Analysis of promoter activity of the early nodulin Enod40 in Lotus japonicus

Microbial Biotechnolog

Lotus japonicus contains two distinct ENOD40 genes that are expressed in symbiotic, nonsymbiotic, and embryonic tissues

ENOD40, an early nodulin gene, has been postulated to play a significant role in legume root nodule ontogenesis, We have isolated two distinct ENOD40 genes from Lotus japonicus, The transcribed regions of the two ENOD40 genes share 65% homology, while the two promoters showed no significant homology, Both transcripts encode a putative dodecapeptide similar to that identified in other legumes forming determinate nodules, Both ENOD40 genes are coordinately expressed following inoculation of roots with Mesorhizobium loti or treatment with purified Nod factors. In the former case, mRNA accumulation could be detected up to 10 days following inoculation while in the latter case the accumulation was transient. High levels of both ENOD40 gene transcripts were found in nonsymbiotic tissues such as stems, fully developed flowers, green seed pods, and hypocotyls, A relatively lower level of both transcripts was observed in leaves, roots, and cotyledons. In situ hybridization studies revealed that, in mature nodules, transcripts of both ENOD40 genes accumulate in the nodule vascular system; additionally, in young seed pods strong signal is observed in the ovule, particularly In the phloem and epithelium, as well as in globular stage embryos.Animal science

Tissue-specific down-regulation of LjAMT1;1 compromises nodule function and enhances nodulation in Lotus japonicus

Plant ammonium transporters of the AMT1 family are involved in N-uptake from the soil and ammonium transport, and recycling within the plant. Although AMT1 genes are known to be expressed in nitrogen-fixing nodules of legumes, their precise roles in this specialized organ remain unknown. We have taken a reverse-genetic approach to decipher the physiological role of LjAMT1;1 in Lotus japonicus nodules. LjAMT1;1 is normally expressed in both the infected zone and the vascular tissue of Lotus nodules. Inhibition of LjAMT1;1 gene expression, using an antisense gene construct driven by a leghemoglobin promoter resulted in a substantial reduction of LjAMT1;1 transcript in the infected tissue but not the vascular bundles of transgenic plants. As a result, the nitrogen-fixing activity of nodules was partially impaired and nodule number increased compared to control plants. Expression of LjAMT1;1-GFP fusion protein in plant cells indicated a plasma-membrane location for the LjAMT1;1 protein. Taken together, the results are consistent with a role of LjAMT1;1 in retaining ammonium derived from symbiotic nitrogen fixation in plant cells prior to its assimilation

Absolute quantification of Medicago truncatula sucrose synthase isoforms and N-metabolism enzymes in symbiotic root nodules and the detection of novel nodule phosphoproteins by mass spectrometry

Mass spectrometry (MS) has become increasingly important for tissue specific protein quantification at the isoform level, as well as for the analysis of protein post-translational regulation mechanisms and turnover rates. Thanks to the development of high accuracy mass spectrometers, peptide sequencing without prior knowledge of the amino acid sequence—de novo sequencing—can be performed. In this work, absolute quantification of a set of key enzymes involved in carbon and nitrogen metabolism in Medicago truncatula ‘Jemalong A17’ root nodules is presented. Among them, sucrose synthase (SuSy; EC 2.4.1.13), one of the central enzymes in sucrose cleavage in root nodules, has been further characterized and the relative phosphorylation state of the three most abundant isoforms has been quantified. De novo sequencing provided sequence information of a so far unidentified peptide, most probably belonging to SuSy2, the second most abundant isoform in M. truncatula root nodules. TiO2-phosphopeptide enrichment led to the identification of not only a phosphorylation site at Ser11 in SuSy1, but also of several novel phosphorylation sites present in other root nodule proteins such as alkaline invertase (AI; EC 3.2.1.26) and an RNA-binding protein

Vitamins A & D Inhibit the Growth of Mycobacteria in Radiometric Culture

The role of vitamins in the combat of disease is usually conceptualized as acting by modulating the immune response of an infected, eukaryotic host. We hypothesized that some vitamins may directly influence the growth of prokaryotes, particularly mycobacteria. complex).Vitamins A and D cause dose-dependent inhibition of all three mycobacterial species studied. Vitamin A is consistently more inhibitory than vitamin D. The vitamin A precursor, β-carotene, is not inhibitory, whereas three vitamin A metabolites cause inhibition. Vitamin K has no effect. Vitamin E causes negligible inhibition in a single strain.We show that vitamin A, its metabolites Retinyl acetate, Retinoic acid and 13-cis Retinoic acid and vitamin D directly inhibit mycobacterial growth in culture. These data are compatible with the hypothesis that complementing the immune response of multicellular organisms, vitamins A and D may have heretofore unproven, unrecognized, independent and probable synergistic, direct antimycobacterial inhibitory activity

Chlorine isotope behavior in subduction zone settings revealed by olivine-hosted melt inclusions from the Central America Volcanic Arc

The isotopic composition of Cl, a highly hydrophilic and incompatible element, can provide new insights into the processes of element recycling in subduction zone settings. Samples from 13 localities in Guatemala, El Salvador, Nicaragua and Costa Rica, representing a ca. 1000 km long NW-SE segment along the Central American Volcanic Arc (CAVA), were selected. Ninety-seven melt inclusions, hosted by olivine Fo(90-70), were measured for Cl isotope ratios and trace element concentrations. Melt inclusions from samples from Guatemala to northwest Nicaragua have a restricted range of delta Cl-37 values (range 1%o, up to 3.8%o) and do not show any systematic variation along the arc. For some samples, the delta Cl-37 in the melt inclusions is shifted by up to 2%o to higher values compared to bulk rock data from the same volcanic center, for which the extent of Cl degassing is not known. The combination of delta Cl-37 values in melt inclusions with trace elements and the existing knowledge about the slab contributions along the arc allows us to elucidate the Cl isotope composition of different endmembers in this subduction zone. From Guatemala to northwest Nicaragua, a fluid component, originating from serpentinite, has a delta Cl-37 value close to +0.6%o. This value, similar to lithospheric serpentinites, confirms that despite the aqueous fluid migration through the entire slab, Cl isotopes do not fractionate significantly during transport. A melt-like component, present in the southern part of the arc, has negative delta Cl-37, possibly down to -2.5%o. This component has lower delta Cl-37 than values of the oceanic crust but similar to sediments currently subducting beneath CAVA. Finally, a common component, most likely amphibole-bearing metasomatized mantle, is identified in samples with the highest delta Cl-37 values (up to +3.0%o). The melting of amphibole, a mineral concentrating 37Cl over 35Cl, could explain the high delta Cl-37 values. The difference between melt inclusions and bulk rock delta Cl-37 in some volcanic centers probably results from late-stage processes such as mixing of different batches of magma at shallower levels after melt inclusions entrapment. Melt inclusions thus give a more comprehensive picture of Cl isotope systematics along the CAVA and in primitive subduction-related magmas

Thermal Stability of F-Rich Phlogopite and K-Richterite During Partial Melting of Metasomatized Mantle Peridotite With Implications for Deep Earth Volatile Cycles

Phlogopite and K-richterite constitute important carrier phases for H and F in Earth's lithosphere and mantle. The relative importance depends on their stabilities at high pressure and temperature, which in turn depends on bulk composition. Most previous experimental studies focused on the thermal stability of phlogopite and K-richterite were conducted using simplified chemical compositions. Here, partial melting experiments on metasomatized and carbonated, OH ± F-bearing near-natural peridotite were performed at high pressures (2 and 5 GPa) and temperatures (1,100–1,350°C) to assess the thermal stability of F-free versus F-bearing phlogopite and K-richterite. Experimental results demonstrate that the thermal stability of F-bearing phlogopite is increased by >55°C/wt.% F, relative to F-free phlogopite, whereas K-richterite is absent in all experiments with significant degrees of melting (>2%). The thermal stability of phlogopite containing several wt.% F exceeds continental and oceanic geotherms within the upper 150 km. Fluorine-rich phlogopite would therefore be stable in virtually all of the continental lithosphere, only to be decomposed during large, regional melting events such as continental break-up, thereby acting as a major long-term sink for F and/or H. This could even be the case for the oceanic asthenosphere, depending on the oceanic geotherm of the area of interest.ISSN:2169-9313ISSN:0148-0227ISSN:2169-935

Alterations in the rumen particle-associated microbiota of goats in response to dietary supplementation levels of schizochytrium spp

Fat rich microorganisms, such as microalgae Schizochytrium spp., are potential biotechno-logical tools in the modulation of rumen microbiome towards ecofriendly and high nutritional value end-products. However, limited in vivo trials have been reported on the topic. The aim of this study was to contribute to the knowledge on the effect of fat rich microalgae on the methanogenic and feed degrading particle-associated microbes in goats’ rumen content. For the trial, twenty-four goats were divided into four homogenous clusters (six goats/treatment) according to their fat corrected (4%) milk yield, body weight and age and individually were fed with alfalfa hay and concentrate feeds (F/C = 50/50). The concentrate of the control group (CON) contained no microalgae, while those of the treated groups were supplemented daily with 20 (ALG20), 40 (ALG40), and 60 (ALG60) g of Schizochytrium spp./goat. The relative abundances of total Archaea, methanogens, Methanomassili-icoccales, Methanobrevibacter spp., Methanosphaera stadmanae and Methanobacterium formicicum were significantly (p < 0.05) decreased in microalgae-fed goats compared to the CON ones. Moreover, a significant decline in the relative abundances of Firmicutes, Ruminococcus flavefaciens, Butyrivibrio fi-brosolvents, and Neocallimastigales in the rumen particle-associated microbiota of microalgae supplemented goats were observed. In conclusion, goats’ diets supplementation with Schizochytrium spp., could be considered a sustainable nutritional strategy for methanogens inhibition in their rumen particle-associated microbiota

Integrating omics technologies for improved quality and safety of seafood products

An essential aspect of product quality of aquatic foods is the rapid and accurate identification of bacterial species. From this perspective omics technologies prove to be very useful in the assessment of the quality and safety of seafood products. Such technologies can identify and detect low levels of contamination by pathogenic and spoilage bacteria and can be used to study the effects caused by processing and storage of seafood products. The integration of food processing with the monitoring of the microbial characteristics using conventional microbiological assays, coupled to molecular techniques may establish the baseline for the development of quicker and more sensitive and reliable methods for seafood safety screening. The use of combined omics technologies, including metagenomics, proteomics and metabolomics, coupled to conventional quality indices such as colour, texture and flavour offer a new tool for novel processing optimization to ensure seafood quality. The aim of this brief review is to outline how omics technologies can generate novel tools for integration into seafood processing and quality control. Considering that the main aspect of the review is the improvement of safety and quality of the final product, from production to consumption, emphasis is given to microbial identification and metabolite detection, the evaluation of the allergenic capacity of fish and seafood and optimization of postharvest processing. Deployment of omics for identification of potential microalgal products of relevance to seafood quality and safety is also considered

Characterization of Neisseria gonorrhoeae strains with decreased susceptibility to fluoroquinolones isolated in Greece from 1996 to 1999

Of the 331 Neisseria gonorrhoeae strains isolated in Greece from 1996 to 1999, 39 (11.8%) exhibited decreased susceptibility to quinolones due to gyrA and parC mutations. Conventional typing and pulsed-field gel electrophoresis showed that 34 of these isolates were clonally related. Epidemiological data indicated that the epidemic clone was sustained in a group of high-frequency transmitters