Spa32 regulates a switch in substrate specificity of the type III secreton of Shigella flexneri from needle components to Ipa proteins

Type III secretion systems (TTSS) are essential virulence determinants of many gram-negative bacteria and serve, upon physical contact with target cells, to translocate bacterial proteins directly across eukaryotic cell membranes. The Shigella TTSS is encoded by the mxi/spa loci located on its virulence plasmid. By electron microscopy secretons are visualized as tripartite with an external needle, a transmembrane domain, and a cytoplasmic bulb. In the present study, we generated a Shigella spa32 mutant and studied its phenotype. The spa32 gene shows low sequence homology to Salmonella TTSS1 invJ/spaN and to flagellar fliK. The spa32 mutant, like the wild-type strain, secreted the Ipas and IpgD, which are normally secreted via the TTSS, at low levels into the growth medium. However, unlike the wild-type strain, the spa32 mutant could neither be induced to secrete the Ipas and IpgD instantaneously upon addition of Congo red nor penetrate HeLa cells in vitro. Additionally, the Spa32 protein is secreted in large amounts by the TTSS during exponential growth but not upon Congo red induction. Interestingly, electron microscopy analysis of the spa32 mutant revealed that the needle of its secretons were up to 10 times longer than those of the wild type. In addition, in the absence of induction, the spa32 mutant secreted normal levels of MxiI but a large excess of MxiH. Taken together, our data indicate that the spa32 mutant presents a novel phenotype and that the primary defect of the mutant may be its inability to regulate or control secretion of MxiH.     

Genetic variation of salt-stressed durum wheat (Triticum turgidum subsp. durum Desf.) genotypes under field conditions and gynogenetic capacity

Agriculture has new challenges against the climate change: the preservation of genetic resources and the rapid creation of new varieties better adapted to abiotic stress, specially salinity. In this context, the agronomic performance of 25 durum wheat (Triticum turgidum subsp. durum Desf.) genotypes (nineteen landraces and six improved varieties), cultivated in two semi-arid regions in the center area of Tunisia, were assessed. These sites (Echbika, 2.2?g?l^?1; Barrouta, 4.2?g?l^?1) differ by their degree of salinity of the water irrigation. The results showed that most of the agronomic traits (e.g. spike per meter square, thousand kernels weight and grain yield) were reduced by salinity. Durum wheat landraces, Mahmoudi and Hmira, and improved varieties, Maali and Om Rabia showed the widest adaptability to different quality of irrigation water. Genotypes including Jneh Kotifa and Arbi were estimated as stable genotypes under adverse conditions. Thereafter, salt-tolerant (Hmira and Jneh Khotifa) and the most cultivated high-yielding (Karim, Razzak and Khiar) genotypes were tested for their gynogenetic ability to obtain haploids and doubled haploid lines. Genotypes with good induction capacity had not necessarily a good capacity of regeneration of haploid plantlets. In our conditions, Hmira and Khiar exhibited the best gynogenetic ability (3.1% and 2.9% of haploid plantlets, respectively).     

Cutting edge: small molecule CD40 ligand mimetics promote control of parasitemia and enhance T cells producing IFN-gamma during experimental Trypanosoma cruzi infection

Host resistance to Trypanosoma cruzi infection depends on a type 1 response characterized by a strong production of IL-12 and IFN-gamma. Amplifying this response through CD40 triggering results in control of parasitemia. Two newly synthesized molecules (<3 kDa) mimicking trimeric CD40L (mini CD40Ls(-1) and (-2)) bind to CD40, activate murine dendritic cells, and elicit IL-12 production. Wild-type but not CD40 knockout mice exhibited a sharp decrease of parasitemia and mortality when inoculated with T. cruzi mixed with miniCD40Ls. Moreover, the immunosuppression induced by T. cruzi infection was impaired in mice treated with miniCD40Ls, as shown by proliferation of splenic lymphocytes, percentage of CD8(+) T cells, and IFN-gamma production. Mice surviving T. cruzi infection in the presence of miniCD40L(-1) were immunized against a challenge infection. Our results indicate that CD40L mimetics are effective in vivo and promote the control of T. cruzi infection by overcoming the immunosuppression usually induced by the parasites.     

Comparative performance of δ13C,ion accumulation and agronomic parameters for phenotyping durum wheat genotypes under various irrigation water salinities

The use of efficient selection traits for screening under contrasting irrigation water salinity is a challenge for breeders. To identify patterns, grain yield (GY) and yield components (kernels m^?2, thousand kernels weight), growth traits (plant height, biomass), flag leaf ion accumulation (Na⁺ and K⁺), carbon isotope composition (δ¹³C_grain) and nitrogen concentration (N_grain) of grains were assessed on 25 durum wheat genotypes (G) in two consecutive growing seasons (2010 and 2011), in three semi‐arid locations in Tunisia. Each location differed in their irrigation water salinity as measured by electrical conductivity: Echbika (S1, 6 dS m^?1), Barrouta (S2, 12 dS m^?1) and Sidi Bouzid (S3, 18 dS m^?1). GY was shown to be negatively correlated to N_grain as well as to δ¹³C_grain. This is confirmed by a multiple linear regression analysis that showed that both δ¹³C_grain and N_grain were the major determinant components for GY variability under S3. A high genotypic variability was observed and the improved genotype Maali exhibited the most stable GY under the three irrigation water salinities and the two cropping seasons. Maali showed the lowest δ¹³C_grain. This indicates that tolerance in durum wheat is likely to be correlated to the ability of maintaining a high stomatal conductance. According to our data suggests δ¹³C_grain can be used for an efficient screening of salt tolerant durum wheat. Under our experimental conditions, N_grain was shown to be highly correlated to δ¹³C_grain and can therefore be easier‐to‐use trait to assess the tolerance to salinity.     

Delivery of biologically active anti-inflammatory cytokines IL-10 and IL-1ra in vivo by the Shigella type III secretion apparatus

Pathogenicity of many Gram-negative bacteria relies on a type III secretion (T3S) apparatus, which is used for delivery of bacterial effectors into the host cell cytoplasm allowing the bacteria to manipulate host cell cytoskeleton network as well as to interfere with intracellular signaling pathways. In this study, we investigated the potential of the Shigella flexneri T3SA as an in vivo delivery system for biologically active molecules such as cytokines. The anti-inflammatory cytokines IL-10 and IL-1 receptor antagonist (IL-1ra) were genetically fused to the first 30 or 60 residues of the Shigella T3S effector IpaH9.8 or to the first 50 residues of the Yersinia enterocolitica effector YopE and the recombinant fusion proteins were expressed in S. flexneri. YopE(50)-IL-10, IpaH(60)-IL-10, and IpaH(60)-IL-1ra were efficiently secreted via the T3S apparatus of Shigella. Moreover, these recombinant proteins did not impair the invasive ability of the bacteria in vitro. In a murine model, Shigella strains expressing YopE(50)-IL-10, IpaH(60)-IL-10, and IpaH(60)-IL-1ra induced a lower mortality in mice that was associated with reduced inflammation and a restricted localization of bacteria within the lung tissues as compared with wild-type Shigella. Moreover, the level of TNF-alpha and IL-1beta mRNA were reduced in the lungs following infection by IL-10- and IL-1ra-secreting Shigella, respectively. These findings demonstrate that the Shigella T3S apparatus can deliver biologically active cytokines in vivo, thus opening new avenues for the use of attenuated bacteria to deliver proteins for immunomodulation or gene therapy purposes.     

New resource assessment characterization factors for rare earth elements: applied in NdFeB permanent magnet case study

The International Journal of Life Cycle Assessment - Rare earth elements (REEs) are among the most critical raw materials with a high supply risk. Despite their criticality, REEs are not covered by...     

Involvement of Carbohydrates in Response to Preconditioning Flooding in Two Clones of <Emphasis Type="Italic">Populus deltoides</Emphasis> Marsh. × <Emphasis Type="Italic">P.</Emphasis> <Emphasis Type="Italic">nigra</Emphasis> L.

Poplar, a fast-growing species widely used outside its original area of distribution, was evaluated in the present study to verify its tolerance and hardening to waterlogging in anticipation of its implementation in rehabilitation practices for marginal lands subjected to flooding. Three water regimes were applied to the plants, grown in pots, with two clones (I-488 and D-64) with different sensitivities to flooding. These plants included a lot consisting of control plants (C), which were irrigated with good drainage, a non-preconditioned lot (NPr), and a third lot that was preconditioned to flooding (Pr). Furthermore, flooding was imposed on NPr and Pr plants by submerging pots up to 5 cm above the collar for 60 days followed by a 40-day recovery period. At the end of these two periods, shoot dry mass, foliar concentrations of certain ions, soluble sugars, starch and proline, and the relative electrolyte leakage were evaluated. The preconditioning treatment conferred different degrees of hardness on the treated plants compared with NPr plants. Our results revealed that high carbohydrate availability (soluble sugars and starch) is suggested to participate in sustaining membrane integrity which may affect the recuperative potential of Pr plants, notably those of clone I-488, from flooding damage.     

Effects of mycorrhizal fungi inoculation and soil amendment with hydrogel on leaf anatomy,growth and physiology performance of olive plantlets under two contrasting water regimes

Abiotic stresses present a real environmental problem in agriculture field. In our paper, we examine the significance of arbuscular mycorrhizal fungi (AMF) and soil amendment with water retaining superpolymers (hydrogel) on growth and physiology performance of olive plantlets. Our experiment was carried out in nursery conditions, to test the impact of hydrogel (TH) and mycorrhizal fungi (TM), used individually or combined (THM), and compare them with non inoculated plants (TC), to understand and reduce the water stress damage in olive plantlets (cv. Chemlali). We also evaluate interactions between hydrogel, mycorrhizal treatments and water regimes. Results of mycorrhization (M%) show that roots colonized by Rhizophagus irregularis of well-watered plants were about 40.87%. In combined treatment (THM), M% was about 32.14%. Compared to TC treatment, TM treatment enhances significantly the dry weights of the whole plant under the two water regimes. The TM treatment had the highest relative water content (66.50%) and Chl (a?+?b) (0.83 mg g^??1) in stressed conditions. We found also that under water stress, the maximal quantum efficiency of the photosystem II measurements in leaves were significantly improved by 50.70% in TH treatment compared to control. For phenolic contents, TH treatment decreased significantly total phenols by 50.10% compared to TC. Our study gives evidence that the use of AMF and the hydrogel separately or in combination may enhance the capacity to avoid drought damages of olive plantlets and improve olive performances.     

Effects of nitrogen rates on grain yield and nitrogen agronomic efficiency of durum wheat genotypes under different environments

Durum wheat is an important staple food crop in Tunisia and other Mediterranean countries and is grown in various climatic conditions. Production and yield are however severely limited not only by drought events but also by reduced levels of nitrogen fertilisation. A study was carried out at two locations in the sub‐humid area of Tunisia: Mateur in 2009–10 and 2010–11 and Beja in 2011–12 and 2012–13 under rainfed conditions. Four durum wheat genotypes (landraces: Bidi, Azizi; improved: Om Rabia, Khiar) were evaluated for nitrogen agronomic efficiency and related agronomic traits under various nitrogen rates: 0, 50, 100, 150, 200 and 250 kg N ha^?1, with three replications. There was a significant interaction effect (P ≤ 0.001) environments × genotypes × N treatments for grain yield (GY), biomass yield (BY), harvest index (HI), partial factor productivity of applied nitrogen (PFP_N) and nitrogen agronomic use efficiencies (NAE). GY was the most affected trait by nitrogen applied showing an increase of 94% under high N treatment (250 kg N ha^?1) compared to control plots without N treatments. A significant linear regression exists between GY (0 N) and GY for the different N rates (r = 0.70; P < 0.001). This effect was more pronounced for improved genotypes than landraces for all parameters excepting BY and NAE_BY. BY showed +11% increase in landraces than improved genotypes. PFP_N showed an average decrease of 65% under high‐N fertilisation with 10% prevalence for improved genotypes. Landraces tend to promote vegetative growth while grain filling efficiency was higher for improved genotypes.