Synthesis and phytotoxic activity of ozonides

The [4 + 3] cycloaddition of the proper furans with the oxyallyl cation, generated in situ from 2,4-dibromopentan-3-one, produced a series of 8-oxabicyclo [3.2.1]oct-6-en-3-ones. Exposure of the oxabicycles to ozone afforded the corresponding 8,9,10,11-tetraoxatricyclo[5.2.1.1 (2,6)]undecan-4-ones in variable yields (7-100%). The phytotoxic properties of these ozonides (or 1,2,4-trioxolanes) and their oxabicycle precursors were evaluated as the ability to interfere with the growth of Sorghum bicolor and Cucumis sativus seedlings. Among oxabicycles, the highest inhibitory activity was shown by compounds possessing a alpha,beta-unsaturated carbonyl moiety. A differential sensitivity of the two crops was evident with ozonides. The most active compounds were also tested against the weed species Ipomoea grandifolia and Brachiaria decumbens. To the best of our knowledge, this is the first article describing ozonides as potential herbicides.     

Interactions of three s-triazines with humic acids of different structure

The adsorption of three s-triazines (atrazine, terbutylazine, and simazine) on three different humic acids (HAs), before and after acidic hydrolysis, was studied at four diverse pH values from 3 to 4.5. The Freundlich sorption affinity (K(f)) and intensity (1/n) of s-triazines were related to the chemical and structural composition of HA and used in a multivariate statistical analysis. At low pH, the sorption values for s-triazines were not directly justified by the content of carboxyl groups in original HA, while only an increase of phenolic carbons in hydrolyzed HA supported the increased s-triazine adsorption. The structural composition of both s-triazines and humic samples explained 86% of the significance in multivariate analyses, whereas the role of pH remained hidden in only 14% of the statistical significance. Adsorption of s-triazines was mainly related to carbon content, hydrophobicity, and aromaticity of HA, thereby implying a predominant binding role of weak dispersive forces. Steric properties explained the larger adsorption affinity of atrazine and terbutylazine with respect to simazine. The occurrence of charge-transfer interactions between atrazine and mainly hydrolyzed HA was further suggested by the increased number of free radicals detected in atrazine-HA complexes at different pH values. Charge-transfer interactions were postulated to occur when HA conformations became progressively controlled by aromatic components. This work indicates that humic matter rich in hydrophobic and aromatic constituents is more likely to adsorb s-triazines and reduce their environmental mobility.     

Optimization of solid-liquid extraction of resveratrol and other phenolic compounds from milled grape canes (Vitis vinifera)

Optimization of the solid-liquid extraction conditions for trans-resveratrol, trans--viniferin, ferulic acid, and total phenolics from milled grape canes has been investigated. The temperature and ethanol concentration were found to be major process variables for all responses, whereas the solvent to solid ratio was found not to be significant for any of the responses studied. The yields of trans-resveratrol, trans--viniferin, and total phenolics increased with increasing temperature. Maximum yields of trans-resveratrol (4.25 mg/g dw), trans--viniferin (2.03 mg/g), and total phenolics (9.28 mg/g dw) were predicted from the combination of a moderate ethanol concentration (50-70%) and the highest temperature (83.6 degrees C), whereas an ethanol concentration of 35% at the lowest temperature studied (16.4 degrees C) was optimal for the extraction of ferulic acid (1.05 mg/g dw). Effective diffusivity values of resveratrol in the solid phase, D eff for different extraction conditions, were calculated by fitting the experimental results to a model derived from the Fick's second law. Effective diffusivity of resveratrol in the solid phase varied from 3.1 x 10 (-13) to 26.6 x 10 (-13) m (2) s (-1) with changing extraction conditions. The increase in effective diffusivity of resveratrol was observed with increasing temperature, and the highest predicted level was obtained when using 54% ethanol/water mixture at 83.6 degrees C. The increase in ethanol concentration exhibited the favorable effect up to 50-55%, thereafter effective diffusivity decreased with a further increase in concentration.     

Phytochemical changes induced by different nitrogen supply forms and radiation levels in two leafy Brassica species

The effect of three different nitrogen (N) supply forms differing in their ammonium-to-nitrate (NH(4):NO(3)) ratio (100% NH(4), 50% NH(4) + 50% NO(3), 100% NO(3)) under three different levels of daily photosynthetic active radiation (PAR) (low, 5.0; medium, 6.8; high, 9.0 mol m(-2) day(-1)) on a range of desirable health-promoting phytochemicals in Brassica rapa subsp. nipposinica var. chinoleifera and Brassica juncea was determined. The 100% NH(4) supply under medium PAR levels led to the highest concentration of glucosinolates based on a low nitrogen/sulfur ratio as well as high levels of carotenoids in the leaves of both Brassica species. However, the 100% NH(4) supply under low and medium PAR levels resulted in low concentrations of flavonoids based on high N concentration in the leaves. Thus, the data provided here have strong implications for crop management strategies aimed at optimizing both the concentration and composition of a range of phytochemicals.     

Uncommonly thorough hydrolysis of peptides during ripening of Ragusano cheese revealed by tandem mass spectrometry

Ragusano is a pasta filata cheese produced from raw milk in Sicily. The proteolysis was extensively analyzed after stretching (day 0), at 4 and 7 months of ripening through soluble nitrogen, urea-PAGE, and peptide identification by tandem mass spectrometry. After stretching, 123 peptides were identified: 72 arising from β-casein, 34 from α(s1)-casein, and 17 from α(s2)-casein. The main protein splitting corresponded to the action of plasmin, chymosin, cathepsin D, cell envelope proteinase, and peptidase activities of lactic acid bacteria. Unlike other types of cheeses, <10% residual β- and α(s)-caseins remained intact at 7 months, indicating original network organization based on large casein fragments. The number of identified soluble peptides also dramatically decreased after 4 and 7 months of ripening, to 47 and 25, respectively. Among them, bioactive peptides were found, that is, mineral carrier, antihypertensive, and immunomodulating peptides and phosphopeptides.     

Development of new fungicides against Magnaporthe grisea: synthesis and biological activity of pyrazolo[3,4-d][1,3]thiazine,pyrazolo[1,5-c][1,3,5]thiadiazine,and pyrazolo[3,4-d]pyrimidine derivatives

Some pyrazolo[3,4-d]pyrimidin-4(5H)-thione, pyrazolo[3,4-d][1,3]thiazin-4-one/thione, and pyrazolo[1,5-c][1,3,5]thiadiazine-4-one/thione derivatives were synthesized and screened for antifungal activity against the causal agent of rice blast disease, Magnaporthe grisea. In all cases a remarkable inhibition of fungal growth was found in the range from 10 to 200 microg x mL(-1). Several compounds were able to control mycelium growth at a rate of 10 microg x mL(-1), a concentration at which the reference compound tricyclazole was completely ineffective. At least in the case of the most active substance, at the same dose the growth of seedlings or cultured cells of rice was substantially unaffected. Results allowed definition of structural requirements either to maintain or to enhance mycotoxic activity.     

Enhancement of alkalinity tolerance in two cucumber genotypes inoculated with an arbuscular mycorrhizal biofertilizer containing Glomus intraradices

The aim of the present study was to determine whether arbuscular mycorrhizal (AM) inoculation with a biofertilizer containing clays as granular carriers, leek root pieces and Glomus intraradices spores could improve alkalinity tolerance of two cucumber genotypes, and to study the changes induced by AM at agronomical and physiological level. A greenhouse experiment was carried out to determine yield, growth, fruit quality, net photosynthesis (A_CO2), electrolyte leakage, and mineral composition of two cucumber (Cucumis sativus L.) genotypes (hybrid “Ekron” or open-pollinated variety “Marketmore”) with inoculated and noninoculated arbuscular mycorrhizal biofertilizer. Plants were supplied with nutrient solutions at two pH values (6.0 or 8.1). The high pH nutrient solution had the same basic composition with an additional 10 mM NaHCO₃ and 0.5 g l⁻¹ CaCO₃. The percentage root colonization was higher in “Marketmore” (21.8%) than “Ekron” (12.7%). Total and marketable yield and total biomass were significantly higher by 189%, 213%, and 77%, respectively, with Ekron in comparison to those recorded with Marketmore. The highest crop performance with Ekron in comparison to Marketmore was due to the improved nutritional status (higher N, P, K, Ca, Mg, Fe, Mn, and B), higher leaf area, and net photosynthesis. Increasing the concentration of NaHCO₃ from 0 to 10 mM in the nutrient solution significantly decreased yield, plant growth, A_CO2, N, P, Fe, Cu, Zn, Mn, and B concentration in leaf tissue, whereas the electrolyte leakage increased. The inoculated plants under alkaline conditions had higher total, marketable yield, and total biomass than noninoculated plant. Mycorrhizal cucumber plants grown under alkaline conditions had a higher macronutrient concentration in leaf tissue compared to noninoculated plants. The highest yield and biomass production in inoculated plants seems to be related to the capacity of maintaining higher net A_CO2 and to a better nutritional status (high P, K, Mg, Fe, Zn, and Mn and low Na accumulation) in response to bicarbonate stress with respect to −AM plants.     

Regulation of mycorrhiza development in durum wheat by P fertilization: Effect on plant nitrogen metabolism

The aim of this work was to study the effect of arbuscular mycorrhizal fungus Glomus mosseae on growth and nitrogen (N) metabolism of durum wheat (Tritcum durum) under various P soil contents. The analyses were extended to macro and micronutrient tissue concentrations, nitrate reductase and glutamine synthetase activities, as well as protein, aminoacids, pyridine dinucleotides and adenine nucleotides. Arbuscular mycorrhiza increased wheat growth in soil in which P availability was low and nitrate was the dominant N form. The root colonization occurred at the highest level in plants grown in limiting soil P and was inversely related to soil P content. The micorrhizal wheat plants contained also the highest concentrations of macro (P, K, Ca, N) and micronutrients (Fe, Zn, Mn) as well as free amino acids, protein, NAD, NADP, AMP, ADP, ATP in roots and leaves. In particular, the micronutrient tissue concentrations (Zn, Mn) supported that mycorrhiza actively modulated their uptake limiting interferences and optimizing growth better than the plant roots, like a very efficient “rootstock”. Control plants grown at the highest soil P did not reach the same concentration as the mycorrhizal plants. Nitrate reductase activities in the roots of mycorrhizal plants were higher than in the control ones, while glutamine synthetase activities were highest in the leaves. Protein and amino acids concentrations, as well as AMP, ADP, ATP, NAD(P), and NAD(P)H were also higher than in the control. Among the free amino acids in the roots, the high levels of glutamine, asparagine, arginine, support the view that ammonium was transferred through the arbuscules to the root cells where it was re‐assimilated in the cortical cells, forming high N : C ratio‐amino acids. They were transferred to the leaves where all the other N compounds could be largely synthesized using the carbon skeletons supplied by photosynthesis.     

Yucca gloriosa: a source of phenolic derivatives with strong antioxidant activity

On the basis of the biological activities exhibited by the phenolic constituents of Yucca schidigera, the antioxidant activity of the methanol extract of Yucca gloriosa roots was evaluated in the TEAC assay. The strong activity exerted by this extract prompted investigation of its phenolic constituents, yielding three new phenolic derivatives, gloriosaols C, D, and E, along with gloriosaols A and B previously isolated from Y. gloriosa roots and yuccaols C-E isolated from Y. schidigera. ESIMS and NMR data of gloriosaols C-E closely resembled those reported for gloriosaols A and B, two diasteroisomers characterized by unusual spirostructures. Careful inspection of ROESY spectra revealed that gloriosaols C-E are diastereoisomers of gloriosaols A and B. A possible assignment of the relative configuration of gloriosaols C-E, derived according to an integrated NMR-quantum mechanical (QM) approach, which was already applied to the determination of the stereostructures of gloriosaols A and B, is also proposed. Gloriosaols A-E exhibited potent antioxidant activity measured by the TEAC assay, showing the potential use of Y. gloriosa as a source of antioxidant principles.     

Synthesis of 3-(4-Bromobenzyl)-5-(aryl methylene)-5H-furan-2-ones and their activity as inhibitors of the photosynthetic electron transport chain

A series of 12 3-(4-bromobenzyl)-5-(arylmethylene)-5 H-furan-2-one lactones, designed using the naturally occurring toxin nostoclides as a lead structure, were synthesized and screened as potential inhibitors of photosynthetic electron transport. The structures were confirmed by (1)H and (13)C NMR, MS, and IR analyses. Their biological activity was evaluated both in vitro, as the ability to interfere with light-driven reduction of ferricyanide by isolated spinach chloroplasts, and in vivo, as the capability to inhibite the oxygen production by intact Chlorella cells. Some of the compounds exhibited inhibitory properties in the micromolar range against basal and phosphorylating electron flow from water to K 3[Fe(CN) 6], with no effect on uncoupled electron flow. Thus, they seem to behave as energy-transfer inhibitors. Although poor solubility in water may limit their effectiveness, the active derivatives could present structures to be exploited for the design of new substances endowed with herbicidal activity.