Multivariate analysis of tronchuda cabbage (Brassica oleracea L. var. costata DC) phenolics: influence of fertilizers

A field experiment was carried out to investigate the effect of fertilization level on the phenolic composition of tronchuda cabbage ( Brassica oleracea L. var. costata DC) external and internal leaves. Eight different plots were constituted: a control without fertilization, one with organic matter, and six experiments with conventional fertilizers (nitrogen, boron, and sulfur, two levels each). The phenolic compounds were analyzed by reversed-phase HPLC-DAD. External and internal leaves revealed distinct qualitative composition. In the internal leaves were found 15 phenolics (5 kaempferol and 10 cinnamic acid derivatives), whereas the external leaves presented 3- p-coumaroylquinic acid and 13 kaempferol derivatives. Principal component analysis (PCA) was applied to assess the relationships between phenolic compounds, agronomical practices, and harvesting time. Samples obtained with conventional practices were quite effectively separated from organic samples, for both types of leaves. In general, samples developed without any fertilization presented the highest phenolics amounts: external and internal leaves contained 1.4- and 4.6-fold more phenolic compounds than the ones that received conventional fertilizer, respectively, and the internal leaves presented 2.4 times more phenolics than the ones grown with organic amendment. Additionally, samples from organic production exhibited higher total phenolics content than those from conventional practices, collected at the same time. Samples harvested first were revealed to be distinct from the ones collected later. The results show that it is possible to grow tronchuda cabbage without excess fertilizers, with highest amounts of phenolics and reduced environment contamination.     

Tronchuda cabbage (Brassica oleracea L. var. costata DC): scavenger of reactive nitrogen species

The ability of tronchuda cabbage ( Brassica oleracea L. var. costata DC) to act as a scavenger of the reactive nitrogen species nitric oxide and peroxynitrite was investigated. The aqueous extracts obtained from tronchuda cabbage seeds and from its external and internal leaves exhibited a concentration dependent scavenging capacity. The antioxidant potential observed against the two reactive species was as follows: seeds > external leaves > internal leaves. In order to establish a possible correlation with the chemical composition of the extracts, the activity of ascorbic and sinapic acids and kaempferol 3- O-rutinoside was also studied. Among the compounds tested, sinapic acid showed the strongest antioxidant activity against both species.     

Free amino acids of tronchuda cabbage (Brassica oleracea L. Var. costata DC): influence of leaf position (internal or external) and collection time

The free amino acid profile of 18 samples of tronchuda cabbage ( Brassica oleracea L. var. costata DC) leaves, harvested at three different months, was determined by HPLC/UV-vis. The tronchuda cabbage leaves total free amino acid content varied from 3.3 to 14.4 g/kg fresh weight. Generally, arginine was the major compound, followed by proline, threonine, glutamine, cysteine, and glutamic acid. This study indicates that free amino acids are not similarly distributed: in external leaves, proline and arginine were the major free amino acids, while in internal ones, arginine was the main free amino acid, followed by threonine, glutamine, and cysteine. Significant differences were observed for valine, proline, arginine, leucine, cysteine, lysine, histidine, and tyrosine contents. The levels of some free amino acids were significantly affected by the collection period. In external leaves, this occurred with glutamic acid, serine, valine, leucine, cysteine, and ornithine contents, while in internal leaves, it occurred with aspartic acid, arginine, and total contents.     

Influence of two fertilization regimens on the amounts of organic acids and phenolic compounds of tronchuda cabbage (Brassica oleracea L. Var. costata DC)

A phytochemical study was undertaken on tronchuda cabbage (Brassica oleracea L. var. costata DC) cultivated under conventional and organic practices and collected at different times. Six organic acids (aconitic, citric, ascorbic, malic, shikimic, and fumaric acids) were identified and quantified by HPLC-UV. Qualitative and quantitative differences were noted between internal and external leaves. Analysis of the phenolics of the internal leaves was achieved by HPLC-DAD, and the phenolic profile obtained was revealed to be distinct from that of the external leaves. By this means were identified and quantified 11 compounds: 3-p-coumaroylquinic acid, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-(caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(sinapoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside, two isomeric forms of 1,2-disinapoylgentiobiose, 1-sinapoyl-2-feruloylgentiobiose, 1,2,2'-trisinapoylgentiobiose, and 1,2'-disinapoyl-2-feruloylgentiobiose. In general, internal leaves exhibited more constant chemical profiles.     

Chemical composition of cabbage (Brassica oleracea L. var. capitata) grown on acid sulfate soils

The purpose of this study was to investigate the influence of soil geochemistry on the concentrations of Ca, K, Mg, P, Co, Ni, Zn, Mn, Cu, and Fe in cabbage (Brassica oleracea L. var. capitata) grown on acid sulfate (AS) soils in Western Finland. A total of 11 topsoil (0–20 cm) and corresponding cabbage samples and three whole‐soil profiles (≈ 0–260 cm) were collected on three agricultural fields. The concentrations of Co and Zn in cabbage were correlated with the NH₄Ac‐extractable (easily available) concentrations in the topsoil, indicating that the uptake of these elements in cabbage is largely governed by soil geochemistry. Yet, the concentrations of Co and Zn in cabbage were not in general elevated relative to that of Finnish average values, although some AS soils showed enriched concentrations of these metals in the soil and cabbage. Significant geochemical differences (e.g., oxidation depth, organic‐matter and S content, pH) were observed among the studied AS soils, while, on the other hand, the concentrations of Ca, K, Mg, P, Ni, Mn, Cu, and Fe in cabbage were relatively similar. The hydroxylamine‐extractable concentrations of these elements in the topsoil were not correlated to those in cabbage, suggesting that uptake is not governed by the oxide‐bound fraction of these elements in the soil. Similarly, the easily available concentrations of Ca, P, Ni, Mn, Cu, and Fe in the topsoil were not correlated to those in cabbage, indicating that uptake is independent of the easily available concentrations in the soil. Hence, it is suggested that cabbage can regulate and thus optimize its concentrations of Ca, P, Ni, Mn, Cu, and Fe. Oxidation depth affected neither the easily available concentrations of Co, Ni, Zn, and Mn in the topsoil nor the concentrations in cabbage. However, the subsoil with a lower oxidation depth, which is to a smaller extent affected by leaching, may partly be enriched in these metals. Nevertheless, these showed no increased concentrations in cabbage. Based on these findings, it is suggested that the large amounts of metals mobilized in AS soils are easily lost to drains, subsequently contaminating nearby waterways and estuaries whereas they are only partly enriched in cabbage and other previously studied crops (oat).     

Enantioselective residue dissipation of hexaconazole in cucumber (Cucumis sativus L.), head cabbage (Brassica oleracea L. var. caulorapa DC.), and soils

In this study, the enantioselective dissipation behavior of hexaconazole was investigated in cucumber fruit, head cabbage, and two different types of agricultural soils. The dissipation kinetics was determined by reverse-phase liquid chromatography-tandem mass spectrometry on a cellulose tris (3-chloro-4-methylphenylcarbamate) chiral column. Dissipation rates of hexaconazole enantiomers followed first-order kinetics; the residues of (+)-enantiomer decreased more rapidly than (-)-enantiomer in cucumber and head cabbage, resulting in relative enrichment of the (-)-form, while the two enantiomers showed similar degradation rates in the tested soils. These results indicate substantial enantioselectivity in the residue dissipation of hexaconazole enantiomers in cucumber and head cabbage; however, nonenantioselective dissipation was observed in the tested soils.     

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.     

Erratum: Chemical composition of cabbage (Brassica oleracea L. var. capitata) grown on acid sulfate soils

The purpose of this study was to investigate the influence of soil geochemistry on the concentrations of Ca, K, Mg, P, Co, Ni, Zn, Mn, Cu, and Fe in cabbage (Brassica oleracea L. var. capitata) grown on acid sulfate (AS) soils in Western Finland. A total of 11 topsoil (0–20 cm) and corresponding cabbage samples and three whole‐soil profiles (≈ 0–260 cm) were collected on three agricultural fields. The concentrations of Co and Zn in cabbage were correlated with the NH₄Ac‐extractable (easily available) concentrations in the topsoil, indicating that the uptake of these elements in cabbage is largely governed by soil geochemistry. Yet, the concentrations of Co and Zn in cabbage were not in general elevated relative to that of Finnish average values, although some AS soils showed enriched concentrations of these metals in the soil and cabbage. Significant geochemical differences (e.g., oxidation depth, organic‐matter and S content, pH) were observed among the studied AS soils, while, on the other hand, the concentrations of Ca, K, Mg, P, Ni, Mn, Cu, and Fe in cabbage were relatively similar. The hydroxylamine‐extractable concentrations of these elements in the topsoil were not correlated to those in cabbage, suggesting that uptake is not governed by the oxide‐bound fraction of these elements in the soil. Similarly, the easily available concentrations of Ca, P, Ni, Mn, Cu, and Fe in the topsoil were not correlated to those in cabbage, indicating that uptake is independent of the easily available concentrations in the soil. Hence, it is suggested that cabbage can regulate and thus optimize its concentrations of Ca, P, Ni, Mn, Cu, and Fe. Oxidation depth affected neither the easily available concentrations of Co, Ni, Zn, and Mn in the topsoil nor the concentrations in cabbage. However, the subsoil with a lower oxidation depth, which is to a smaller extent affected by leaching, may partly be enriched in these metals. Nevertheless, these showed no increased concentrations in cabbage. Based on these findings, it is suggested that the large amounts of metals mobilized in AS soils are easily lost to drains, subsequently contaminating nearby waterways and estuaries whereas they are only partly enriched in cabbage and other previously studied crops (oat).     

Metabolism of the insecticide metofluthrin in cabbage (Brassica oleracea)

The metabolic fate of metofluthrin [2,3,5,6-tetrafluoro-4-(methoxymethyl)benzyl (E,Z)-(1R,3R)-2,2-dimethyl-3-(prop-1-enyl)cyclopropanecarboxylate] separately labeled with (14)C at the carbonyl carbon and the α-position of the 4-methoxymethylbenzyl ring was studied in cabbage ( Brassica oleracea ). An acetonitrile solution of (14)C-metofluthrin at 431 g ai ha(-1) was once applied topically to cabbage leaves at head-forming stage, and the plants were grown for up to 14 days. Each isomer of metofluthrin applied onto the leaf surface rapidly volatilized into the air and was scarcely translocated to the untreated portion. On the leaf surface, metofluthrin was primarily degraded through ozonolysis of the propenyl side chain to produce the secondary ozonide, which further decomposed to the corresponding aldehyde and carboxylic acid derivatives. In the leaf tissues, the 1R-trans-Z isomer was mainly metabolized to its dihydrodiol derivative probably via an epoxy intermediate followed by saccharide conjugation in parallel with the ester cleavage, whereas no specific metabolite was dominant for the 1R-trans-E isomer. Isomerization of metofluthrin at the cyclopropyl ring was negligible for both isomers. In this study, the chemical structure of each secondary ozonide derivative was fully elucidated by the various modes of liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) spectroscopy together with cochromatography with the synthetic standard, and their cis/trans configuration was examined by the nuclear Overhauser effect (NOE) difference NMR spectrum.     

Neglected landraces of collard (Brassica oleracea L. var. viridis) from the Carolinas (USA)

A common garden crop grown in the coastal plain region of North and South Carolina (United States) is the non-heading, leafy green type of Brassica oleracea L. known as collard (B. oleracea L. subsp. oleracea convar. acephala (DC.) Alef. var. viridis L.). Predominantly a fall and winter vegetable in this region, collard is often the only green planting to be found in the yard or garden of a rural home during these cool seasons. Historically, the traditional collard patch and even commercial fields were planted with unique varieties perpetuated by individual seed savers, and collectively, the regional diversity for this crop was probably very significant for well over a century. Genetic erosion of this collard germplasm pool has been severe in recent decades as commercial hybrids have been adopted by both large-scale producers and home gardeners. Although a significant number of collard landraces are being perpetuated to this day, existing diversity among landraces still grown in the region is now in the hands of an aging population of seed savers who maintain germplasm through on-farm preservation. From 2003 to 2006, we explored the coastal plain region of North and South Carolina in search of collard gardens containing traditional landraces. Exploration trips were conducted mid-winter to early spring. About 90 samples of collard were obtained from seed savers during the course of this exploration. Observations of morphological differences of these landraces indicate that significant diversity exists in this group. Obtained landraces are being deposited into the U.S. plant introduction collection and will be available for future use. This preserved collection could prove to be an important new source of genes for B. oleracea improvement.     

Brassica oleracea L. Var. costata DC and Pieris brassicae L. aqueous extracts reduce methyl methanesulfonate-induced DNA damage in V79 hamster lung fibroblasts

Brassica oleracea L. var. costata DC leaves and Pieris brassicae L. larvae aqueous extracts were assayed for their potential to prevent/induce DNA damage. None of them was mutagenic at the tested concentrations in the Ames test reversion assay using Salmonella His(+) TA98 strains, with and without metabolic activation. In the hypoxanthine-guanine phosphoribosyltransferase mutation assay using mammalian V79 fibroblast cell line, extracts at 500 μg/mL neither induced mutations nor protected against the mutagenicity caused by methyl methanesulfonate (MMS). In the comet assay, none of the extracts revealed to be genotoxic by itself, and both afforded protection, more pronounced for larvae extracts, against MMS-induced genotoxicity. As genotoxic/antigenotoxic effects of Brassica vegetables are commonly attributed to isothiocyanates, the extracts were screened for these compounds by headspace-solid-phase microextraction/gas chromatography-mass spectrometry. No sulfur compound was detected. These findings demonstrate that both extracts could be useful against damage caused by genotoxic compounds, the larvae extract being the most promising.     

Changes in broccoli (Brassica oleracea L. Var. italica) health-promoting compounds with inflorescence development

Changes in phenolic compounds, total glucosinolates, and vitamin C were monitored during the productive period along five inflorescence development stages of three broccoli commercial cultivars (Marathon, Monterrey, and Vencedor). In an attempt to identify differences due to agronomic factors, broccoli cultivars were grown under different sulfur fertilization with poor (15 kg/ha) and rich (150 kg/ha) rates. Phenolic compounds and vitamin C concentrations showed, in all broccoli cultivars, a rising trend from the first stage until the over-maturity stage, both for rich and poor sulfur fertilization. Significant differences were detected in the first two stages between rich and poor sulfur fertilization in total glucosinolates for all broccoli cultivars, where the highest concentration was always observed in the second development stage (used as minimally processed product) during poor fertilization. With regard to the last three stages, the glucosinolate concentration in the poor sulfur fertilization started to slope down until the over-maturity stage. Where rich sulfur fertilization is concerned, the highest level was reached during the third stage (used as minimally processed product also), and after that, glucosinolate concentration decreased until the fifth stage.     

Valorization of cauliflower (Brassica oleracea L. var. botrytis) by-products as a source of antioxidant phenolics

The present study reports the development of two extraction protocols, with potential industrial applicability, to valorize cauliflower (Brassica oleracea L. var. botrytis) byproducts as a source of antioxidant phenolics. In addition, the nonionic polystyrene resin Amberlite XAD-2 was used to obtain purified extracts. The extract yield, phenolic content, phenolic yield, and correlation between the antioxidant activity and the phenolic content were studied. The water and ethanol protocols yield a phenolic content of 33.8 mg/g freeze-dried extract and 62.1 mg/g freeze-dried extract, respectively. This percentage increased considerably when the extracts were purified using Amberlite XAD-2 yielding a phenolic content of 186 mg/g freeze-dried extract (water extract) and 311.1 mg/g freeze-dried extract (ethanol extract). Cauliflower byproduct extracts showed significant free radical scavenging activity (vs both DPPH(*) and ABTS(*)(+) radicals), ferric reducing ability (FRAP assay), and capacity to inhibit lipid peroxidation (ferric thiocyanate assay). In addition, the antioxidant activity was linearly correlated with the phenolics content. The results obtained indicate that the cauliflower byproducts are a cheap source of antioxidant phenolics very interesting from both the industrial point of view and the possible usefulness as ingredients to functionalize foodstuffs.     

Biochemical characterization of cystine lyase from broccoli (Brassica oleracea var. italica)

Cystine lyase is the enzyme responsible for off-aroma deterioration in fresh unblanched broccoli. In this research, cystine lyase purification from broccoli has been optimized. Only one protein peak with cystine lyase activity was found during purification. Broccoli cystine lyase was purified 100-fold to homogeneity. L-Cystine, L-cysteine-S-sulfate, L-djenkolic acid, and some S-alkyl-L-cysteines and their sulfoxides are substrates, but the enzyme had negligible activity with L-cystathionine. A K(m) value of 81.2 microM was found for L-cystine. Inhibition and K(i) determinations indicated that L-cysteine is a linear noncompetitive inhibitor with a K(i) of 5 mM and DL-homocysteine is a competitive inhibitor with a K(i) of 1.5 mM. The molecular weight of cystine lyase was determined to be 100 kDa by three methods, with two subunits of 48 kDa each and a carbohydrate content of 3%. Further characterization included cofactor quantification, the effects of temperature and pH on activity and stability, and amino acid composition.     

Kinetics of changes in glucosinolate concentrations during long-term cooking of white cabbage (Brassica oleracea L. ssp. capitata f. alba)

Brassica vegetables are the predominant dietary source of glucosinolates (GLS) that can be degraded in the intestinal tract into isothiocyanates, which have been shown to possess anticarcinogenic properties. The effects of pilot-scale long-term boiling on GLS in white cabbage (Brassica oleracea L. ssp. capitata f. alba cv. 'Bartolo') was experimentally determined and mathematically modeled. Cabbage was boiled, resulting in a dramatic decrease of 56% in the total GLS levels within the plant matrix during the first 2 min. After 8-12 min of boiling, the decrease progressed to over 70%. Progoitrin had an exceptionally higher decline rate in comparison to all other GLS. As boiling progressed the concentration of all GLS continued to decrease at a lower rate for the remaining cooking period. A mathematical model was used to describe the concentration profile of the GLS in the plant matrix, based on leaching of GLS to the water phase due to cell lysis and thermal degradation of the GLS both in the plant matrix and in the water phase. The model described the concentration profiles very well. Estimated lysis and degradation rate constants for white cabbage differed from those reported in the literature for red cabbage. The degradation rate constants found were significantly higher in the plant matrix when compared to those in the water phase for all GLS. Identification of the kinetics of decline of GLS during cooking can aid in designing processing and preparation methods and determining the conditions for the optimal effects of ingestion of Brassicaceae toward cancer prevention.     

effect of dose size on bioavailability of acylated and nonacylated anthocyanins from red cabbage (Brassica oleracea L. Var. capitata)

Recent studies indicate that anthocyanin intake conveys a variety of health benefits, which depend on absorption and metabolic mechanisms that deliver anthocyanins and their bioactive metabolites to responsive tissues. The anthocyanin bioavailability of red cabbage (Brassica oleracea L. var. capitata) was evaluated as reflected by urinary excretion of anthocyanins and anthocyanin metabolites. Twelve volunteers consumed 100, 200, and 300 g of steamed red cabbage (containing 1.38 micromol of anthocyanins/g of cabbage) in a crossover design. Anthocyanin concentration in cabbage extract and urine was measured by HPLC-MS/MS. Six nonacylated and 30 acylated anthocyanins were detected in red cabbage, and 3 nonacylated anthocyanins, 8 acylated anthocyanins, and 4 metabolites were present in urine. Mean 24 h excretion of intact anthocyanins increased linearly from 45 (100 g dose) to 65 nmol (300 g dose) for acylated anthocyanins and from 52 (100 g dose) to 79 nmol (300 g dose) for nonacylated anthocyanins. Urinary recovery of intact anthocyanins (percent of anthocyanin intake) decreased linearly from 0.041% (100 g dose) to 0.020% (300 g dose) for acylated anthocyanins and from 0.18% (100 g dose) to 0.09% (300 g dose) for nonacylated anthocyanins. Anthocyanin metabolites consisted of glucuronidated and methylated anthocyanins. The results show that red cabbage anthocyanins were excreted in both intact and metabolized forms and that recovery of nonacylated anthocyanins in urine was >4-fold that of acylated anthocyanins.     

Variation and population structure at microsatellite and isozyme loci in wild cabbage ( Brassica oleracea L.) in Dorset (UK)

We examined variation at four isozyme and seven microsatellite loci in natural populations of Brassica oleracea L. on the coast of Dorset. All loci were polymorphic, and the diversity index of isozyme loci was similar to that of the microsatellites. Both microsatellites and isozymes showed significant spatial differentiation of genetic variation, but the scale differed between the marker types. Significant non-random mating at isozyme loci was detected within small groups of about five plants, in which adjacent individuals were separated by less than 5 m. On the other hand, non-random mating at microsatellite loci was only detected within the Dorset region as a whole. F_ST was significant for all loci and there was evidence of isolation by distance at both microsatellite and isozyme loci. From these data we infer that there is low but significant amounts of gene flow among B. oleracea populations in Dorset. The differences in the genetic structure between the two types of marker may be due to higher mutation rates at the microsatellite loci; however, the data suggest that stepwise gain or loss of single repeat units is not the principal mechanism.     

花椰菜种质资源萌发期耐盐性综合评价

以98份花椰菜自交系资源为试材,测定了盐胁迫下种子的发芽率(GP)、发芽势(GR)、发芽指数(GI)、活力指数(VI)、苗高(SH)和根长(RL)等指标的耐盐系数,采用隶属函数法进行了耐盐性的综合评价。结果表明:花椰菜种质材料萌发期的耐盐性强弱评价结果受多个指标的影响。运用模糊数学隶属函数法,并赋予测定指标以相应的权重,计算出种质材料耐盐性强弱的综合评价D值,对98份种质材料耐盐性的强弱进行了综合评价和排序。综合评价D值与GP、GR、GI、VI、SH、RL的耐盐系数隶属函数值的相关性均达极显著水平(r=0.910**、0.921**、0.955**、0.972**、0.585**、0.686**),综合评价D值可以全面反映供试种质材料的耐盐性。分别对综合评价D值、GP、GR、GI、VI、SH、RL的隶属函数值进行聚类分析比较,发现GR、GI、VI、RL可以作为花椰菜种质材料萌发期耐盐性筛选的指标,而GP、SH不宜直接作为花椰菜种质材料萌发期耐盐性筛选的指标。基于综合评价D值聚类分析,可以将98份种质材料的耐盐性分为强、中、弱、差四大群类,其中有7份材料属于强耐盐性群类的种质,可供花椰菜耐盐性品种选育改良利用及耐盐性机制、耐盐遗传机理等方面研究。