Contrasting Performance of Two Winter Wheat Varieties Susceptible to Leaf Rust under Diverse Pathogen Pressure, Fungicide Application, and Cultivation Practices.

This study investigated the relationship between yield, thousand kernel weight (TKW), and crude protein of soft white winter wheat-club variety (Barbee) and soft white winter wheat common variety (Zvezdana) susceptible to leaf rust and powdery mildew under different cultivation practices. Results revealed divergence in associations between yield, TKW, and crude protein loss of winter wheat varieties susceptible to obligate pathogens. Under the same level of leaf rust infection, N-input limited yield loss of the two varieties but not to the same extent. TKW loss was affected only by variety×cultivation practice and was significantly correlated with yield loss (r = -0.727, p = 0.011) and crude protein loss (r = -0.600, p = 0.05) only in club winter wheat. We suspected that Ninput affects the difference in the relationship between yield and TKW loss among varieties. Crude protein and yield loss had a low association (R2 = 18%, p = 0.05). Finally, this study indicated that more attention should be paid to the determination of pathogen pressure that triggers yield loss. It also pointed out that yield, TKW, and crude protein response to fungicides could differ in susceptible varieties. The contribution of fungicide to yield enhancement was highly associated with the specific reaction of the variety to pathogen infection rather than solely the disease level itself.

Comprehensive analysis of oxidative stability and nutritional values of germinated linseed and sunflower seed oil.

Germination of seeds is known to affect the nutritional composition of cold-pressed oils. This study focused on the effects of germination on the antioxidants and oxidative stability of linseed and sunflower seed oil. As hypothesized, germination led to increased antioxidant activities and tocopherol, chlorophyll and carotenoid content. Analysis revealed a 37.2 ± 3.5-fold and 11.6 ± 1.5-fold increase in polyphenol content in linseed and sunflower seed oil from germinated seeds, respectively. Using LC-HRMS/MS, profiles with up to 69 polyphenolic substances were identified in germinated seed oils for the first time. Germination promoted lipid hydrolysis, as evidenced by NMR, with overall significant decreases in triacylglycerol content leading to increased diacylglycerol and free fatty acid values. Rancimat measurements predicted a 4.10 ± 0.52-fold longer shelf-life for germinated linseed oil. This study successfully demonstrated the potential of germination to develop PUFA-rich oils with enhanced antioxidant capacity and oxidative stability.

Molecular Epidemiology of Xanthomonas euvesicatoria Strains from the Balkan Peninsula Revealed by a New Multiple-Locus Variable-Number Tandem-Repeat Analysis Scheme.

Bacterial spot of pepper and tomato is caused by at least three species of Xanthomonas, among them two pathovars of Xanthomonas euvesicatoria, which are responsible for significant yield losses on all continents. In order to trace back the spread of bacterial spot pathogens within and among countries, we developed the first multilocus variable number of tandem repeat analyses (MLVA) scheme for pepper- and tomato-pathogenic strains of X. euvesicatoria. In this work, we assessed the repeat numbers by DNA sequencing of 16 tandem repeat loci and applied this new tool to analyse a representative set of 88 X. euvesicatoria pepper strains from Bulgaria and North Macedonia. The MLVA-16 scheme resulted in a Hunter-Gaston Discriminatory Index (HGDI) score of 0.944 and allowed to resolve 36 MLVA haplotypes (MTs), thus demonstrating its suitability for high-resolution molecular typing. Strains from the different regions of Bulgaria and North Macedonia were found to be widespread in genetically distant clonal complexes or singletons. Sequence types of the variable number of tandem repeats (VNTR) amplicons revealed cases of size homoplasy and suggested the coexistence of different populations and different introduction events. The large geographical distribution of MTs and the existence of epidemiologically closely related strains in different regions and countries suggest long dispersal of strains on pepper in this area.

Multilocus Genotyping Reveals New Molecular Markers for Differentiating Distinct Genetic Lineages among "Candidatus Phytoplasma Solani" Strains Associated with Grapevine Bois Noir.

Grapevine Bois noir (BN) is associated with infection by "Candidatus Phytoplasma solani" (CaPsol). In this study, an array of CaPsol strains was identified from 142 symptomatic grapevines in vineyards of northern, central, and southern Italy and North Macedonia. Molecular typing of the CaPsol strains was carried out by analysis of genes encoding 16S rRNA and translation elongation factor EF-Tu, as well as eight other previously uncharacterized genomic fragments. Strains of tuf-type a and b were found to be differentially distributed in the examined geographic regions in correlation with the prevalence of nettle and bindweed. Two sequence variants were identified in each of the four genomic segments harboring hlyC, cbiQ-glyA, trxA-truB-rsuA, and rplS-tyrS-csdB, respectively. Fifteen CaPsol lineages were identified based on distinct combinations of sequence variations within these genetic loci. Each CaPsol lineage exhibited a unique collective restriction fragment length polymorphism (RFLP) pattern and differed from each other in geographic distribution, probably in relation to the diverse ecological complexity of vineyards and their surroundings. This RFLP-based typing method could be a useful tool for investigating the ecology of CaPsol and the epidemiology of its associated diseases. Phylogenetic analyses highlighted that the sequence variants of the gene hlyC, which encodes a hemolysin III-like protein, separated into two clusters consistent with the separation of two distinct lineages on the basis of tufB gene sequences. Alignments of deduced full protein sequences of elongation factor-Tu (tufB gene) and hemolysin III-like protein (hlyC gene) revealed the presence of critical amino acid substitutions distinguishing CaPsol strains of tuf-type a and b. Findings from the present study provide new insights into the genetic diversity and ecology of CaPsol populations in vineyards.

Bioactive compounds and "in vitro" antioxidant activity of some traditional and non-traditional cold-pressed edible oils from Macedonia.

The bioactive compounds and "in vitro" antioxidant activity measured by three antioxidant assays of some traditional and non-traditional cold-pressed edible oils from Macedonia were object of this study. The fatty acid composition showed dominance of monounsaturated oleic acid in "sweet" and "bitter" apricot kernel oils with percentages of 66.7 ± 0.5 and 57.8 ± 0.3%, respectively. The most dominant fatty acid in paprika seed oil was polyunsaturated linoleic acid with abundance of 69.6 ± 2.3%. The most abundant tocopherol was γ-tocopherol with the highest quantity in sesame seed oil (57.6 ± 0.1 mg/100 g oil). Paprika seed oil, sesame seed oil and sweet apricot oil were the richest source of phytosterols. DPPH assay was the most appropriate for the determination of the antioxidant activity of cold-pressed sunflower oil due to high abundance of α-tocopherol with a level of 22.8 ± 1.1 mg/100 g of oil. TEAC assay is the best for the determination of the antioxidant activity of sesame seed oil and paprika seed oils as the richest sources of phenolic compounds. ß-carotene assay was the most suitable assay for oils obtained from high pigmented plant material. Triacylglycerols and phytosterol profiles can be used as useful markers for the origin, variety and purity of the oils.

Groundwater quality in alluvial and prolluvial areas under the influence of irrigated agriculture activities.

The aim of this study was to investigate the groundwater pollution from alluvial aquifers lying under surface agriculture activities in two geologically different areas: alluvial and prolluvial. The groundwater in investigated areas is neutral to alkaline (pH 7.05-8.45), and the major dissolved ions are bicarbonate and calcium. Groundwater samples from the alluvial area are characterized by nitrate concentration above the national maximum concentration limit (MCL) at 20.5% of samples [mean value (Me) 6.31 mg/L], arsenic concentrations greater than national MCL at 35.6% of investigated samples (Me 12.12 µg/L) and elevated concentrations of iron (Me 202.37 µg/L) and manganese (Me 355.22 µg/L) at 22.7% and 81% of investigated samples, respectively. Groundwater samples from the prolluvial area did not show significantly elevated concentrations of heavy metals, but the concentration of nitrate was considerably higher (Me 65.06 mg/L). Factor analysis positively correlates As with Mn and Fe, suggesting its natural origin. Nitrate was found in positive correlation with SO42- and Ni but in negative with NH4+, suggesting its anthropogenic origin and the relationship of these ions in the process of denitrification. The t-test analysis showed a significant difference between nitrate pollution of groundwater from alluvial and prolluvial areas. According to the chemical composition of groundwater, the process of denitrification is considered to be the main reason for the reduced presence of nitrate in the groundwater lying under alluvial deposits represented by chalk and sandstones. Denitrification in groundwater lying under prolluvial deposits represented by magmatic and metamorphic rock formations was not observed.

New insights into the chemistry of Coenzyme Q-0: A voltammetric and spectroscopic study.

Coenzyme Q-0 (CoQ-0) is the only Coenzyme Q lacking an isoprenoid group on the quinoid ring, a feature important for its physico-chemical properties. Here, the redox behavior of CoQ-0 in buffered and non-buffered aqueous media was examined. In buffered aqueous media CoQ-0 redox chemistry can be described by a 2-electron-2-proton redox scheme, characteristic for all benzoquinones. In non-buffered media the number of electrons involved in the electrode reaction of CoQ-0 is still 2; however, the number of protons involved varies between 0 and 2. This results in two additional voltammetric signals, attributed to 2-electrons-1H(+) and 2-electrons-0H(+) redox processes, in which mono- and di-anionic compounds of CoQ-0 are formed. In addition, CoQ-0 exhibits a complex chemistry in strong alkaline environment. The reaction of CoQ-0 and OH(-) anions generates several hydroxyl derivatives as products. Their structures were identified with HPLC/MS. The prevailing radical reaction mechanism was analyzed by electron paramagnetic resonance spectroscopy. The hydroxyl derivatives of CoQ-0 have a strong antioxidative potential and form stable complexes with Ca(2+) ions. In summary, our results allow mechanistic insights into the redox properties of CoQ-0 and its hydroxylated derivatives and provide hints on possible applications.

Hydroxylated derivatives of dimethoxy-1,4-benzoquinone as redox switchable earth-alkaline metal ligands and radical scavengers.

Benzoquinones (BQ) have important functions in many biological processes. In alkaline environments, BQs can be hydroxylated at quinoid ring proton positions. Very little is known about the chemical reaction leading to these structural transformations as well as about the properties of the obtained hydroxyl benzoquinones. We analyzed the behavior of the naturally occurring 2,6-dimethoxy-1,4-benzoquinone under alkaline conditions and show that upon substitution of methoxy-groups, poly-hydroxyl-derivatives (OHBQ) are formed. The emerging compounds with one or several hydroxyl-substituents on single or fused quinone-rings exist in oxidized or reduced states and are very stable under physiological conditions. In comparison with the parent BQs, OHBQs are stronger radical scavengers and redox switchable earth-alkaline metal ligands. Considering that hydroxylated quinones appear as biosynthetic intermediates or as products of enzymatic reactions, and that BQs present in food or administered as drugs can be hydroxylated by enzymatic pathways, highlights their potential importance in biological systems.

Development of a rapid and simple voltammetric method to determine total antioxidative capacity of edible oils.

In this work we report on a new, rapid and simple voltammetric method to determine the total antioxidant capacity (TAC) of the edible oils. The method explores the ABTS radical (2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid)) assay as a redox probe and it relays on measuring catalytic voltammetric currents. The electrocatalysis comprises redox regeneration of the electrochemically created ABTS(+) radical either by Trolox (6-hydroxy-2,5,7,8-tetramethychroman-2-carboxylic acid) or by antioxidants present in studied oils. The detection limit of the method is determined to be 0.5 mg/L of Trolox equivalent, being a slightly lower than the corresponding UV-VIS spectrophotometric method. Applying the proposed voltammetric method the total antioxidant capacity of three types of commercially available cold-pressed edible oils are determined, and the results are found to be in a very good agreement with those obtained by UV-VIS spectrophotometry. The reported voltammetric method is cheap, rapid and simple, and it can be used as a sustainable alternative to the UV-VIS methods for the determination of total antioxidant capacitance of oils and other liquid lipophilic nutrients. Potent antioxidant capacity of studied oils was also confirmed by electron paramagnetic resonance spectroscopy of superoxide anion produced by macrophages.

Assessment of the genotoxicity of heavy metals in Phaseolus vulgaris L. as a model plant system by Random Amplified Polymorphic DNA (RAPD) analysis.

Impact assessments of environmental pollutants are important in eco-genotoxicology. A random amplified polymorphic DNA (RAPD) technique was used to detect genotoxicity-induced DNA damage in Phaseolus vulgaris L. from heavy metals at two different concentrations. The results from six 10-base pair (bp) random RAPD primers with 60-70% GC content used, showed a total of 295 RAPD fragments of 700-4000 bp in molecular size in the seedlings of untreated and treated samples, of which only 163 fragments were polymorphic. Polymorphisms became evident as the disappearance and/or appearance of DNA fragments in treated samples compared to the control. A dendrogram constructed using the Numerical Taxonomy and Multivariate Analysis System (NTSYSps), showed that the control group merged with groups treated with CuSO(4)·5H(2)O (150 mg L(-1)) and MnSO(4)·H(2)O (150 mg L(-1)) in a separate cluster. These groups were linked with all of the other samples treated with metals at concentrations of 150 mg L(-1) and CuSO(4)·5H(2)O and Cd(NO(3))(2) at concentrations of 350 mg L(-1). Finally, the samples treated with metals at concentrations of 350 mg L(-1) together with NiSO(4) at the concentration of 150 mg L(-1), clustered separately. The DNA polymorphism detected by RAPD analysis offered a useful biomarker assay for the detection of toxic chemicals genotoxicity in plant model systems.