Detection and Elimination of Viruses in Callus, Somatic Embryos and Regenerated Plantlets of Grapevine

The distribution of some grapevine viruses in flower explants, embryogenic and non-embryogenic calli, single somatic embryos and plants regenerated from embryogenic cultures was investigated by RT-PCR and ELISA. Immature anthers and ovaries of the cultivars Grignolino infected by GRSPaV, GLRaV-1 and GVA, Müller-Thurgau infected by GRSPaV and GLRaV-3 and Bosco infected by GRSPaV were cultivated on media inducing indirect somatic embryogenesis. Viruses were detected both in anthers and ovaries. Four months after culture initiation 65.6% of tested calli were infected by at least one virus; high percentages of virus infection were found in calli originating from ovaries. No virus was detected in calli tested 8 months after culture initiation, as well as in single somatic embryos or in embryo-derived plantlets. Somatic embryogenesis confirmed its effectiveness in eliminating phloem-limited grapevine viruses. Regeneration of RT-PCR negative plantlets occurred even when at least a sector of the callus was still infected: the mechanism whereby somatic embryos are freed of some viruses could be related to the rapid proliferation of embryogenic cells within the callus or to the origin of the embryogenic callus from virus-free cells within the original explant.     

Titanium dioxide photoinduced degradation of some pesticide/fungicide precursors

Five-membered nitrogen heterocycles (pyrrole, imidazole and 1,2,4-triazole) have been degraded using titanium dioxide and simulated solar radiation at pH = 8. The degradations followed a simple Langmuir-Hinshelwood mechanism. Accordingly, the adsorption equilibrium constants K of the heterocycles on the titanium dioxide surface and the rate constants k of degradation of the heterocycle-catalyst adduct have been obtained experimentally. While the K values decrease with heterocycle pKa, the k values increase with increasing pKa. Therefore, apparently, the rate constant depends on the availability of the electron pair on nitrogen, but at the same time the electron pair repulsion induced by the negatively charged titanium dioxide surface at pH = 8 causes a reverse effect in the adsorption equilibrium constant. Only in the case of imidazole, where the adsorption equilibrium constant is low enough (K = 0.013 M(-1)), can the rate constant be approximated to a pseudo-first-order rate expression: k(obs) = Kk. In all other cases, k(obs) = Kk/(1 + K(heterocycle)).     

Changes in compact bone microstructure of rats subchronically exposed to cadmium

Background

Chronic exposure to cadmium (Cd), even at low concentrations, has an adverse impact on the skeletal system. Histologically, primary and secondary osteons as basic structural elements of compact bone can also be affected by several toxicants leading to changes in bone vascularization and mechanical properties of the bone. The current study was designed to investigate the effect of subchronic peroral exposure to Cd on femoral bone structure including histomorphometry of the osteons in adult male rats.In our study, 20 one-month-old male Wistar rats were randomly divided into two experimental groups. In the first group, young males received a drinking water containing 30 mg of CdCl₂/L, for 90 days. Ten one-month-old males without Cd intoxication served as a control group. After 90 days of daily peroral exposure, body weight, femoral weight, femoral length, cortical bone thickness and histological structure of the femora were analysed.

Results

We found that subchronic peroral application of Cd had no significant effect on body weight, femoral length and cortical bone thickness in adult rats. On the other hand, femoral weight was significantly increased (P < 0.05) in Cd-intoxicated rats. These rats also displayed different microstructure in the middle part of the compact bone where vascular canals expanded into central area of substantia compacta and supplied primary and secondary osteons. Additionally, a few resorption lacunae which are connected with an early stage of osteoporosis were identified in these individuals. Histomorphometrical evaluations showed that all variables (area, perimeter, maximum and minimum diameter) of the primary osteons’ vascular canals, Haversian canals and secondary osteons were significantly decreased (P < 0.05) in the Cd group rats. This fact points to alterations in bone vascularization.

Conclusions

Subchronic peroral exposure to Cd significantly influences femoral weight and histological structure of compact bone in adult male rats. It induces an early stage of osteoporosis and causes reduced bone vascularization. Histomorphometrical changes of primary and secondary osteons allow for the conclusion that the bone mechanical properties could be weakened in the Cd group rats. The current study significantly expands the knowledge on damaging action of Cd on the bone.     

Evidence for genetic diversity of Toxoplasma gondii in selected intermediate hosts in Serbia

To contribute to the insight into the worldwide population structure of Toxoplasma gondii, we genetically characterized a total of eight strains isolated from intermediate hosts including humans, sheep and pigeons in Serbia. Although parasite DNA was detected in 28.2% (60/213) of the human samples from 162 patients serologically suspected of active toxoplasmosis, as well as in 5/7 seropositive pigeons and in 2/12 seropositive sheep examined, multilocus PCR-RFLP genotyping, using SAG1, 5′SAG2, 3′SAG2, GRA6, 5′GRA7 and 3′GRA7 as markers, was successful in only four human isolates (of which one was isolated from both the bronchoalveolar lavage fluid and blood samples of a single patient), one sheep and three pigeons. Of the eight isolates, five were type II (62.5%), one was type III, one was atypical, and one had a type I allele at GRA6 as the single locus genotyped. Although type II, as elsewhere in Europe, predominated, these results may suggest a higher genetic diversity of T. gondii in Serbia, reflecting local environmental contamination and also the geographical position of the country in South-East Europe.     

Morpho-pomological and chemical characterization of pomegranate (Punica granatum L.) genotypes in Apulia region,Southeastern Italy

A recent increasing demand in Western countries for pomegranate products by consumers is especially supported for the nutritional and medicinal characteristics, due to the antioxidant properties of this fruit. Some studies have been published on the morphological and biochemical characteristics of pomegranate fruits in some Mediterranean countries, but little information is available about the genotypes present in Italy and in particular in Apulia (Southeastern region of Italy). This study (2008–2009) evaluated morpho-pomological and chemical parameters of eight pomegranate genotypes localized in private small orchards. Significant differences were observed among the pomegranate genotypes for many of the parameters investigated. In particular, fruit weight ranged from 168.9 g (SouMol) to 574.9 g (SouOst), °Brix from 14.7 (ComTri) to 18.0 (SouMol), titratable acidity from 5.4 (ComMol) to 25.0 (SouTri) g/L. SouMol showed the highest polyphenols (97.1 mg/L) and vitamin C (236.3 mg/L) contents. Oil content of the seeds was between 5.90% and 10.30%, no differences have been observed for the fatty acid composition with conjugated linoleic acid (CLA) isomers as the most abundant fraction (81.23%). Considering all the evaluated parameters, and especially those referring to the organoleptic characteristics and antioxidants content, it must be stated that the best genotypes worthy to be considered from agricultural and industrial points of view were AdeSgi for fresh market and SouOst for the juice industry.     

Determination of Plant-Available Nutrients in Two Wood Ashes: The Influence of Combustion Conditions

Wood ashes were traditionally used as a fertilizing agent from the very beginning of working land but the fertilizing quality varies according to the combusted biomass and also the industrial burning equipment. The differences between wood ash originated from fluid and classic boiler were investigated by X-ray powder diffraction (XRD), X-ray fluorescence spectrometry (XRF), differential thermal analysis (DTA), thermo-gravimetric analysis (TGA), scanning electron microscopy (SEM), and Mehlich 3 and deionized water extraction procedures in relation to extraction time. The results, especially of the SEM and chemical extractions, have demonstrated that the type of burning boiler determines the portions of the leachable nutrient elements. The silica and alumina ash content conducted by the elevated temperature transforms the main proportion of calcium and potassium into the insoluble feldspars. Due to this effect only a small amount of nutrients could be released and served as a fertilizing matter.     

First report of Pseudomonas tolaasii on Agaricus bisporus in Serbia

Symptoms of brown blotch were observed on cultivated Agaricus bisporus on several mushroom farms in Serbia. Subsequently, samples were collected from nine localities during 2006?C2010 and fluorescent Gram-negative bacterial strains were isolated. All strains caused sunken brown lesions on A. bisporus tissue blocks after artificial inoculation. Isolated strains showed the LOPAT characteristics of the group Va being negative for levan production; the ability to produce a hypersensitivity reaction in tobacco; were oxidase and arginine dihydrolase positive; and lacked pectolytic activity. The results from other tests were as follows: oxidative metabolism of glucose; catalase activity positive; aesculin hydrolisis negative; Tween 80 hydrolysis positive; casein hydrolysis positive; gelatine hydrolysis and nitrate reduction negative; mannitol, erythritol, sorbitol, inositol and trehalose were used as a carbon source but not sucrose, arabinose and D(-) tartrate. Based on these features bacterial strains were identified as Pseudomonas tolaasii. The identity of isolated strains was confirmed by polymerase chain reaction. This is the first report of P. tolaasii in Serbia.     

Trichomonas gallinae, in comparison to Tetratrichomonas gallinarum, induces distinctive cytopathogenic effects in tissue cultures

In the present study the interaction of three genetically different clonal cultures of Trichomonas gallinae and Tetratrichomonas gallinarum with a permanent chicken liver (LMH) and a permanent quail fibroblast (QT35) cell culture was studied. Proliferation of T. gallinae cells was associated with a disintegration of the cell monolayer. The initial lesions on the LMH monolayer consisted of a progressive accumulation of the flagellate, forming clumps attached to the monolayer. A prolonged incubation time was characterized by appearance of holes in the cell monolayer with accumulation of trichomonads at their periphery. According to the severeness of the monolayer disruption differences among three tested T. gallinae clones were noticed. Furthermore, filtrates obtained either from axenic cultures of T. gallinae or from infected cell cultures produced a cytopathogenic effect similar to the protozoal cells, on both types of cell cultures. However, the destructive effect of the flagellates and their cell-free filtrates was much more pronounced on the LMH monolayer in comparison with the QT35 cells. Furthermore, freshly seeded LMH and QT35 cells suspended in cell-free filtrates of T. gallinae were unable to form a confluent monolayer. In comparison to T. gallinae, clonal cultures of T. gallinarum or their cell-free filtrates produced no effect on both types of monolayers. Interestingly, the cell-free filtrates obtained from both trichomonad species had an effect on the viability of both cell cultures. However, the cytotoxic effect of T. gallinarum filtrates was less severe than that recorded by T. gallinae. Consequently, for the first time a destruction of specified monolayers induced by T. gallinae-free filtrates could be demonstrated.     

Eco-geographical distribution of <Emphasis Type="Italic">Lactuca aculeata</Emphasis> natural populations in northeastern Israel

The wild lettuce, Lactuca aculeata Boiss. et Ky., is closely related and fully interfertile with cultivated lettuce, Lactuca sativa L. Altogether, 155 accessions of this wild species, which is rare conserved within the world’s germplasm collections, were collected from 12 localities throughout northeastern Israel. Ten of these localities represent different sites across the Golan Heights, while two, high-density populations were suprisingly found at the Hula Plain (first report of this species in this region). Lactuca aculeata was recorded at various elevations (222–968 m a.s.l.) and habitats. The taxonomic status of 30 L. aculeata accessions was morphologically validated during ex situ seed regeneration of 31 random accessions representing all 12 localities. Characterization of 12 traits showed that the Israeli populations of L. aculeata do not exhibit broad morphological variability, but more great levels of variation were obtained for developmental traits. Relatively little variability was observed in the morphology of cauline leaves or in the distribution of anthocyanin pigmentation in bracteae. In two cases, L. aculeata plants lacked trichomes in the inflorescence, a typical feature in the majority of plants. These unique collections of wild lettuce may carry novel sources of genetic variation for a wide range of traits and, thus, should be of interest for careful evaluation and exploitation in lettuce breeding.     

Comparison of Antioxidant Components and Activity of Buckwheat and Wheat Flours

Plant phenolics and tocopherols content were determined in light and wholegrain buckwheat and wheat flour. Antioxidant activity of flours were comparatively assessed by scavenging activity on 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH^•), hydroxyl (^•OH), and superoxide anion (O₂^•–) radicals, reducing activity, and chelating activity on Fe²⁺. Rutin, quercetin, and ferulic acid were quantified in both buckwheat flours, while ferulic acid was quantified in wholegrain wheat flour. Significantly higher content of phenolics and tocopherols was found in buckwheat than in wheat flours. Tocopherols in buckwheat flours were present in the order: γ‐ > α‐>> δ‐tocopherol, and in wheat flours: α‐ > γ‐ >> δ‐tocopherol. Buckwheat flours possessed better scavenging abilities on DPPH^•, ^•OH and O₂^•– radicals, as well as better reducing activity, while wheat flours showed better chelating activity on Fe²⁺, according to IC₅₀ values. Results suggest the possibility of improving the antioxidant properties of wheat‐based food products through addition of buckwheat flour.