Nitrous oxide emission factors for urine and dung from sheep fed either fresh forage rape (Brassica napus L.) or fresh perennial ryegrass (Lolium perenne L.)

In New Zealand, agriculture is predominantly based on pastoral grazing systems and animal excreta deposited on soil during grazing have been identified as a major source of nitrous oxide (N₂O) emissions. Forage brassicas (Brassica spp.) have been increasingly used to improve lamb performance. Compared with conventional forage perennial ryegrass (Lolium perenne L.), a common forage in New Zealand, forage brassicas have faster growth rates, higher dry matter production and higher nutritive value. The aim of this study was to determine the partitioning of dietary nitrogen (N) between urine and dung in the excreta from sheep fed forage brassica rape (B. napus subsp. oleifera L.) or ryegrass, and then to measure N₂O emissions when the excreta from the two different feed sources were applied to a pasture soil. A sheep metabolism study was conducted to determine urine and dung-N outputs from sheep fed forage rape or ryegrass, and N partitioning between urine and dung. Urine and dung were collected and then used in a field plot experiment for measuring N₂O emissions. The experimental site contained a perennial ryegrass/white clover pasture on a poorly drained silt-loam soil. The treatments included urine from sheep fed forage rape or ryegrass, dung from sheep fed forage rape or ryegrass, and a control without dung or urine applied. N₂O emission measurements were carried out using a static chamber technique. For each excreta type, the total N₂O emissions and emission factor (EF3; N₂O–N emitted during the 3- or 8-month measurement period as a per cent of animal urine or dung-N applied, respectively) were calculated. Our results indicate that, in terms of per unit of N intake, a similar amount of N was excreted in urine from sheep fed either forage rape or ryegrass, but less dung N was excreted from sheep fed forage rape than ryegrass. The EF3 for urine from sheep fed forage rape was lower compared with urine from sheep fed ryegrass. This may have been because of plant secondary metabolites, such as glucosinolates in forage rape and their degradation products, are transferred to urine and affect soil N transformation processes. However, the difference in the EF3 for dung from sheep fed ryegrass and forage rape was not significant.     

Organoselenides from Nicotiana tabacum genetically modified to accumulate selenium

Nicotiana tabacum L. (tobacco) plants were transformed to overexpress a selenocysteine methyltransferase gene from the selenium hyperaccumulator Astragalus bisulcatus (Hook.) A. Gray (two-grooved milkvetch), and an ATP-sulfurylase gene from Brassica oleracea L. var. italica (broccoli). Solvent extraction of leaves harvested from plants treated with selenate revealed five selenium-containing compounds, of which four were identified by chemical synthesis as 2-(methylseleno)acetaldehyde, 2,2-bis(methylseleno)acetaldehyde, 4-(methylseleno)-(2E)-nonenal, and 4-(methylseleno)-(2E,6Z)-nonadienal. These four compounds have not previously been reported in nature.     

含硫、硒化合物在油菜中的积累及其对硫甙水平的影响

温室条件下,对硫胁迫与供硫充足的油菜植株,分别用Ｌ－甲硫氨酸和硒酸盐（ＳｅＯ＾２－４）代替营养液中的硫酸协以及增加ＳＯ＾２－４供应浓度的方法,探讨了不同处理对油菜植株硫甙合成积累的影响,结果说明,增加供硫浓度可明显促进硫甙的合成速率,且在４８ｈ内以两个供硫水平的植株皆呈密切的二次回归递增趋势,ＳｅＯ＾２－４对植株甙的合成积累具有强烈的阻抑效应,并对供硫充足的植株影响更大,４８ｈ内呈直线下降,Ｌ－甲     

Beneficial endophytic microorganisms of Brassica – A review

Brassica species display enormous diversity and subsequently provide the widest assortment of products used by man from a single plant genus. Many species are important for agriculture, horticulture, in bioremediation, as medicines, soil conditioners, composting crops, and in the production of edible and industrial oils such as liquid fuels and lubricants. Many wild Brassica relatives possess a number of useful agronomic traits, including beneficial microbial endophytes that could be incorporated into breeding programs. Endophytes of Brassica, and/or their metabolites, have been demonstrated to improve and promote plant growth; increase yield; reduce disease symptoms caused by plant pathogens; reduce herbivory from insect pests; remove contaminants from soil; improve plant performance under extreme conditions of temperature and water availability; solubilise phosphate and contribute assimilable nitrogen to their hosts. Brassica napus (oilseed rape) and Brassica oleracea var. botrytis (broccoli and cauliflower) are the most economically important species of Brassica worldwide. These commercial crops are attacked by a wide range of pathogens and insect pests that are responsible for millions of dollars in lost revenue, with current control options offering little mitigation. No alternative control products are available for the Brassica industry, although it has been well documented in the literature that the use of endophytic microorganisms can offer beneficial traits to their host plants, including pest and disease resistance. The aim of this review is to describe the literature concerning beneficial microbial endophytes and their prospects to enhance or provide additional traits to their Brassica host species.     

Glucosinolates in <Emphasis Type="Italic">Brassica</Emphasis> foods: bioavailability in food and significance for human health

Glucosinolates are sulphur compounds that are prevalent in Brassica genus. This includes crops cultivated as vegetables, spices and sources of oil. Since 1970s glucosinolates and their breakdown products, have been widely studied by their beneficial and prejudicial biological effects on human and animal nutrition. They have also been found to be partly responsible for the characteristic flavor of Brassica vegetables. In recent years, considerable attention has been paid to cancer prevention by means of natural products. The cancer-protective properties of Brassica intake are mediated through glucosinolates. Isothyocianate and indole products formed from glucosinolates may regulate cancer cell development by regulating target enzymes, controlling apoptosis and blocking the cell cycle. Nevertheless, variation in content of both glucosinolates and their bioactive hydrolysis products depends on both genetics and the environment, including crop management practices, harvest and storage, processing and meal preparation. Here, we review the significance of glucosinolates as source of bioactive isothiocyanates for human nutrition and health and the influence of environmental conditions and processing mechanisms on the content of glucosinolate concentration in Brassica vegetables. Currently, this area is only partially understood. Further research is needed to understand the mechanisms by which the environment and processing affect glucosinolates content of Brassica vegetables. This will allow us to know the genetic control of these variables, what will result in the development of high quality Brassica products with a health-promoting activity.     

Lambs Fed Fresh Winter Forage Rape (Brassica napus L.) Emit Less Methane than Those Fed Perennial Ryegrass (Lolium perenne L.), and Possible Mechanisms behind the Difference

The objectives of this study were to examine long-term effects of feeding forage rape (Brassica napus L.) on methane yields (g methane per kg of feed dry matter intake), and to propose mechanisms that may be responsible for lower emissions from lambs fed forage rape compared to perennial ryegrass (Lolium perenne L.). The lambs were fed fresh winter forage rape or ryegrass as their sole diet for 15 weeks. Methane yields were measured using open circuit respiration chambers, and were 22-30% smaller from forage rape than from ryegrass (averages of 13.6 g versus 19.5 g after 7 weeks, and 17.8 g versus 22.9 g after 15 weeks). The difference therefore persisted consistently for at least 3 months. The smaller methane yields from forage rape were not related to nitrate or sulfate in the feed, which might act as alternative electron acceptors, or to the levels of the potential inhibitors glucosinolates and S-methyl L-cysteine sulfoxide. Ruminal microbial communities in forage rape-fed lambs were different from those in ryegrass-fed lambs, with greater proportions of potentially propionate-forming bacteria, and were consistent with less hydrogen and hence less methane being produced during fermentation. The molar proportions of ruminal acetate were smaller and those of propionate were greater in forage rape-fed lambs, consistent with the larger propionate-forming populations and less hydrogen production. Forage rape contained more readily fermentable carbohydrates and less structural carbohydrates than ryegrass, and was more rapidly degraded in the rumen, which might favour this fermentation profile. The ruminal pH was lower in forage rape-fed lambs, which might inhibit methanogenic activity, shifting the rumen fermentation to more propionate and less hydrogen and methane. The significance of these two mechanisms remains to be investigated. The results suggest that forage rape is a potential methane mitigation tool in pastoral-based sheep production systems.     

Effect of zinc on the biosynthesis of indole glucosinolates glucobrassicin and neoglucobrassicin in etiolated seedlings of rape (Brassica napus var.arvensis (Lam.) Thell)

The effect of Zn²⁺ ions (in the form of ZnCl₂) in the ceoncentration range 10^?3 to 10^?6 M on the content and biosynthesis of indole glucosinolates glucobrassicin and neoglucobrassicin has been studied on etiolated seedlings of rape (Brassica napus var.arvensis (Lam.) Thell). In the “long-term” experiment zine ions influenced the seedlings during eight days of germination, whereas in the “short-term” experiment zinc ions acted only 72 h on seven days old intact seedlings. The biosynthesis of indole glucosinolates has been followed by the incorporation of³⁵S from Na₂ ³⁵SO₄ into both glucosinolates in experiments with, hypocotyl segments of the rape seedlings. Zinc ions at chronic “long-term” application increased the glucobrassicin and neoglucobrassicin level in the seedlings. The neoglucobrassicin content especially was increased. A “short-term” application of zinc ions increased the level of both glucosinolates at higher and lower concentrations, whereas medium concentrations (10^?4 and 10^?5 M) lowered their level. Zn²⁺ ions lowered absorption of³⁵SO₄ ^?2 ions by hypocotyl segments and simultaneously lowered the incorporation of³⁵S into glucobrassicin. On the contrary, the incorporation of³⁵S into neoglucobrassicin and proteins was stimulated. Zinc ions do exhibit a specific effect on neoglucobrassicin biosynthesis, on membrane permeability as against sulphate ions and on the incorporation of sulphur into proteins.     

Toxicity of hydrolysis volatile products of Brassica plants to Sclerotinia sclerotiorum,in vitro

Oilseed rape stem rot disease caused by Sclerotinia sclerotiorum causes serious yield losses worldwide. Glucosinolates as specific secondary metabolites of Brassicaceae are produced in various parts of the host plants. Their enzymatic hydrolysis releases chemical components, particularly isothiocyanates, with fungitoxic activity and volatile characteristics. To investigate the effect of volatiles derived from Brassica tissues, the pathogen was exposed to hydrolysis products of Brassica shoot parts as sources of glucosinolates including oilseed rape varieties and two species, black and white mustard. The results showed significant differences in inhibition of S. sclerotiorum growth between varieties and species. All tissues of black mustard inhibited completely the exposed colonies of the pathogen and oilseed rape varieties Dunkeld, Oscar and Rainbow had significant inhibitory effect on the fungus. The genotypes demonstrated significant differences for the production of toxic volatiles, indicating that GSL contents in Brassica species and even cultivars have different potentials for toxic products.     

Photosynthesis and crop growth of spring oilseed rape and broccoli under elevated tropospheric ozone

The impact of a season-long exposure to moderately elevated tropospheric O₃ concentrations on the canopy growth and photosynthetic capacity of two important Brassica crops, spring oilseed rape and broccoli, was studied during three consecutive growing seasons (2007–2009). Brassica napus L. cv. Ability and Brassica oleracea L. cv. Monaco were exposed to non-filtered ambient air (NF) and non-filtered air with addition of 20 (NF+) and/or 40 ppb O₃ (NF++) in open-top chambers. Light saturated CO₂ assimilation (A_sat), stomatal conductance (g_st), maximum and actual quantum yield of photosystem II (F_v/F_m, F_v′/F_m′), performance index (PI) and leaf area index (LAI) were monitored on a weekly basis from emergence or planting until harvest. Before flowering, elevated O₃ did not have an influence on LAI nor on the photosynthetic capacity of the upper canopy leaves of either crops. This corresponded with the absence of a reduction of aboveground biomass of oilseed rape at maximum leaf area (MLA) and of broccoli plants harvested before flowering. After flowering, which coincided with MLA, the oilseed rape canopy showed a faster decline of LAI and of the chlorophyll content in NF+ compared to NF. In the NF++ treatment, this effect was intensified with an additional decrease of A_sat, g_st, F_v/F_m, F_v′/F_m′ and PI. In broccoli these detrimental O₃ effects were only detected in the lower canopy leaves. The changes in canopy development and photosynthetic performance of the upper canopy leaves can unravel the underlying mechanisms leading to the contrasting yield effects of O₃ on broccoli and spring oilseed rape that were previously reported (De Bock et al., 2011).     

Nutritional and phytochemical value of Brassica crops from the agri‐food perspective

Brassica foods are among the top 10 economic crops in the world (i.e. broccoli, kale, cauliflower and Chinese cabbage). These vegetables have been identified as important components of a healthy diet because of their high levels of nutrients and health‐promoting phytochemicals (i.e. phenolics, glucosinolates, vitamins and minerals). Epidemiological studies have shown that increased consumption of Brassica foods is strongly associated with a reduced risk of degenerative diseases, cancer, cardiovascular disease and immune dysfunction. Nevertheless, the nutritional content and profile in Brassica vegetables have been reported to vary considerably during the growth period due to agronomical factors including light, temperature, water availability and soil fertility among others. Moreover, the conditions of postharvest processing and cooking are also important factors on food quality. A better understanding of specific preharvest and postharvest conditions is essential to improve cultivars with value‐added nutritional quality. Thus, in this article are going to be addressed the effects of the most common crop management strategies and processes on the variation of nutritive compounds present within Brassica from the agri‐food perspective.     

Interaction of oxidized glutathione with allyl isothiocyanate

Isothiocyanates formed from glucosinolates in Brassica species have a strong affinity for amino acids and proteins, especially for their thiol, sulphide and terminal amino groups. To investigate the action of isothiocyanate on cystine residues in proteins and peptides, the present study on the interaction between allyl isothiocyanate and oxidized glutathione under physiological conditions was undertaken. Oxidized glutathione was oxidatively cleaved to some modified glutathiones by the attack of allyl isothiocyanate on its disulphide bond. Two new modified products were isolated from the reaction mixture by gel chromatography and HPLC, and their structures were determined by NMR and mass spectral analyses as glutathionyl N-allyldithiocarbaramate and its allyl thiohydantoin derivative. The formation of these products indicated oxidative cleavage of the disulphide bond in the cystine residue; the electrophilic attack of the isothiocyanate on the sulphur atom must cleave the disulphide bond oxidatively to dithiocarbamate and sulphenate, as in the case of cystine.     

Seed glucosinolates of fourteen wild Brassica species

Seed glucosinolates have been determined for 14 wild Brassica species, by micro-scale GC analysis of silylated derivatives. Of these, 12 were investigated for the first time. The majority of taxa exhibited high alkenylglucosinolate levels, although prop-2-enylglucosinolate appears to be generally absent. Other known methionine-derived glucosinolates predominate in B. tournefortii, B. elongata and B. deflexa. Phenylalanine-derived 4-hydroxybenzylglucosinolate is characteristic of section Brassicaria plants and represents the first finding of this glucosinolate in authenticated Brassica material.     

Changes in glucosinolates during crop development in single- and double-low genotypes of winter oilseed rape (Brassica napus): I. Production and distribution in vegetative tissues and developing pods during development and potential role in the recycling of sulphur within the crop

The distributions of glucosinolates and sulphur were measured in the vegetative and reproductive tissues in a series of single- and double-low cultivars of oilseed rape (Bienvenu, Ariana, Cobra and Capricorn) grown on a sulphur-sufficient soil at Rothamsted in 1987/88, and in crops of the cv. Libravo grown with none or 40 kg/ha of sulphur on a sulphur-deficient soil at Woburn in 1990/91. The glucosinolate measurements demonstrated large differences in the abilities of single- and double-low cultivars to synthesise glucosinolates, and showed that the biosynthetic differences were associated more with the developing pods than the vegetative tissues. It indicated that potential contribution of intact glucosinolates from vegetative tissues to the seed was likely to be small, but did not preclude the possibility that the vegetative tissues were a source of glucosinolate precursors. The sulphur measurements showed that the glucosinolates contained only a small proportion of the crop's total sulphur and that they were unlikely to be a major source of recyclable sulphur, even under conditions of severe sulphur deficiency.     

Ring oxygenated indole glucosinolates of Brassica species

Detailed chemical, degradative and spectroscopic analysis of two ring oxygenated indole glucosinolates isolated from Brassica species has confirmed these to be substituted in the 4- rather than the 5-position, although the latter had been suggested on biosynthetic grounds.     

Chemical composition and ultrastructure of the epicuticular wax in three lines of Brassica napus (L)

The epicuticular wax in three lines of Brassica napus (rape) has been investigated and the detailed chemistry and ultrastructure of the waxes examined. A distinct chemical make-up has been found for all three waxes which is correlated with three distinct crystallite structures. A tentative scheme for classification of Brassica wax mutants is described in which the two newly analysed rape mutants can be placed. Mass spectral analysis of all wax components confirms and extends previous ideas about the chemistry of Brassica waxes.     

Changes in the profile of flavonoid accumulation in Medicago truncatula leaves during infection with fungal pathogen Phoma medicaginis

Medicago truncatula is a model species for the study of the unique secondary metabolism in legumes. LC/MS/MS analysis was used to identify and profile flavonoid glycoconjugates and free aglycones in leaves of M. truncatula (ecotype R108-1) infected with the fungal pathogen Phoma medicaginis. Use of a high resolution analyzer with a collision induced dissociation tandem mass spectrometer (CID MS/MS) permitted structural elucidation of target secondary metabolites and four new acylated flavone glycosides have been identified. Changes in the phytoalexin medicarpin and its isoflavone precursors were quantitatively monitored at various time points after fungal spore application. Application of spores induced disease symptoms in the leaves of infected plants and resulted in an increase in the medicarpin precursors formononetin 7-O-glucoside and malonylated formononetin 7-O-glucoside between one and three days post-infection. Relative concentrations of medicarpin were highest five days post-infection. The rapid increase of these molecules was clearly positively correlated to the infection process as certain of them were absent in uninfected leaves, suggesting that the relative rate of their synthesis is tightly related with the infection process.     

Sulphur Deficiency Causes a Reduction in Antimicrobial Potential and Leads to Increased Disease Susceptibility of Oilseed Rape

The reduction of atmospheric sulphur dioxide pollution is causing increasing problems of sulphur deficiency in sulphur‐demanding crop plants in northern Europe. Elemental sulphur and many sulphur containing compounds such as cysteine‐rich antifungal proteins, glucosinolates (GSL) and phytoalexins play important roles in plant disease resistance. The aim of this work was to analyse the effect of inadequate sulphur supply on disease resistance of oilseed rape (Brassica napus). Compared with fertilized oilseed rape, healthy looking S‐deficient plants showed increased susceptibility to the blackleg fungus Leptosphaeria maculans, to the generalist necrotroph Botrytis cinerea and to the oomycete Phytophthora brassicae. To analyse possible causes of the increased disease susceptibility of S‐deficient plants, protein extracts and methanolic extracts of secondary metabolites of plants grown with and without adequate sulphur supply were tested for antimicrobial activity. None of the protein extracts showed antimicrobial activity. However, extracts containing secondary metabolites from normally grown plants showed a strong antimicrobial activity in in vitro tests with various fungal and bacterial pathogens. This activity was almost totally lost in extracts derived from S‐deficient plants. The antimicrobial activity did not appear to be based on the activity of phytoalexins because it was present in healthy plants and was not increased by a previous inoculation with Botrytis cinerea. The loss of antifungal activity in S‐deficient plants correlated with a strong reduction of various GSL, thus suggesting a reduced level of GSL as a possible cause of the reduced antimicrobial potential. However, limited tests of commercially available GSL or their degradation products did not demonstrate a causal link. Our results show that S‐deficiency of oilseed rape negatively affects disease resistance and suggest that this effect is at least partially caused by a reduction of sulphur‐dependent phytoanticipins.     

Development of an LC/MS/MS Method for Simultaneous Detection of 11 Polyphenols in Rat Plasma and Its Pharmacokinetic Application after Oral Administration of Coreopsis tinctoria Extract

Eleven polyphenols, classified as flavonoid glycosides, flavonoid aglycones, and phenolic acids, are important bioactive components in the capitula of Coreopsis tinctoria (CCT). Nevertheless, their full pharmacokinetic profiles have not been demonstrated simultaneously. Therefore, a liquid chromatography – tandem mass spectrometry (LC/MS/MS) method was developed in the present work and used it to study the pharmacokinetics of these 11 compounds. We performed LC/MS/MS with a gradient mobile phase composed of water containing 0.1 % formic acid and acetonitrile containing 0.1 % formic acid on a Proshell 120 SB C18 column (2.1 mm×100 mm, 2.7 μm). We achieved a good chromatographic peak shape, resolution, and mass signal response, and multiple reaction monitoring facilitated the simultaneous detection of 11 analytes. In addition, we validated the selectivity, correlation coefficient, precision, extraction recovery, matrix effects, and stability of the LC/MS/MS method to be acceptable for 11 analytes in rat plasma. Subsequently, rats were orally administered with 50 % ethanol eluent of CCT (ECCT). Nine of 11 polyphenols were absorbed quickly (except for QCD and TCA), and their plasma levels peaked within 40 min. The exposure and C_max values of flavonoid glycosides and phenolic acids were lower than those of flavonoid aglycones. This is the first report to demonstrate the pharmacokinetics of 11 polyphenols in ECCT, which may play an important role in future studies of the bioactive components of ECCT and their bioactive mechanisms.     

Development of isothiocyanate-enriched broccoli,and its enhanced ability to induce phase 2 detoxification enzymes in mammalian cells

Broccoli florets contain low levels of 3-methylsuphinylpropyl and 4-methylsulphinylbutyl glucosinolates. Following tissue disruption, these glucosinolates are hydrolysed to the corresponding isothiocyanates (ITCs), which have been associated with anticarcinogenic activity through a number of physiological mechanisms including the induction of phase II detoxification enzymes and apoptosis. In this paper, we describe the development of ITC-enriched broccoli through the introgression of three small segments of the genome of Brassica villosa, a wild relative of broccoli, each containing a quantitative trait locus (QTL), into a broccoli genetic background, via marker-assisted selection and analysis of glucosinolates in the florets of backcross populations. Epistatic and heterotic effects of these QTLs are described. The ITC-enriched broccoli had 80-times the ability to induce quinone reductase (a standard assay of phase II induction potential) when compared to standard commercial broccoli, due both to an increase in the precursor glucosinolates and a greater conversion of these into ITCs.