
plots was also significantly higher than that from control
plots and was statistically non-significant from that
obtained in chemically protected plots. Female butterflies
showed distinct oviposition preference for B. carinata over
B. juncea and the larvae reared on B. carinata completed
development in shorter period and grew bigger and heavier
than those reared on B. juncea. Results indicated that B.
carinata has potential to be used as trap crop to manage
P. brassicae.

Brassica oleracea L. crops including kales, cabbages, and Tronchuda cabbages are widely grown in northwestern Spain and Portugal but little information is available on leaf glucosinolate content of these crops. The objectives were to determine the diversity for the total glucosinolate content and profile on leaves in a collection of 153 kales, 26 cabbages, and three Tronchuda cabbages varieties grown at two growing seasons and to determine the seasonal variation of glucosinolates in cabbages and Tronchuda cabbage varieties. Sinigrin, glucoiberin, and glucobrassicin were the major glucosinolates found in kales. Glucoiberin was the most common glucosinolate in Tronchuda cabbages in both planting seasons and in cabbages sown in fall season whereas glucobrassicin and glucoiberin were the most common glucosinolates in cabbages in spring season. In kales the total glucosinolate content ranged from 11.0 to 53 lmol g À1 dw, with a mean value of 26.3 lmol g À1 dw. Four kale varieties (MBG-BRS0468, MBG-BRS0476, MBG-BRS0060 and MBG-BRS0223) showed the highest total sinigrin or glucobrassicin contents. So, they could be good candidates for future breeding programs. In cabbages, the total glucosinolate content ranged from 10.9 to 27 g À1 dw. Total glucosinolate concentration during spring sowing (22 lm g À1 dw) was higher than those in fall sowing (13 lm g À1 dw). Regarding both high glucosinolate content and the agronomic value, MBG-BRS0057 and MBG-BRS0074 could be good sources of beneficial glucosinolates. The presence of high concentrations of sinigrin, glucoiberin, and glucobrassicin warrant further search into their potential use to enhance the level of these important phytochemicals in these edible crops.

Ethnopharmacological relevance: The aerial parts of Sisymbrium officinale Scop. are commonly used to treat airway ailments, moreover in antiquity the herbal drug was reputed to possess anticancer properties. The results obtained in present work support the traditional use and the properties ascribed to Sisymbrium officinale. Aim of the study: In order to give a scientific basis to the traditional uses of Sisymbrium officinale, this study was aimed to evaluate in vitro the myorelaxant activity, the antimicrobial properties and the antimutagenic effect of an aqueous dry extract of the aerial parts of the plant. A phytochemical characterization of the extract was also performed. Materials and methods: The myorelaxant activity was studied against the contractions induced by carbachol, histamine and leukotriene C 4 , in isolated guinea-pig trachea. The antimicrobial activity was tested against six bacteria and one yeast. The Ames test, performed by the preincubation method, was used to study the antimutagenic activity of the extract by its capability to inhibit the mutagenic effect of 2-nitrofluorene, sodium azide, methyl methanesulfonate and 2-aminoanthracene, in Salmonella typhimurium TA98, Salmonella typhimurium TA100 and Escherichia coli WP2uvrA strains. The chemical composition of the extract was analyzed by TLC and HPLC. Results: Sisymbrium officinale showed to reduce the chemically-induced contractions of isolated guineapig trachea with major potency against leukotriene C 4 and histamine. The extract did not show any antibacterial activity. The Ames test showed a strong antimutagenic activity against 2-aminoanthracene, in Escherichia coli WP2uvrA and in Salmonella typhimurium TA98 strains. The phytochemical study highlighted the presence of putranjivine, the glucosinolate marker of Sisymbrium officinale, and of proline. Conclusions: The myorelaxant activity of Sisymbrium officinale offers a scientific basis to its use in traditional medicine. The strong antimutagenic effect suggests further studies to evaluate its possible chemopreventive activity.

Weeds with allelopathic properties can influence vegetational patterns in agroecosystems. Wild mustard (Brassica kaber), has the potential to form and release biologically-toxic isothiocyanates into the soil. The effect of mustard management on the incidence of soil-borne pathogens on corn was investigated. Under field conditions a significant negative correlation was found between plant-parasitic nematode numbers and mustard biomass. The most abundant parasitic nematode genera were: Meloidogyne, Tylenchorhynchus, Helicotylenchus, and Pratylenchus. The incidence of root- pathogenic fungi (Fusarium spp., Rhizoctonia solani, and Pythium aphanidermatum) in corn was low. No differences in soil fertility or moisture were found. Greenhouse experiments confirmed these results and showed no differences in corn-root biomass that could explain the differences in nematode populations. Secondary compounds exuded by mustard roots were hypothesized to explain the phenomenon. Neither Water and acetone extracts of mustard roots or carbofuran alone affected nematodes movement. However, after exposed to root extracts carbofuran had a highly toxic effect on nematodes of the same genera as mentioned above. Allyl, benzyl and phenylethyl isothiocyanates, either as volatiles or in solution, were also toxic to the same nematodes genera. Isothiocyanates volatiles were more toxic to nematodes than they were in sterile soil. Soil microorganisms seem to enhance the toxic effect on nematodes. Volatiles of isothiocyanates, singly and mixed, inhibited hyphal growth of three isolates of Pythium aphanidermatum in a dose-dependent response. Pythium isolates were obtained, one each, from corn and mustard roots growing together in greenhouse, and a third one from an area near Mexico City where the wild mustard is not common. Isothiocyanate mixtures had additive toxic effect on isolates from corn and mustard, and a synergistic toxic effect on the Mexican isolate. When fully inhibited by allyl isothiocyanate, a bacterial colony, identified as Pseudomonas, grew from all Pythium inocula. The nature of the infection of P. aphanidermatum by Pseudomonas was assessed. All results are discussed in the context of controlling soil-borne pathogens through the management of weeds with the ability to release allelochemicals into the soil.