Genetic diversity analysis of North Africa’s barley using SSR markers

It was demonstrated that some North Africa barley accessions have diverse tolerance sources for abiotic stresses and a good nutritional quality, but the studies done were incomplete since they were realized separately in each country apart.To implement a more complete analysis, 31 barley accessions originated from North Africa (Algeria, Tunisia and Egypt) were analyzed using 11 SSR markers selected from the seven barley linkage groups for studying the genetic diversity among these chosen barley accessions.Over the 11 SSR markers, a total of 478 reproducible bands were scored with an average of 2.13 alleles/primer and the average polymorphism information content of 0.5.Genetic distance analysis of the 31 accessions showed a large genetic diversity and high number of different groups. The most accessions are clustered according to their eco-geographical origin, according to their pedigree and agronomic characters or according to the caryopsis character (hulled or naked caryopsis). This high number of obtained groups is an invaluable aid in crop improvement strategies and confirms the opinion suggesting that North Africa could be a secondary center of origin of barley. The various growing conditions and the multiple uses of barley in each country may be the cause of the large variability of the barley germplasm in each region.     

Redox-related gene expression and sugar accumulation patterns are altered in the edible inflorescence produced by the cultivated form of pacaya palm (Chamaedorea tepejilote)

Background and AimsThe pacaya palm is a dioecious neotropical palm species that is exploited in Latin America for its male inflorescence, which is edible when immature. It is cultivated, in a non-intensive manner, in Guatemala, where a morphotype occurs that produces much larger, more highly branched inflorescences compared with wild palms. We sought to identify molecular factors underlying this phenotypic divergence, which is likely to be a product of domestication.MethodsWe performed RNA-seq-based studies on immature pacaya palm male inflorescences in order to identify genes that might be directly or indirectly affected in their expression in relation to domestication. We also measured the accumulation of a range of soluble sugar molecules to provide information on the biochemical status of the two different types of material.Key ResultsA total of 408 genes were found to display significantly different expression levels between the wild and cultivated morphotypes. Three different functional categories were found to be enriched in the gene set that was upregulated in the cultivated morphotype: redox balance; secondary metabolism; and transport. Several sugars were found to accumulate at higher levels in inflorescences of the cultivated morphotype, in particular myo-inositol, fructose and glucose.ConclusionsThe observed upregulation of redox-related genes in the cultivated morphotype is corroborated by the observation of higher myo-inositol accumulation, which has been shown to be associated with enhanced scavenging of reactive oxygen species in other plants and which may affect meristem activity.     

Environmental Fate of Two Organophosphorus Insecticides in Soil Microcosms under Mediterranean Conditions and Their Effect on Soil Microbial Communities

Assessing dissipation is an integral part of determining pesticide risk. The adsorption and dissipation characteristics of two model insecticides, chlorpyrifos (CHP) and dimethoate (DMT), in a Mediterranean soil were investigated in order to evaluate soil microbial toxicity and to study their soil bioavailability for the purpose of managing pesticide residue with potential bioremediation of contaminated soil. The aim of this study was also to define novel methods for assessing the ecotoxicity of CHP and DMT on microorganisms in the soil. Koc values ranged between 33420–91601 cm³/g and 129–184 cm³/g for CHP and DMT, respectively, indicating that the former is characterized by a strong adsorption affinity, whereas, the latter has a weaker one. In the dissipation study, the half-life (T_1/2) of CHP in top soil was 11.55days; whereas, when dissipation was studied in the same soil sterilized, the half-life was 13.86 days, showing a relatively important abiotic degradation effect. For DMT, however, T_1/2 was 17.32 days and 13.86 days in sterilized soil and non-sterilized soil, respectively, illustrating partial biotic degradation. In terms of leaching behavior, the groundwater ubiquity scores calculated for CHP and DMT were 0.85 and 1.95, respectively, indicating that CHP is a non-leacher, while DMT can be considered a transition insecticide.     

Ca2+-ATPases in non-failing and failing heart: evidence for a novel cardiac sarco/endoplasmic reticulum Ca2+-ATPase 2 isoform (SERCA2c)

We recently documented the expression of a novel human mRNA variant encoding a yet uncharacterized SERCA [SR (sarcoplasmic reticulum)/ER (endoplasmic reticulum) Ca2+-ATPase] protein, SERCA2c [Gélébart, Martin, Enouf and Papp (2003) Biochem. Biophys. Res. Commun. 303, 676-684]. In the present study, we have analysed the expression and functional characteristics of SERCA2c relative to SERCA2a and SERCA2b isoforms upon their stable heterologous expression in HEK-293 cells (human embryonic kidney 293 cells). All SERCA2 proteins induced an increased Ca2+ content in the ER of intact transfected cells. In microsomes prepared from transfected cells, SERCA2c showed a lower apparent affinity for cytosolic Ca2+ than SERCA2a and a catalytic turnover rate similar to SERCA2b. We further demonstrated the expression of the endogenous SERCA2c protein in protein lysates isolated from heart left ventricles using a newly generated SERCA2c-specific antibody. Relative to the known uniform distribution of SERCA2a and SERCA2b in cardiomyocytes of the left ventricle tissue, SERCA2c was only detected in a confined area of cardiomyocytes, in close proximity to the sarcolemma. This finding led us to explore the expression of the presently known cardiac Ca2+-ATPase isoforms in heart failure. Comparative expression of SERCAs and PMCAs (plasma-membrane Ca2+-ATPases) was performed in four nonfailing hearts and five failing hearts displaying mixed cardiomyopathy and idiopathic dilated cardiomyopathies. Relative to normal subjects, cardiomyopathic patients express more PMCAs than SERCA2 proteins. Interestingly, SERCA2c expression was significantly increased (166+/-26%) in one patient. Taken together, these results demonstrate the expression of the novel SERCA2c isoform in the heart and may point to a still unrecognized role of PMCAs in cardiomyopathies.     

Microbial Succession in a Compost-packed Biofilter Treating Benzene-contaminated Air

Air artificially contaminated with increasing concentrations of benzene was treated in a laboratory scale compost-packed biofilter for 240 days with a removal efficiency of 81–100%. The bacterial community in the packing material (PM) at different heights of the biofilter was analysed every 60 days. Bacterial plate counts and ribosomal intergenic spacer analysis (RISA) of the isolated strains showed that the number of cultivable aerobic heterotrophic bacteria and the species diversity increased with benzene availability. Identification of the isolated species and the main bands in denaturing gradient gel electrophoresis (DGGE) profiles from total compost DNA during the treatment revealed that, at a relatively low volumetric benzene load (1.2≤VBL≤6.4 g m⁻³ _PM h⁻¹), besides low G+C Gram positive bacteria, originally present in the packing compost, bacteroidetes and β- and γ-proteobacteria became detectable in the colonising population. At the VBL value (24.8 g m⁻³ _PM h⁻¹) ensuring the maximum elimination capacity of the biofilter (20.1 g m⁻³ _PM h⁻¹), strains affiliated to the genus Rhodococcus dominated the microflora, followed by β-proteobacteria comprising the genera Bordetella and Neisseria. Under these conditions, more than 35% of the isolated strains were able to grow on benzene as the sole carbon source. Comparison of DGGE and automated RISA profiles of the total community and isolated strains showed that a complex bacterial succession occurred in the reactor in response to the increasing concentrations of the pollutant and that cultivable bacteria played a major role in benzene degradation under the adopted conditions.     

NKT cells in the induced sputum of severe asthmatics

To determine whether there was a specific inflammatory process in severe asthmatics, the phenotypic characteristics of induced sputum immune cells were analysed among patients with severe asthma. Twenty-two induced sputa (10 severe asthmatics) were studied. Flow cytometric analysis was performed using immune cells of the sputum and monoclonal antibodies to CD3, CD4, CD8, CD56, CD25, and TCRgammadelta. The number of NKT (CD3(+) CD56(+)) cells was significantly higher in the sputum of severe asthmatics compared with mild asthmatic and healthy control groups (P < .05). CD8(+)CD56(+) cells were the predominant subtype of the increased NKT cells in severe asthmatics. CD3(+)CD56(+)Valpha24(+), TCRgammadelta(+) CD56(+), and CD4(+)CD25(+) T cells were significantly increased in severe asthmatic patients. These results suggest that the immunopathogenesis of severe asthmatics vary between severe and mild asthmatics, and that CD8(+)CD56(+) NKT cells may play an important role in the immunopathogenesis of severe asthma.     

Caldimonas hydrothermale sp. nov., a novel thermophilic bacterium isolated from roman hot bath in south Tunisia

A polyphasic approach was used to characterize a bacterium, HAN-85^T, isolated from thermal water in natural thermal spring at Tozeur, an oasis in southwest Tunisia. The novel isolate was thermophilic, strictly aerobic and amylolytic bacterium, which stained Gram negative. Cells were short rods motile by means of a single polar flagellum. Their optimum temperature and pH required for growth were 55°C and pH 7, respectively. Comparative 16S rRNA gene sequence analyses showed that strain HAN-85^T belonged to the genus Caldimonas, with highest sequence similarity to the type strains Caldimonas manganoxidans and Caldimonas taiwanensis. DNA–DNA hybridization measurements revealed low DNA relatedness (35.2–44.5%) between the novel isolate and its closest relative, C. manganoxidans. The major cellular fatty acid components were 16:0, 17:0 cyclo and summed feature 3. The DNA G+C content was 68.3 mol%. Taken together, the results of DNA–DNA hybridization, fatty acids profile, physiological tests and biochemical analyses have allowed the genotypic and phenotypic differentiation of the isolate from currently recognized Caldimonas species. Therefore, we suggest that this isolate is a novel species within the genus Caldimonas and propose that it should be named Caldimonas hydrothermale sp. nov. The type strain is HAN-85^T (=DSM 18497^T =LMG 23755^T). The Gen Bank/Embl/DDBJ accession number for the 16S rRNA gene sequence of strain DSM 18497^T is AM283038.     

<Emphasis Type="Italic">Planomonospora algeriensis</Emphasis> sp. nov., an actinobacterium isolated from a Saharan soil of Algeria

A filamentous actinobacterium, designated strain PM3^T, was isolated from a Saharan soil sample collected from Béni-Abbès, Béchar (South-West Algeria). A polyphasic taxonomic study was carried out to establish the status of strain PM3^T. The isolate was found to have morphological and chemotaxonomical properties associated with members of the genus Planomonospora. The new isolated microorganism developed cylindrical sporangia arranged in double parallel rows on aerial mycelium, each one containing a motile single sporangiospore. The cell wall of the strain was found to contain meso-diaminopimelic acid. Whole-cell hydrolysates were found to contain madurose, glucose, mannose and ribose. The predominant menaquinone was identified as MK-9(H₂) (69.6%). The polar lipids detected were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidylhydroxyethanolamine and glucosamine-containing lipids. The major fatty acids were found to be C_17:1ω9c (38.6%) and C_17:0 (24.2%). Results of 16S rRNA gene sequence comparison revealed that strain PM3^T shared a high degree of 16S rRNA gene sequence similarity with Planomonospora sphaerica DSM 44632^T (99.3%), Planomonospora parontospora subsp. parontospora DSM 43177^T (99.2%) and P. parontospora subsp. antibiotica DSM 43869^T (99.0%). DNA–DNA hybridization values between strain PM3^T and the type strains of the closely related species were between 58.4 and 70.1%. The combination of phylogenetic analysis, DNA–DNA relatedness data, phenotypic characteristics and chemotaxonomic data support the conclusion that strain PM3^T represents a novel species of the genus Planomonospora, for which the name Planomonospora algeriensis sp. nov. is proposed. The type strain is PM3^T (=DSM 46752^T = CECT 9047^T).     

Influence of freezing stress on morphological alteration and biofilm formation by Listeria monocytogenes: relationship with cell surface hydrophobicity and membrane fluidity

The morphological changes and adhesive property of three Listeria monocytogenes strains submitted to freezing stress (?20 °C) were studied. The atomic force micrographs showed a reduction in the cell size and an evolution to coccoid shape. The phenotypic slime production of L. monocytogenes and the expression of the adhesive gene were investigated before and after 10 months of incubation in salmon at ?20°. Our results showed that after ten months, stressed stains become more adherent and able to produce slime. In addition, we noted that this pathogen presents same physiological changes to adapt to starvation conditions. The cellular fatty acids composition of adhered and floating cells of three L. monocytogenes strains was taken into consideration. The stressed strains presented different chain lengths and therefore an increase in the hydrophobicity level. Moreover, we noted that the adhesive property of L. monocytogenes strains affects the Benzalkonium chloride bacterial sensitivity which increased after biofilm formation.     

Variation of the Chemical Composition and Antimicrobial Activity of the Essential Oils of Natural Populations of Tunisian Daucus carota L. (Apiaceae)

The essential oils of Daucus carota L. (Apiaceae) seeds sampled from ten wild populations spread over northern Tunisia were characterized by GC‐FID and GC/MS analyses. In total, 36 compounds were identified in the D. carota seed essential oils, with a predominance of sesquiterpene hydrocarbons in most samples (22.63–89.93% of the total oil composition). The main volatile compounds identified were β‐bisabolene (mean content of 39.33%), sabinene (8.53%), geranyl acetate (7.12%), and elemicin (6.26%). The volatile composition varied significantly across the populations, even for oils of populations harvested in similar areas. The chemometric principal component analysis and the hierarchical clustering identified four groups, each corresponding to a composition‐specific chemotype. The in vitro antimicrobial activity of the isolated essential oils was preliminarily evaluated, using the disk‐diffusion method, against one Gram‐positive (Staphylococcus aureus) and two Gram‐negative bacteria (Escherichia coli and Salmonella typhimurium), as well as against a pathogenic yeast (Candida albicans). All tested essential oils exhibited interesting antibacterial and antifungal activities against the assayed microorganisms.