Effects of varying nitrogen fertilization on crop yield and grain quality of emmer grown in a typical Mediterranean environment in central Italy

Experiments were carried out to study the effects of N fertilizer rates and timing of application on the yield and grain quality of a rainfed emmer crop (Triticum dicoccum Shübler) under Mediterranean conditions. The following parameters were analyzed: hulled and net grain yield, hulled index, spikes m^?2, spikelets per spike, kernels m^?2, thousand-kernel weight, biomass, plant height, lodging, grain protein and ash content. In the first experiment, different N rates (30, 60 and 90 kg N ha^?1 plus a control not fertilized) were split at three phenological stages (seeding 20%, tillering 40% and stem elongation 40%). In the second experiment, three N doses (30, 60 and 90 kg N ha^?1) were applied to three crop stages (seeding, tillering and stem elongation). In the third experiment, the rate of 90 kg N ha^?1 was distributed in different amounts (90-0-0, 0-90-0, 0-0-90, 45-45-0, 45-0-45, 0-45-45, 30-30-30) at the three mentioned crop stages. Increasing N rates resulted in higher hulled and net grain yield, as well as protein content. Fertilization (from 60 to 90 kg N ha^?1) applied to tillering maximized hulled and net grain yield. Fertilization (90 kg N ha^?1) applied to stem elongation gave the highest grain protein content (%) while splitting application (30 kg N ha^?1 each) at three phenological stages maximized protein yield per hectare. Application of half or one-third of 90 kg N ha^?1 to stem elongation improved grain protein content in comparison with applications at sowing, or at both sowing and tillering. The main factor determining higher yields with increasing N rates in this emmer crop was the number of kernels m^?2. None of the yield components accounted for differences in grain yield when timing and splitting application were varied.     

Chromatographic methods for metabolite profiling of virus- and phytoplasma-infected plants of Echinacea purpurea

This study was focused on the effects of virus and phytoplasma infections on the production of Echinacea purpurea (L.) Moench secondary metabolites, such as caffeic acid derivatives, alkamides, and essential oil. The identification of caffeic acid derivatives and alkamides was carried out by means of high-performance liquid chromatography-diode array detection (HPLC-DAD), HPLC-electrospray ionization-mass spectrometry (ESI-MS), and MS(2). Quantitative analysis of these compounds was carried out using HPLC-DAD. The results indicated that the presence of the two pathogens significantly decreases (P < 0.05) the content of cichoric acid, the main caffeic acid derivative. Regarding the main alkamide, dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, a significant decrease (P < 0.05) in the content of this secondary metabolite was observed in virus-infected plants in comparison with healthy plants, while in the phytoplasma-infected sample the variation of this secondary metabolite was not appreciable. The % relative area of the E/Z isomers of this alkamide was also found to change in infected samples. The gas chromatography (GC) and GC-MS analysis of E. purpurea essential oil enabled the identification of 30 compounds. The main significant differences (P < 0.05) in the semiquantitative composition were observed for three components: limonene, cis-verbenol, and verbenone. The results indicate that the presence of virus and phytoplasma has an appreciable influence on the content of E. purpurea secondary metabolites, which is an important issue in defining the commercial quality, market value, and therapeutic efficacy of this herbal drug.     

The effect of organic supplementation of solarized soil on the quality of tomato fruit

Soil solarization, used to control weeds and soil-borne pathogens in hot climates, has not yet been widely adapted as a commercial practice because of its lack of efficacy. Experiments were carried out in southern Italy over two growing seasons to study the effect of three levels (0, 0.35 and 0.7 kg m⁻²) of organic supplementation of the soil prior to solarization on soil mineral availability and fruit quality attributes. Soil temperature and chemical properties were monitored, together with changes in the physical characteristics and chemical composition of tomato fruits grown under commercial greenhouse conditions. Organic supplementation increased the soil temperature achieved through solarization by 3.9 to 5.5 °C. Organic supplementation increased (P ≤ 0.05) the soil concentration of NO₃⁻-N, exchangeable K₂O, Ca²⁺, Na⁺ and Mg²⁺ and the level of electrical conductivity in the soil extract. Physical characteristics of tomato fruits were improved by supplementation, with fresh and dry weight enhanced up to 11 and 21%, respectively, mesocarp thickness up to 19%, firmness up to 36% and skin redness (a^*/b^* ratio) up to 24%. As the supplementation rate was raised from 0 to 0.7 kg m⁻², the fruit content of reducing sugars increased (P ≤ 0.01) from 1.75 to 2.14 g per 100 g f.w., ash from 0.49 to 0.62%, soluble solid from 5.12 to 6.18 °Brix, titratable acidity from 0.16 to 0.19%, and ascorbic acid from 25.1 to 32.5 mg 100 mL⁻¹. We concluded that organic supplementation appears to be a valuable and environmentally friendly way to improve the mineral availability in the soil and improve fruit quality of tomato.     

Effect of grazing and dietary protein on eating quality of Podolian beef

Twenty-four young Podolian bulls were used to evaluate the effect of rearing system (extensive vs. intensive) in relation to postmortem aging (11 and 18 d) on the eating quality of the meat, with the diet of outdoor animals adjusted to the protein content of the indoor system (15% CP) or to the minimum protein content required for satisfactory growth (12% CP). At 415 ± 9.35 (SE) d of age, with a mean BW of 337.5 ± 16.51 (SE) kg, animals were allotted to 3 groups for the finishing period (172 d): 1) indoor group receiving a diet at 15% CP; 2) grazing animals receiving a diet at 15% CP of DM; and 3) grazing animals receiving a diet at 12% CP of DM. Longissimus dorsi lumborum muscle sampled from the right half and divided longitudinally into 2 sections was aged in vacuum packaging at 4°C until 11 and 18 d postmortem, respectively. Rearing system did not affect (P > 0.05) color, Warner-Bratzler shear force, texture profile, water-holding capacity, and most of the sensory attributes of the beef steaks. However, sensory tenderness was less in the meat from outdoor animals receiving a diet with 15% CP than in meat from outdoor animals receiving a diet with 12% CP or from the indoor group (P < 0.05). Meat aged 18 d showed decreased Warner-Bratzler shear force (P < 0.001), hardness (P < 0.001), cohesiveness (P < 0.05), springiness (P < 0.05), gumminess (P < 0.01), chewiness (P < 0.01), and thawing loss (P < 0.01) compared with meat aged for 11 d. Prolonging the aging time up to 18 d significantly increased b* (yellowness; P < 0.05), cooking losses (P < 0.001), and the intensity of all the texture sensory attributes, namely, juiciness and fatness (P < 0.05), chewiness, tenderness, and flavor (P < 0.01). Significant correlations were found between instrumental and sensory variables (range of r = -0.55 to -0.85, P < 0.05 to 0.001). Overall, in the present study, the rearing system did not markedly affect meat sensory and physical properties. Thus, we conclude that an outdoor system, even with reduced protein supplementation, may represent a valid farming system for local breeds in Mediterranean areas characterized by poor-quality pastures. However, an extended aging period is suggested to improve the main factor limiting the quality of this product, namely, reduced tenderness.     

Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial‐like cells

In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD‐MSCs) to characterize and differentiate them into endothelial‐like cells. AD‐MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony‐forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM‐2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial‐like cells were characterized by the evaluation of morphological changes and gene expression analysis for endothelial markers (CD31, CD144, CD146). Characterization of AD‐MSCs showed their ability to form clones, to differentiate in vitro and the OCT‐4, SOX‐2, NANOG genes expression. Immunophenotypic characterization showed the CD90 presence and the CD45 absence. The endothelial‐like cells showed morphological changes, the expression of CD31, CD144, CD146 genes and the presence of CD31 membrane receptor. Matrigel assay showed their ability to form network and vessels‐like structures. This study lays the foundations for future evaluation of the potential AD‐MSCs pro‐angiogenic and therapeutic role.     

Duplex real-time polymerase chain reaction for simultaneous detection and quantification of Anaplasma marginale and Anaplasma centrale

Anaplasma marginale and Anaplasma centrale are rickettsial pathogens responsible for acute disease and mild infections, respectively, in cattle herds. A duplex real-time polymerase chain reaction (PCR) assay with probes labeled with different fluorophores was developed for simultaneous detection and quantification of A. marginale and A. centrale DNA in bovine blood samples. The assay was able to detect as few as 10(1) and 10(2) DNA copies for A. marginale and A. centrale, respectively, with optimal specificity and reproducibility. Analysis by real-time and nested PCR carried out on 54 samples previously tested by reverse line blot hybridization showed that the established duplex real-time PCR assay can detect and quantify the 2 Anaplasma spp., even if present simultaneously in the same blood samples. Such an assay could be used in pathogenesis studies on bovine acute anaplasmosis.     

Selection-pressure effects of medium pH during regeneration on successive performances of leaf-derived Tomuri’ and ‘Hayward’ kiwifruit (Actinidia deliciosa) somaclones cultured on proliferation culture media with variable pH

Summary

The aims of the work were in vitro selecting for tolerance to high culture pH (CpH) in kiwifruit and evaluating the effects of selection pressure exerted by pH variations during regeneration. Leaf-derived ‘Tomuri’ and ‘Hayward’ kiwifruit (Actinidia deliciosa) somaclones, which had been regenerated at pH 5.7, 7 and 7.5, were compared to each other and to controls (i.e. cultures derived from ‘Tomuri’ and ‘Hayward’ mother plants through standard micropropagation techniques) for shoot growth, proliferation and appearance when cultured at up to pH 8. The proliferation rate (PR), the number of nodes (NN) produced by each shoot and axillary shoot length (ASL) decreased in all ‘Tomuri’ and ‘Hayward’ somaclones and in controls as culture pH increased. At CpH 8, ‘Tomuri’ somaclones regenerated at pH 7.0 generally tended to have higher average PR and NN than the other somaclones and control; their ASL also exceeded that of the other somaclones and was just slightly higher than in control. Regardless of CpH, ‘Hayward’ somaclone 7.5-1 tended to have higher PR, NN and ASL than other female somaclones. When shoot growth and appearance (tissue hyperhydricity, leaf yellowing or browning) were considered together through a multivariate data analysis, most ‘Tomuri’ and ‘Hayward’ somaclones which appeared less sensitive than controls to high CpH had been regenerated at pH 7 and 7.5. These regeneration and culture methods, especially the selection pressure exerted by the high pH during regeneration, thus seem to be effective in obtaining ‘Tomuri’ and ‘Hayward’ variants tolerant to high culture pH.     

Evaluation of the activity of aqueous extracts of some organic waste materials on in vitro-cultured shoots of ‘M9’ apple rootstock

Summary

The present work investigated the effects of different aqueous extracts of organic waste compounds on growth, proliferation and photosynthetic activity in ‘M9’ (Malus domestica Borkh.) shoot cultures, with the aim of determining the feasibility of using in vitro cultures as a tool for the rapid evaluation of organic amendments in agriculture. Aqueous extracts of the following organic waste compounds: cow manure (CM), sugarbeet industrial waste (SB), mixed grape, poultry and municipal solid waste (GPM), and citrus pruning and industrial waste (CPI) were prepared at a rate of 1:10 (w/v) compound:distilled water. The basal media used in the proliferation phase were: (i) PM1, modified Murashige and Skoog (MS) enriched with 4.4 µM 6-benzyladenine (BA); (ii) PM2, as PM1 but with a reduced cytokinin concentration (1 µM BA) to evaluate possible hormone effects; and (iii) PM3, 4.4 µM BA with reduced salt strength (0.33 MS) to induce nutrient deficiency. Hormone-free medium with half-strength MS salts was used for rooting. All media were enriched with each extract at 0, 0.2, 2, 20 or 200 ml l^–1. Photosynthetic activity was measured on PM3 medium enriched with SB or CM. Standard culture conditions were 22° ± 2°C, with a 16 h photoperiod at 30 µmoles photosynthetically active radiation (PAR) m^–2 s^–1, but at 80 µmoles PAR m^–2 s^–1 to determine photosynthetic activity. Shoot weight increase in PM1 was not affected by the GPM and CPI extracts, while the growth trends of CM- and SB-treated shoots were described by a second-degree function with maxima at 2 ml l^–1 and 0.2 ml l^–1, respectively. Shoot proliferation for SB was represented by a quadratic curve (maximum at 2 ml l^–1), was linearly reduced as GPM increased, but was not affected by CM or CPI. Treatments did not significantly affect rooting percentage and root length; however root number was increased by SB at 2 ml l^–1.CO₂ fixation increased linearly with both SB and CM, despite reduced growth at the highest levels of extract.     

Differential in Gel Electrophoresis (DIGE) Comparative Proteomic Analysis of Macrophages Cell Cultures in Response to Perthamide C Treatment

Secondary metabolites contained in marine organisms disclose diverse pharmacological activities, due to their intrinsic ability to recognize bio-macromolecules, which alter their expression and modulate their function. Thus, the identification of the cellular pathways affected by marine natural products is crucial to provide important functional information concerning their mechanism of action at the molecular level. Perthamide C, a marine sponge metabolite isolated from the polar extracts of Theonella swinhoei and endowed with a broad and interesting anti-inflammatory profile, was found in a previous study to specifically interact with heat shock protein-90 and glucose regulated protein-94, also disclosing the ability to reduce cisplatin-mediated apoptosis. In this paper, we evaluated the effect of this compound on the whole proteome of murine macrophages cells by two-dimensional DIGE proteomics. Thirty-three spots were found to be altered in expression by at least 1.6-fold and 29 proteins were identified by LC ESI-Q/TOF-MS. These proteins are involved in different processes, such as metabolism, structural stability, protein folding assistance and gene expression. Among them, perthamide C modulates the expression of several chaperones implicated in the folding of proteins correlated to apoptosis, such as Hsp90 and T-complexes, and in this context our data shed more light on the cellular effects and pathways altered by this marine cyclo-peptide.