Ralstonia syzygii, the Blood Disease Bacterium and some Asian R. solanacearum strains form a single genomic species despite divergent lifestyles

The Ralstonia solanacearum species complex includes R. solanacearum, R. syzygii, and the Blood Disease Bacterium (BDB). All colonize plant xylem vessels and cause wilt diseases, but with significant biological differences. R. solanacearum is a soilborne bacterium that infects the roots of a broad range of plants. R. syzygii causes Sumatra disease of clove trees and is actively transmitted by cercopoid insects. BDB is also pathogenic to a single host, banana, and is transmitted by pollinating insects. Sequencing and DNA-DNA hybridization studies indicated that despite their phenotypic differences, these three plant pathogens are actually very closely related, falling into the Phylotype IV subgroup of the R. solanacearum species complex. To better understand the relationships among these bacteria, we sequenced and annotated the genomes of R. syzygii strain R24 and BDB strain R229. These genomes were compared to strain PSI07, a closely related Phylotype IV tomato isolate of R. solanacearum, and to five additional R. solanacearum genomes. Whole-genome comparisons confirmed previous phylogenetic results: the three phylotype IV strains share more and larger syntenic regions with each other than with other R. solanacearum strains. Furthermore, the genetic distances between strains, assessed by an in-silico equivalent of DNA-DNA hybridization, unambiguously showed that phylotype IV strains of BDB, R. syzygii and R. solanacearum form one genomic species. Based on these comprehensive data we propose a revision of the taxonomy of the R. solanacearum species complex. The BDB and R. syzygii genomes encoded no obvious unique metabolic capacities and contained no evidence of horizontal gene transfer from bacteria occupying similar niches. Genes specific to R. syzygii and BDB were almost all of unknown function or extrachromosomal origin. Thus, the pathogenic life-styles of these organisms are more probably due to ecological adaptation and genomic convergence during vertical evolution than to the acquisition of DNA by horizontal transfer.     

Solute carrier 11 cation symport requires distinct residues in transmembrane helices 1 and 6

Ubiquitous solute carriers 11 (SLC11) contribute to metal-ion homeostasis by importing Me(2+) and H(+) into the cytoplasm. To identify residues mediating cation symport, Escherichia coli proton-dependent manganese transporter (MntH) was mutated at five SLC11-specific transmembrane (TM) sites; each mutant activity was compared with wild-type MntH, and the biochemical results were tested by homology threading. Cd(2+) and H(+) uptake kinetics were analyzed in whole cells as a function of pH and temperature, and right-side out membrane vesicles were used to detail energy requirements and to probe site accessibility by Cys replacement and thiol modification. This approach revealed that TM segment 1 (TMS1) residue Asp(34) couples H(+) and Me(2+) symport and contributes to MntH forward transport electrogenicity, whereas the TMS6 His(211) residue mediates pH-dependent Me(2+) uptake; MntH Asn(37), Asn(250), and Asn(401) in TMS1, TMS7, and TMS11 participate in transporter cycling and/or helix packing interactions. These biochemical results fit the LeuT/SLC6 structural fold, which suggests that conserved peptide motifs Asp(34)-Pro-Gly (TMS1) and Met-Pro-His(211) (TMS6) form antiparallel "TM helix/extended peptide" boundaries, lining a "pore" cavity and enabling H(+)-dependent Me(2+) import.     

How Subchronic and Chronic Health Effects can be Neglected for GMOs, Pesticides or Chemicals

Chronic health effects are increasing in the world such as cancers, hormonal, reproductive, nervous, or immune diseases, even in young people. During regulatory toxicological subchronic tests to prevent these on mammalian health, prior commercialization of chemicals, including pesticides and drugs, or GMOs, some statistically significant findings may be revealed. This discussion is about the need to investigate the relevant criteria to consider those as biologically significant. The sex differences and the non linear dose or time related effects should be considered in contrast to the claims of a Monsanto-supported expert panel about a GMO, the MON 863 Bt maize, but also for pesticides or drugs, in particular to reveal hormone-dependent diseases and first signs of toxicities.     

Effect of organic amendment on soil fertility and plant nutrients in a post-fire Mediterranean ecosystem

Backgrounds and aims

In Mediterranean frequently burnt areas, the decrease of soil fertility leads to regressive vegetation dynamics. Organic amendments could help to accelerate post-fire ecosystem resilience, by improving soil properties and plant nutrition. This study was conducted to assess the potential of a composted biosolid to restore an early post-fire shrubland.

Methods

About 50 Mg.ha^?1 of fresh co-composted sewage sludge and green wastes were surface applied 7 months after fire on a silty-clayey soil. We monitored over a 2-year period organic matter and nutrient transfers to soil, nutrient responses of dominant plant species, and ecosystem contamination by potentially toxic trace elements.

Results

Over the experimental survey, compost rapidly and durably improved soil P₂O₅, MgO and K₂O content, and temporarily increased N-(NO₃ ^? + NO₂ ^?) content. Plant nutrition was improved more or less durably depending species. The most positive compost effect was on plant and soil phosphorus content. Plant nutrient storage was not improved 2 years after amendment, suggesting luxury consumption. No contamination by trace elements was detected in soil and plant.

Conclusions

The use of compost after fire could help for rapidly restoring soil fertility and improving plant nutrition. The increase of soil nutrient pools after amendment emphazised the diversity of plant nutritional traits. Eutrophication risk could occur from high compost and soil P₂O₅ content.     

The macrophage-specific membrane protein Nramp controlling natural resistance to infections in mice has homologues expressed in the root system of plants

In mice, natural resistance or susceptibility to infection with Mycobacteria, Salmonella, and Leishmania is controlled by a gene named Bcg. Bcg regulates the capacity of macrophages to limit intracellular replication of the ingested parasites, and is believed to regulate a key bactericidal mechanism of this cell. Recently, we have cloned the Bcg gene and shown that it encodes a novel macrophage-specific membrane protein designated Nramp. A routine search of the public databases for sequences homologous to Nramp identified 3 expressed sequence tags (EST) that show strong similarities to the mammalian protein.We report the identification and cloning of a full-length cDNA clone corresponding to a plant homologue (OsNramp1) of mammalian Nramp. Predicted amino acid sequence analysis of the plant protein indicates a remarkable degree of similarity (60% homology) with its mammalian counterpart, including identical number, position, and composition of transmembrane domains, glycosylation signals, and consensus transport motif, suggesting an identical overall secondary structure and membrane organization for the two proteins. This high degree of structural similarity indicates that the two proteins may be functionally related, possibly through a common mechanism of transport. RNA hybridization studies and RT-PCR analyses indicate that OsNramp1 mRNA is expressed primarily in roots and only at very low levels in leaves/stem. DNA hybridization studies indicate that OsNramp1 is not a single gene, but rather forms part of a novel gene family which has several members in all plants tested including cereals such as rice, wheat, and corn, and also in common weed species. The striking degree of conservation between the macrophage-specific mammalian Nramp and its OsNramp1 plant homologue is discussed with respect to possible implications in the metabolism of nitrate in both organisms.     

Nramp1 phagocyte intracellular metal withdrawal defense

Natural resistance-associated macrophage proteins (Nramp) are multispecific symporters facilitating proton-dependent import of divalent metals. Nramp1 restricts microbial access to essential micro-nutrients such as iron and manganese within professional phagosomes. Increased understanding of Nramp1 roles in human phagocytes will be useful for future therapeutic approaches against selected infectious and immune diseases.     

Regulation of Iron Homeostasis in Arabidopsis thaliana by the Clock Regulator Time for Coffee

In plants, iron homeostasis is tightly regulated to supply sufficient amounts of this metal for an optimal growth while preventing excess accumulation to avoid oxidative stress. To identify new regulators of iron homeostasis, a luciferase-based genetic screen using the Arabidopsis AtFer1 ferritin promoter as a target was developed. This screen identified TIME FOR COFFEE (TIC) as a regulator of AtFer1 gene expression. TIC was previously described as a nuclear regulator of the circadian clock. Mutants in the TIC gene exhibited a chlorotic phenotype rescued by exogenous iron addition and are hypersensitive to iron during the early stages of development. We showed that iron overload-responsive genes are regulated by TIC and by the central oscillator of the circadian clock. TIC represses their expression under low iron conditions, and its activity requires light and light/dark cycles. Regarding AtFer1, this repression is independent of the previously characterized cis-acting element iron-dependent regulatory sequence, known to be involved in AtFer1 repression. These results showed that the regulation of iron homeostasis in plants is a major output of the TIC- and central oscillator-dependent signaling pathways.     

Gene organization and expression of the divalent cation transporter Nramp in the protistan parasite Perkinsus marinus

Trophozoites of the protistan parasite Perkinsus marinus reside and proliferate inside phagosomelike structures of hemocytes from the host, the eastern oyster Crassostrea virginica. In a murine model, it has been proposed that the outcome of intracellular parasite-host interactions is determined, at least in part, by the activity of the host's divalent cation transporter natural resistance-associated macrophage protein 1 (Nramp1). Although nucleotide sequences from members of the Nramp family in protozoan parasites have recently become available in public databases, little is known about their molecular, structural, and functional aspects that may relate to the parasite's survival of intracellular killing by the host. The complementary DNA (cDNA) sequence of the Nramp from P. marinus (PmNramp) was obtained by polymerase chain reaction amplification with degenerated primers, followed by rapid amplification of cDNA ends. The 2,082-bp cDNA sequence encoded a predicted protein of 558 amino acids. PmNramp is a single-copy gene composed of 7 exons and 6 short introns (44-61 bp) with the canonical splicing signal (GT/AG). A phylogenetic analysis indicates that P. marinus and apicomplexan Nramp genes derive from a common "archetype" Nramp ancestor. However, the apicomplexan Nramps are highly divergent from the P. marinus sequence and the rest of the archetype Nramp group. Preliminary studies suggest that expression of PmNramp in in vitro-cultured P. marinus trophozoites is modulated by metals and by exogenous oxidative stress.