Effect of nitrate and humic substances of different molecular size on kinetic parameters of nitrate uptake in wheat seedlings

The kinetic parameters of nitrate uptake (I_max, Km and C_min) were evaluated in young seedlings of Triticum durum L., cv. Appulo, exposed to nitrate and/or to soil‐extracted humic acids (HAs) of different molecular weight. The uptake was enhanced after induction at low levels of nitrate (50 μM KNO₃), while it was inhibited after induction at higher concentrations (2000 μM). The kinetic parameters of uptake were selectively influenced by pre‐treatment with HAs: total (TE) and, at a greater extent, low (LMS, < 3500 Da) molecular size humic fraction increased either the nitrate uptake rate (I_max) and the efficiency of the whole transport system (low Km and C_min), while an opposite result was evidenced in high molecular size (HMS, > 3500 Da)‐treated plants. An additive effect was shown when nitrate and humic substances were provided simultaneously: the uptake rate was enhanced in TE‐ and LMS‐treated plants, but was strongly delayed in HMS‐treated plants. Removal of nitrate and/or humic fractions de‐induced the system and NO₃^— uptake rate decreased. Exposure to HAs was not able to induce nitrate reductase activity in root and leaf tissues. Inhibitors of protein synthesis p‐fluorophenylalanine and cycloheximide reversed the positive effect of LMS fraction on nitrate uptake. This would support the hypothesis of a promoting effect of HAs on the molecular expression of proteins of the nitrate transport system.     

Characterization of polyphenol oxidase and peroxidase and influence on browning of cold stored strawberry fruit

Polyphenol oxidase and peroxidase were extracted from two different varieties of strawberry fruit (Fragaria x ananassa D, cv. 'Elsanta' and Fragaria vesca L, cv. 'Madame Moutot') and characterized using reliable spectrophotometric methods. In all cases, the enzymes followed Michaelis-Menten kinetics, showing different values of peroxidase kinetics parameters between the two cultivars: Km = 50.68 +/- 2.42 mM ('Elsanta') and 18.18 +/- 8.79 mM ('Madame Moutot') mM and Vmax = 0.14 +/- 0.03 U/g ('Elsanta') and 0.05 +/- 0.01 U/g ('Madame Moutot'). The physiological pH of fruit at the red ripe stage negatively affected the expression of both oxidases, except polyphenol oxidase from 'Madame Moutot' that showed the highest residual activity (68% of the maximum). Peroxidase from both cultivars was much more thermolable as compared with PPO, losing over 60% of relative activity already after 60 min of incubation at 40 degrees C. The POD activation energy was much lower than the PPO activation energy (DeltaE = 97.5 and 57.8 kJ mol-1 for 'Elsanta' and 'Madame Moutot', respectively). Results obtained from d-glucose and d-fructose inhibition tests evidenced a decreasing course of PPO and POD activities from both cultivars as the sugar concentration in the assay medium increased. Changes in CIE L*, a*, b*, chroma, and hue angle values were taken as a browning index of the samples during storage at 4 degrees C. A decrease in L* was evident in both cultivars but more marked in 'Elsanta'. PPO and POD activities from cv. 'Elsanta' were very well-correlated with the parameter L* (r2=0.86 and 0.89, respectively) and hue angle (r2=0.85 and 0.93, respectively). According to these results, the browning of the fruit seemed to be in relation to both oxidase activities.     

Quantifying plant species diversity in a Natura 2000 network: Old ideas and new proposals

Assessing the effects of the spatial components on species diversity in a network of protected areas represents an important step for assessing its conservation “capacity”. A clear evaluation on how α-, β-, and γ-diversity are partitioned among and within spatial scales can help to drive manager decisions and provide method for monitoring species diversity. Moving from these concepts, a probabilistic sample of plant species composition was here applied for quantifying plant species diversity within the Sites of Community Importance (SCIs) of the Natura 2000 network in the Siena Province. All analyses were performed separately for all species and those species defined as “focal” (included in regional, national or continental “red” lists). The results indicated that species richness of the SCIs differed from one location to another one independently from the sampling efforts. Diversity partitioning indicated that most of the flora diversity within the network was given by larger-scale β-diversity, i.e. the differences in species composition among SCIs. β-diversity was then decomposed in two components: β_Area (due to the differences in area among SCIs) and β_Replacement (due to the compositional differences across SCIs). β_Area was particularly important for all species, while β_Replacement was the most important factor for focal species. The consequent implications for monitoring and nature conservation strategies are discussed.     

Monitoring chemical changes of dry-cured Parma ham during processing by surface autofluorescence spectroscopy

Parma hams at various processing stages were investigated by surface autofluorescence spectroscopy. Fluorescence "landscapes" of raw meat and salted (3 months), matured (11 and 12 months), and aged (15 and 18 months) Parma hams were obtained, and a three-dimensional data array (sample x emission x excitation) was used to develop a PARAFAC model including five components, which all exhibited characteristics of pure fluorophores regarding both excitation and emission spectra. The relative amount of each component related strongly to the processing stage, and sample age showed good correlation to fluorescence data (R = 0.98), with a relative error of prediction of approximately 1 month. Fluorescence measurements from samples of either semimembranosus or biceps femoris were used to predict chemical or sensory reference data, yielding good correlation for biceps femoris data, thereby enabling moisture content, sensory and instrumental color, and proteolysis value to be fairly well predicted. Overall, surface autofluorescence of Parma hams proved to hold relevant information, relating to major chemical/physical changes during processing. It is concluded that fluorescence spectroscopy has potential as an innovative method of quality control in dry-cured ham.     

Characterization of the anthocyanin fraction of sicilian blood orange juice by micro-HPLC-ESI/MS

Anthocyanins of red orange juices were analyzed by micro-HPLC coupled on-line with an MS detector equipped with an ESI source. The use of microcolumn HPLC greatly enhanced detection performance, allowing direct identification of the anthocyanins present in the orange juices. The use of a soft ionization technique allows detection of the molecular ions of the aglycons. Eight components were identified, five of them for the first time in red orange juice. Three additional anthocyanins were detected, of which only the aglycon was identified.     

β-cyclodextrin and caffeine complexes with natural polyphenols from olive and olive oils: NMR, thermodynamic, and molecular modeling studies

Complexes of β-cyclodextrin (β-CD) and caffeine (Caf) with biophenols present in olive and olive oil (tyrosol, hydroxytyrosol, homovanillic acid, 3,4-dihydroxyphenylacetic acid, and protocatechuic acid) were investigated by NMR spectroscopy and thermodynamical-molecular dynamic studies to verify the formation of supermolecular aggregates. The obtained results indicated that the investigated biophenols form inclusion complexes with β-CD in a molar ratio of 1:1 in aqueous solution having binding constant values from 10- to 40-fold bigger than those of the corresponding complexes with Caf. Then, β-CD preferentially encloses the biophenol molecule, decreasing its bitter taste and, at the same time, preserving it against chemical and physical decomposition reactions that occur during storage.     

Differences in the chemical composition of dissolved organic matter from waste material of different sources

The chemical composition of waste-material-derived dissolved organic matter (DOM) was characterized by chemolytic analyses and ¹H, ¹³C and ³¹P nuclear magnetic resonance (NMR) spectroscopy. Dissolved organic matter was extracted by water from an aerobic fermented urban waste compost, a sewage sludge and a pig slurry and then fractionated using the XAD-8 method. The amount of water-extractable dissolved organic carbon (DOC) ranged from 3% in the sewage sludge to 22% in the pig slurry. Dissolved organic matter isolated from pig slurry was equally distributed between hydrophilic and hydrophobic DOC, whereas in the sewage-sludge-derived material the hydrophobic fraction was predominant. Dissolved organic C from the urban waste compost was mainly within the hydrophilic fraction. Wet-chemical analysis and ¹H- and ¹³C-NMR spectra showed that both DOM fractions from the urban waste compost were low in neutral, acidic and amino sugars as well as in lignin-derived compounds. In turn, the materials were rich in low-molecular-weight aliphatic compounds. The chemical structure of both fractions is probably the result of the intensive transformation of urban waste compost during its fermentation. The hydrophilic fractions of DOM from sewage sludge and pig slurry contained considerable amounts of carbohydrates but were also rich in low-molecular-weight aliphatics. The respective hydrophobic fractions had the largest contents of CuO-extractable phenols which may in part derive from sources other than lignin. By contrast with the other materials, the hydrophobic fraction from the pig slurry seemed to contain polymeric rather than low-molecular-weight material. The ³¹P-NMR spectrum of the hydrophilic DOM fraction from urban waste compost did not show signals of inorganic or organic P compounds while the spectrum of the hydrophobic fraction revealed traces of monoester P, diester P, and orthophosphate. ³¹P-NMR spectroscopy suggested that both the hydrophobic and hydrophilic fractions from pig slurry did not contain organic P. The hydrophilic DOM fraction from sewage sludge contained orthophosphate, organic monoester P and a little pyrophosphate. The hydrophobic fraction contained mainly organic diester P and smaller amounts of teichoic acids and organic monoester P. Considering that water-soluble fractions of urban waste compost contained no easily plant-available P and a low content of labile organics, we conclude that this material contains less labile nutrients and is more refractory than the soluble constituents of pig slurry and sewage sludge.