Cruciferous Seeds Research Articles

Bioaccessibility of glucosinolates, isothiocyanates and inorganic micronutrients in cruciferous vegetables through INFOGEST static in vitro digestion model

Green tissues and seeds from cruciferous vegetables growing in conventional and ecological conditions (Brassica carinata; Brassica rapa; Eruca vesicaria and Sinapis alba) were analyzed to determine their contents of glucosinolates, isotihiocyanates (ITCs) and inorganic micronutrients (Ca, Cr, Cu, Fe, Mn, Ni, Se and Zn), and the bioaccessibility of these compounds. Regarding total contents and bioaccessibility values of these compounds, no clear difference was found between the organic and conventional systems. Glucosinolates bioaccessibility present in green tissues were high, with values around 60–78%. In additon, it was quantified in bioaccessible fraction ITCs concentrations such as Allyl – ITC; 3 – Buten – 1 – yl – ITC and 4 – Penten – 1 – yl – ITC. Trace elements bioaccessibility in green tissues was also high for Ca (2.26–7.66 mg/g), Cu (0.60–2.78 µg/g), Se (9.93–74.71 µg/Kg) and Zn (12.98–20.15 µg/g). By contrast, the bioaccessibility of glucosinolates and trace elements in cruciferous seeds was extremely low. With the exception of Cu, these bioaccessibility percentages did not exceed 1% in most cases.

Determination of Gamma-aminobutyric Acid (GABA) Content in Grains and Cruciferous Vegetable Seeds

Gamma aminobutyric acid (GABA) has great physiological functions, mainly as a major inhibitory neurotransmitter in the brain, which makes it important for the development of functional foods. This study detected GABA in grains and cruciferous vegetable seeds by using HPLC after pre-column on-line derivatization with diode array detection (DAD) and fluorescence detection (FLD). The limit of quantitation was 2.94 and 2.86 µg/mL with DAD and FLD, respectively. GABA recoveries ranged from 98.8 to 111.2% on both detectors. Intra and inter-day precision showed relative standard deviations, generally, less than 10% for both DAD and FLD. GABA was determined in different grains (flaxseeds, white quinoa seeds, and buckwheat) and cruciferous vegetable seeds (broccoli, kale, daikon radish, mustard, cabbage, and brussels sprouts). Organic broccoli seeds contained the highest amount and mustard seeds the least amount of GABA in the Brassica family with none being detected in organic white quinoa and flaxseeds. A statistically significant difference (p < 0.05) exists between the various lots of the broccoli seeds. GABA is important as a natural source in functional foods.

Benzyl isothiocyanate ameliorates lipid accumulation in 3T3-L1 preadipocytes during adipocyte differentiation.

Benzyl isothiocyanate (BITC) is an organosulfur compound derived from cruciferous vegetables and papaya seeds. In this study, we investigated the effect of BITC on the lipid accumulation in 3T3-L1 preadipocytes during adipocyte differentiation. The treatment of BITC during the differentiation-inducing stage significantly ameliorated the lipid accumulation, whereas it had no inhibitory effect during the differentiation-maintaining stage. BITC also significantly suppressed the mRNA expression of the adipocyte-specific markers, such as CCAAT/enhancer-binding protein α (C/EBPα), C/EBPβ, C/EBPδ and peroxisome proliferator-activated receptor γ. BITC significantly inhibited the phosphorylation of extracellular signal-regulated kinase phosphorylation, whereas it enhanced that of AMP-activated protein kinase. Furthermore, BITC significantly suppressed the intracellular 2-deoxyglucose uptake as well as glucose transporter 4 expression. These results suggest that inhibition of the adipocyte differentiation and glucose uptake may mainly contribute to the inhibitory effect of BITC on the lipid accumulation in 3T3-L1 preadipocytes. Abbreviations: PPARγ: peroxisome proliferator-activated receptor γ; CEBP: CCAAT/enhancer-binding protein; GLUT4: glucose transporter 4; AMPK: AMP-activated protein kinase; ERK1/2: extracellular signal-regulated kinase 1/2; MAPK: a mitogen-activated protein kinase; ITCs: isothiocyanates; BITC: benzyl isothiocyanate; FBS: fetal bovine serum; CS: calf serum; AITC: allyl ITC; IBMX: 3-isobutyl-1-methylxanthine; LDH: lactate dehydrogenase; KRH: Krebs-Ringer-Hepes-bicarbonate; 2-DG: 2-deoxy-d-glucose.

Low-Pressure Plasma Application for the Inactivation of the Seed-borne Pathogen Xanthomonas campestris.

The aim of this study was to investigate the effect of low-pressure plasma treatment on seed disinfection and the possible mechanisms underlying this effect. Seed-borne disease refers to plant diseases that are transmitted by seeds; seed disinfection is an important technique for prevention of such diseases. In this study, the effectiveness of low-pressure plasma treatment in the inactivation of the seed-borne plant pathogenic bacterium, Xanthomonas campestris, inoculated on cruciferous seeds, was evaluated. The highest inactivation effect was observed when the treatment voltage and argon gas flow rate were 5.5 kV and 0.5 L/min, respectively. The viable cell number of X. campestris was 6.6 log cfu/seed before plasma treatment, and decreased by 3.9 log after 5 min of treatment and by 6.6 log after 40 min. Ethidium monoazide treatment and quantitative real-time PCR results indicated that both the cell membrane and target DNA region were damaged following 5 min of plasma treatment. Although both heat and ozone were generated during the plasma treatment, the contribution of both factors to the inactivation of X. campestris was small by itself in our low-pressure plasma system. Overall, we have shown that our low-pressure plasma system has great applicability to controlling plant pathogenic bacterium contamination of seeds.

Seed-borne fungi on genebank-stored cruciferous seeds from Japan

Seed-borne fungi on genebank-stored cruciferous seeds from Japan

Manipulation of sinapine, choline and betaine accumulation in Arabidopsis seed: Towards improving the nutritional value of the meal and enhancing the seedling performance under environmental stresses in oilseed crops

Sinapoylcholine (sinapine) is the most abundant antinutritional phenolic compound in cruciferous seeds. The quaternary ammonium compounds, choline, betaine and N,N-dimethylglycine, reside along a biosynthetic pathway linked to the synthesis of membrane phospholipids and neurotransmitters with various biological functions. In chicken, choline intake is required for optimal egg-laying performance and a choline supplement in diet is positively correlated with weight gains. A key step in sinapine biosynthesis is catalyzed by sinapoylglucose: choline sinapoyltransferase (SCT; EC 2.3.1.91) to form an ester linkage with sinapoylglucose and choline. The objective of this work was to reduce the sinapine content and simultaneously enhance free choline levels in cruciferous seeds. We report here the characterization of an Arabidopsis T-DNA insertion mutant lacking SCT activity in the seed. The sct mutant seeds contain less than 1% of sinapine and a more than 2-fold increase in free choline compared with wild type. We further expressed a choline oxidase (COX; EC 1.1.3.17) gene from Arthrobacter pascens in the Arabidopsis sct mutant and wild-type background using a napin gene promoter to convert free choline into betaine, an effective stress-alleviating compound in plants. Betaine was not detected in WT or sct mutant seeds. The sct+COX seeds contain nearly 2-fold greater levels of betaine relative to WT+COX seeds, demonstrating a positive correlation between endogenous choline and betaine production. In contrast, stable comparable levels of free choline were detected between sct+COX and WT+COX plants suggesting choline homeostasis likely prevent high levels of betaine production in the seed of transgenic COX plants.

Prediction of the kind of sprouts of Cruciferae family based on artificial neural network analysis

The aim of this work was to show that artificial neural networks (ANNs) are a convenient tool for predicting the kind of sprouts originated from <i>Cruciferae<i> family. For this purpose, the known contents of bioactive compounds of small radish, radish, white mustard, and rapeseed seeds and sprouts were used for the prediction of the kind of a specific sprout. The input data reflected the contents of the following compounds in cruciferous seeds in the course of germination: soluble proteins (SP), ascorbic acid (AH<sub>2</sub>), total glucosinolates (GLS), reduced glutathione (GSH), and tocopherols (α-T, β-T, γ-T, δ-T), expressed in their biological activity. The ANN used was trained on the learning set. The ability of the utilised ANN to generalise the gained knowledge based on the learning set was verified by the validating and testing sets. The trained and validated ANN was able to classify, with complete accuracy, the kind of sprouts out of the four kinds used. It can be concluded that ANN can be used as a useful tool for determining the identity of cruciferous seeds and sprouts based on the determined levels of their bioactive components.

Inositol phosphate content and trypsin inhibitor activity in ready-to-eat cruciferous sprouts

Four cruciferous seeds (small radish, radish, white mustard and rapeseed) were germinated in order to study the fate of inositol hexaphosphate (IP-6, phytic acid) and activity of trypsin inhibitor (TIA). Reduction in the content of phytic acid depended on the time of germination. After four days of germination, when sprouts were ready-to-eat, the amount of this compound was about 50% lower in three out of the four seeds evaluated. Next, a sharp reduction in phytic acid occurred during thermal treatments (pasteurization and sterilization) of germinated rapeseed and radish sprouts. In thermal processes, a decrease in inositol hexaphosphate content was accompanied by the appearance of lower forms of inositol phosphates: IP-5, IP-4 and IP-3. The analysis of TIA, in rapeseed and radish seeds, in four-day sprouts and in these sprouts after thermal treatment, showed that only thermal processes caused complete disappearance this activity.

Allelopathic Effect of Some Cruciferous Seeds on Rhizoctonia solani kuhn and Gossypium barbadense L.

Allelopathic Effect of Some Cruciferous Seeds on Rhizoctonia solani kuhn and Gossypium barbadense L.

Artificial neural networks for prediction of antioxidant capacity of cruciferous sprouts

The aim of this work was to show that artificial neural networks (ANNs) are the convenient tool for modeling the biological properties of food. For this reason, as a good example, known contents of bioactive compounds of cruciferous sprouts were taken for the prediction of their trolox equivalent antioxidant capacity–one of the most important biological properties of food. The input data reflected the contents of the following compounds in cruciferous seeds in the course of germination: total phenolic compounds (TPC), inositol hexaphosphate (InsP6), glucosinolates (GLS), soluble proteins (SP), ascorbic acid (AH2), and total tocopherols (Ttot). Additionally, the trolox equivalent antioxidant capacity of germinated cruciferous seeds (TEACexp) was determined. The used ANN which was trained on the learning set, generalized the obtained prediction ability in respect to the data contained in validating and testing sets. The predicted TAEC values calculated by ANN were satisfactory correlated with experimental TAEC values. Therefore, the ANN seems to find application in the quality analysis of functional properties of food of plant origin for the prediction of the trolox equivalent antioxidant capacity.

Isolation and biochemical characterization of a basic myrosinase from ripe Crambe abyssinica seeds, highly specific for epi-progoitrin.

On the basis of previous studies on the mechanism-based inhibition, activation, and active site structure of myrosinase(s) isolated from Sinapis alba and other cruciferous seeds, crambe myrosinase shows uncommon properties and behavior. For this reason homogeneous crambe myrosinase was isolated and investigated to establish the most important physicochemical features, including kinetic properties determined with the epimers progoitrin (R) and epi-progoitrin (S) as substrates, with and without ascorbate as an activator. The results of this study demonstrate that crambe myrosinase is highly specific for epi-progoitrin due to a better stabilization of the enzyme-substrate complex. This stabilization is caused by additional hydrogen bonding that only epi-progoitrin can set up between its hydroxyl group and a suitable residue in the hydrophobic pocket where the "docking" of the glucosinolates side chain takes place.

Isolation and inhibition of a trypsin-like activity from larvae of corn borer (Ostrinia nubilalis) using reverse micelles

Endoproteinase(s) was isolated from a freeze-dried powder of larvae of Ostrinia nubilalis using reverse micellar solutions. The inhibition of proteinase was studied in reverse micelles with commercial Bowman-Birk soybean trypsin inhibitor and three trypsin inhibitors recently isolated from ripe cruciferous seeds.

A New Approach to the Study of Glucosinolates by Isocratic Liquid Chromatography. Part I. Rapid Determination of Desulfated Derivatives of Rapeseed Glucosinolates

Liquid chromatographic (LC) analysis of desulfated derivatives of rapeseed glucosinolates has been carried out under isocratic elution conditions with different CN-bonded stationary phases. The effects of the eluant composition (water, acetonitrile, and methanol) with the stationary phase (Zorbax CN, Lichrospher CN, and Ultrasphere CN) and temperature (20 and 50 degrees C) are described. An isocratic LC method performed at room temperature using a Lichrospher CN column and water as mobile phase is proposed. The chromatographic analysis can be done in less than 12 min, and it is easier and less expensive than the traditional gradient mode. Four commercial samples of rapeseed containing various quantities of other cruciferous seeds (wild mustard and stinkweed) as an admixture have been analyzed to determine the total glucosinolate content. Relative standard deviations of repeatability of the isocratic and gradient LC methods ranged from 0.4 to 1.7% and from 2.7 to 4.7%, respectively. Comparison of the results showed good agreement between the 2 methods (beter than 98%).

Isolation of Sinigrin and Glucotropaeolin from Cruciferous Seeds

AbstractA procedure is described which allows the isolation of ca. 1 g Sinigrin (K‐salt) or glucotropaeolin (tetramethylammonia‐(TMA)‐salt) from seeds of Brassica nigra and Lepidium sativum, respectively within one day as crystalline substances. Both compounds possess excellent long term stabilities. In contrast to sinigrin the TMA salt of glucotropaeolin does not contain water of crystallization.

Biocontrol of seed-borne Alternaria raphani and A. brassicicola

Forty-two microorganisms were tested as biological control agents against Alternaria raphani and A. brassicicola. Tests were conducted for in vitro antagonistic ability, for ability to control the pathogens on naturally infected seeds germinated on moistened blotters, and in planting mix in growth chamber studies, and for their ability to reduce pod infection. The organisms tested were obtained from cruciferous seeds or were strains already identified as being effective against soil-borne Pythium species. The blotter test indicated that six organisms increased both the number of healthy seedlings and the number of seedlings produced from A. raphani infected radish seeds. An additional seven strains improved either germination or increased the number of healthy seedlings. Twenty-nine organisms increased the number of healthy cabbage seedlings from A. brassicicola infected seeds, but total germination was not modified by any treatment. Experiments in planting mix showed that five antagonists (Chaetomium globosum, two strains of Trichoderma harzianum, T. koningii, and Fusarium sp.) increased the number of healthy plants in both radish samples tested, while four additional antagonists provided a significant increase in only one of the samples tested. The five antagonists that consistently increased numbers of healthy radish seedlings also decreased pod infection by A. raphani. None were as effective as iprodrone, however. Several effective antagonists were found to be mycoparasitic against Alternaria spp. Some strains of Trichoderma previously found to be effective against Pythium spp. were also effective against Alternaria spp., indicating that these strains have a wide host range.

Biochemistry of Fern Spore Germination: Globulin Storage Proteins in Matteuccia struthiopteris L.

Two globulin storage proteins have been identified in spores of the ostrich fern, Matteuccia struthiopteris (L.) Todaro. The two proteins comprise a significant amount of the total spore protein, are predominantly salt-soluble, and can be extracted by other solvents to a limited extent. The large 11.3 Svedberg unit (S) globulin is composed of five polypeptides with molecular weights of 21,000, 22,000, 24,000, 28,000 and 30,000. Each polypeptide has several isoelectric point (pI) variants between pH 5 and 7. The small 2.2S storage protein has a pI > 10.5 and is composed of at least two major polypeptides of 6,000 and 14,000 M(r). The amino acid composition of both storage proteins reveals that the 11.3S protein is particularly rich in aspartic and glutamic acid, while the 2.2S protein has few acidic amino acids. During imbibition and germination the globulin fraction declines rapidly, with a corresponding degradation of individual polypeptides of each protein. Polyclonal antibodies against each of the two proteins were produced and used for immunolocalization to determine the site of storage protein deposition within the quiescent spore. The proteins were sequestered in protein bodies of 2 to 10 micrometers, that are morphologically similar to those found in the seeds of flowering plants. The results suggest that spore globulins are biochemically similar to seed globulins, especially those found in some cruciferous seeds.

Determination of Total Glucosinolates in Cruciferous Seeds by Semimicro‐gravimetry and Spectrophotometry with Tetrachloropalladate

AbstractFor the determination of the glucosinolate content in cruciferous seeds by spectrophotometry with tetrachloropalladate it is necessary to classify the samples according to their different absorption spectra of the corresponding Pd‐complexes into groups. Standard samples are recommended for the preparation of the different calibration curves, which have to be analysed by photometry as well as by a reference method. In order to eliminate the influence of the colour of the extracts and the Pd‐reagent solution on the analytical results double‐cell blank cuvettes have been used. A modified gravimetric method on a semimicro scale was employed as reference method. This latter method is recommended as sole procedure if only few samples have to be analysed. Using the proposed alterations of the Pd‐method improvements of the accuracy and the precision of the results and the sensitivity of the method can be achieved.

Glucosinolates and lipid content in developing and germinating cruciferous seeds

The changes in the glucosinolate content and its relations with oil and fatty acid composition was determined in developing and germinating Brassica seeds. During development there was a gradual increase in the glucosinolate content in Brassica napus, B. juncea and Eruca sativa and their amount was higher in seed meals over the pod shells; the difference was more marked at the later stages of seed development. The glucosinolate content showed a positive correlation with oil, erucic acid and total unsaturated fatty acid contents, significant in E. sativa. During germination, the accumulation of glucosinolates was higher in the epicotyl than in the hypocotyl portion of the seedlings. The amount of vitamin C was found positively correlated with glucosinolate and negatively with the oil content.

Complex‐Formation between Glucosinolates and Tetrachloropalladate(II) and its Utilization in Plant Breeding

AbstractA method is described which allows a fast determination of the content of glucosinolates in cruciferous seeds. Basically the reaction between glucosinolates and tetrachloropalladate (II) is used. For the quantitative measurement of the resulting coordination complexes an 8‐channel photometer is recommended. Furthermore an analytical grist‐mill has been constructed which allows a fast preparation of the samples.

The fatty acid composition of various cruciferous seeds

AbstractSeeds from 26 Cruciferae species in 7 genera have been investigated for fat content and fatty acid composition of the oil. The GLC retention data have been verified by mass spectrometry. The oil from Cardamine graeca contained 54% of cis‐15‐tetracosenoic acid; it is the highest content of this acid so far reported in any seed fat.