Catastrophic drought in the Afro-Asian monsoon region during Heinrich event 1

Between 15,000 and 18,000 years ago, large amounts of ice and meltwater entered the North Atlantic during Heinrich stadial 1. This caused substantial regional cooling, but major climatic impacts also occurred in the tropics. Here, we demonstrate that the height of this stadial, about 16,000 to 17,000 years ago (Heinrich event 1), coincided with one of the most extreme and widespread megadroughts of the past 50,000 years or more in the Afro-Asian monsoon region, with potentially serious consequences for Paleolithic cultures. Late Quaternary tropical drying commonly is attributed to southward drift of the intertropical convergence zone, but the broad geographic range of the Heinrich event 1 megadrought suggests that severe, systemic weakening of Afro-Asian rainfall systems also occurred, probably in response to sea surface cooling.     

Oleaster oil positively modulates plasma lipids in humans

The olive tree had been domesticated during the early Neolithic in the Near East, and more than 1000 different cultivars have been identified to date. However, examples of wild olive trees (Olea europaea oleaster) can still be found in the Mediterranean basin. Evidence of oleaster use for oil production can be found in historical and sacred texts, such as the Odyssey, the Holey Koran, and the Holey Bible. While the nutritional and healthful properties of olive oil are actively being explored, there are no data on the human actions of oleaster oil. Therefore, we investigated the effect of prolonged, i.e., 1 month, consumption of oleaster oil on the lipid profile of a 40 healthy Algerian subjects (aged 27.9 ± 3.85 years), as compared to nonconsumers from the same area. Plasma urea, creatinine, and uric acid concentrations and glycemia did not significantly differ, at the end of the study, between controls and oleaster-oil-supplemented subjects. Conversely, we recorded significant decreases of plasma triglyceride concentration (-24.8%; p < 0.05), total cholesterol (-12.13%; p < 0.05), and low-density lipoprotein-cholesterol (LDL-C) (-24.39%; p < 0.05) in oleaster-oil-treated subjects. Concomitantly, high-density lipoprotein-cholesterol (HDL-C) concentrations were significantly increased (17.94%; p < 0.05) by oleaster oil administration. In conclusion, we show that oil obtained from feral olive trees, i.e., oleasters, improves the plasma lipid profile of healthy volunteers.     

Influence of shearing on oxidative stress and some physiological parameters in ewes

The aim of the present study was to evaluate the effect of shearing on physiological and oxidative stress parameters in ewes. Twenty Comisana ewes were used and divided into two groups. Ten ewes were left unshorn as a control group (Group A) and 10 ewes were shorn (Group B). All measurements were taken before and after shearing, and repeated 8 h after shearing and 1, 2, 3, 4, 5, 10 and 15 days after shearing. Reactive oxygen species (dROMs), antioxidant barrier (oxy‐adsorbent), thiol antioxidant barrier (SHp) and packed cell volume (PCV) were assessed in blood samples collected by means of jugular venipuncture. Rectal temperature (RT), respiratory rate (RR) and heart rate (HR) were also measured. Two‐way repeated measures analysis of variance (ANOVA), followed by Bonferroni's test, was used for the assessment of significant effects due to shearing and time. The statistical analysis showed significant increases (P < 0.01) of dROMs, oxy‐adsorbent, SHp, and a significant decrease (P < 0.01) of RT and RR associated with time and shearing. Our results indicate that shearing causes a change in the ewe's homeostatic balance that leads to oxidative stress.     

sunTILL: a TILLING resource for gene function analysis in sunflower

Background

Cultivated sunflower (Helianthus annus L.) is a globally important oilseed crop, subjected to intensive genetic and genomic studies. Although classical mutagenesis has successfully been applied to Helianthus genus in the past, we have developed the first sunflower TILLING resource.

Results

To balance the maximum mutation density with an acceptable plant survival rate, a 'kill curve' analysis was first conducted with different ethylmethanesulfonate (EMS) dosages and different exposure times. According to the germination rate, a treatment with 0.7% EMS for 6 h was chosen. An M₂ progeny of 3,651 fertile plants was obtained. Totally, 4.79% of the whole population showed clear aberrant phenotypes. A microsatellite analysis on a representative sample of the original seed stock and mutant lines confirmed the uniformity of the genetic background of plant material. The TILLING procedure was successfully applied to sunflower genome, initially by a CelI-nuclease mismatch cleavage assay coupled with a DNA-pooling level test. To investigate the efficiency of the mutagenic treatment, a pilot screening was carried out on 1,152 M₂ lines focusing on four genes, three involved in the fatty acid biosynthetic pathway and one for downy mildew resistance. A total of 9 mutant lines were identified and confirmed by sequencing; thereby, the estimated overall mutation frequency for the pilot assay resulted to be 1/475 kb.

Conclusion

A first TILLING population for a high throughput identification of EMS-induced point mutations in sunflower genome has been successfully obtained. This represents a powerful tool to a better understanding of gene function in sunflower.     

Clavibacter michiganensis subsp. michiganesis: Tracking strains using their genetic differentiations by ISSR markers in Southern Turkey

There is no available data published related to the dissemination of bacterial canker caused by Clavibacter michiganensis subsp. michiganensis (Cmm) and its genetic diversity in Turkey. It is important to control new introduced inoculum sources by commercial seeds and seedlings. Pathogens were identified by morphological features and the identity was confirmed by PCR amplification using a specific primer PSA-4, PSA-R in addition to microbiological tests. ISSR markers showing high polymorphism ratios were selected and used to characterize Cmm strains. The collected strains were classified into different groups on the basis of ISSR-PCR fingerprints, which showed remarkable genetic specificity and diversity not previously identified in Cmm, suggesting that genetic differences are related to dissemination of the pathogen in the region. This is the first ever study carried out on the characterization of Cmm using ISSR. The selected ISSR primers to characterize Cmm can be used to determine genetic differences in further studies.     

Agronomic Practices at Low Environmental Impact for Durum Wheat in Mediterranean Conditions

The application of conservative agricultural practices such as crop rotation, shallow tillage, and organic fertilizer could usefully sustain crop yield and increase soil fertility, thus playing an important role in the sustainable agriculture. This study was conducted to determine the effects of conservative agronomic practices on yield and quality of wheat. The effects of these practices on soil fertility were further investigated in this four-year study (2005–2008). Two cropping systems, durum wheat in continuous cropping, and in two-year rotation with leguminous crops, were investigated at Foggia (Southern Italy) in rain-fed conditions. Within each cropping system, two levels of crop management were compared: i) conventional, characterized by a higher soil tillage management and mineral fertilizers application; ii) conservative, with a lower soil tillage management and organic-mineral fertilizers. The seasonal weather greatly affected the wheat yield and quality, inducing lower production in years that were characterized by unfavorable climatic conditions. This trend was found when the conventional treatment was applied, both in continuous cropping and rotations. The effects of cropping systems and crop management pointed out the positive role played by the leguminous crops (common vetch and chickpea) in crop rotation. This introduction improved wheat yield in rotation (6.47% compared to the continuous cropping), improved grain protein content (5.88%), and reduced the productive gap between conventional and conservative treatments (9.24 and 14.14% of the wheat in rotation and continuous cropping, respectively). Conversely, the effects of cropping systems and crop management on soil fertility were not very high, since the differences found at the end of the study in total nitrogen values were poor. However, total organic carbon (16.04 and 17.58% for cropping system and crop management, respectively) and available phosphorus values (11.30 and 7.43%) depend on root organic matter contribution, plant biomass residues, and fertilizations. The suitable crop rotation and the sustainable crop management appear important agronomical practices to improve yield and quality of wheat, and may reduce the environmental risks resulting from conventional intensive cropping systems.     

High levels of genetic differentiation between Wolbachia-infected and non-infected populations of Folsomia candida (Collembola, Isotomidae)

The Gram-negative -Proteobacterium Wolbachia pipientis has been described as an obligate endosymbiont in many arthropod species, where it induces a variety of reproductive alterations, including parthenogenesis. Recently, this microorganism has also been detected in the parthenogenetic collembolan Folsomia candida. Here, we confirm the occurrence of the endosymbiont also in two Italian parthenogenetic populations of F. candida using ultrastructural (electron microscopy) and molecular (PCR screening on two bacterial genes) evidence. The strain isolated in the Italian populations has almost-identical gene sequences compared to that previously isolated in other populations of F. candida. In addition, we discovered a population of Folsomia cf. candida, which showed the presence of both males and females. This population is not infected by Wolbachia. A screening of two mitochondrial genes (COI and COII) showed that the bisexual population has high levels of genetic divergence in comparison with the parthenogenetic ones, even suggesting the possibility that it belongs to a different species. Furthermore, the remarkably high levels of genetic divergence between the two parthenogenetic populations suggests a possible influence of Wolbachia on inducing such differentiation, and, in the long term, speciation.     

Determination of choline in milk, milk powder, and soy lecithin hydrolysates by flow injection analysis and amperometric detection with a choline oxidase based biosensor

A fast-response and interference-free amperometric biosensor based on choline oxidase immobilized onto an electropolymerized polypyrrole film for flow injection determination of choline in milk, milk powder, and soy lecithin hydrolysates is described. The sensor displayed an Imax value of 1.9 +/- 0.2 microA and an apparent Michaelis-Menten constant, k'M, equal to 1.75 +/- 0.07 mM. Detection limits of 0.12 microM could be obtained. Because even a slight deterioration of the anti-interference membrane can adversely affect measurement accuracy, a real time monitoring of the biosensor selectivity has been achieved by a dual Pt electrode flow-through cell where the enzyme modified electrode is coupled to an enzyme-free electrode in a parallel configuration. Finally, bracketing technique (alternate injections of sample and standards) allows a two-point calibration to be performed in real-time, correcting for any drift in sensor response.     

Phenolic constituents and antioxidant properties of Xanthosoma violaceum leaves

An extract of Xanthosoma violaceum leaves was subjected to a polyphenol profile determination, including total polyphenols, and antioxidant activity evaluation. Analysis of the extract resulted in the isolation of a new flavone C-glycoside, apigenin 6-C-beta-D-glucopyranosyl-8-C-beta-D-apiofuranoside (1), as well as known flavone C-glycosides, including vitexin (2), isovitexin (3), isovitexin 4'-O-rhamnopyranoside (4), apigenin 6-C-[beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside] (5), and apigenin 6,8-diC-beta-D-glucopyranoside (6). The antioxidant activity of the extract was assessed by means of two different in vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and peroxidation induced by the water-soluble radical initiator 2,2'-azobis(2-amidinopropane) hydrochloride, on mixed dipalmitoylphosphatidylcholine/linoleic acid unilamellar vesicles (LP-LUV test). In both tests used, the extract and a fraction II showed a significant antioxidant/free-radical scavenging effect (fraction II, EC(50) = 11.6 microg/mL) in comparison to alpha-tocopherol (EC(50) = 10.1 microg/mL).     

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.