Phenolic constituents and antioxidant properties of Xanthosoma violaceum leaves

An extract of Xanthosoma violaceum leaves was subjected to a polyphenol profile determination, including total polyphenols, and antioxidant activity evaluation. Analysis of the extract resulted in the isolation of a new flavone C-glycoside, apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-apiofuranoside (1), as well as known flavone C-glycosides, including vitexin (2), isovitexin (3), isovitexin 4'-O-rhamnopyranoside (4), apigenin 6-C-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside] (5), and apigenin 6,8-diC-beta-D-glucopyranoside (6). The antioxidant activity of the extract was assessed by means of two different in vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and peroxidation induced by the water-soluble radical initiator 2,2'-azobis(2-amidinopropane) hydrochloride, on mixed dipalmitoylphosphatidylcholine/linoleic acid unilamellar vesicles (LP-LUV test). In both tests used, the extract and a fraction II showed a significant antioxidant/free-radical scavenging effect (fraction II, EC(50) = 11.6 microg/mL) in comparison to alpha-tocopherol (EC(50) = 10.1 microg/mL).     

Multielement composition of wines and their precursors including provenance soil and their potentialities as fingerprints of wine origin

The influence of the provenance soil and vinification process on the wine multielemental composition was investigated. For this purpose, two different vineyards from the Douro wine district, Portugal, were selected. Monovarietal grapes from a 10 year old vineyard were used to produce a red table wine, in a very modern winery. Polyvarietal grapes from a 60-70 year old vineyard were used to produce a red fortified wine, similar to Port, through a traditional vinification process. The multielement compositions (Al, As, B, Ba, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, Hf, Li, Mn, Mo, Nb, Ni, Pb, Rb, Sb, Sc, Sr, Ti, Th, Tl, U, V, W, Y, Zn, Zr, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, and Lu) of soil, grape juices (prepared in the laboratory), and samples collected in the different steps of each winemaking process were measured. Inductively coupled plasma mass spectrometry was used, after suitable pretreatment of the samples (by UV irradiation for liquid samples and high-pressure microwave digestion for soil). Both vinification processes influenced the multielement composition of the wines. Most of the elements presented similar or even lower concentrations in the wine as compared to that observed in the respective grape juice, probably as a result of precipitation or coprecipitation with suspended particles during fermentation and/or wine aging. Evidence of effective contamination during grape pressing, fermentation, and/or fining of wines (depending on the element) was observed for Cd, Cr, Cu, Fe, Ni, Pb, V, and Zn in the fortified wine and Al, Cr, Fe, Ni, Pb, and V in the table wine. Nevertheless, significant correlations were obtained between the multielement composition of the wine and the respective grape juice (R = 0.997 and 0.979 for the fortified and table wines, respectively, n = 31, P < 0.01), as well as between that in the wine (median of the two studied wines) and the provenance soil (R = 0.994, n = 19, P < 0.01), for the set of elements determined in common in the different types of samples. These results are promising concerning the usefulness of the elemental patterns of both soil and wine as fingerprints of the origin of the studied wines. Nevertheless, more wines from the same and other wine districts must be studied in order to consolidate this conclusion. The multielement compositions of the studied wines were compared with those of wines of different characteristics and origins, as well as with the respective legal threshold limit values, when available. Relatively low metal levels, below their threshold limit values, were found in all cases.     

Semiarid soils submitted to severe drought stress: influence on humic acid characteristics in organic-amended soils

Purpose  

The objective of this work was to assess the effect of severe drought conditions on the characteristics of semiarid soil humic acids (HAs) as well as the effect of organic amendment on such changes.     

Comparison between phenetic characterisation using RAPD and ISSR markers and phenotypic data of cultivated chestnut (Castanea sativa Mill.)

Patterns of phenotypic and phenetic variability in sixPortuguese cultivars of chestnut (Castanea sativa Mill.) are evaluated. Morphological characterisationwas based on the quantification of seventeen traits. Varianceanalysis showed significant differences among cultivars, and cultivar× year for all the traits studied, and trees within cultivarsshowed also some significant differences for some of themorphological variables. A significant correlation was obtainedbetween length of the leaf blade and the percentage of unisexual andandrogynic inflorescence with the effective thermal index,accumulated rainfall from April to October and from July to October,or the accumulated temperature below seven during the dormant period.Principal Component and cluster analysis were performed to group thecultivars, according to their similarity coefficients. For molecularcharacterisation, 125 RAPD and 157 ISSR polymorphic markers wereamplified using 28 and 7 primers respectively. High level ofcongruence among the two marker systems (r =90.5%) was obtained from comparison of pheneticsimilarities based on the percentage of shared fragments. ISSRmarkers revealed important advantages over RAPDs, due to a higheffective multiplex ratio (12.5 for ISSR compared with 2.2 forRAPD analysis) and reproducibility. Although morphological andmolecular results are comparable, slight differences are showed incluster analysis UPGMA dendrograms. Molecular analysis explainedhomonym situations among 'Martainha' and'Longal' cultivars in Portugal.     

Real-time polymerase chain reaction based assays for quantitative detection of barley, rice, sunflower, and wheat

Quality assurance is a major issue in the food industry. The authenticity of food ingredients and their traceability are required by consumers and authorities. Plant species such as barley (Hordeum vulgare), rice (Oryza sativa), sunflower (Helianthus annuus), and wheat (Triticum aestivum) are very common among the ingredients of many processed food products; therefore the development of specific assays for their specific detection and quantification are needed. Furthermore, the production and trade of genetically modified lines from an increasing number of plant species brings about the need for control within research, environmental risk assessment, labeling/legal, and consumers' information purposes. We report here the development of four independent real-time polymerase chain reaction (PCR) assays suitable for identification and quantification of four plant species (barley, rice, sunflower, and wheat). These assays target gamma-hordein, gos9, helianthinin, and acetyl-CoA carboxylase sequences, respectively, and were able to specifically detect and quantify DNA from the target plant species. In addition, the simultaneous amplification of RALyase allowed bread from durum wheat to be distinguished. Limits of detection were 1 genome copy for barley, sunflower, and wheat and 3.3 copies for rice real-time PCR systems, whereas limits of quantification were 10 genome copies for barley, sunflower, or wheat and approximately 100 haploid genomes for rice real-time PCR systems. Real-time PCR cycling conditions of the four assays were stated as standard to facilitate their use in routine laboratory analyses. The assays were finally adapted to conventional PCR for detection purposes, with the exception of the wheat assay, which detects rye simultaneously with similar sensitivity in an agarose gel.     

Application of para-nitrophenol (pNP) enzyme assays in degraded tropical soils

Enzyme activities have been used as indicators of soil quality and changes in biogeochemical function due to management or perturbations. The objective of this study was to answer a number of methodological questions regarding sampling schemes, sample handling recommendations, and assay procedures to facilitate the use of enzyme assays in the tropical highlands of East Africa. We used para-nitrophenol (pNP) based substrates for five enzymes: β-glucosidase, cellobiohydrolase, chitinase, acid phosphatase, and alkaline phosphatase. In the first experiment, we examined sampling procedures and compared the results of determining enzyme activities on a plot using composite or discrete samples. Composite samples usually had higher activities than the means of individual cores (P<0.05), but relative ranking of sites was the same if analyses were based on composite or discrete samples. In the second experiment, we examined the effects of storage time and conditions on enzyme activity. Enzyme activity degraded rapidly in frozen samples, but was better maintained in samples stored at 4 °C. Phosphatase and cellobiohydrolase activity declined after 14 days of storage, while the activity of the other enzymes remained close to the values of fresh samples for 28 or more days. In the third experiment, we examined the effect of the addition of an antiseptic, toluene, to prevent bacterial growth during the assay. We found no consistent toluene effect (P>0.4), probably because the assays were of short duration and microbial growth was minimized. Finally, we looked at the incubation time necessary to produce reliable results. Phosphatases, with relatively high activities could reliably be determined in 2 h incubations, but the other enzymes had much lower activities and required longer incubation times for reliable determination. For the enzymes we looked at, 4 h was a good standard time for determining the activity of even the lowest activity enzymes. The results of this study provide practical guidelines for applying these enzyme assays in the degraded tropical soils.     

Validation of a mass spectrometry method to quantify oak ellagitannins in wine samples

Detection and individual quantification of oak wood ellagitannins in oak barrel aged red wine samples are difficult mainly due to their low levels and the similarity between their structures. In this work, a quantification method using mass spectrometry has been developed and validated to quantify wine ellagitannins after sample fractionation with a previously reported method. The use of an internal standard is a requirement to correct mass signal variability. (-)-Gallocatechin, among the different tested compounds, was the only one that proved to be a suitable internal standard making possible the accurate and individual quantification of the main oak wood ellagitannins. The developed methodology has been used to detect and quantify these ellagitannins in different Spanish commercial wines, proving its usefulness.     

Late nitrogen fertilization affects nitrogen remobilization in wheat

A pot experiment with wheat plants was carried out to study how late application of nitrogen (N) fertilizer affects the use of pre‐anthesis N reserves during the grain‐filling period. Increasing doses of N fertilizer were applied (0, 40, and 52 mg N plant^–1), either in two amendments (growth stages GS20 and GS30, according to Zadoks scale) or in three amendments (GS20, GS30, and GS37). The experiment was arranged in a complete randomized three‐block design with 129 plants per treatment. The plants were watered daily, harvested every 2 d between anthesis and maturity, and were separated into roots, leaf sheaths, leaf blades, and ears for further N determination. Grain N concentration improved due to a late N application in GS37 by 14% (higher N dose) and by 7% (further splitting the same N‐fertilizer dose, respectively). The higher the N‐fertilizer dose applied, the greater was the amount of pre‐anthesis reserves in vegetative organs, these reserves became later available for remobilization. Although splitting the same N dose in three amendments did not increase the N reserves, these reserves were more efficiently remobilized allowing an improvement in grain N concentration. The fertilizer management did not change the temporary pattern of N accumulation in the ear, but did induce a change in the amount of N remobilized and in the contribution of each organ (root, leaf sheath, leaf blade) to this remobilization. Late N amendment allowed a greater N availability of leaf blades and ear N reserves (from 20% up to 26% and from 19% up to 22%, respectively) for remobilization towards the grain, decreasing the root contribution from 28% down to 15%, while the contribution of leaf sheaths was maintained around 35% irrespective of the N applied.