Molecular phylogenetic,morphological, and pathogenic analyses reveal a single clonal population of Septoria lycopersici with a narrower host range in Brazil

Septoria leaf spot, caused by Septoria lycopersici, is considered one of the most important diseases of tomato in Brazil. Despite its importance, the disease agent is still poorly studied. Septoria isolates collected from different production regions of Brazil were characterized by molecular, morphological, and pathogenic methods. A set of 104 isolates was sequenced for the DNA Tub, Cal, and EF1-α loci. Ten isolates were selected, according to geographical region of origin and type of leaf lesion (typical or atypical), for morphological characterization and for evaluation of aggressiveness on tomato cultivar Santa Clara. To evaluate the pathogen host range, cultivated and wild Solanaceae plants were inoculated with four selected isolates. The results showed that all isolates grouped with the type isolate of S. lycopersici in maximum likelihood and Bayesian inference trees. The isolates were morphologically similar. All isolates selected for pathogenicity testing on tomato were able to induce typical symptoms of the disease, but differed in their aggressiveness. A total of eight species of Solanaceae were also identified as potential alternative hosts for S. lycopersici. This information will provide a more accurate assessment of the risks involved with the introduction of new crops, especially of the genus Solanum, in areas where the species is already present. In addition, it will provide the basis for the establishment of more efficient methods in the management of Septoria leaf spot of tomatoes in natural conditions and in the different production systems.     

Chromosome and plasmid-encoded N-acyl homoserine lactones produced by Agrobacterium vitis wildtype and mutants that differ in their interactions with grape and tobacco

Agrobacterium vitis causes crown gall disease on grapevines. It also induces a specific necrosis on grape roots and a hypersensitive response (HR) on tobacco that are regulated by a complex quorum-sensing regulatory system. Strain F2/5 produces at least six N-acyl-homoserine lactones (AHLs) that function as signal molecules in quorum-sensing. The AHLs differ in acyl side chain length (8–16 carbons) as determined by gas chromatography/mass spectrometry and electrospray ionization tandem mass spectrometry. Mutant derivatives of F2/5 differ in ability to cause necrosis and the HR and show variable AHL profiles as determined by a thin-layer chromatography/biosensor assay. All wildtype A. vitis strains revealed the presence of long-chain AHLs regardless of tumorigenicity or ability to cause the HR. Whereas genes encoding long-chain AHLs are predicted to reside on the F2/5 chromosome, the determinants for short-chain AHLs were shown to be located on conjugal plasmids.     

Contrasting effects of heat shock during in vitro maturation on development of in vitro‐fertilized and parthenogenetic bovine embryos

This study investigated the influence of heat shock during in vitro maturation on embryo development following in vitro fertilization (IVF) or parthenogenesis (Part). Immature bovine cumulus–oocyte complexes were exposed to heat shock (41.0°C) during the first 12 hr of in vitro maturation (IVM), followed by 12 hr at 38.5°C. Control group consisted of in vitro maturation for 24 hr at 38.5°C. Oocytes were in vitro‐fertilized or activated with ionomycin and cultured in vitro for 192 hr post‐in vitro insemination or parthenogenetic activation (hpia). There was an interaction (p < .01) between temperature of IVM and method of oocyte activation (IVF or Part) for cleavage at 48 hpia. Heat shock had a negative impact (p < .01) on cleavage of IVF embryos, whereas no (p > .05) effect was found in the Part embryos. Embryo development towards blastocyst stage at 168 and 192 hpia decreased in both IVF and Part embryos derived from heat‐shocked oocytes. Heat shock increased (p < .05) the apoptotic index in Part blastocysts, but no effect (p > .05) was found in IVF counterparts. Heat shock also down‐regulated the expression of AQP3 (p < .01) and up‐regulated the expression of HSP70.1 (p < .01) in Part blastocysts, whereas it down‐regulated the expression of ATP1A1 (p < .05) in IVF blastocysts. In conclusion, the effects of heat shock during IVM on early embryo cleavage and blastocyst apoptosis are influenced by the method of oocyte activation and expression of some genes can be disturbed in embryos derived from heat‐shocked oocytes.     

Conventional analyses of data from dietary validation studies may misestimate reporting accuracy: illustration from a study of the effect of interview modality on children's reporting accuracy

OBJECTIVE: To compare two approaches to analysing energy- and nutrient-converted data from dietary validation (and relative validation) studies - conventional analyses, in which the accuracy of reported items is not ascertained, and reporting-error-sensitive analyses, in which reported items are classified as matches (items actually eaten) or intrusions (items not actually eaten), and reported amounts are classified as corresponding or overreported. DESIGN: Subjects were observed eating school breakfast and lunch, and interviewed that evening about that day's intake. For conventional analyses, reference and reported information were converted to energy and macronutrients; then t-tests, correlation coefficients and report rates (reported/reference) were calculated. For reporting error-sensitive analyses, reported items were classified as matches or intrusions, reported amounts were classified as corresponding or overreported, and correspondence rates (corresponding amount/reference amount) and inflation ratios (overreported amount/reference amount) were calculated. SUBJECTS: Sixty-nine fourth-grade children (35 girls) from 10 elementary schools in Georgia (USA). RESULTS: For energy and each macronutrient, conventional analyses found that reported amounts were significantly less than reference amounts (every P < 0.021; paired t-tests); correlations between reported and reference amounts exceeded 0.52 (every P < 0.001); and median report rates ranged from 76% to 95%. Analyses sensitive to reporting errors found median correspondence rates between 67% and 79%, and that median inflation ratios, which ranged from 7% to 17%, differed significantly from 0 (every P < 0.0001; sign tests). CONCLUSIONS: Conventional analyses of energy and nutrient data from dietary reporting validation (and relative validation) studies may overestimate accuracy and mask the complexity of dietary reporting error.     

Composition rather than viscosity modifies the aroma compound retention of flavored stirred yogurt

The influence of thickening agents (modified starch/pectin mixture 0 and 7 g/L) and mechanical treatment (low, medium, and high) on the retention of esters (pentyl acetate and ethyl pentanoate), aldehydes (hexanal and (E)-2-hexenal), and a lactone (gamma-octalactone) in low-fat flavored stirred yogurts were investigated under equilibrium conditions. In the range studied, the thickening agent and mechanical treatment had little influence on aroma compound retention compared to the decreasing effect of increasing dairy protein concentration on aldehyde retention and the "salting out" effect of carbohydrates on esters. Moreover, experiments in dynamic mode (study of the release of hexanal when yogurts were heated) showed, in the conditions studied, that heat and mass transfer coefficients were not influenced by any of the studied factors (thickening agents and mechanical treatment). These results under static and dynamic conditions are not related to the significant decreasing effect of thickening agents on apple sensory scores associated with hexanal, observed in a previous sensory study. Thus, this sensory effect of thickening agents may be due to sensory interactions between perceptions rather than physicochemical interactions.     

Bioactive constituent production in St. John's Wort in vitro hairy roots. Regenerated plant lines

A wild strain of Agrobacterium rhizogenes was used to regenerate twelve in vitro plant lines from different hairy roots of H. perforatum (St. John's Wort). The production of the main bioactive constituents was observed even though their yields varied in the different plant lines. Two lines were selected for the hyperoside production (4.9-4.6 mg/gdw) while nine were characterized by significant yields of chlorogenic acid (ranged from 0.47 to 1.09 mg/gdw). Furthermore, one out of twelve lines showed a 10-fold higher hypericin content (0.25 mg/gdw) than that reported for the in vitro shoots in the literature. Morphological and phytochemical features were determined in order to select H. perforatum genotypes enriched in valuable bioactive compounds.     

Use of electrochemical biosensor and gas chromatography for determination of dichlorvos in wheat

Two methods for the determination of dichlorvos in durum wheat by electrochemical assay and gas chromatography, respectively, have been developed. Dichlorvos, an organophosphorus anticholinesterase pesticide, was extracted from wheat with hexane, and the filtered extract was directly analyzed by gas chromatography with nitrogen-phosphorus flame detection (NPD). Recoveries of dichlorvos from milled wheat spiked at 0.25-1.5 microg/g ranged from 96.5 to 100.9%, and the limit of detection was 0.02 microg/g. The electrochemical assay was based on the detection of choline, the acetylcholinesterase product, via a choline oxidase biosensor. An aliquot of the filtered hexane extract was partitioned with phosphate buffer solution, and the organic layer was evaporated prior to electrochemical analysis. A limit of detection of 0.05 microg/g of dichlorvos was obtained with mean recoveries of 97-103% at spiking levels of 0.25-1.5 microg/g. A good correlation (r = 0.9919) was found between the results obtained with the electrochemical and those obtained with the gas chromatographic methods. The electrochemical method was peer-validated in two laboratories that analyzed 10 blind samples (5 duplicates), including a blank and 4 spiked samples with dichlorvos at levels of 0.25, 0.60, 1.00, and 1.50 microg/g. Within-laboratory repeatability (RSDr) and between-laboratory reproducibility (RSDR) ranged from 5.5 to 7.8% and from 9.9 to 17.6%, respectively.     

Soil chemical and biochemical properties of a salt-marsh alluvial Spanish area after long-term reclamation

Marisma, one of the largest salt-marsh alluvial areas in SW Spain, has been reclaimed since 1970 by artificial drainage and amendment with phosphogypsum (PG) so as to reduce Na⁺ saturation. Within the reclaimed area, two 250- × 20-m plots were treated as follows: (1) amendment with 25 Mg/ha of PG every 2 to 3 years between 1979 and 2003 (plot PY); (2) like PY but PG treatment stopped after 1997 (plot DR). A contiguous virgin Marisma salt-marsh plot (MV), neither drained nor amended, was the control. In MV, soil microbial biomass C, most enzyme activities and total organic C content were much greater than in PY and DR soils, despite the salinity stress. The decrease in soil organic matter content in PY and DR soils was likely due to cotton-cropping practices, which favoured the organic C mineralisation and nutrients removal by crops. Microbial activity of MV soil, probably stimulated by the rhizodepositions of the natural vegetation, did not suffer from the osmotic effect due to the raising of soil solution ionic strength. Microbial quotient could be ranked as MV > PY > DR, whereas the metabolic quotient had an opposite trend. Thus, both quotients suggested that the interruption of PG amendment was not favouring microbial activity. Principal component analysis clearly identified the chemical and biochemical soil properties mostly affected by the reclamation and also indicated the longer PG amendment in PY plot. Stepwise discriminant analysis identified two physiologically different types of soil microflora, one less active present in the MV virgin soil and another more active present in the reclaimed PY and DR soils.     

Effects of fly attack (Bactrocera oleae) on the phenolic profile and selected chemical parameters of olive oil

The phenolic fraction of virgin olive oil influences both its quality and oxidative stability. One of the principal threats of the quality of olive fruit is the olive fly ( Bactrocera oleae) as it alters the chemical composition. The attack of this olive pest has been studied in order to evaluate its influence on the quality of virgin olive oil (free acidity, peroxide value, fatty acid composition, water content, oxidative stability, phenols, and antioxidant power of phenolic fraction). The study was performed using several virgin olive oils obtained from olives with different degrees of fly infestation. They were acquired in different Italian industrial mills from the Abruzzo region. Qualitative and quantitative analyses of phenolic profiles were performed by capillary electrophoresis-diode array detection, and electrochemical evaluation of the antioxidant power of the phenolic fraction was also carried out. These analyses demonstrated that the degree of fly attack was positively correlated with free acidity ( r = 0.77, p < 0.05) and oxidized products ( r = 0.58, p < 0.05), and negatively related to the oxidative stability index ( r = -0.54, p < 0.05) and phenolic content ( r = -0.50, p < 0.05), mainly with secoiridoid compounds. However, it has been confirmed that the phenolic fraction of olive oil depends on several parameters and that a clear correlation does not exist between the percentages of fly attack and phenolic content.