Variation of glucosinolates in vegetable crops of Brassica oleracea

Glucosinolates were evaluated in 5 groups and 65 accessions of Brassica oleracea (50 broccoli, 4 Brussels sprouts, 6 cabbage, 3 cauliflower, and 2 kale) grown under uniform cultural conditions. Glucosinolates and their concentrations varied among the different groups and within each group. The predominant glucosinolates in broccoli were 4-methylsulfinylbutyl glucosinolate (glucoraphanin), 3-butenyl glucosinolate (gluconapin), and 3-indolylmethyl glucosinoate (glucobrassicin). Glucoraphanin concentration in broccoli ranged from 0.8 micromol g(-1) DW in EV6-1 to 21.7 micromol g(-1) DW in Brigadier. Concentrations of the other glucosinolates in broccoli varied similarly over a wide range. In Brussels sprouts, cabbage, cauliflower, and kale, the predominant glucosinolates were sinigrin (8.9, 7.8, 9.3, and 10.4 micromol g(-1) DW, respectively) and glucobrassicin (3.2, 0.9, 1.3, and 1.2 micromol g(-1) DW, respectively). Brussels sprouts also had significant amounts of gluconapin (6.9 micromol g(-1) DW). Wide variations in glucosinolate content among genotypes suggest differences in their health-promoting properties and the opportunity for enhancement of their levels through genetic manipulation.     

Variation of glucosinolates in wild radish (Raphanus raphanistrum) accessions

Glucosinolate composition was determined in wild radish accessions from eight states in the northeastern and southern United States to determine the variability of production among accessions. Glucosinolates were evaluated from roots, leaves, flowers, primary, and secondary branches. Seventeen glucosinolates were identified, with glucoerucin, glucoraphenin, glucobrassicin, and gluconasturtiin contributing 90% to 100% of the total glucosinolates. Flowers contained the highest glucosinolate concentrations, 12.07 to 55.36 μmol/g, but flowers contributed only 5.3 to 21.3% to the total glucosinolates. Of the eight accessions, the Mississippi accession produced significantly higher levels of total glucosinolates and glucosinolates which can be degraded to isothiocyanates per plant, totals of 618.97 and 563.53 μmol/plant, respectively. Total plant biomass did not differ between accessions indicating a difference in the ability of the Mississippi accession to produce glucosinolates. Further studies are needed to determine if this accession would consistently produce higher glucosinolate levels under different environmental conditions.     

花椰菜主要生物活性物质及其抗氧化能力分析

分析了花椰菜不同器官(花球、茎、叶和根)中芥子油苷等主要生物活性物质及其抗氧化能力,发现烯丙基芥子油苷是花椰菜中主要的芥子油苷,不同器官中芥子油苷的组分与含量差异显著,根系中的总芥子油苷、总脂肪类、总芳香类以及烯丙基芥子油苷和4-甲氧基-吲哚-3-甲基芥子油苷的含量均显著高于其他器官,叶片中吲哚类芥子油苷的含量最高,花球中总芥子油苷、烯丙基芥子油苷和吲哚-3-甲基芥子油苷含量次之,茎中的总芥子油苷含量最低;叶片中的叶绿素、类胡萝卜素、维生素C和多酚含量及抗氧化能力显著高于其他器官。这些结果表明花椰菜的叶片和根系中生物活性物质含量丰富,具有进一步开发和综合利用潜力。     

Factors affecting the glucosinolate content of kale (Brassica oleracea acephala group)

Kales (Brassica oleracea acephala group) are important vegetable crops in traditional farming systems in the Iberian Peninsula. They are grown throughout the year to harvest their leaves and flower buds. The glucosinolate content of kales is dependent upon the environmental factors, plant part examined, phenological stage of plant growth, and level of insect damage. The objectives of this study were to evaluate the changes in the total and individual glucosinolate concentrations during plant development and to determine if significant variation of glucosinolate levels can be explained by insect pests attack and other environmental factors in four locations in northwestern Spain. The total glucosinolate concentration in leaves of B. oleracea increased with plant age from seedling to early flowering stages. At that stage, the aliphatic glucosinolate content in leaves of B. oleracea declined drastically over time as the content in the flower buds increased. The highest contents of indolyl glucosinolate (glucobrassicin) and of the aromatic glucosinolate occurred in leaves harvested at the optimum consumption stage while flower buds contained the highest concentration of aliphatic glucosinolates, especially sinigrin. Sinigrin is reported to have anticarcinogenic properties. There appears to be a loss of total and individual glucosinolate concentrations related to pest attack. Leaves damaged by lepidopterous pests contained a lower total glucosinolate content (25.8 micromol g-1 dw) than undamaged leaves (41 micromol g-1 dw). The amounts of sinigrin, glucoiberin, and glucobrassicin were also lowest in insect-damaged leaves. Environmental factors such as soil properties and temperature appear to influence the glucosinolate content in leaves although more research on this subject is needed.     

Soluble and total myrosinase activity in defatted Crambe abyssinica meal

Crambe defatted meal contains 4-6% w/w of glucosinolates, with epiprogoitrin accounting for >90% of the total. This feature limits the use of the meal as feed due to the antinutritional properties of myrosinase-glucosinolate breakdown products. In this context, myrosinase activity assumes particular importance. In this study the total and soluble myrosinase activities have been evaluated directly on defatted meals of eight Crambe abyssinica varieties. The pH-stat method, which is the most suitable for assays in heterogeneous solid-water systems, was used. The total myrosinase activity in C. abyssinica varieties, determined using epiprogoitrin as substrate, ranged from 288 to 653 units g(-1). These activity values were up to 26 times higher than those obtained using other substrates, namely, sinigrin, glucosinalbin, glucotropaeolin, progoitrin, and glucoraphenin. Crambe myrosinase is unusual in that, unlike other Brassicaceae containing a typical main glucosinolate, it does not show the same specificity toward its natural substrates.     

Changes in glucosinolate concentrations, myrosinase activity, and production of metabolites of glucosinolates in cabbage (Brassica oleracea Var. capitata) cooked for different durations

In cabbage, glucosinolates such as sinigrin are hydrolyzed by plant myrosinase to allyl isothiocyanate (AITC), allyl cyanide, and, in the presence of an epithiospecifier protein, 1-cyano-2,3-epithiopropane (CEP). Isothiocyanates have been implicated in the cancer-protective effects of Brassica vegetables. The effect of processing on the hydrolysis of glucosinolates was investigated in cabbage. Cabbage was steamed or microwaved for six time durations over 7 min. Glucosinolate concentrations were slightly reduced after microwave cooking (P < 0.001) but were not influenced after steaming (P < 0.05). Myrosinase activity was effectively lost after 2 min of microwave cooking and after 7 min of steaming. Hydrolysis of residual glucosinolates following cooking yielded predominantly CEP at short cooking durations and AITC at longer durations until myrosinase activity was lost. Lightly cooked cabbage produced the highest yield of AITC on hydrolysis in vitro, suggesting that cooking Brassica vegetables for a relatively short duration may be desirable from a health perspective.     

Flavor-active compounds potentially implicated in cooked cauliflower acceptance

The aim of the present study was to determine the flavor-active compounds responsible for the "sulfur" and "bitter" flavors of cooked cauliflower potentially implicated in cauliflower rejection by consumers. Eleven varieties of cauliflower were cooked and assessed by a trained sensory panel for flavor profile determination. Among the 13 attributes, the varieties differed mainly according to their "cauliflower odor note" and their "bitterness". Various glucosinolates were quantified by HPLC and correlated with bitterness intensity. The results showed that neoglucobrassicin and sinigrin were responsible for the bitterness of cooked cauliflower. Application of Dynamic Headspace GC-Olfactometry and DH-GC-MS showed that allyl isothiocyanate (AITC), dimethyl trisulfide (DMTS), dimethyl sulfide (DMS), and methanethiol (MT) were the key odorants of cooked cauliflower "sulfur" odors. Moreover, these volatile compounds corresponded to the main compositional differences observed between varieties. Finally, AITC, DMTS, DMS, MT, sinigrin, and neoglucobrassicin were shown to be potential physicochemical determinants of cooked cauliflower acceptance.     

Health-promoting compounds in broccoli as influenced by refrigerated transport and retail sale period

Total aliphatic and indole glucosinolates, phenolic compounds (flavonoids and hydroxycinnamoyl derivatives), and vitamin C contents were evaluated in freshly harvested broccoli (Brassica oleracea L., var. italica, cv. Marathon) inflorescences. These were film-wrapped and stored for 7 days at 1 degrees C to simulate a maximum period of commercial transport and distribution. After cold storage, inflorescences were kept for 3 days at 15 degrees C to simulate a retail sale period. For wrapping, low-density polyethylene (LDPE) of 11 microm thickness was used. Gas composition was about 17% O(2) and 2% CO(2) during cold storage and about 16% O(2) and 3-4% CO(2) during shelf life within packages. The predominant glucosinolates were 4-methylsulfinylbutyl-glucosinolate (glucoraphanin), 3-indolylmethyl-glucosinolate (glucobrassicin), and 1-methoxy-3-indolylmethyl-glucosinolate (neoglucobrassicin). The predominant hydroxycinnamoyl derivatives were identified as 1,2,2'-trisinapoylgentiobiose, 1,2-diferuloylgentiobiose, 1,2'-disinapoyl-2-feruloylgentiobiose, and 3-O-caffeoyl-quinic (neochlorogenic acid). Results showed major losses at the end of both periods, in comparison with broccoli at harvest. Thus, the respective losses, at the end of cold storage and retail periods, were 71-80% of total glucosinolates, 62-59% of total flavonoids, 51-44% of sinapic acid derivatives, and 73-74% caffeoyl-quinic acid derivatives. Slight differences in all compound concentrations between storage and retail sale periods were detected. Distribution and retail periods had minimal effects on vitamin C. Weight loss was monitored at the end of both periods.     

Development and validation of a spectrophotometric method for quantification of total glucosinolates in cruciferous vegetables

Given their putative role in chemoprevention, validated methods are needed for quantification of total glucosinolates. Based on the colorimetric reaction of ferricyanide with 1-thioglucose, released by alkaline hydrolysis of glucosinolates, we developed a simple and sensitive method for spectrophotometric quantification of total glucosinolates in cruciferous vegetables. Lyophilized and ground vegetables are extracted with 80% boiling methanol. Extracted glucosinolates are isolated using a strong anion exchange column and then hydrolyzed with 2 N NaOH to release 1-thioglucose. Ferricyanide is added, and the decrease in absorbance is measured at 420 nm, with final values adjusted for background. Recovery of internal standard (sinigrin) was 107%. Intra- and interassay coefficients of variation were 5.4% and 15.8%, respectively. Dose response was linear with sinigrin and amount of plant material extracted (R(2) ≥ 0.99). Using sinigrin, the lower limit of quantification was 0.6 mg. This straightforward method may be an alternative to time-consuming and costly chromatographic methods.     

Genetic variation in leaf and stem glucosinolates in resynthesized lines of winter rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

Glucosinolates are secondary components characteristic for the Brassicaceae with complex biological functions. Glucosinolates in the seeds are anti-nutritive when feeding animals and their inheritance have been extensively investigated. Much less is known about the genetics of glucosinolates in leaves and stems, which may attract some insects, while repelling others. They may also inhibit bacterial processes of importance when using green biomass for the production of biogas. The objective of this study was to analyse the genetic variation of total and individual glucosinolates in the green material of rapeseed. For this 28 resynthesized winter rapeseed lines were tested at two locations. There was a large variation in leaf glucosinolate content between 0.10 and 4.75 μmol/g dry matter. The predominant leaf glucosinolates are the alkenyle glucosinolates progoitrin, gluconapin and glucobrassicanapin. The line R53 is exceptional, while combining a relative high content of the indole glucosinolate glucobrassicin with low alkenyle glucosinolates in the leaves. The total glucosinolate concentration in the stems and leaves is not correlated with the seed glucosinolate concentrations. Heritabilities are above h2 = 0.60 for progoitrin, h2 = 0.65 for gluconapin, h2 = 0.30 for glucobrassicanapin and h2 = 0.52 for total glucosinolate content in the leaves. In conclusion, resynthesized rapeseed is an important genetic resource to modify the leaf glucosinolate content and composition of rapeseed.     

Glucosinolate Profiles of Arabidopsis thaliana in Response to Cadmium Exposure

The influence of cadmium on growth, cadmium accumulation, composition, and content of glucosinolates was investigated in Arabidopsis thaliana after 4 weeks of growth in hydroponics. Accumulation of 3,820 and 321 μg Cd g^?1 dry weight in the roots and leaves of A. thaliana, respectively, exposed to 50 μM Cd. Cadmium treatment significantly decreased the total concentration of glucosinolates both in the leaves and roots. Cd-induced alteration of total glucosinolate content in the roots was mainly due to the decrease of indolyl-glucosinolates. In the Cd treatment leaves, significant decreases were, respectively, detected for glucoibervirin and 4-methoxyglucobrassicin (P?<?0.01), while other glucosinolate levels did not decrease significantly. In response to cadmium, the three indolyl-glucosinolates all showed significant decreases in the roots. The distinctive influence of cadmium on glucosinolate profiles in Cd-sensitive A. thaliana may be of great ecological importance, decreasing the resistance to phytophage attack. Taken together, our data is discussed in relation to jasmonic acid and salicylic acid as possible molecules that modulate the alteration of glucosinolate profiles in response to cadmium. The similar effects of Cd treatment on the levels of individual glucosinolates in leaves and roots were observed at higher-concentration cadmium treatment (100 μM Cd).     

喷施蔗糖和葡萄糖对小白菜硫代葡萄糖苷含量的影响

以小白菜"上海青"为试验材料,研究了喷施蔗糖和葡萄糖对小白菜生长和硫代葡萄糖苷含量的影响。结果显示,蔗糖和葡萄糖喷施对小白菜地上部鲜重和干重均没有显著影响,喷施蔗糖均能够显著增加小白菜总脂肪族硫代葡萄糖苷、总吲哚族硫代葡萄糖苷和总硫代葡萄糖苷含量,其中,0.1mol/L蔗糖喷施显著提高了3-丁烯基硫代葡萄糖苷和吲哚-3-甲基硫代葡萄糖苷含量,0.2mol/L蔗糖喷施显著增加了吲哚-3-甲基硫代葡萄糖苷和4-甲氧基吲哚-3-甲基硫代葡萄糖苷含量。0.2mol/L葡萄糖喷施显著增加总硫代葡萄糖苷及单个硫代葡萄糖苷含量。蔗糖和葡萄糖喷施均有增加总吲哚族硫代葡萄糖苷相对百分含量的趋势。研究表明外源喷施糖对小白菜硫代葡萄糖苷含量和组成具有显著的影响。     

High-performance liquid phase separation of glycosides. 5. Determination of individual glucosinolates in cabbage and rapeseed by laser-induced fluorescene capillary electrophoresis via the enzymatically released isothiocyanate aglycon.

A capillary electrophoresis (CE) method was developed for the profiling and determination of individual glucosinolates (GSs) via their isothiocyanate degradation products upon myrosinase digestion. The resulting isothiocyanates, the structures of which are reflective of the parent GS's, were then converted to their corresponding amines via base hydrolysis or reaction with 1, 2-benzenedithiol. Subsequently, the amines were fluorescently labeled to allow their sensitive detection by laser-induced fluorescence (LIF). The CE method involved the use of in situ charged micelles for the separation of isothiocyanates and their corresponding fluorescently labeled amines by micellar electrokinetic capillary chromatography (MECC). The term "in situ charged micelles" refers to micelles formed by complexing the polar hydroxyl groups of glycosidic surfactants with borate. The MECC method with on-column LIF detection was applied to the determination of GSs in white cabbage, rapeseed leaves, and rapeseed roots.     

Use of near-infrared spectroscopy for screening the individual and total glucosinolate contents in Indian mustard seed (Brassica juncea L. Czern. & Coss.)

The potential of near-infrared spectroscopy (NIRS) for screening the sinigrin, gluconapin, 4-hydroxyglucobrassicin, and total glucosinolate contents of Indian mustard (Brassica juncea L. Czern. & Coss.) seed was assessed. Intact seed samples of this species were analyzed by NIRS and their reference values regressed against different spectral transformations by modified partial least-squares (MPLS) regression. The coefficients of determination (r (2)) for sinigrin, gluconapin, 4-hydroxyglucobrassicin, and total glucosinolate contents were, respectively, 0.86, 0.95, 0.33, and 0.82. The standard deviation to standard error of prediction (SEP) ratio, and SEP to standard error of laboratory ratio were for these constituents as follows: sinigrin, 2.59 and 2.70; gluconapin, 4.16 and 2.08; 4-hydroxyglucobrassicin, 1.18 and 1.40; and total glucosinolates, 2.18 and 1.60. By comparison of commercial sinigrin spectrum with the first MPLS loadings of the sinigrin equation, it can be concluded that the molecule of sinigrin has a specific signal in the seed spectrum of Brassica.     

Behavior of glucosinolates in pickling cruciferous vegetables

Crucifer species, which include widely consumed vegetables, contain glucosinolates as secondary metabolites. Cruciferous vegetables are consumed in Japan in salt-preserved or pickled form as well as cooked and raw fresh vegetables. In this study, changes in contents of glucosinolates during the pickling process were investigated. 4-Methylthio-3-butenyl glucosinolate, a major glucosinolate in the root of Japanese radish, daikon (Raphanus sativus L.), was detected in pickled products with a short maturation period but not in those with a long maturation period. As a model pickling experiment, fresh watercress (Nasturtium officinale) and blanched watercress were soaked in 3% NaCl solution for 7 days. The results showed that the ratio of indole glucosinolates to total glucosinolates increased during the pickling process, whereas total glucosinolates decreased. Myrosinase digestion of glucosinolates in nozawana (Brassica rapa L.) indicated that indole glucosinolates, especially 4-methoxyglucobrassicin, were relatively resistant to the enzyme. The effect of pickling on glucosinolate content and the possible mechanism are discussed in view of degradation by myrosinase and synthetic reaction in response to salt stress or compression during the pickling process.     

Effects of intact glucosinolates and products produced from glucosinolates in myrosinase-catalyzed hydrolysis on the potato cyst nematode (Globodera rostochiensis Cv. Woll)

The potato cyst nematode (Globodera rostochiensis cv. Woll) is responsible for large yield losses in the potato crop, and opportunities for reducing the attack of these plant nematode species are, therefore, important. This study has been devoted to the testing of the in vitro effects on the potato cyst nematode of eight glucosinolates [prop-2-enyl-, but-3-enyl-, (R)-4-methylsulfinylbut-3-enyl-, benzyl-, phenethyl-, 4-hydroxybenzyl-, (2S)-2-hydroxybut-3-enyl-, and (2R)-2-hydroxy-2-phenylethylglucosinolate] as well as the effects of the products of this myrosinase-catalyzed hydrolysis. The glucosinolates were used at three concentrations, 0.05, 0.3, and 1.0 mg/mL, in the presence or absence of the enzyme myrosinase. The effects of the compounds on the mortality were monitored every 8 h for a 72 h period. No effects were found for any of the intact glucosinolates. However, when active myrosinase was included with 1 mg/mL phenethylglucosinolate at pH 6.5, 100% mortality was observed within just 16 h. A similar effect was achieved at the same concentration of benzyl- and prop-2-enylglucosinolates in the myrosinase-containing solutions, although longer exposures were required (24 and 40 h, respectively). The main aglucone products released from the glucosinolates with pronounced effects on the nematodes were shown to be the corresponding isothiocyanates. The results suggest that mixtures of these specific glucosinolates and active myrosinase or autolysis of plant materials containing these enzymes and glucosinolates might be used to control the potato cyst nematode in the soil.     

核盘菌侵染拟南芥的过程及其芥子油苷的变化

研究了核盘茵侵染拟南芥哥伦比亚生态型(Col-O)过程中,拟南芥的发病症状、活性氧和芥子油苷组分和含量的变化,及其抗性相关基因和芥子油苷生物合成相关基因表达的变化.结果表明,在侵染过程中,拟南芥的受侵染部位出现黄色水渍状病斑,随发病时间的延长病斑逐渐扩大到整株,其相对含水量逐渐下降,相对电导率不断上升,活性氧(过氧化氢...     

Bioactive compounds, myrosinase activity, and antioxidant capacity of white cabbages grown in different locations of Spain

The influence of two Spanish growing locations with well-differentiated climatic conditions (northern and eastern areas) on the main bioactive compounds, glucosinolates (GLS), total phenolic compounds (TPC), and vitamin C, as well as myrosinase activity and antioxidant capacity in five white cabbage ( Brassica oleracea L. var. capitata) cultivars was investigated. Cabbages with the highest concentration of total GLS presented the highest vitamin C level (r = 0.75, P ≤ 0.05) and the lowest antioxidant capacity (r = -0.76, P ≤ 0.05). The cultivars with the highest vitamin C content had the lowest myrosinase activity (r = -0.89, P ≤ 0.05) and antioxidant capacity (r = -0.86, P ≤ 0.05), whereas those with the largest TPC amount showed the highest antioxidant capacity (r = 0.71, P ≤ 0.05). Cabbage cultivars grown in the northern area of Spain with low temperatures and radiation led to higher mean values of myrosinase activity (29.25 U/g dm), TPC (10.0 GAE mg/g dm), and antioxidant capacity (81.6 μmol Trolox/g dm), whereas cultivars grown in the eastern area with high temperature and radiation led to larger mean values of GLS (14.3 μmol/g dm) and vitamin C (5.3 mg/g dm). The results of this investigation provide information regarding the most suitable Spanish growing location to produce white cabbage with an optimized content of health-promoting compounds.     

Glucosinolate profiles in the seeds,leaves and roots of rocket salad (Eruca sativa Mill.) and anti-oxidative activities of intact plant powder and purified 4-methoxyglucobrassicin

Abstract

Six desulfo-glucosinolates (DS-GSLs), DS-glucoraphanin, DS-4-(β-d-glucopyranosyldisulfanyl)butyl GSL, DS-glucoerucin, DS-glucobrassicin, DS-dimeric 4-mercaptobutyl GSL and DS-4-methoxyglucobrassicin, were isolated from rocket salad (Eruca sativa Mill.) By using high-pressure liquid chromatography (HPLC) and identified by their retention times on HPLC and electrospray ionization mass spectrometry (ESI-MS) techniques. Total GSL contents were 125, 11 and 22 µmol g^?1 dry weight (DW) in seeds, leaves and roots, respectively. Two DS-GSLs, namely DS-glucoraphanin and DS-glucoerucin, were the major GSLs in seeds and roots, while DS-glucoraphanin, DS-glucoerucin and DS-dimeric 4-mercaptobutyl GSL were the predominant GSLs in leaves (> 1 µmol g^?1 DW or > 10% of the total GSL content). In addition, the anti-oxidant activity (n = 3) of intact leaf and root powders, whole DS-GSL eluates and purified DS-4-methoxyglucobrassicin were measured using the XYZ-dish method. DS-4-methoxyglucobrassicin showed the greatest anti-oxidant activity value (3.8 unit g^?1 DW), and the value for whole DS-GSL eluates (1.6) was higher than that of intact leaves (1.5) and roots (0.87).     

Glucosinolates and myrosinase activity in red cabbage (Brassica oleracea L. var. Capitata f. rubra DC.) after various microwave treatments

Total and individual levels of glucosinolates (GSs) were measured in red cabbage after various microwave treatments varying in time and intensity of the treatments. Furthermore, the myrosinase enzyme activity of the microwave-heated vegetables was determined. The retention of GSs in the cabbage and the residual activity of the hydrolytic enzyme as a result of microwave preparation were compared with untreated cabbage. In general, high total GS levels were observed for all of the applied microwave treatments. Strikingly, many of the time/energy input combinations resulted in levels exceeding the total GS content of the untreated cabbage material. Moreover, the increase in levels seems to be associated with the energy input applied. A possible explanation for this behavior is an increased extractability of GS from heat-treated cabbage as compared to raw cabbage. Substantial myrosinase activity was retained in cabbage treated at low (24 min, 180 W) and intermediate microwave powers (8 min, 540 W) while microwave cooking for 4.8 min at 900 W (259.2 kJ energy input) resulted in a complete loss of hydrolytic activity. In this respect, differences in observed temperature profiles of the various microwave treatments play an important role. Higher retention of GSs and controllable amounts of active myrosinase can offer increasing health-promoting properties of microwave-prepared Brassica vegetables.