Phenolic compounds in external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC)

Glycosylated kaempferol derivatives from the external leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC) characterized by reversed-phase HPLC-DAD-MS/MS-ESI were kaempferol 3-O-sophorotrioside-7-O-glucoside, kaempferol 3-O- (methoxycaffeoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside-7-O-sophoroside, kaempferol 3-O-sophoroside-7-O-sophoroside, kaempferol 3-O-tetraglucoside-7-O-sophoroside, kaempferol 3-O-(sinapoyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl/caffeoyl)sophoroside-7-O-glucoside, kaempferol 3-O-sophorotrioside, kaempferol 3-O-(sinapoyl)sophoroside, kaempferol 3-O-(feruloyl)sophorotrioside, kaempferol 3-O-(feruloyl)sophoroside, kaempferol 3-O-sophoroside, and kaempferol 3-O-glucoside. These acylated derivatives are reported for the first time in nature, with the exception of kaempferol 3-O-(sinapoyl)sophoroside. Quantification of the identified compounds was achieved by HPLC-DAD and carried out in samples cultivated under conventional or organic practices and collected at different times. In general, samples from organic production exhibited higher total phenolics content than those from conventional practices collected in the same period.     

Protein from red cabbage (Brassica oleracea) seeds with antifungal, antibacterial, and anticancer activities

A 30 kDa antifungal protein was purified from red cabbage ( Brassica oleracea ) seeds. It exhibited a molecular mass and N-terminal amino acid sequence disinct from those of previously isolated Brassica antifungal proteins. The protocol used entailed ion exchange chromatography on Q-Sepharose and SP-Sepharose followed by fast protein liquid chromatography on Mono S. The protein hindered mycelial growth in Mycosphaerella arachidicola (with an IC50=5 μM), Setospaeria turcica, and Bipolaris maydis. It also inhibited the yeast Candida albicans with an IC50=96 μM. It exerted its antifungal action by permeabilizing the fungal membrane as evidenced by staining with Sytox green. The antifungal activity was stable from pH 3 to 11 and from 0 to 65 °C. It manifested antibacterial activity against Pseudomonas aeruginosa (IC50=53 μM). Furthermore, after 48 h of culture, it suppressed proliferation of nasopharyngeal cancer and hepatoma cells with IC50=50 and 90 μM, respectively.     

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.     

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).     

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.     

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.     

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).     

Use of human urine fertilizer in cultivation of cabbage (Brassica oleracea)--impacts on chemical, microbial, and flavor quality

Human urine was used as a fertilizer in cabbage cultivation and compared with industrial fertilizer and nonfertilizer treatments. Urine achieved equal fertilizer value to industrial fertilizer when both were used at a dose of 180 kg N/ha. Growth, biomass, and levels of chloride were slightly higher in urine-fertilized cabbage than with industrial-fertilized cabbage but clearly differed from nonfertilized. Insect damage was lower in urine-fertilized than in industrial-fertilized plots but more extensive than in nonfertilized plots. Microbiological quality of urine-fertilized cabbage and sauerkraut made from the cabbage was similar to that in the other fertilized cabbages. Furthermore, the level of glucosinolates and the taste of sauerkrauts were similar in cabbages from all three fertilization treatments. Our results show that human urine could be used as a fertilizer for cabbage and does not pose any significant hygienic threats or leave any distinctive flavor in food products.     

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.     

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.     

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.     

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.     

Characterisation of wild cabbage (Brassica oleracea L.) based on isoenzyme data. Considerations on the current status of this taxon in Spain

As a part of a wide project to determine the conservation status and collecting expeditions of wild plant genetic resources of genus Brassica (species n = 9) of the Iberian Peninsula, twenty-nine wild populations of Brassica oleracea L. were characterised using the isoenzymatic profiles of seeds collected from their natural habitats. The zymograms, obtained using bulking seeds, identified samples faster than other methodologies, which could be useful in seed bank management as well as the regular monitoring of the status of in situ conservation. Five isoenzymatic systems were analysed considering the electrophoretic phenotype of the whole population (not individuals). Using enzymatic bands as qualitative characters, data were evaluated using phenetic clustering methods unweighted pair group method using arithmetic averages. The enzyme systems showed high polymorphism levels, and each studied population of B. oleracea showed a characteristic zymogram in each of the assayed enzyme systems. The clustering analysis using the Dice coefficient showed that most of the studied populations can be considered significantly different from each other. This study proposes a rapid screening strategy for obtaining the electrophoretic profiles of samples and comparing population patterns. Two main of conclusions can be drawn from the results of this work: (1) compared to other molecular techniques and the efficiency and reliability of isoenzyme analyses, this method appears to be a useful tool for a rapid characterisation of samples; (2) in situ observations of populations suggest that conservation status of wild B. oleracea in their natural habitat in Spain can be considered satisfactory.     

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.     

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.     

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.     

Critical Levels of Boron in Soils for Cauliflower (Brassica oleracea Var. Botrytis)

Influence of boron (B) application to cauliflower (Brassica oleracea var. botrytis) was investigated in a pot experiment taking 15 Inceptisols with four levels of B. The critical levels of B for deficiency, adequacy and toxicity in soil and in cauliflower plant were also determined. Hot-calcium chloride (CaCl₂) extractable B in these soils varied from 0.33 to 0.78 mg kg^-1 and its content for deficiency to cauliflower was 0.48 mg kg^-1. Boron application significantly increased cauliflower yield, plant B concentration and uptake of B. The critical plant B concentrations for deficiency, sufficiency and toxicity varied with the growth stages and the values being 26, 31 and 48 mg kg^-1 at 50 days of growth and 17, 24 and 35.5 mg kg^-1 at harvest, respectively. The study also recommends application of fertilizer B at the rate 0.9–4.5 kg ha^-1 for optimum B nutrition to cauliflower in Inceptisols of the Gangetic plains of India.     

Protective effect of selenium in Broccoli (Brassica oleracea) plants subjected to cadmium exposure

The protective effect of selenium against the cadmium-induced oxidative effect in broccoli ( Brassica oleracea) plants was studied. Plants grown in hydroponic culture were supplied with selenium [as Se(IV)] and cadmium [as Cd(II)], individually or simultaneously. Cadmium accumulation in roots was noticeably higher than in the aerial parts of the plants, and this effect was even more acute when selenium was simultaneously added. Cadmium phytotoxicity was evidenced by an increase in the malondialdehyde (MDA) concentration in the roots and a decrease of photosynthetic pigment and tocopherol concentration in the aerial parts of the plant. The simultaneous addition of selenium alleviated cadmium-induced stress in the roots after 40 days of exposition. In the leaves, a more remarkable decrease of tocopherol and chlorophyll concentration was observed in the cadmium-enriched plants after 10 days of exposure. The results provided evidence that selenium supplementation helps the plant to minimize the cadmium oxidant effect. Tocopherol concentration in broccoli fruit of cadmium-supplied plants was not affected in comparison to control. However, the proportion of alpha-tocopherol increases with the addition of selenium. This response is important not only for the protective effect against oxidative damage in the plant but also in terms of human nutrition.