Effects of elicitors on the production of resveratrol and viniferins in cell cultures of Vitis vinifera L. cv Italia

Methyl jasmonate, jasmonic acid and chitosan were tested as elicitors on cell suspension cultures obtained from Vitis vinifera cv Italia to investigate their effect on stilbene production. Stilbene accumulation in the callus, grown under nonelicited conditions, was also investigated. Calli and cell suspensions were obtained in a B5 culture medium supplemented with 0.2 mg L(-1) NAA and 1 mg L(-1) KIN. Stilbene determination was achieved by HPLC/DAD/MS. Whereas callus biosynthesized only piceid, cell suspensions elicited with jasmonates produced several stilbenes, mainly viniferins. In suspended cells, methyl jasmonate and jasmonic acid were the most effective in stimulating stilbene biosynthesis, whereas chitosan was less effective; in fact, the amount of stilbenes obtained with this elicitor was not significantly different from that obtained for the control cells. The maximum production of total stilbenes was at day 20 of culture with 0.970 and 1.023 mg g(-1) DW for MeJA and JA, respectively.     

Mulberry leaf polyphenols possess antiatherogenesis effect via inhibiting LDL oxidation and foam cell formation

Oxidized low-density lipoprotein (ox-LDL) and its uptake by machrophage are the hallmark in atherogenesis. In the present study, we aimed to investigate the antiatherogenic effect of mulberry leaf extracts (MLE) and the polyphenolic extracts (MLPE), which contained polyphenols including quercetin (11.70%), naringenin (9.01%) and gallocatechin gallate (10.02%). Both MLE and MLPE inhibited the oxidation and lipid peroxidation of LDL, while MLPE was shown to be more potent. As 1.0 mg/mL MLE reduced 30% of ox-LDL-generated ROS, 0.5 mg/mL MLPE decreased 46% of the ROS and was shown to be more potent on elevating SOD-1 and GPx in macrophages. At the same dose of 0.5 mg/mL, MLPE exhibited 1.5-fold potency than MLE in decreasing the formation of foam cells. Both MLE and MLPE reduced the expression of PPARγ, CD36 and SR-A, implicating the molecular regulation on ox-LDL uptake. These results suggested that MLPE potentially could be developed as an antiatherogenic agent and deserve further investigation.     

Plant secondary metabolites, biphenyl, and hydroxypropyl-β-cyclodextrin effects on aerobic polychlorinated biphenyl removal and microbial community structure in soils

Biphenyl and several plant secondary metabolites (PSM) (carvone, isoprene, limonene, naringin, and coumarin) and surfactant (hydroxypropyl-β-cyclodextrin, (HP-β-CD)) have been shown to improve aerobic polychlorinated biphenyl (PCB) removal by several bacterial species. The objective of this study was to determine whether these treatments also affect PCB removal and microbial community structure in a high organic matter soil (Pahokee soil series with 67% organic matter) and low organic matter soil (Woolper soil series with 6% organic matter), as determined by monitoring changes in PCB levels and phospholipid fatty acids (PLFA) profiles in laboratory microcosms amended with these compounds. Biphenyl enhanced di-chlorinated and tri-chlorinated biphenyl removal in both soils, but PSM did not improve removal of these congeners. On the contrary, HP-β-CD decreased PCB removal when used in combination with biphenyl. Two-way analysis of variance indicated that HP-β-CD significantly increased tetra- and penta-chlorinated biphenyl removal from the high organic matter soil, but not from the low organic matter soil. Principal components analysis of PLFA data indicated that HP-β-CD increased proportions of 18:1ω7c associated with Gram-negative bacteria, but decreased 10me16 and 10me17 lipid associated with Gram-positive bacteria, while biphenyl and PSMs had no detectable effects on soil microbial communities. PCB removal was not correlated to any PLFA. In conclusion, PSM previously shown to enhance PCB removal in soil-free systems were not effective in two divergent soils evaluated in this study, and HP-β-CB had increase, decrease, or no effect on PCB removal depending on types of PCB congeners, soils, and co-amendments.     

中国耕地等别分异特性及其对耕地保护的影响

为了深刻地认识耕地质量,科学地进行耕地质量保护与建设,基于中国耕地质量等级调查与评定工作成果及1:50万国家级农用地分等汇总数据库,对比分析了中国省域的耕地质量等别分异特性。该文分析确定了“优等地分布范围小,低等地分布范围广,省际间及省内耕地等别分异规律复杂”的中国耕地等别基本特征;应用标准亩的概念,以全国1等耕地和广东省平均等5.4等耕地为基准,核算出中国标准耕地总量分别为45?750.7?khm2和68?390.2?khm2;并结合全国土地利用总体规划给定的2020年各省基本农田保护面积,提出了划定永久基本农田要统筹考虑基本农田保护率的法律要求和各省耕地质量等别实际。基于上述研究结论,建议在土地整治规划、基本农田调整划定、高标准基本农田建设和永久基本农田保护中都要坚持数量质量并重,以耕地等别提升和产能建设为核心,建立动态监测和报告制度。     

Protein haze formation in white wines: effect of saccharomyces cerevisiae cell wall components prepared with different procedures

Cell wall material was extracted by five different methods from an oenological strain of Saccharomyces cerevisiae. Enzyme preparations containing beta-glucanase activity (Zymolyase, Glucanex, and Finizym 250L) allowed a better extraction yield compared to that of dithiothreitol (DTT) and EDTA. The yeast extracts were only soluble in part in wine. The wine-soluble fraction (WSF) of the five extracts, differing in both protein and sugar contents, when added in increasing amounts to white wine differently affected protein haze formation, as determined by the heat test, giving dose/response curves of different shapes. The curves obtained with the WSF derived from DDT and Zymolyase extracts showed a plateau value corresponding to 90% and 80% of wine haze reduction, respectively. In contrast, addition of the WSF derived from the other extracts resulted in increased turbidity with respect to the original wine. The mannoproteins (MP), isolated from each yeast extract by Concanavalin-A chromatography, gave dose/response curves showing shapes more similar among them than those obtained with the whole WSFs. The best wine stabilization was obtained with the MP of the DTT and Zymolyase extracts. The supernatants obtained after heating the MP of the different extracts were also tested. The stabilizing effect of the heat-stable MP (HSMP) was always larger than that of the corresponding total (un-heated) MP. The HSMP obtained starting from the DTT and Zymolyase extracts showed the best haze-protecting effect, which was, however, lower than that obtained with their corresponding WSF. This result suggests that wine protein stabilization by compounds of the yeast cell wall could be related, in addition to the action of the MP, also to the presence of other substances of different nature.     

Growth and anthraquinone production of Morinda elliptica cell suspension cultures in a stirred-tank bioreactor

The effects of medium strategy, number of impellers, aeration mode, and mode of operation on Morinda elliptica cell suspension cultures in a stirred-tank bioreactor are described. A lower number of impellers and continuous aeration contributed toward high cell growth rate, whereas a higher number of impellers reduced cell growth rate, although not anthraquinone yield. The semicontinuous mode could indirectly imitate the larger scale version of production medium strategy and improved anthraquinone production even with 0. 012% (v/v) antifoam addition. Production medium promoted both growth (maximum dry cell weight of 24.6 g/L) and anthraquinone formation (maximum content of 19.5 mg/g of dry cell weight), without any necessity for antifoam addition. Cultures in production medium or with higher growth rate and anthraquinone production were less acidic than cultures in growth medium or with lower growth rate and anthraquinone production. Using the best operating variables, growth of M. elliptica cells (24.6 g/L) and anthraquinone yield (0.25 g/L) were 45% and 140%, respectively, lower than those using a shake flask culture after 12 days of cultivation.     

Influence of abiotic stress on the GSH/GSSG system of plant cell cultures

Cell suspension cultures of various crop plants have been used for studying salt and drought stress on the GSH/GSSG-system, As already recorded for whole plants, cell cultures of different species vary in their glutathione levels. In general, cell cultures of species with relatively high glutathione content are more tolerant to abiotic stress. A high glutathione content alone is not adequate to prevent cell damage and a decrease in cell growth. E.g. the addition of sulphur to stressed cell cultures slightly enhances the GSH content but does not enhance the adaptation of these cells to stress. Thus, the role of glutathione in the adaptation of cells to stress situations seems to be only one component in a more complex response mechanism.     

Inhibitory effect of yuzu essential oil on the formation of N-nitrosodimethylamine in vegetables

The inhibitory effect of yuzu (Citrus junos Tanaka) essential oil on the formation of N-nitrosodimethylamine (NDMA) in the presence of vegetables (31 species) or saliva was investigated by HPLC. Most vegetable extracts enhanced the formation of NDMA. However, the formation ratio of NDMA in vegetable extracts was decreased by yuzu oil in the range of 59 to 22%. In the presence of yuzu oil and saliva, its ratio ranged between 62 and 24%. These results indicated that yuzu oil inhibited the formation of NDMA even in vegetables and saliva. The contents of ascorbic acid, nitrate, and nitrite in the 31 vegetable species were 0.3-65 mg/100 g, 3-581 mg/100 g, and 10-750 microg/100 g, respectively. Ascorbic acid and nitrite had little effect on the inhibition or formation of NDMA at their intact levels. Nitrate accelerated the formation of NDMA, and the addition of saliva further enhanced it. The mechanism of inhibition of NDMA formation by alpha-terpinene was studied. It was assumed from the results of LC-MS that a new compound formed by the reaction of alpha-terpinene with nitrite would be a derivative of alpha-terpinene with dinitroso groups. The molecular weight of this compound was 194. It is suggested that terpene hydrocarbons in citrus essential oils would contribute to the decrease of NDMA formation.     

Differential effect of afforestation on nitrogen-fixing and denitrifying communities and potential implications for nitrogen cycling

Denitrification and nitrogen (N) fixation are two major processes which drive N transformation in ecosystems. We investigated the effect of afforestation of pasture in New Zealand on denitrifying and N-fixing communities using molecular techniques. We optimised and applied multiplex-terminal restriction fragment length polymorphism for simultaneous analysis of denitrifying and N-fixing communities. Soil samples from four pine (Pinus radiata) forests of various age and adjacent pastures were analysed for microbial communities, denitrification rate and some environmental variables. Analysis of variance revealed that the denitrifying community was mainly influenced by site while the N-fixing community was influenced by both site and land-use types. Among environmental variables, moisture and porosity had the strongest impact on denitrifying communities while C:N ratio explained most of the variability in the N-fixing community. Denitrification rate as measured by nitrous oxide (N₂O) flux, was significantly lower in older forest soils compared to adjacent pasture soils. Simple and step-wise regression analyses revealed that the N₂O flux rate was strongly linked to the denitrifying community composition, providing evidence for the influence of community composition on denitrification. Moreover, we explored the relationship between denitrifying and N-fixing communities using various statistical approaches. Our results for the first time showed that denitrifying and N-fixing communities are related in situ, and suggest that changes in land-use may have a significant impact on N-transformations and ecosystem processes.     

Impact of harvest year on amino acids and sugars in potatoes and effect on acrylamide formation during frying

Acrylamide is formed via the Maillard reaction between reducing sugars and asparagine in a number of carbohydrate-rich foods during heat treatment. High acrylamide levels have been found in potato products processed at high temperatures. To examine the impact of harvest year, information on weather conditions during growth, that is, temperature, precipitation, and light, was collected, together with analytical data on the concentrations of free amino acids and sugars in five potato clones and acrylamide contents in potato chips (commonly known as crisps in Europe). The study was conducted for 3 years (2004-2006). The contents of acrylamide precursors differed between the clones and the three harvest years; the levels of glucose were up to 4.2 times higher in 2006 than in 2004 and 2005, and the levels of fructose were 5.6 times higher, whereas the levels of asparagine varied to different extents. The high levels of sugars in 2006 were probably due to the extreme weather conditions during the growing season, and this was also reflected in acrylamide content that was approximately twice as high as in preceding years. The results indicate that acrylamide formation is dependent not only on the content and relative amounts of sugars and amino acids but also on other factors, for example, the food matrix, which may influence the availability of the reactants to participate in the Maillard reaction.     

Effect of chemical systemic acquired resistance elicitors on avenanthramide biosynthesis in oat (Avena sativa)

Oats produce a group of phenolic antioxidants termed avenanthramides. These metabolites are, among food crops, unique to oats and have shown, in experimental systems, certain desirable nutritional characteristics such as inhibiting atherosclerotic plaque formation and reducing inflammation. Avenanthramides occur in both the leaves and grain of oat. In the leaves they are expressed as phytoalexins in response to crown rust (Puccina coronata) infection. The experiments reported here demonstrate that avenanthramide levels in vegetative tissue can be enhanced by treatment with benzothiadiazole (BTH), an agrochemical formulated to elicit systemic acquired resistance (SAR). The response to BTH was dramatically stronger than those produced with salicylic acid treatment. The roots of BTH treated plants also showed a smaller but distinct increase in avenanthramides. The dynamics of the root avenanthramide increase was substantially slower than that observed in the leaves, suggesting that avenanthramides might be transported from the leaves.     

Inhibitory effect of fruit extracts on the formation of heterocyclic amines

Natural extracts have attracted considerable attention for development into effective inhibitors against the formation of genotoxic heterocyclic amines (HAs) in processed foods. In this study, four fruit extracts (apple, elderberry, grape seed, and pineapple) were evaluated for their effects on HA formation in fried beef patties. Apple and grape seed extracts were found to be the most effective in both the degree of inhibition in the formation of individual HAs (2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx), and 2-amino-1-methyl-6-henylimidazo [4,5-b]pyridine (PhIP)) and in the reduction of total HA content (approximately 70% relative to the control). Activity-guided analysis of apple extract using model systems (PhIP- and MeIQx-producing models) showed that the proanthocyanidins, phloridzin and chlorogenic acid were responsible for reducing the amount of HAs formed. Proanthocyanidins were identified as the dominant inhibitors because they were strongly active against HA formation in both the PhIP and MeIQx model systems. For phloridzin, the inhibitory effect was observed only on the formation of PhIP. In contrast, chlorogenic acid, although effective against the formation of MeIQx, significantly enhanced the formation of PhIP. This is the first report showing the inhibitory activities of apple phenolics on the formation of heterocyclic amines. The findings provide valuable information for the development of effective strategies to minimize HA content of cooked meats and to identify several new natural products that may have new applications in the food industry.     

Effect of environmental conditions on polychlorinated biphenyl transformations and bacterial communities in a river sediment

Purpose  

The aim of this study was to evaluate polychlorinated biphenyl (PCB) removal in relation to the associated bacterial community composition in Ohio River sediments (USA) using field and laboratory approaches.     

The influence of phosphorus nutrition on growth,chlorophyll formation and 14CO2 fixation of chlorophyllous carrot callus cultures

Freshly isolated carrot root sec. phloem explants were cultured in nutrient solution plus inositol, IAA and kinetin and a varied supply of inorganic phosphorus. ¹⁴CO₂ fixation experiments with the greened cultures were carried out under steady state conditions at the age of 21 days in light and darkness for four minutes. The results show that photosynthetic CO₂ fixation by the Calvin cycle is greatly reduced in phosphorus deficient cultures whereas CO₂ fixation via PEPC is enhanced. Under phosphorus deficiency the export of primary photosynthetates from the chloroplasts seems to be suppressed while the radioactivity of the labelled insoluble fraction, which represents mainly chloroplastic starch after this short time of ¹⁴CO₂ fixation, was increased. The results are discussed with respect to the influence of phosphorus nutrition on photosynthetic CO₂ fixation and PEP carboxylation in carrot tissue cultures.     

Biosynthesis and characterization of 14C-enriched flavonoid fractions from plant cell suspension cultures

A range of radiolabeled anthocyanins, proanthocyanidins, and other flavonoids were accumulated by cell suspension cultures of two plant species, ohelo (Vaccinium pahalae) and grape (a Vitis hybrid, Bailey Alicant A), after providing uniformly labeled [(14)C]sucrose to the medium. Approximately 15% of administered label was recovered in a series of flavonoid-rich fractions varying in composition. Anthocyanins, and monomers to oligomers of proanthocyanidins, were labeled effectively and characterized from both species. Most of the proanthocyanidin oligomers were based on the flavan-3-ols (+)-catechin and (-)-epicatechin. Cyanidin and peonidin glycosides were the dominant forms of anthocyanins in both species. Whereas the predominant form of flavonoids identified from ohelo cell cultures was proanthocyanidins, grape cell cultures produced mostly anthocyanins. The labeled phytochemicals were produced for use in subsequent in vivo animal feeding studies to gauge their bioavailability and accumulation in target organs.