Analysis of puroindoline a and b sequences from Triticum aestivum cv. `Penawawa' and related diploid taxa

Flanking sequences of the puroindoline a(pinA) and puroindoline b (pinB) genes from Triticum monococcum, T. urartu, Aegilops speltoides and A. tauschii were obtained by inverse PCR. Two accessions from each of these related diploid taxa were sequenced, and the sequences compared to those of the soft hexaploid wheat cultivar `Penawawa'. As expected from the origin of the D-genome in hexaploid wheat, the highest sequence identity was observed for the A. tauschii sequences, whereas the sequences for A.speltoides were the most divergent. The promoter sequences were further analyzed for putative regulatory elements, and the possible significance of several highly conserved sequence motifs is discussed. Western blots of Triton X-114 extracted proteins separated by SDS-PAGE confirmed the accumulation of pinA and pinB gene products in the endosperm of all the germplasm lines studied. The deduced amino acid sequences for the related diploid taxa do not imply any major structural changes as compared to the wild-type T. aestivum puroindolines, and is therefore in agreement with the soft endosperm texture of these species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Productive and biochemical responses of durum wheat to UV filtration

A UV exclusion experiment was conducted on durum wheat (Triticum durum Desf. var. Claudio) grown in pots. Plants were grown under three different radiative treatments in greenhouses covered with plastic filters: Teflon, transparent to the entire region of natural UV‐visible sunlight (TEF); polyester, transparent above 312 nm (MYL, excluding UVB) and Lee, transparent above 400 nm (LEE, excluding both UVA and UVB). Analyses have been carried out to determine the concentration of photosynthetic pigments in leaves, UV‐absorbing compounds, nitrogen and carbon in leaves, culms and spikes and proteins and gluten in grains. In particular, plants grown under UV exclusion showed a reduction in protein and dry gluten content (consequently without variation in the ratio dry gluten/protein), but at the same time, a significant increase of gluten index, which is a parameter to define the quality of gluten, was observed. The results highlighted the influence played by UV radiation on some biochemical parameters, mainly UV‐absorbing compounds, leaf nitrogen and grain protein characteristics of durum wheat cultivated under Mediterranean conditions. In particular, natural level of UV in the Mediterranean improves the characteristics of durum wheat flour important for pasta production (high dry gluten level), while the UV exclusion could improve characteristics of flour important for bread production (high gluten index).     

Molecular markers for improving control of soil-borne pathogen <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> in sugar beet

Fusarium spp. cause severe damage in many agricultural crops, including sugar beet, with Fusarium oxysporum historically being considered as the most damaging of all species. Sugar beet needs to be protected from this class of soil-borne pathogens in order to ensure an optimal sugar yield in the field. Genetic control of the disease is crucial in managing these pathogens. Identification of single nucleotide polymorphism (SNP) markers linked to resistance can be a powerful tool for the introgression of valuable genes needed to develop Fusarium-resistant varieties. A candidate gene approach was carried out to identify SNP markers linked to putative Fusarium resistance sources in sugar beet. Five resistant analogue genes (RGAs) were screened by means of high resolution melting (HRM) analysis in a set of sugar beet lines, considered as resistant and susceptible to Fusarium oxysporum. HRM polymorphisms were observed in 80% of amplicons. Two HRM polymorphisms were significantly associated with Fusarium resistance (P < 0.05). The amplicons that showed association were sequenced and two SNPs were identified. The association was further validated on 96 susceptible and 96 resistant plants using competitive allele-specific PCR (KASPar) technology. The selected SNPs could be used for marker-assisted breeding of Fusarium resistance in sugar beet.     

Effect of Transglutaminase on Protein Electrophoretic Pattern of Rice,Soybean, and Rice‐Soybean Blends

The interactions taking place in composite dough containing rice flour and soybean proteins (5% w/w) in the presence of transglutaminase, an enzyme with cross‐linking activity, were studied using different electrophoretic analyses. The interaction between rice proteins and soybean proteins was intensified by the formation of new intermolecular covalent bonds catalyzed by transglutaminase and the indirect formation of disulfide bonds among proteins. The main protein fractions involved in those interactions were both β‐conglycinin and glycinin of soybean and the glutelins of the rice flour, although albumins and globulins were also cross‐linked. The addition of soybean proteins to rice flour improves the amino acid balance and they also might play an important role on the rice dough properties because soybean proteins interact with rice proteins, yielding protein aggregates of high molecular weight.     

Use of an in vitro digestion model to study the bioaccessibility of 4-hydroxy-2-nonenal and related aldehydes present in oxidized oils rich in omega-6 acyl groups

Mixtures of either sunflower oil or thermodegraded sunflower oil and a standard meal were submitted to an in vitro digestion model. The same experiment was carried out with fluid deep-frying fat and thermodegraded fluid deep-frying fat. The thermodegradation of the oil and fat was provoked by submitting them to 190 degrees C with aeration in a convection oven, and the presence in the headspace of the thermodegraded oil and fat of oxygenated alpha,beta-unsaturated aldehydes (OalphabetaUAs), such as 4-hydroxy-2-nonenal (HNE), 4-oxo-2-nonenal (ONE), and 4,5-epoxy-2-decenal (EDE), was monitored by solid phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS). The digestion products were separated by centrifugation in a lipidic phase, an aqueous phase, and a pellet phase. The headspace of these three phases was also studied by SPME/GC-MS to check if the toxic and very reactive OalphabetaUAs above-mentioned remained unaltered after the in vitro digestion process or if they had reacted with the various compounds present in the digestion products, so disappearing from the samples. With the same aim the extract in ethyl acetate of the aqueous and pellet phases, and of the lipidic phase after dilution, were studied by GC-MS. All results obtained showed that a certain proportion of the toxic OalphabetaUAs remains unaltered after digestion, dispersed in the three phases above-mentioned, and thus are bioaccessible in the gastrointestinal tract and so could reach the systemic circulation. Compounds that may originate in Maillard type reactions (2-pentylpyridine) are found among digestion products, proving that these reactions are possible in this process if adequate substrates are present. In addition, it has been shown that toxic metabolites from the synthetic antioxidant BHT, present in fat before digestion, remain unaltered after this process and could reach the systemic circulation.     

Environmental correlates of two macro-decapods distribution in Central Italy: Multi-dimensional ecological knowledge as a tool for conservation of endangered species

Using a multi-dimensional ecological design, this study aimed first to analyse whether local environmental conditions can account for the spatial segregation of two Italian native decapods, the crayfish Austropotamobius italicus and the river crab Potamon fluviatile, in Central Italy freshwater ecosystems. Second, we aimed to analyse which environmental variables were more closely associated with the presence/absence of the two decapods in specific sites within their distribution area. Following a factorial design, a total of 32 sites were selected in two neighbouring geographic areas, one occupied by crayfish and one by crabs. Within each distribution area, eight streams where the decapod was present and eight where it was not present were selected. At each site, macro-invertebrate community composition and 16 abiotic variables were recorded and analysed with multi-variate methods. Variations in physical (minimum and maximum temperatures), chemical (calcium, oxygen, nitrate and nitrite) and geomorphological (substrate composition) parameters explained spatial segregation of P. fluviatile and A. italicus in the study area. The occurrence of crayfish reflected variations of chemistry (such as pH, calcium, nitrate and nitrite concentrations), temperature, water depth and substrate composition. On the contrary, the presence of the river crab, within its occurrence zone, was not associated to any biotic and abiotic parameters and was probably affected by anthropogenic pressure and uncontrolled harvesting. These findings provide fundamental ecological data for the maintenance of the two decapod natural populations as well as for the selection of areas and streams adequate for their reintroduction.     

Aerobic Biological Treatment of Chestnut Processing Wastewater

Chestnut agro-industrial companies consume a high volume of water for washing and processing fruit, generating a large volume of wastewater. This work studied the biodegradation of chestnut processing wastewater through aerobic assays, varying substrate, and biomass concentrations. In general, this wastewater presents a good biodegradability, especially in experiments with relatively low chemical oxygen demand (COD) (0.4 and 0.6?g O₂ L^?1) allowing a COD removal of 85?C90?%. The best results were obtained in the reactor initially loaded with 2?g?L^?1 of biomass and 0.4 or 0.6?g O₂ L^?1 of COD. These experiments also showed high COD removal rates: 4.25 and 3.88?g COD g^?1 volatile suspended solids (VSS) h^?1, respectively. The sedimentation rate, evaluated for different initial values of biomass (1, 2, and 3?g?L^?1), always presented higher values in the experiments with 2 and 3?g?L^?1 of biomass, regardless of the initial COD value used. After comparing different kinetic models (Monod, Contois, and Haldane), it was observed that the Haldane inhibition model satisfactorily describes the COD biodegradation. AQUASIM software allowed calculating the kinetic constant ranges: K _s, 1.59?C6.99?g COD L^?1; ?? _max, 25?C40?g COD g^?1 VSS day^?1; and K _i values, 0.07?C0.11. These kinetic constants corresponds to maximum rates (??*) between 1.48 and 4.25?g COD g^?1 VSS day^?1 for substrate concentrations (S*) from 0.38 to 0.88?g COD L^?1.     

Combined effect of sourdough lactic acid bacteria and additives on bread firmness and staling

The effect of various sourdoughs and additives on bread firmness and staling was studied. Compared to the bread produced with Saccharomyces cerevisiae 141, the chemical acidification of dough fermented by S. cerevisiae 141 or the use of sourdoughs increased the volume of the breads. Only sourdough fermentation was effective in delaying starch retrogradation. The effect depended on the level of acidification and on the lactic acid bacteria strain. The effect of sourdough made of S. cerevisiae 141-Lactobacillus sanfranciscensis 57-Lactobacillus plantarum 13 was improved when fungal alpha-amylase or amylolytic strains such as L. amylovorus CNBL1008 or engineered L. sanfranciscensis CB1 Amy were added. When pentosans or pentosans, endoxylanase enzyme, and L. hilgardii S32 were added to the same sourdough, a greater delay of the bread firmness and staling was found. When pentosans were in part hydrolyzed by the endoxylanase enzyme, the bread also had the highest titratable acidity, due to the fermentation of pentoses by L. hilgardii S32. The addition of the bacterial protease to the sourdough increased the bread firmness and staling.     

Endocrine disruption in a terrestrial isopod under exposure to bisphenol A and vinclozolin

Background, aim, and scope  

In the past decade there has been an increasing awareness about the possible consequences of human and wildlife exposure to endocrine disrupting compounds (EDCs). Bisphenol A (BPA) and vinclozolin (Vz) are EDCs which impacts on vertebrates have been largely investigated. Nevertheless, research on invertebrate effects, especially on soil organisms, are still largely under-represented. This work aims to extend the limited ecotoxicological datasets available and to provide tools to assess the effects of EDCs on the terrestrial species, using Porcellio scaber (Crustacea: Isopoda) as a model organism.     

土壤添加沼渣对温室气体排放及小麦生长的影响

Digestate, the product obtained after anaerobic digestion of organic waste for biogas production, is rich in plant nutrients and might be used to fertilize crops. Wheat(Triticum spp. L.) was fertilized with digestate, urea, or left unfertilized and cultivated in the greenhouse for 120 d. Emissions of greenhouse gasses(carbon dioxide(CO_2), methane(CH_4), and nitrous oxide(N_2O)) were monitored and plant growth characteristics were determined at harvest. The digestate was characterized for heavy metals, pathogens, and C and N mineralization potential in an aerobic incubation experiment. No Salmonella spp., Shigella spp., or viable eggs of helminths were detected in the digested pig slurry, but the number of faecal coliforms was as high as 3.6 × 10~4colony-forming units(CFU) g-~(1)dry digestate. The concentrations of heavy metals did not surpass the upper limits established by US Environmental Protection Agency(EPA). After 28 d, 17% of the organic C(436 g kg~(-1)dry digestate) and 8% of the organic N(6.92 g kg~(-1)dry digestate)were mineralized. Emissions of CO_2 and CH_4 were not significantly affected by fertilization in the wheat-cultivated soil, but digestate significantly increased the cumulative N_2O emission by 5 times compared to the urea-amended soil and 63 times compared to the uncultivated unfertilized soil. It could be concluded that digestate was nutrient rich and low in heavy metals and pathogens, and did not affect emissions of CH_4 and CO_2 when applied to a soil cultivated with wheat, but increased emission of N_2O.