Search

Abstract Leaf mustard (Brassica juncea L.) is explored for its biofumigant properties, derived from its secondary metabolites, particularly allyl isothiocyanate (AITC), produced during the enzymatic breakdown of glucosinolates like sinigrin. The research examines eight leaf mustard cultivars developed in Yeosu city, South Korea, focusing on their genetic characteristics, AITC concentration and nitriles formation rates from glucosinolates. Results indicate that the allelopathic effects, largely dependent on AITC concentration and enzymatic activity, vary across cultivar. Sinigrin and AITC constitute 79% and 36%, respectively, of glucosinolate and its hydrolysis products. The cultivar 'Nuttongii' demonstrates significant potential for inhibiting weeds, exhibiting the highest AITC concentration at 27.47 ± 6.46 µmole g−1 These outcomes highlight the importance of selecting mustard cultivars for biofumigation based on their glucosinolate profiles and hydrolysis product yields. The study also identifies a significant genetic influence on AITC and nitrile formation, suggesting that epithiospecifier protein modulation could enhance both allelopathic and other beneficial effects. Collectively, the research underscores the promise of mustard as a sustainable, environmentally friendly alternative to traditional herbicides.

In order to obtain active ingredient sinigrin with high efficiency and high yield from seeds of Thlaspi arvense, the enzymolysis assisted-ultrasonic extraction method was used. The content of sinigrin was determined by high performance liquid chromatography (HPLC). Based on the single factor experiment, enzymolysis time, enzymolysis temperature and enzyme addition amount were the key factors, and the yield of sinigrin was taken as the response value. The Design-Expert 11 software was applied and the Both-Behnken Design response surface methodology was used to optimize the enzymolysis assisted ultrasonic extraction process of sinigrin. The yield of sinigrin was compared with that of ultrasonic and Soxhlet extraction. The results showed that Agilent HC-C18 column (25 mmx4.6 mmx5 µm), mobile phase A methanol, mobile phase B 0.5% phosphoric acid aqueous solution, detection wavelength 215 nm, column temperature 25 °C, flow rate 1 mL⋅min-1, sample size 10 µL, and the same degree elution for 10 min, the separation effect and peak shape were better. Using the screened alkaline protease as enzyme preparation, the optimal extraction conditions were that the enzymolysis temperature was 49 °C, the enzyme dosage was 2.90%, the enzymolysis time was 18 min, the yield of sinigrin was 2.679 mg⋅g-1 under optimal conditions, which was no significant difference with the theoretical value (2.687 mg⋅g-1). The yield of sinigrin was significantly higher than that of ultrasonic extraction (2.036 mg⋅g-1) and Soxhlet extraction (1.702 mg⋅g-1) (P<0.05). The enzymolysis assisted-ultrasonic extraction process of sinigrin from T. arvense was simple, energy saving, ecofriendly, high efficiency and high yield, and it could be applied to the production and utilization of sinigrin from T. arvense, which laid a theoretical foundation for the in-depth development of T. arvense. [ABSTRACT FROM AUTHOR]

Glucosinolates is a class of secondary metabolites with anti-inflammatory and anti-obesity properties and an intense bitter taste in tumorous stem mustard (TSM), which are used to produce a famous pickle named Zhacai. Spectrophotometric, chromatographic, or infrared techniques showed capacity to evaluate glucosinolate level. The aim of current study was to improve a spectrophotometric methodology to quantify total glucosinolates content (TGC) in TSM. Microwave treatment at medium temperature for 20 s was used to improve the previous method of water bath in enzyme inactivation. Absorbance should be tested at 420 nm for measuring TGC, while not 450 nm. The optimized extraction conditions of glucosinolates were that solid-liquid ratio was 1:22; temperature was 60°C; time was 30 min. The regression curve used for TGC calculation was made using a variety of individual glucosinolate. Recovery rate and variable coefficient of TGC quantitation in TSM were 103% and 5.98%. These results obtained by this method have high consistency within 3 days. In addition, we also introduced the limitations of this method. [ABSTRACT FROM AUTHOR]

Sinigrin is one type of glucosinolates present in cruciferous plants which can be enzymatically hydrolyzed by myrosinase (MYR) to produce allyl isothiocyanate (AITC) with anti-neoplastic potency. This study demonstrates the anticancer activity of silver nanoparticles (AgNPs) green synthesized using sinigrin as a reducing and capping agent. The synthesized sinigrin-coated silver nanoparticles (SIN-AgNPs) characterized by analytical instruments (UV–Vis spectrometry, dynamic light scattering, transmission electron microscopy, and Raman spectroscopy) reveal a mean size of ⁓20 nm with good polydispersity, zeta-potential of -42 mV, and functional groups of sinigrin anchored on the surface of AgNPs. The cDNA of myrosinase was cloned in pcDNA3-eGFP plasmid and transfected to A549 lung adenocarcinoma cells to overexpress myrosinase. The average IC50 values of SIN-AgNPs against parental and myrosinase-expressed A549 cells using the MTT assay were determined to be 3 and 20 µg/ml, respectively. The apoptotic level of myrosinase-expressed A549 cells treated with SIN-AgNPs was further confirmed to be higher than the parental A549 cells. [ABSTRACT FROM AUTHOR]

Cerebral ischaemia‐reperfusion (CIR) injury occurs in stroke patients after the restoration of cerebral perfusion. Sinigrin, a phytochemical found in cruciferous vegetables, exhibits strong antioxidant activity. This study investigated the role of sinigrin in oxidative stress using a CIR injury model. The effects of sinigrin were studied in middle cerebral artery occlusion (MCAO) rats and oxygen–glucose deprivation/reoxygenation (OGD/R)‐injured SH‐SY5Y cells. Sinigrin treatment improved brain injury and neurological deficits induced by MCAO surgery in rats. Sinigrin inhibited apoptosis in brain tissues and SH‐SY5Y cells following OGD/R induction. Additionally, sinigrin elevated the levels of superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GSH‐Px) while reducing malondialdehyde (MDA) levels. Furthermore, sinigrin inhibited the toll‐like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88) signalling pathway. The anti‐apoptotic and antioxidant activities of sinigrin in OGD/R‐injured SH‐SY5Y cells were reversed by TLR4 overexpression. In conclusion, sinigrin inhibits oxidative stress in CIR injury by suppressing the TLR4/MyD88 signalling pathway. [ABSTRACT FROM AUTHOR]

Cruciferous plants manufacture glucosinolates (GSLs) as special and important defense compounds against insects. However, how insect feeding induces glucosinolates in Brassica to mediate insect resistance, and how plants regulate the strength of anti-insect defense response during insect feeding, remains unclear. Here, mustard (Brassica juncea), a widely cultivated Brassica plant, and beet armyworm (Spodoptera exigua), an economically important polyphagous pest of many crops, were used to analyze the changes in GSLs and transcriptome of Brassica during insect feeding, thereby revealing the plant–insect interaction in Brassica plants. The results showed that the content of GSLs began to significantly increase after 48 h of herbivory by S. exigua, with sinigrin as the main component. Transcriptome analysis showed that a total of 8940 DEGs were identified in mustard challenged with beet armyworm larvae. The functional enrichment results revealed that the pathways related to the biosynthesis of glucosinolate and jasmonic acid were significantly enriched by upregulated DEGs, suggesting that mustard might provide a defense against herbivory by inducing JA biosynthesis and then promoting GSL accumulation. Surprisingly, genes regulating JA catabolism and inactivation were also activated, and both JA signaling repressors (JAZs and JAMs) and activators (MYCs and NACs) were upregulated during herbivory. Taken together, our results indicate that the accumulation of GSLs regulated by JA signaling, and the regulation of active and inactive JA compound conversion, as well as the activation of JA signaling repressors and activators, collectively control the anti-insect defense response and avoid over-stunted growth in mustard during insect feeding. [ABSTRACT FROM AUTHOR]

Introduction: Memory dysfunction has remained a challenging issue globally. Nootropics have proven fruitful in managing cognitive dysfunction but because of their side effects, opportunities exist to explore alternatives. White cabbage is a cost-effective natural source of phytochemicals without side effects and has remained uninvestigated as a nootropic agent. This study sought to identify secondary metabolites in white cabbage extract (WCE) and to predict the molecular interaction between the phytochemical constituents of cabbage and phosphodiesterase-1B (PDE1B) using in silico studies. Methods: The WCE was prepared by macerating crushed fresh white cabbage with ethanol for 24 h with continuous stirring. The phytochemical profile of WCE was analyzed using thin layer chromatography (TLC)-densitometry, and molecular docking studies were performed to predict the underlying mechanism action of the phytochemicals with PDE1B. Results: The TLC-densitometry analysis showed that WCE was a rich source of sinigrin, whereas quercetin, chlorogenic acid, and rutin were not detected. In silico studies identified neobrassicin as having the highest affinity (∆Gbind: −19.3358 kcal/mol) for PDE1B. However, quercetin (∆Gbind: −13.1813 kcal/mol) and chlorogenic acid (∆Gbind: −14.8706 kcal/mol) exhibited moderate interaction with PDE1B. Conclusion: These results suggest that WCE has the potency to improve memory function by blocking PDE1B, and this preliminary study implies upcoming in vitro and in vivo research.

The research is devoted to the study on adaptation, verification and implementation of the method for determining sinigrin in mustard seeds and products of its processing, as well as in other types of food raw materials of plant origin. The relevance of the research is determined, on the one hand, by the need to control sinigrin in the technological processes of mustard processing, and, on the other hand, by the lack of effective standardized methods for determining sinigrin in the Russian Federation. The aim of the research is to modify and (or) adapt the most effective methods for the identification and quantification of sinigrin, presented in the publications of foreign researchers. The objectives of the study included construction of calibration graphs; modification of the sample preparation technique to reduce the complexity and duration of the analysis while maintaining the required parameters of accuracy and reproducibility; adaptation of the analysis method using high performance liquid chromatography; verification and approbation of the proposed method using real production samples, i.e. mustard seeds.High performance liquid chromatography has been used as the basic method.As a result of the research, the sample preparation technique has been modified. It has made it possible to reduce labor intensity and reduce the time for its implementation while maintaining the required metrological characteristics. The calibration dependence has been determined and a method for the identification and quantitative analysis of sinigrin in raw materials of plant origin using a high-performance liquid chromatograph equipped with a diode array and mass spectrometric detectors proposed. As a result of verification, the metrological characteristics of the proposed method have been determined: the measurement range is from 0.0005 to 10%, the accuracy index is ± 0.01%. The method has been tested on real objects, mustard seeds, which are the main sources of sinigrin. The proposed method for the identification and quantification of sinigrin can be recommended for use in research and production laboratories equipped with the necessary measuring instruments and testing equipment.

Cruciferous plants manufacture glucosinolates (GSLs) as special and important defense compounds against insects. However, how insect feeding induces glucosinolates in Brassica to mediate insect resistance, and how plants regulate the strength of anti-insect defense response during insect feeding, remains unclear. Here, mustard (Brassica juncea), a widely cultivated Brassica plant, and beet armyworm (Spodoptera exigua), an economically important polyphagous pest of many crops, were used to analyze the changes in GSLs and transcriptome of Brassica during insect feeding, thereby revealing the plant–insect interaction in Brassica plants. The results showed that the content of GSLs began to significantly increase after 48 h of herbivory by S. exigua, with sinigrin as the main component. Transcriptome analysis showed that a total of 8940 DEGs were identified in mustard challenged with beet armyworm larvae. The functional enrichment results revealed that the pathways related to the biosynthesis of glucosinolate and jasmonic acid were significantly enriched by upregulated DEGs, suggesting that mustard might provide a defense against herbivory by inducing JA biosynthesis and then promoting GSL accumulation. Surprisingly, genes regulating JA catabolism and inactivation were also activated, and both JA signaling repressors (JAZs and JAMs) and activators (MYCs and NACs) were upregulated during herbivory. Taken together, our results indicate that the accumulation of GSLs regulated by JA signaling, and the regulation of active and inactive JA compound conversion, as well as the activation of JA signaling repressors and activators, collectively control the anti-insect defense response and avoid over-stunted growth in mustard during insect feeding.

The diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), is an important insect pest of cruciferous crops. Understanding its preference patterns can lead to more efficient management methods, such as trap crops. Several strains of P. xylostella were used to test whether there were differences in oviposition preference in a four-choice setting, on abaxial versus adaxial leaf surfaces in 28 different plant species, and on substrates with different concentrations of sinigrin (allylglucosinolate). Additionally, the larval preference of P. xylostella was studied with 17 plant species of known glucosinolate content that were compared to Arabidopsis thaliana L. in two-choice tests. Our research shows that the diet on which P. xylostella has fed hardly affects multiple-choice host-plant preference, abaxial and adaxial oviposition preference, or oviposition response to pure glucosinolates. Our study also shows that glucosinolate content affects larval preference, which together with the known correlation between glucosinolate content and P. xylostella oviposition, indicates that crops with high glucosinolate content could be more susceptible to damage by P. xylostella than crops with low glucosinolate content. These findings are discussed in regards to their significance in the management of P. xylostella. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is the most important causal agent of head blight in wheat, and stalk and ear rot in maize. A field experiment was conducted to investigate the effect of incorporation of Brassicaceae cover crops on Fusarium graminearum in a wheat-maize rotation. Five species belonging to Brassicaceae (Brassica juncea, Eruca sativa, Raphanus sativus, B. carinata, B. oleracea var. caulorapa L.) were used in the field experiment to investigate their potential to suppress F. graminearum inoculum in soil, disease incidence in maize and to reduce subsequent mycotoxin contamination in maize. Brassica juncea was found to contain the highest glucosinolate concentration in shoots (31 µmol g−1). Severity of ear rot and stalk rot in maize was not significantly reduced in the amended plots. Incorporation of R. sativus 'Terranova' significantly decreased the amount of F. graminearum DNA by 58% compared with the cultivated fallow treatment, however the DNA concentration was not significantly different to fallow uncultivated. Fusarium graminearum DNA and deoxynivalenol in maize was 50% lower after incorporation of B. oleracea var. caulorapa L. compared to after fallow treatment but the difference was not significant. The brassica crops used in the present field experiment were not effective in suppressing F. graminearum, therefore further studies to optimise the current approach are recommended. [ABSTRACT FROM AUTHOR]

Alliaria petiolata (garlic mustard) is a wild Eurasian brassica that produces the secondary compound sinigrin, a type of glucosinolate, that defends against herbivores and pathogens and is toxic to native plants and butterflies outside of garlic mustard's home range. While it is well-established that sinigrin produced by garlic mustard negatively impacts native species in North America, whether and how different growing conditions affect sinigrin concentration is not well understood. Here, we conducted field surveys and greenhouse experiments with garlic mustard to evaluate sinigrin concentration and growth in response to intraspecific density and abiotic environmental conditions. We found greater levels of sinigrin in smaller individual plants regardless of intraspecific density, soil fertility or light conditions. Our findings suggest that leaf concentrations of sinigrin are not controlled directly by abiotic factors but rather by plant size. [ABSTRACT FROM AUTHOR]

Multi-resistant Staphylococcus aureus (S. aureus) infection is a significant global health concern owing to its high mortality and morbidity rates. Coagulase (Coa), a key enzyme that activates prothrombin to initiate host coagulation, has emerged as a promising target for anti-infective therapeutic approaches. This study identified sinigrin as a potent Coa inhibitor that significantly inhibited S. aureus -induced coagulation at concentration as low as 32 mg/L. Additionally, at a higher concentration of 128 mg/L, sinigrin disrupted the self-protection mechanism of S. aureus. Thermal shift and fluorescence-quenching assays confirmed the direct binding of sinigrin to the Coa protein. Molecular docking analysis predicted specific binding sites for sinigrin in the Coa molecule, and point mutation experiments highlighted the importance of Arg-187 and Asp-222 as critical binding sites for both Coa and sinigrin. In vivo studies demonstrated that the combination of sinigrin with oxacillin exhibited greater antibacterial efficacy than oxacillin alone in the treatment of S. aureus -induced pneumonia in mice. Furthermore, sinigrin was shown to reduce bacterial counts and inflammatory cytokine levels in the lung tissues of S. aureus -infected mice. In summary, sinigrin was shown to directly target Coa, resulting in the attenuation of S. aureus virulence, which suggests the potential of sinigrin as an adjuvant for future antimicrobial therapies. [ABSTRACT FROM AUTHOR]

Encapsulation of sinigrin in liposomes with the proliposomal method was performed in order to evaluate the effect of this process on in vitro simulated digestion and antioxidant potential of sinigrin. The recovery of sinigrin after simulated gastric and duodenal digestion of its free and liposomal forms was determined with HPLC-UV using human digestive juices. The antioxidant potential of sinigrin and sinigrin-loaded liposomes was determined with the Rancimat test as their ability to prolong oxidative stability of edible oil. The efficiency of 62% was obtained by encapsulating sinigrin in liposomes. The values of mean diameter, polydispersity index and zeta potential showed satisfactory size uniformity and physical stability of the liposomes containing sinigrin. Liposomes were shown to inhibit the digestion of sinigrin in both human gastric and intestinal juices, clearly enabling its prolonged release. Moreover, sinigrin in the liposomal form significantly prolonged the induction time of edible oil oxidation compared to its free form. The results obtained are encouraging from the point of view of a possible incorporation of the sinigrin-loaded liposomes in real functional food systems or their use as nutraceuticals.

Endophytic fungi of crops can promote plant growth through various mechanisms of action (i.e., improve nutrient uptake and nutrient use efficiency, and produce and modulate plant hormones). The genus Brassica includes important horticultural crops, which have been little studied in their interaction with endophytic fungi. Previously, four endophytic fungi were isolated from kale roots (Brassica oleracea var. acephala), with different benefits for their host, including plant growth promotion, cold tolerance, and induction of resistance to pathogens (Xanthomonas campestris) and pests (Mamestra brassicae). In the present work, the molecular and morphological identification of the four different isolates were carried out, describing them as the species Acrocalymma vagum, Setophoma terrestris, Fusarium oxysporum, and the new species Pyrenophora gallaeciana. In addition, using a representative crop of each Brassica U’s triangle species and various in vitro biochemical tests, the ability of these fungi to promote plant growth was described. In this sense, the four fungi used promoted the growth of B. rapa, B. napus, B. nigra, B. juncea, and B. carinata, possibly due to the production of auxins, siderophores, P solubilization or cellulase, xylanase or amylase activity. Finally, the differences in root colonization between the four endophytic fungi and two pathogens (Leptosphaeria maculans and Sclerotinia sclerotiorum) and the root glucosinolate profile were studied, at different times. In this way, how the presence of progoitrin in the roots reduces their colonization by endophytic and pathogenic fungi was determined, while the possible hydrolysis of sinigrin to fungicidal products controls the colonization of endophytic fungi, but not of pathogens. [ABSTRACT FROM AUTHOR]

Myrosinase can hydrolyze glucosinolates to generate isothiocyanates, which have cancer prevention and anti-cancer properties. The main sources of myrosinase are cruciferous plants. To further improve the efficiency of isothiocyanates preparation, it is necessary to explore novel sources of myrosinases. In this study, we described a bacterium, Shewanella baltica Myr-37, isolated from marine mud, capable of producing a novel myrosinase (Smyr37) with a molecular weight of 100 kDa. The crude enzyme of Smyr37 showed the highest activity at 50 °C and pH 8.0. The sinigrin- and glucoraphanin-hydrolyzing activities of Smyr37 were 6.95 and 5.87 U/mg, respectively. Moreover, when the reaction temperature was 40 °C and pH was 7.0, the crude enzyme of Smyr37 could efficiently degrade glucoraphanin into sulforaphane within 25 min with a yield of 0.57 mg/mL. The corresponding conversion efficiency of sulforaphane from glucoraphanin was 89%. In summary, S. baltica Myr-37 myrosinase Smyr37, a novel myrosinase, can be used in the preparation of isothiocyanates. [ABSTRACT FROM AUTHOR]

Diabetes mellitus (DM) is a common metabolic disorder which arises due to the improper carbohydrate metabolism, decreased secretion/activity of insulin, and genetic abnormalities, which result in the increased blood glucose level generally known as hyperglycemia. Diabetes holds an increased global prevalence in each year and is responsible for increased morbidity and mortality rates. Hence, the current investigation focusses to assess the antidiabetic potential of sinigrin on diabetic animal model through the suppression of inflammation. Diabetes was initiated to the animals via administering streptozotocin (STZ) and supplemented with the sinigrin at 25- and 50-mg/kg dose via oral route. The diabetic rats demonstrated the elevated glucose, food and water intake, kidney and liver weights, and reduced bodyweight and depleted insulin status. The sinigrin treatment remarkably improved and modulated these changes in diabetic animals. Additionally, the sinigrin supplementation also modulated the changes in glucose-6-phosphatase; fructose 1,6-bisphosphatase; AST; ALT; creatinine; and inflammatory mediators in the STZ-provoked diabetic animals. The levels of hexokinase, protein, and antioxidants also improved by the sinigrin treatment. The histological investigations of pancreas also witnessed the therapeutic actions of sinigrin, which is supported by the findings of biochemical examinations. Therefore, it was clear that the sinigrin supplementation displayed remarkable antidiabetic effect on STZ-initiated diabetic animals via modulating inflammation and other biochemical changes, which recommends that sinigrin could be a talented candidate for diabetes management in the future. [ABSTRACT FROM AUTHOR]

Allyl isothiocyanate (AITC) is a phytochemical that is abundantly present in cruciferous vegetables of the Brassicaceae family, such as cabbage, broccoli, mustard, wasabi, and cauliflower. The pungent taste of these vegetables is mainly due to the content of AITC present in these vegetables. AITC is stored stably in the plant as its precursor sinigrin (a type of glucosinolate), which is physically separated from myrosin cells containing myrosinase. Upon tissue disruption, myrosinase gets released and hydrolyzes the sinigrin to produce AITC and by-products. AITC is an organosulfur compound, both an irritant and toxic, but it carries pharmacological properties, including anticancer, antibacterial, antifungal, and anti-inflammatory activities. Despite the promising anticancer effectiveness of AITC, its clinical application still possesses challenges due to several factors, i.e., low aqueous solubility, instability, and low bioavailability. In this review, the anticancer activity of AITC against several cancer models is summarized from the literature. Although the mechanism of action is still not fully understood, several pathways have been identified; these are discussed in this review. Not much attention has been given to the delivery of AITC, which hinders its clinical application. However, the few studies that have demonstrated the use of nanotechnology to facilitate the delivery of AITC are addressed. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is a globally important cereal pathogen, causing head blight in wheat, resulting in yield losses and mycotoxin contamination. Currently, triazole fungicides are used to suppress Fusarium graminearum, however, the declining effectiveness of triazoles and concerns over the safety of pesticides have led to the pursuit of safe alternative crop protection strategies such as biofumigation. In the present study, species belonging to Brassicaceae (Brassica juncea, Raphanus sativus, Eruca sativa) were assessed for their biofumigation potential against F. graminearum and the glucosinolate profile of the brassicas was determined. In Petri dishes, mycelial plugs of Fusarium graminearum were exposed to frozen/defrosted leaf discs of brassicas collected at early-leaf, stem-extension, and early-bud stages. Additionally, F. graminearum inoculum was incubated in soil amended with chopped tissues of brassicas in a closed jar experiment. Glucosinolate analysis of the leaf tissue of brassicas revealed that the total glucosinolate concentration of B. juncea 'Brons' increased with advancing growth stage (24.5–51.9 µmol g−1). Brassica juncea leaf discs were effective against mycelial growth, while the sinigrin content in the leaf tissue corresponded to the level of suppression. At the stem-extension and early-bud stages, B. juncea 'Brons' showed 87–90% suppression with four leaf discs, and 100% suppression with eight leaf discs. Brassica juncea 'Caliente Rojo' leaf discs collected at the stem-extension stage showed 94% inhibition with eight discs. In the closed jar experiment, each brassica species significantly suppressed F. graminearum inoculum by 41–55%. The findings suggest that the brassica species investigated in the present study could be effective in reducing the inoculum of F. graminearum in soil prior to cereal production. [ABSTRACT FROM AUTHOR]

Mainly found in brussels sprouts, broccoli, and black mustard seeds, sinigrin (2-propenyl glucosinolate) has enjoyed some attention currently for its effects on health and disease prevention. The present research design is aimed at investigating the effects of sinigrin on inflammation, oxidative stress (OS) and memory. Randomly, six groups of male Wistar rats were categorized into the control and experimental groups. The experimental groups were treated with sinigrin (10 and 20 mg/kg, orally). The control positive group was given the pentylenetetrazole (PTZ) treatment and the control negative one was given normal saline. All groups were kindled by the sub-threshold dose (35 mg/kg, i.p.) of PTZ for 12 times in one month. When the kindling procedure was done, the seizure behaviors and the behavioral function were evaluated. For cognitive parameters, the shuttle box test was employed. When the experiment was terminated, the rats were euthanized and their blood serum as well as brain samples were isolated for respective measuring of OS and gene expression parameters. The treatment with sinigrin significantly delayed the appearance of the seizure symptoms in comparison to that of the PTZ group. It also significantly increased the memory parameters like retention latency and the total time having been spent in the light compartment in the epileptic rats. In addition, sinigrin increased the superoxide dismutase and catalase levels. Treatment with sinigrin suppressed the Il1b and Nlrp3 gene expression at hippocampal level. In sum, sinigrin prevents inflammation, OS and memory impairment against the PTZ-kindling epilepsy in rats. [ABSTRACT FROM AUTHOR]

Endophytic fungi of crops can promote plant growth through various mechanisms of action (i.e., improve nutrient uptake and nutrient use efficiency, and produce and modulate plant hormones). The genus Brassica includes important horticultural crops, which have been little studied in their interaction with endophytic fungi. Previously, four endophytic fungi were isolated from kale roots (Brassica oleracea var. acephala), with different benefits for their host, including plant growth promotion, cold tolerance, and induction of resistance to pathogens (Xanthomonas campestris) and pests (Mamestra brassicae). In the present work, the molecular and morphological identification of the four different isolates were carried out, describing them as the species Acrocalymma vagum, Setophoma terrestris, Fusarium oxysporum, and the new species Pyrenophora gallaeciana. In addition, using a representative crop of each Brassica U’s triangle species and various in vitro biochemical tests, the ability of these fungi to promote plant growth was described. In this sense, the four fungi used promoted the growth of B. rapa, B. napus, B. nigra, B. juncea, and B. carinata, possibly due to the production of auxins, siderophores, P solubilization or cellulase, xylanase or amylase activity. Finally, the differences in root colonization between the four endophytic fungi and two pathogens (Leptosphaeria maculans and Sclerotinia sclerotiorum) and the root glucosinolate profile were studied, at different times. In this way, how the presence of progoitrin in the roots reduces their colonization by endophytic and pathogenic fungi was determined, while the possible hydrolysis of sinigrin to fungicidal products controls the colonization of endophytic fungi, but not of pathogens.

Field pennycress, Thlaspi arvense L., is a new oil seed cover crop being commercialized as a source of biofuel in the United States. Along the path to commercialization, pennycress seed and meal will be marketed as animal feed. As a member of the Brassicacaeae family, pennycress seed is abundant in glucosinolates which are anti-nutritional factors that make the seed and meal less palatable to animals. More specifically, sinigrin has been identified as the only glucosinolate present in pennycress seed. As work continues to develop varieties low in glucosinolates, a rapid assay is needed to determine the sinigrin level in pennycress seed. The objective of this study was to develop and utilize a high throughput, rapid assay to screen pennycress seeds from wild populations for total sinigrin content. Pennycress seeds were ground in potassium phosphate buffer with endogenous myrosinase to catalyze the enzymatic hydrolysis of sinigrin to equimolar amounts of glucose. Glucose content was determined using a blood glucose meter and test strips. From the molar concentration of glucose, the molar concentration of sinigrin was known and the sinigrin level in seeds was calculated in µmol/g. The results were confirmed by HPLC/MS analysis. A rapid and inexpensive method was developed and used to screen seeds of 429 wild pennycress lines for sinigrin. Sinigrin levels in seed ranged from 25 to 85 μmol/g in the pennycress lines screened. Environmental factors also seemed to contribute to the levels of sinigrin in seeds found within the line 'tt8-t/ARV1' grown at ten different locations in the Midwest, USA. This rapid and inexpensive assay will allow breeders to efficiently screen lines and select for varieties that produce seeds with low sinigrin levels for improved feed value. • Introduces new rapid inexpensive assay for detecting glucosinolates in pennycress seed. • Provides method development and validation for determining glucose levels in seed. • Screened 429 wild populations of pennycress for glucosinolate levels. • Evaluated glucosinolate levels across growing locations and years. [ABSTRACT FROM AUTHOR]

As the major and abundant type of glucosinolates (GL) in plants, sinigrin has potential functions in promoting health and insect defense. The final step in the biosynthesis of sinigrin core structure is highly representative in GL compounds, which corresponds to the process from 3-methylthiopropyl ds-GL to 3-methylthiopropyl GL catalyzed by sulfotransferase (SOT). However, due to the lack of the crystallographic structure of SOT complexed with the 3-methylthiopropyl GL, little is known about this sulfonation process. Fortunately, the crystal structure of SOT 18 from Arabidopsis thaliana (AtSOT18) containing the substance (sinigrin) similar to 3-methylthiopropyl GL has been determined. To understand the enzymatic mechanism, we employed molecular dynamics (MD) simulation and quantum mechanics combined with molecular mechanics (QM/MM) methods to study the conversion from ds-sinigrin to sinigrin catalyzed by AtSOT18. The calculated results demonstrate that the reaction occurs through a concerted dissociative mechanism. Moreover, Lys93, Thr96, Thr97, Tyr130, His155, and two enzyme peptide chains (Pro92-Lys93 and Gln95-Thr96-Thr97) play a role in positioning the substrates and promoting the catalytic reaction by stabilizing the transition state geometry. Particularly, His155 acts as a catalytic base while Lys93 acts as a catalytic acid in the reaction process. The presently proposed concerted dissociative mechanism explains the role of AtSOT18 in sinigrin biosynthesis, and could be instructive for the study of GL biosynthesis catalyzed by other SOTs. [ABSTRACT FROM AUTHOR]

Allopolyploid Brassica juncea is particularly enriched in sinigrin, a kind of 3C aliphatic glucosinolates (GSLs), giving rise to characteristic taste after picking. However, the molecular mechanism underlying 3C aliphatic GSLs biosynthesis in this species remains unknown. In this study, we genome-widely identified GSLs metabolic genes, indicating different evolutionary rate of GSLs metabolic genes between subgenomes of B. juncea. Eight methythioalkylmalate synthase (MAMs) homologs were identified from B. juncea, in which six MAM1s were located in chloroplast and the other two were not detected with any expression. Furthermore, BjMAM1-4, BjMAM1-5, and BjMAM1-6 displayed higher expression levels in leaves than other tissues. Silenced expression analysis revealed that BjMAM1-4 and BjMAM1-6 function in 3C and 4C aliphatic GSLs accumulation. The specificity of the substrate selection for the second cycle reaction is much lower than that of the first cycle, suggesting these genes may preferentially catalyze 3C aliphatic GSLs biosynthesis. Our study provides insights into the molecular mechanism underlying the accumulation of 3C aliphatic GSLs, thereby facilitating the manipulation of aliphatic GSLs content in B. juncea.

Fusarium graminearum is the most important causal agent of head blight in wheat, and stalk and ear rot in maize. A field experiment was conducted to investigate the effect of incorporation of Brassicaceae cover crops on Fusarium graminearum in a wheat-maize rotation. Five species belonging to Brassicaceae (Brassica juncea, Eruca sativa, Raphanus sativus, B. carinata, B. oleracea var. caulorapa L.) were used in the field experiment to investigate their potential to suppress F. graminearum inoculum in soil, disease incidence in maize and to reduce subsequent mycotoxin contamination in maize. Brassica juncea was found to contain the highest glucosinolate concentration in shoots (31 µmol g−1). Severity of ear rot and stalk rot in maize was not significantly reduced in the amended plots. Incorporation of R. sativus ‘Terranova’ significantly decreased the amount of F. graminearum DNA by 58% compared with the cultivated fallow treatment, however the DNA concentration was not significantly different to fallow uncultivated. Fusarium graminearum DNA and deoxynivalenol in maize was 50% lower after incorporation of B. oleracea var. caulorapa L. compared to after fallow treatment but the difference was not significant. The brassica crops used in the present field experiment were not effective in suppressing F. graminearum, therefore further studies to optimise the current approach are recommended.

Encapsulation of sinigrin in liposomes with the proliposomal method was performed in order to evaluate the effect of this process on in vitro simulated digestion and antioxidant potential of sinigrin. The recovery of sinigrin after simulated gastric and duodenal digestion of its free and liposomal forms was determined with HPLC-UV using human digestive juices. The antioxidant potential of sinigrin and sinigrin-loaded liposomes was determined with the Rancimat test as their ability to prolong oxidative stability of edible oil. The efficiency of 62% was obtained by encapsulating sinigrin in liposomes. The values of mean diameter, polydispersity index and zeta potential showed satisfactory size uniformity and physical stability of the liposomes containing sinigrin. Liposomes were shown to inhibit the digestion of sinigrin in both human gastric and intestinal juices, clearly enabling its prolonged release. Moreover, sinigrin in the liposomal form significantly prolonged the induction time of edible oil oxidation compared to its free form. The results obtained are encouraging from the point of view of a possible incorporation of the sinigrin-loaded liposomes in real functional food systems or their use as nutraceuticals. [ABSTRACT FROM AUTHOR]

The selection of appropriate food resources by bees is a critical aspect for the maintenance of their populations, especially in the current context of global change and pollinator decline. Wild bees have a sophisticated ability to forage selectively on specific resources, and can assess the quality of pollen using contact chemosensory perception (taste). While numerous studies have investigated the detection of pollen macronutrients in bees and their impact on bee health and reproductive success, only a few studies have described the gustatory responses of bees toward specialized metabolites. In addition, these studies mostly focused on the response to nectar and neglected pollen, which is the main food resource for both bee imagines and larvae. Whether bees have the ability to detect specialized toxic metabolites in pollen and then rapidly adapt their foraging behavior to avoid them is very little studied. In this study, we tested whether pollen specialized metabolites affect bumblebees at both the micro-colony and individual levels (i.e., bioassays using supplemented pollen), and whether foragers detect these specialized metabolites and potentially display an avoidance behavior (i.e., preference tests using supplemented syrup). Bumblebees were fed with either amygdalin-, scopolamine- or sinigrin-supplemented pollen diets in ratios that mimic 50%, 100%, and 200% of naturally occurring concentrations. We found no effect of these specialized metabolites on resource collection, reproductive success and stress response at the micro-colony level. At the individual level, bumblebees fed on 50%-amygdalin or 50%-scopolamine diets displayed the highest scores for damage to their digestive systems. Interestingly, during the preference tests, the solution with 50%-scopolamine displayed a phagostimulatory activity, whereas solution with 50%-amygdalin had a deterrent effect and could trigger an active avoidance behavior in bumblebees, with a faster proboscis retraction. Our results suggest that regulation of toxin intake is not as well-established and effective as the regulation of nutrient intake in bees. Bees are therefore not equally adapted to all specialized pollen metabolites that they can come into contact with.

ABSTRACT The cowpea is subject to both quantitative and qualitative loss during storage due to attack by the insect pest Callosobruchus maculatus (Fabricius) (Coleoptera: Chrysomelidae). There is a growing interest in controlling the pest using insecticides that result from the reactions of secondary metabolites produced by plants, due to their high efficiency, wide availability and selectivity for the target organisms. The present study aimed to evaluate the fumigant activity of allyl isothiocyanate (AITC) on the development of C. maculatus and its effect on the physical properties of the cowpea. To evaluate the instantaneous population growth rate (ri), the insects were exposed to concentrations of 3.81, 5.71, 7.67, 9.41 and 17.85 mL AITC m-3 cowpea (corresponding to LC1, LC10, LC30, LC50 and LC95) for 40 days, after which the physical properties of the cowpea were analyzed. The degree of infestation increased considerably with the reduction in AITC concentration, as did the water content and electrical conductivity. The germination percentage and apparent density decreased with the reduction in AITC concentration. The LC10 sublethal concentration reduced reproduction in C. maculatus and the degree of infestation of the cowpea after 40 days of storage. AITC inhibits the development of C. maculatus and helps maintain the physical properties of the cowpea, thereby representing an alternative to the use of traditional insecticides and showing potential for the control of insect pests in stored grain.

Fusarium graminearum is a globally important cereal pathogen, causing head blight in wheat, resulting in yield losses and mycotoxin contamination. Currently, triazole fungicides are used to suppress Fusarium graminearum, however, the declining effectiveness of triazoles and concerns over the safety of pesticides have led to the pursuit of safe alternative crop protection strategies such as biofumigation. In the present study, species belonging to Brassicaceae (Brassica juncea, Raphanus sativus, Eruca sativa) were assessed for their biofumigation potential against F. graminearum and the glucosinolate profile of the brassicas was determined. In Petri dishes, mycelial plugs of Fusarium graminearum were exposed to frozen/defrosted leaf discs of brassicas collected at early-leaf, stem-extension, and early-bud stages. Additionally, F. graminearum inoculum was incubated in soil amended with chopped tissues of brassicas in a closed jar experiment. Glucosinolate analysis of the leaf tissue of brassicas revealed that the total glucosinolate concentration of B. juncea ‘Brons’ increased with advancing growth stage (24.5–51.9 µmol g−1). Brassica juncea leaf discs were effective against mycelial growth, while the sinigrin content in the leaf tissue corresponded to the level of suppression. At the stem-extension and early-bud stages, B. juncea ‘Brons’ showed 87–90% suppression with four leaf discs, and 100% suppression with eight leaf discs. Brassica juncea ‘Caliente Rojo’ leaf discs collected at the stem-extension stage showed 94% inhibition with eight discs. In the closed jar experiment, each brassica species significantly suppressed F. graminearum inoculum by 41–55%. The findings suggest that the brassica species investigated in the present study could be effective in reducing the inoculum of F. graminearum in soil prior to cereal production.

Allyl isothiocyanate (AITC) is a phytochemical that is abundantly present in cruciferous vegetables of the Brassicaceae family, such as cabbage, broccoli, mustard, wasabi, and cauliflower. The pungent taste of these vegetables is mainly due to the content of AITC present in these vegetables. AITC is stored stably in the plant as its precursor sinigrin (a type of glucosinolate), which is physically separated from myrosin cells containing myrosinase. Upon tissue disruption, myrosinase gets released and hydrolyzes the sinigrin to produce AITC and by-products. AITC is an organosulfur compound, both an irritant and toxic, but it carries pharmacological properties, including anticancer, antibacterial, antifungal, and anti-inflammatory activities. Despite the promising anticancer effectiveness of AITC, its clinical application still possesses challenges due to several factors, i.e., low aqueous solubility, instability, and low bioavailability. In this review, the anticancer activity of AITC against several cancer models is summarized from the literature. Although the mechanism of action is still not fully understood, several pathways have been identified; these are discussed in this review. Not much attention has been given to the delivery of AITC, which hinders its clinical application. However, the few studies that have demonstrated the use of nanotechnology to facilitate the delivery of AITC are addressed.

Myrosinase can hydrolyze glucosinolates to generate isothiocyanates, which have cancer prevention and anti-cancer properties. The main sources of myrosinase are cruciferous plants. To further improve the efficiency of isothiocyanates preparation, it is necessary to explore novel sources of myrosinases. In this study, we described a bacterium, Shewanella baltica Myr-37, isolated from marine mud, capable of producing a novel myrosinase (Smyr37) with a molecular weight of 100 kDa. The crude enzyme of Smyr37 showed the highest activity at 50 °C and pH 8.0. The sinigrin- and glucoraphanin-hydrolyzing activities of Smyr37 were 6.95 and 5.87 U/mg, respectively. Moreover, when the reaction temperature was 40 °C and pH was 7.0, the crude enzyme of Smyr37 could efficiently degrade glucoraphanin into sulforaphane within 25 min with a yield of 0.57 mg/mL. The corresponding conversion efficiency of sulforaphane from glucoraphanin was 89%. In summary, S. baltica Myr-37 myrosinase Smyr37, a novel myrosinase, can be used in the preparation of isothiocyanates.

Glucosinolates (GSLs) are important secondary metabolites in Brassicales related to insect and disease resistance, flavor formation, and human health. Here, we determined the GSL profile with sinigrin as the predominant GSL in Brassica nigra. A total of 184 GSL biosynthetic genes (BniGSLs) were identified in B. nigra by a genome-wide search for orthologs of 82 of the 95 known GSL genes in Arabidopsis thaliana. Transcriptome data demonstrated that at least one BniGSL was highly expressed in stems and leaves at each step of the sinigrin synthesis pathway, which ensured the synthesis of a large amount of sinigrin in B. nigra. Among these key candidates of BniGSLs, the high expression of BniMAM1-2, BniCYP79F1, and BniAOP2-1/2, and the absence of MAM3 and AOP3, may contribute remarkably to the synthesis and accumulation of sinigrin. In addition, the low expression of some key BniGSLs partially explains the low content of indolic and aromatic GSLs in B. nigra. This study provided a genetic explanation for the formation of the unique GSL profile with sinigrin as the main GSL in B. nigra. The results of this study will be valuable for further functional analysis of BniGSLs and genetic improvement of GSLs in B. nigra and other Brassica species. [ABSTRACT FROM AUTHOR]

Application of nitrogen (N) is a common practice used to achieve profitable yields in horticultural crops and N application can be used as a tool to manipulate the enhancement of phytochemicals in vegetable crops to address consumer-oriented quality production. Our previous findings recommended 90 kg ha−1 for certain types of cauliflower varieties without compromising yields. Thus, this study was aimed to investigate the effect of N application on glucosinolates and phenolic acids, at harvest, in varieties ‘Largardo’, ‘Eskimo’ and ‘CF-744’ grown in the field. N was applied as ammonium nitrate (NH4NO3) at concentrations from 0, 60, 90, 120, 150 to 180 kg ha−1. Variety ‘CF-744’ was more sensitive to N supply and at 180 kg ha−1 N it showed the highest accumulation of glucosinolates (sinigrin, glucoiberin, progoitrin, 4-methoxyglucobrassicin) at harvest. However, 90 kg ha−1 N supply demonstrated the highest accumulation of majority of the glucosinolates in varieties ‘Largardo’ and ‘Eskimo’. Different varieties responded differently to N supply and glucosinolate levels in cauliflowers. Also, different varieties responded differently to N supply and antioxidant property. In all three varieties, the N supply at 120 kg ha−1 showed the highest accumulation of protocatechoic acid, 4-hydroxy benzoic acid, ρ-coumaric acid and caffeic acid.

In recent years, Brassica juncea has been selected as a special agricultural crop in Jeongseon, Gangwon-do, Korea, and is actively grown there. However, there have been no studies on B. juncea cultivated in Jeongseon (BJJ). Sinigrin, an index component of cruciferous vegetables, has been reported to have antiobesity effects. In this study, we developed a method for obtaining a BJJ extract with optimized sinigrin content, and investigate the antiobesity properties of the BJJ extract and sinigrin. The optimal extraction conditions for BJJ were found to be with 60% ethanol, at 70°C, for 3 hr. Lipid accumulation and ROS production were significantly suppressed in both the BJJ extract and sinigrin-treated groups. Furthermore, BJJ extract and sinigrin were effectively controlled the expression of proteins that regulate lipid accumulation, fatty acid oxidation, and energy metabolism. Thus, BJJ extract containing sinigrin may be used as a health functional food material. Practical applications: Brassica juncea has been reported to be rich in flavonoids, polyphenols, and glucosinolate, which are secondary vegetable metabolites. In this study, an extraction method to optimize the content of sinigrin in BJJ was established, and the antiobesity mechanism for the extract was confirmed. Lipid accumulation and ROS production were significantly suppressed in both the BJJ extract and sinigrin-treated groups in the study. It was confirmed that the expression of proteins that regulate lipid accumulation, lipid synthesis, fatty acid oxidation, heat generation, and energy metabolism was effectively controlled by the BJJ extract and sinigrin. Therefore, the ethanol extraction method of this study are considered to be useful for the preparation of extracts using cruciferous vegetables, and BJJ extract containing sinigrin have the potential to be used as a health functional food material for obesity. [ABSTRACT FROM AUTHOR]

Introduction: Glucosinolates (1–5) are important secondary metabolites found in Isatis indigotica roots. Due to their high hydrophilic and ionic nature, purified glucosinolates often contain salt impurities and moisture. Accurate assessment of their purities is important for glucosinolates being utilised as chemical markers. Objective: To develop and validate quantitative proton (1H) nuclear magnetic resonance (qHNMR) methods for purity assessments of aliphatic and indole glucosinolates (1–5). Method: Several NMR parameters such as pulse program, relaxation time, and delay time were optimised. Three qHNMR methods were developed using gluconapin (3), neoglucobrassicin (4), and sinigrin (5) for method validation and with maleic acid as internal standard. Results: The quantification was based on the integrated area ratios of an olefinic proton (H‐4 for 1–3; H‐6 for 4; and H‐3 for 5) of the side chain from glucosinolates relative to the olefinic proton from the internal standard using deuterated water (D2O) as the solvent. The qHNMR methods were successfully applied for purity assessments of four aliphatic glucosinolates (1–3 and 5: progoitrin, epiprogoitrin, gluconapin, and sinigrin), and an indole glucosinolate (4: neoglucobrassicin). Conclusion: The purity of glucosinolates isolated from I. indigotica and commercial sinigrin was accurately assessed using the developed qHNMR method. The qHNMR provides a reliable and superior means to determine the purity of glucosinolates. Three qHNMR methods were developed and validated for purity assessment of aliphatic and indole glucosinolates using maleic acid as internal standard. The quantification was based on integrated area ratios of an olefinic proton of the side chain from glucosinolates relative to the olefinic proton from internal standard. The qHNMR method is a simple, rapid, irreplaceable, and accurate approach for purity assessments of glucosinolates. [ABSTRACT FROM AUTHOR]

Wasabi, horseradish and mustard are popular pungent crops in which the characteristic bioactive hydrolysis of specialized glucosinolates (GSLs) occurs. Although the metabolic pathways of GSLs are well elucidated, how plants have evolved convergent mechanisms to accumulate identical GSL components remains largely unknown. In this study, we discovered that sinigrin is predominantly synthesized in wasabi, horseradish and mustard in Brassicaceae. We de novo assembled the transcriptomes of the three species, revealing the expression patterns of gene clusters associated with chain elongation, side chain modification and transport. Our analysis further revealed that several gene clusters were convergently selected during evolution, exhibiting convergent shifts in amino acid preferences in mustard, wasabi and horseradish. Collectively, our findings provide insights into how unrelated crop species evolve the capacity for sinigrin super-accumulation and thus promise a potent strategy for engineering metabolic pathways at multiple checkpoints to fortify bioactive compounds for condiment or pharmaceutical purposes. Jinghua Yang et al. provide the de novo assemblies of the transcriptomes of wasabi, horseradish and mustard to examine the evolution of specialized glucosinolates. This study demonstrates convergent evolution of super-accumulation of sinigrin in these taxa and may allow for potential strategies to engineer metabolic pathways of bioactive compounds for human food sources or pharmaceutical purposes. [ABSTRACT FROM AUTHOR]

Millions of patients suffer from asthma worldwide. However, the first-line drugs used to treat asthma, namely, the beta-adrenergic receptors agonists (β-agonists), are not recommended for use as monotherapy because of their severe dose-related side effects. This limitation has prompted the search for new therapies, which can be used in conjunction with β--agonists so that lower doses can be administered. Sinigrin is a major compound found in many antiasthmatic medicinal plants. In this study, we explored the antiasthmatic activity of sinigrin when used in combination with β-agonists and its underlying mechanism. Sinigrin enhanced the asthma-relieving effects of isoproterenol and reduced the effective isoproterenol dose in an acute-asthma model in guinea pigs. Mechanistically, sinigrin enhanced the cAMP levels induced by β-agonists by inhibiting PDE4. The resulting increase in cAMP levels stimulated the activity of the downstream effector protein kinase A, which would be expected to ultimately induce the relaxation of airway smooth muscle. In conclusion, sinigrin enhances the asthma-relieving effects of β-agonists by regulating the cAMP signaling pathway and represents a potential add-on drug to β-agonists for the treatment of asthma.

Brassica vegetables may modulate cancer risk by regulation of xenobiotic metabolizing enzymes (XMEs). In a randomized crossover study, the effect of kale consumption on CYP1A2, CYP2A6, XO, and NAT2 activity was determined by urinary caffeine metabolite ratios, UGT1A1 activity by serum bilirubin concentrations, and GSTA protein and GST activity in blood by ELISA. Adults (n = 25) consumed a basal diet supplemented with kale and radish for 14 days or control vegetables. The kale diet increased CYP1A2 activity by 16.4% on day 8 and 15.2% on day 15 compared to control. Conjugated bilirubin was reduced by the kale diet, decreasing from 19.4 to 14.3 to 9.5% of total bilirubin on days 1, 8, and 15, respectively, which may be explained by induction of MRP2. Other XMEs were not affected by diet. The implications of these results for cancer risk will be clarified as the functions of these XMEs become better understood.

Sinigrin is present in significant amounts in cruciferous vegetables. Epidemiological studies suggest that the consumption of such vegetables decreases the risk of cancer, and the effect is attributed mainly to allyl isothiocyanate (AITC), a hydrolysis product of sinigrin catalyzed by myrosinase. Anticancer activity of AITC has been previously investigated for several cancer models, but less attention was paid to delivering AITC on the target site. In this study, the gene sequences of core streptavidin (coreSA) and myrosinase (MYR) were cloned in a pET-30a(+) plasmid and transformed into BL21(DE3) E. coli competent cells. The MYR-coreSA chimeric protein was expressed and purified using immobilized metal affinity chromatography and further characterized by gel electrophoresis, Western blot, and enzyme activity assay. The purified MYR-coreSA chimeric protein was tethered on the outer membrane of biotinylated adenocarcinoma A549 cells and then treated with various concentrations of sinigrin. Our results showed that 20 µM of sinigrin inhibited the growth of A549 cells tethered with myrosinase by ~60% in 48 h. Furthermore, the levels of treated cells undertaken apoptosis were determined by Caspase-3/7 activation and Annexin-V. In summary, sinigrin harnessed like a prodrug catalyzed by myrosinase to the production of AITC, which induced cell apoptosis and arrested the growth of lung cancer cells.

Leaf mustard (Brassica juncea var. cernua) is cultivated well in Okinawa Prefecture. The Okinawa region belongs to a subtropical climate and the cultivation of leaf mustard requires sunshade treatment. In this investigation, leaf mustard was cultivated under shading material and chemical components were compared with those with no shading. About 53% of sunshine was shaded by the material. Edible parts were subjected to the analysis of the content of free amino acids, sinigrin and allyl isothiocyanate using high-performance liquid chromatography. Many contents of leaf mustard cultivated with the no shading treatment were higher than those cultivated with shading. The flesh weight and dry matter rate of leaf mustard cultivated with no shading were also higher than those cultivated with shading. This suggested that cultivation of leaf mustard with shading was able to control the texture and content of various flavorful chemical components. [ABSTRACT FROM AUTHOR]

Millions of patients suffer from asthma worldwide. However, the first-line drugs used to treat asthma, namely, the beta-adrenergic receptors agonists (β-agonists), are not recommended for use as monotherapy because of their severe dose-related side effects. This limitation has prompted the search for new therapies, which can be used in conjunction with β--agonists so that lower doses can be administered. Sinigrin is a major compound found in many antiasthmatic medicinal plants. In this study, we explored the antiasthmatic activity of sinigrin when used in combination with β-agonists and its underlying mechanism. Sinigrin enhanced the asthma-relieving effects of isoproterenol and reduced the effective isoproterenol dose in an acute-asthma model in guinea pigs. Mechanistically, sinigrin enhanced the cAMP levels induced by β-agonists by inhibiting PDE4. The resulting increase in cAMP levels stimulated the activity of the downstream effector protein kinase A, which would be expected to ultimately induce the relaxation of airway smooth muscle. In conclusion, sinigrin enhances the asthma-relieving effects of β-agonists by regulating the cAMP signaling pathway and represents a potential add-on drug to β-agonists for the treatment of asthma. [ABSTRACT FROM AUTHOR]

A plant's chemical‐based resistance against insects can be characterized by an accumulation of phytochemicals that can severely harm or deter the generalist and non‐adapted specialist insects. In the absence of an herbivore threat, plants maintain a basic concentration of defensive compounds as a constitutive resistance mechanism. On the other hand, plants are capable of sharply increasing their production of defensive compounds as induced defence in response to herbivore attacks. In this study, we highlight the role of chemical‐based defence by comparing the constitutive and herbivore‐induced resistances of cauliflower (Brassica oleracea var. botrytis L.) and yellow cress (Rorippa dubia (Persoon)) with respect to time‐dependent changes of phytochemicals that directly influence the performance of the generalist herbivore insect, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). The results of the study showed that both plant species were able to switch, albeit different degrees, between constitutive and induced resistances to combat herbivore attack. Brassica oleracea amassed lesser defensive compounds of constitutive resistance (trypsin inhibitor, polyphenol oxidase and sinigrin) than R. dubia. This led to a higher relative growth rate (RGR) of the S. litura larvae fed on cauliflower than in those fed on yellow cress. Brassica oleracea also demonstrated a lower induced response capability than R. dubia in its production of allelochemicals in herbivore‐induced resistances. This was shown by the decrease of the RGR of larvae fed on induced foliage of the cauliflower compared to those fed on yellow cress. These results of the study proved and added to the belief that modern crop variety is not only lower in constitutive resistance but also weaker in induced resistance capability against herbivores, which is likely to be an effect of domestication even Rorippa and Brassica are relatively far from each other in Brassicaceae phylogeny. [ABSTRACT FROM AUTHOR]

Application of nitrogen (N) is a common practice used to achieve profitable yields in horticultural crops and N application can be used as a tool to manipulate the enhancement of phytochemicals in vegetable crops to address consumer-oriented quality production. Our previous findings recommended 90 kg ha−1 for certain types of cauliflower varieties without compromising yields. Thus, this study was aimed to investigate the effect of N application on glucosinolates and phenolic acids, at harvest, in varieties 'Largardo', 'Eskimo' and 'CF-744' grown in the field. N was applied as ammonium nitrate (NH4NO3) at concentrations from 0, 60, 90, 120, 150 to 180 kg ha−1. Variety 'CF-744' was more sensitive to N supply and at 180 kg ha−1 N it showed the highest accumulation of glucosinolates (sinigrin, glucoiberin, progoitrin, 4-methoxyglucobrassicin) at harvest. However, 90 kg ha−1 N supply demonstrated the highest accumulation of majority of the glucosinolates in varieties 'Largardo' and 'Eskimo'. Different varieties responded differently to N supply and glucosinolate levels in cauliflowers. Also, different varieties responded differently to N supply and antioxidant property. In all three varieties, the N supply at 120 kg ha−1 showed the highest accumulation of protocatechoic acid, 4-hydroxy benzoic acid, ρ-coumaric acid and caffeic acid. [ABSTRACT FROM AUTHOR]

Key message: Overexpression of BoMYB29 gene up-regulates the aliphatic glucosinolate pathway in Brassica oleracea plants increasing the production of the anti-cancer metabolite glucoraphanin, and the toxic and pungent sinigrin. Isothiocyanates, the bio-active hydrolysis products of glucosinolates, naturally produced by several Brassicaceae species, play an important role in human health and agriculture. This study aims at correlating the content of aliphatic glucosinolates to the expression of genes involved in their synthesis in Brassica oleracea, and perform functional analysis of BoMYB29 gene. To this purpose, three genotypes were used: a sprouting broccoli, a cabbage, and a wild genotype (Winspit), a high glucosinolate containing accession. Winspit showed the highest transcript level of BoMYB28, BoMYB29 and BoAOP2 genes, and BoAOP2 expression was positively correlated with that of the two MYB genes. Further analyses of the aliphatic glucosinolates also showed a positive correlation between the expression of BoAOP2 and the production of sinigrin and gluconapin in Winspit. The Winspit BoMYB29 CDS was cloned and overexpressed in Winspit and in the DH AG1012 line. Overexpressing Winspit plants produced higher quantities of alkenyl glucosinolates, such as sinigrin. Conversely, the DH AG1012 transformants showed a higher production of methylsulphinylalkyl glucosinolates, including glucoraphanin, and, despite an up-regulation of the aliphatic glucosinolate genes, no increase in alkenyl glucosinolates. The latter may be explained by the absence of a functional AOP2 gene in DH AG1012. Nevertheless, an extract of DH AG1012 lines overexpressing BoMYB29 provided a chemoprotective effect on human colon cells. This work exemplifies how the genetic diversity of B. oleracea may be used by breeders to select for higher expression of transcription factors for glucosinolate biosynthesis to improve its natural, health-promoting properties. [ABSTRACT FROM AUTHOR]

Evolutionary traps arise when organisms use novel, low‐quality or even lethal resources based on previously reliable cues. Persistence of such maladaptive interactions depends not only on how individuals locate important resources, such as host plants, but also the mechanisms underlying poor performance. Pieris macdunnoughii (Remington) (Lepidoptera: Pieridae) lays eggs on a non‐native mustard, Thlaspi arvense (L.) (Brassicaceae), which is lethal to the larvae. We first tested whether larval feeding behavior was affected before (pre‐) ingestion or following (post‐) ingestion of leaf material, indicating activity of feeding deterrents, toxins, or both in this evolutionary trap. Neonates were less likely to start feeding and eventually fed more slowly on T. arvense than on the native host plant Cardamine cordifolia (Gray) (Brassicaceae) in both laboratory and field. Starvation was a primary cause of mortality, indicating the role of a feeding deterrent. Feeding did not differ between larvae from invaded and uninvaded population. Second, T. arvense defensive chemistry is dominated by the glucosinolate sinigrin (allyl or 2‐propenyl glucosinolate). Adding sinigrin to the leaves of T. arvense and native hosts C. cordifolia and Descurainia incana (Bernhardi ex Fischer & Meyer) (Brassicaceae) delayed the onset of feeding, caused larvae to feed more slowly, and decreased survival on the native hosts. This evolutionary trap may not be driven by a novel deterrent, but rather by a change in the concentration of a deterrent found in native hosts. Many insects have adapted to evolutionary traps posed by invasive plants, incorporating the new plant into their diets. Thlaspi arvense remains lethal to P. macdunnoughii, and pre‐ingestive deterrents such as excess sinigrin may contribute to persistent maladaptation. [ABSTRACT FROM AUTHOR]

This study identified glucosinolate (GLS) breakdown products and determined the relationships between such products and parent GLS in boiled Brussels sprouts. This is the first ever study to analyse all groups of GLS breakdown products (aliphatic, aralkyl and indole) in boiled vegetables. In the examined boiled Brussels sprouts, isothiocyanates were not determined in the edible parts or cooking water. The presence of seven breakdown products was determined as indole-3-acetonitrile, indole-3-carbinol, ascorbigen and 3,3'-diindolylmethane released form glucobrassicin, 3-butenylnitrile released from sinigrin, 4-methylsulfinylbutanenitrile released from glucoiberin, and 2-phenylacetonitrile released from gluconasturtiin. In the edible parts, the content of indole-3-acetonitrile and 3-butenylnitrile was particularly high at approximately 30 and 16 µmol/100 g FW, which was equivalent to 54 and 9% of the initial value of the parent GLS, respectively.

Volatile allyl isothiocyanate (AITC) derives from the biodegradation of the glucosinolate sinigrin and has been associated with growth inhibition in several plants, including the model plant Arabidopsis thaliana. However, the underlying cellular mechanisms of this feature remain scarcely investigated in plants. In this study, we present evidence of an AITC-induced inhibition of actin-dependent intracellular transport in A. thaliana. A transgenic line of A. thaliana expressing yellow fluorescent protein (YFP)-tagged actin filaments was used to show attenuation of actin filament movement by AITC. This appeared gradually in a time- and dose-dependent manner and resulted in actin filaments appearing close to static. Further, we employed four transgenic lines with YFP-fusion proteins labeling the Golgi apparatus, endoplasmic reticulum (ER), vacuoles and peroxisomes to demonstrate an AITC-induced inhibition of actin-dependent intracellular transport of or, in these structures, consistent with the decline in actin filament movement. Furthermore, the morphologies of actin filaments, ER and vacuoles appeared aberrant following AITC-exposure. However, AITC-treated seedlings of all transgenic lines tested displayed morphologies and intracellular movements similar to that of the corresponding untreated and control-treated plants, following overnight incubation in an AITC-absent environment, indicating that AITC-induced decline in actin-related movements is a reversible process. These findings provide novel insights into the cellular events in plant cells following exposure to AITC, which may further expose clues to the physiological significance of the glucosinolate-myrosinase system.