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Many legumes have been selected as model plants to degrade organic contaminants with their special associated rhizosphere microbes in soil. However, the function of root nodules during microbe-assisted phytoremediation is not clear. A pot study was conducted to examine phenanthrene (PHE) utilizing bacteria associated with root nodules and the effects of cowpea root nodules on phytoremediation in two different types of soils (freshly contaminated soil and aged contaminated soil). Cowpea nodules in freshly-contaminated soil showed less damage in comparison to the aged-contaminated soil, both morphologically and ultra-structurally by scanning electron microscopy. The study of polycyclic aromatic hydrocarbon (PAH) attenuation conducted by high performance liquid chromatography revealed that more PAH was eliminated from liquid culture around nodulated roots than nodule-free roots. PAH sublimation and denaturation gradient gel electrophoresis were applied to analyze the capability and diversity of PAH degrading bacteria from the following four parts of rhizo-microzone: bulk soil, root surface, nodule surface and nodule inside. The results indicated that the surface and inside of cowpea root nodules were colonized with bacterial consortia that utilized PHE. Our results demonstrated that root nodules not only fixed nitrogen, but also enriched PAH-utilizing microorganisms both inside and outside of the nodules. Legume nodules may have biotechnological values for PAH degradation.

A gram-negative bacterium designated BN30(T), which is motile with a polar flagellum, non-endospores forming, oxidase- and catalase-positive, was isolated from soybean root nodule. The organism is facultative anaerobic and surface-wrinkled rod. It can grow at 10-40 °C, pH 6-8 and 6 % (w/v) NaCl. BLASTn search based on 16S rRNA gene sequence revealed that the strain is closely related to Diaphorobacter aerolatus 8604S-37(T), Alicycliphilus denitrificans K601(T), Simplicispira limi EMB325(T), Diaphorobacter nitroreducens NA10B(T) and Diaphorobacter oryzae RF3(T), which all belonged to the family Comamonadaceae in class Betaproteobacteria. Phylogenetic analysis showed that the strain formed a firm clade with three Diaphorobacter species, being closest to Diaphorobacter aerolatus 8604S-37(T) with similarity of 98.64 %. DNA-DNA relatedness values between strain BN30(T) and five reference strains ranged from 11.5 to 35.9 %. All the results of phylogenetic analysis, chemotaxonomical data (predominant fatty acids are C16:0, sum feature 3, sum feature 8 and C17:0 cyclo; major polar lipids are diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol; major quinone is Q-8; G+C of total DNA is 65.2 %), physiological and phenotypic results supported that BN30(T) represented a novel species within the genus Diaphorobacter. The name Diaphorobacter ruginosibacter was proposed, and the type strain is BN30(T) (=ACCC06116(T) = DSM 27467(T)).

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) is normally limited by their low solubility and poor bioavailability. Prior research suggests that biosurfactants are synthesized as intermediates during the production of mucilage at the root tip. To date the effects of mucilage on PAH degradation and microbial community response have not been directly examined. To address this question, our research compared 3 cowpea breeding lines (Vigna unguiculata) that differed in mucilage production for their effects on phenanthrene (PHE) degradation in soil. The High Performance Liquid Chromatography results indicated that the highest PHE degradation rate was achieved in soils planted with mucilage producing cowpea line C1, inoculated with Bradyrhizobium, leading to 91.6% PHE disappearance in 5 weeks. In root printing tests, strings treated with mucilage and bacteria produced larger clearing zones than those produced on mucilage treated strings with no bacteria or bacteria inoculated strings. Experiments with 14C-PHE and purified mucilage in soil slurry confirmed that the root mucilage significantly enhanced PHE mineralization (82.7%), which is 12% more than the control treatment without mucilage. The profiles of the PHE degraders generated by Denaturing gradient gel electrophoresis suggested that cowpea C1, producing a high amount of root mucilage, selectively enriched the PHE degrading bacteria population in rhizosphere. These findings indicate that root mucilage may play a significant role in enhancing PHE degradation and suggests that differences in mucilage production may be an important criterion for selection of the best plant species for use in phytoremediation of PAH contaminated soils.

Legumes are important for nitrogen cycling in the environment and agriculture due to the ability of nitrogen fixation by rhizobia. In this review, we introduce an important and potential role of legume-rhizobia symbiosis in aiding phytoremediation of some metal contaminated soils as various legumes have been found to be the dominant plant species in metal contaminated areas. Resistant rhizobia used for phytoremediation could act on metals directly by chelation, precipitation, transformation, biosorption and accumulation. Moreover, the plant growth promoting (PGP) traits of rhizobia including nitrogen fixation, phosphorus solubilization, phytohormone synthesis, siderophore release, and production of ACC deaminase and the volatile compounds of acetoin and 2, 3-butanediol may facilitate legume growth while lessening metal toxicity. The benefits of using legumes inoculated with naturally resistant rhizobia or recombinant rhizobia with enhanced resistance, as well as co-inoculation with other plant growth promoting bacteria (PGPB) are discussed. However, the legume-rhizobia symbiosis appears to be sensitive to metals, and the effect of metal toxicity on the interaction between legumes and rhizobia is not clear. Therefore, to obtain the maximum benefits from legumes assisted by rhizobia for phytoremediation of metals, it is critical to have a good understanding of interactions between PGP traits, the symbiotic plant-rhizobia relationship and metals.

Sinorhizobium meliloti CCNWSX0020, isolated from root nodules of Medicago lupulina growing in gold mine tailings in the northwest of China, displayed both copper resistance and growth promotion of leguminous plants in copper-contaminated soil. Nevertheless, the genetic and biochemical mechanisms responsible for copper resistance in S. meliloti CCNWSX0020 remained uncharacterized. To investigate genes involved in copper resistance, an S. meliloti CCNWSX0020 Tn 5 insertion library of 14,000 mutants was created. Five copper-sensitive mutants, named SXa-1, SXa-2, SXc-1, SXc-2, and SXn, were isolated, and the disrupted regions involved were identified by inverse PCR and subsequent sequencing. Both SXa-1 and SXa-2 carried a transposon insertion in lpxXL ( SM0020_18047 ), encoding the LpxXL C-28 acyltransferase; SXc-1 and SXc-2 carried a transposon insertion in merR ( SM0020_29390 ), encoding the regulatory activator; SXn contained a transposon insertion in omp ( SM0020_18792 ), encoding a hypothetical outer membrane protein. The results of reverse transcriptase PCR (RT-PCR) combined with transposon gene disruptions revealed that SM0020_05862 , encoding an unusual P-type ATPase, was regulated by the MerR protein. Analysis of the genome sequence showed that this P-type ATPase did not contain an N-terminal metal-binding domain or a CPC motif but rather TPCP compared with CopA from Escherichia coli . Pot experiments were carried out to determine whether growth and copper accumulation of the host plant M. lupulina were affected in the presence of the wild type or the different mutants. Soil samples were subjected to three levels of copper contamination, namely, the uncontaminated control and 47.36 and 142.08 mg/kg, and three replicates were conducted for each treatment. The results showed that the wild-type S. meliloti CCNWSX0020 enabled the host plant to grow better and accumulate copper ions. The plant dry weight and copper content of M. lupulina inoculated with the 5 copper-sensitive mutants significantly decreased in the presence of CuSO 4 .

Here the spatial distribution of soil enzymatic properties in agricultural land was evaluated on a county-wide (567 km2) scale in Changwu, Shaanxi Province, China. The spatial variations in activities of five hydrolytic enzymes were examined using geostatistical methods. The relationships between soil enzyme activities and other soil properties were evaluated using both an integrated total enzyme activity index (TEI) and the geometric mean of enzyme activities (GME). At the county scale, soil invertase, phosphatase, and catalase activities were moderately spatially correlated, whereas urease and dehydrogenase activities were weakly spatially correlated. Correlation analysis showed that both TEI and GME were better correlated with selected soil physicochemical properties than single enzyme activities. Multivariate regression analysis showed that soil OM content had the strongest positive effect while soil pH had a negative effect on the two enzyme activity indices. In addition, total phosphorous content had a positive effect on TEI and GME in orchard soils, whereas alkali-hydrolyzable nitrogen and available potassium contents, respectively, had negative and positive effects on these two enzyme indices in cropland soils. The results indicate that land use changes strongly affect soil enzyme activities in agricultural land, where TEI provides a sensitive biological indicator for soil quality.

Background Determination of the minimum gene set for cellular life is one of the central goals in biology. Genome-wide essential gene identification has progressed rapidly in certain bacterial species; however, it remains difficult to achieve in most eukaryotic species. Several computational models have recently been developed to integrate gene features and used as alternatives to transfer gene essentiality annotations between organisms. Results We first collected features that were widely used by previous predictive models and assessed the relationships between gene features and gene essentiality using a stepwise regression model. We found two issues that could significantly reduce model accuracy: (i) the effect of multicollinearity among gene features and (ii) the diverse and even contrasting correlations between gene features and gene essentiality existing within and among different species. To address these issues, we developed a novel model called feature-based weighted Naïve Bayes model (FWM), which is based on Naïve Bayes classifiers, logistic regression, and genetic algorithm. The proposed model assesses features and filters out the effects of multicollinearity and diversity. The performance of FWM was compared with other popular models, such as support vector machine, Naïve Bayes model, and logistic regression model, by applying FWM to reciprocally predict essential genes among and within 21 species. Our results showed that FWM significantly improves the accuracy and robustness of essential gene prediction. Conclusions FWM can remarkably improve the accuracy of essential gene prediction and may be used as an alternative method for other classification work. This method can contribute substantially to the knowledge of the minimum gene sets required for living organisms and the discovery of new drug targets.

The FlgM-FliA regulatory circuit plays a central role in coordinating bacterial flagellar assembly. In this study, we identified multiple novel binding partners of FlgM using bacterial two-hybrid screening. Among these binding partners, FliS, the secretion chaperone of the filament protein FliC, was identified to compete with FliA for the binding of FlgM. We further showed that by binding to FlgM, FliS protects it from secretion and degradation, thus maintaining an intracellular pool of FlgM reserved as the FliS-FlgM complex. Consequently, we found that the flagellar late-class promoter activities are significantly increased in the fliS deletion mutant. The fliS mutant is weakly motile and shows significantly increased biofilm formation on biotic surface. Based on the results obtained, we established for the first time the regulatory role of the flagellin chaperone FliS to fine-tune late flagellar assembly by modulating FlgM activity.

The legume species of Astragalus as traditional Chinese medicine source and environmental protection plants showed an extensive distribution in the arid region of northwestern China. However, few rhizobia associating with Astragalus have been investigated in this region so far. In this study, 78 endophytic bacteria were isolated from root nodules of 12 Astragalus species and characterized by the PCR-RFLP of 16S rRNA gene and symbiotic genes together with the phylogenetic analysis. Results showed that the majority (53%) of isolates are non-nodulating Agrobacterium sp. and the rest are Mesorhizobium genomic species (41%), Ensifer spp. and Rhizobium gallicum (6%), respectively. Mesorhizobium genomic species are broadly distributed in the Astragalus symbioses and most of them share similar symbiotic genes. It seems that horizontal gene transfer occurred frequently among different genomic species independent of their original hosts and sites. Astragalus adsurgens is nodulated by a widely range of rhizobial species in the nodulation test, revealing that it could play an important role in diversification of Astragalus symbionts and that might be a reason for its wide adaptation to diverse environments.

BACKGROUND: Legume-rhizobium symbiosis is a complex process that is regulated in the host plant cell through gene expression network. Many nodulin genes that are upregulated during different stages of nodulation have been identified in leguminous herbs. However, no nodulin genes in woody legume trees, such as black locust (Robinia pseudoacacia), have yet been reported. METHODOLOGY/PRINCIPAL FINDINGS: To identify the nodulin genes involved in R. pseudoacacia-Mesorhizobium amorphae CCNWGS0123 symbiosis, a suppressive subtractive hybridization approach was applied to reveal profiling of differentially expressed genes and two subtracted cDNA libraries each containing 600 clones were constructed. Then, 114 unigenes were identified from forward SSH library by differential screening and the putative functions of these translational products were classified into 13 categories. With a particular interest in regulatory genes, twenty-one upregulated genes encoding potential regulatory proteins were selected based on the result of reverse transcription-polymerase chain reaction (RT-PCR) analysis. They included nine putative transcription genes, eight putative post-translational regulator genes and four membrane protein genes. The expression patterns of these genes were further analyzed by quantitative RT-PCR at different stages of nodule development. CONCLUSIONS: The data presented here offer the first insights into the molecular foundation underlying R. pseudoacacia-M. amorphae symbiosis. A number of regulatory genes screened in the present study revealed a high level of regulatory complexity (transcriptional, post-transcriptional, translational and post-translational) that is likely essential to develop symbiosis. In addition, the possible roles of these genes in black locust nodulation are discussed.

The copper tolerance gene of wild-type heavy metal-tolerance Sinorhizobium meliloti CCNWSX0020 was mutated by transposon Tn5-a. The mutant was sensitive up to 1.4mM Cu(2+). Production, components, surface morphology, and functional groups of extracellular polymeric substances (EPS) of the wild-type strains were compared with sensitive mutant in immobilization of Cu(2+). EPS produced by S. meliloti CCNWSX0020 restricts uptake of Cu(2+). The cell wall EPS were categorized based on the compactness and fastness: soluble EPS (S-EPS), loosely bound EPS (LB-EPS), and tightly bound EPS (TB-EPS). LB-EPS played a more important role than S-EPS and TB-EPS in Cu(2+) immobilization. Scanning electron microscopy (SEM) analysis LB-EPS had rough surface and many honeycomb pores, making them conducive to copper entry; therefore, they may play a role as a microbial protective barrier. Fourier transform-infrared (FT-IR) analysis further confirm that proteins and carbohydrates were the main extracellular compounds which had functional groups such as carboxyl (COOH), hydroxyl (OH), and amide (NH), primarily involved in metal ion binding.

A novel actinobacterium, designated strain F22T, was isolated from grassland soil collected from the Ziwuling area on the Loess Plateau, China. The novel strain was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces . Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain F22T belonged to the genus Streptomyces , being most closely related to Streptomyces resistomycificus NBRC 12814T (98.28 % sequence similarity), Streptomyces ciscaucasicus NBRC 12872T (98.14 %), Streptomyces chartreusis NBRC 12753T (98.14 %) and Streptomyces canus NRRL B-1989T (98.14 %). In DNA–DNA hybridizations and comparisons of morphological and phenotypic data, strain F22T could be distinguished from all of its closest phylogenetic relatives. Strain F22T exhibited antibacterial and antifungal activity, especially against Staphylococcus aureus , Bacillus subtilis and Cylindrocarpon destructans. Based on the DNA–DNA hybridization data and morphological, phenotypic and phylogenetic evidence, strain F22T represents a novel species of the genus Streptomyces , for which the name Streptomyces ziwulingensis sp. nov. is proposed. The type strain is F22T ( = CCNWFX 0001T = JCM 18081T = ACCC41875T).

The plant-growth-promoting bacterium Agrobacterium tumefaciens CCNWGS0286, isolated from the nodules of Robinia pseudoacacia growing in zinc-lead mine tailings, both displayed high metal resistance and enhanced the growth of Robinia plants in a metal-contaminated environment. Our goal was to determine whether bacterial metal resistance or the capacity to produce phytohormones had a larger impact on the growth of host plants under zinc stress. Eight zinc-sensitive mutants and one zinc-sensitive mutant with reduced indole-3-acetic acid (IAA) production were obtained by transposon mutagenesis. Analysis of the genome sequence and of transcription via reverse transcriptase PCR (RT-PCR) combined with transposon gene disruptions revealed that ZntA-4200 and the transcriptional regulator ZntR1 played important roles in the zinc homeostasis of A. tumefaciens CCNWGS0286. In addition, interruption of a putative oligoketide cyclase/lipid transport protein reduced IAA synthesis and also showed reduced zinc and cadmium resistance but had no influence on copper resistance. In greenhouse studies, R. pseudoacacia inoculated with A. tumefaciens CCNWGS0286 displayed a significant increase in biomass production over that without inoculation, even in a zinc-contaminated environment. Interestingly, the differences in plant biomass improvement among A. tumefaciens CCNWGS0286, A. tumefaciens C58, and zinc-sensitive mutants 12-2 ( zntA ::Tn 5 ) and 15-6 (low IAA production) revealed that phytohormones, rather than genes encoding zinc resistance determinants, were the dominant factor in enhancing plant growth in contaminated soil.

During a study of the diversity and phylogeny of rhizobia isolated from root nodules of Oxytropis ochrocephala grown in the northwest of China, four strains were classified in the genus Rhizobium on the basis of their 16S rRNA gene sequences. These strains have identical 16S rRNA gene sequences, which showed a mean similarity of 94.4 % with the most closely related species, Rhizobium oryzae. Analysis of recA and glnA sequences showed that these strains have less than 88.1 and 88.7 % similarity with the defined species of Rhizobium, respectively. The genetic diversity revealed by ERIC-PCR fingerprinting indicated that the isolates correspond to different strains. Strain CCNWQLS01T contains Q-10 as the predominant ubiquinone. The major fatty acids were identified as feature 8 (C18: 1ω7c and/or C18: 1ω6c; 67.2 %). Therefore, a novel species Rhizobium qilianshanense sp. nov. is proposed, and CCNWQLS01T (= ACCC 05747T = JCM 18337T) is designated as the type strain.

The mining industry generates huge amounts of wastewater, containing toxic heavy metals. Treatment to remove heavy metals is necessary and recent work has been focused on finding more environmentally friendly materials for removing heavy metals from wastewater. Biosorption can be an effective process for heavy metal removal from aqueous solutions. Our objectives were to investigate the removal of copper (II) from aqueous solutions using dead cells of Mesorhizobium amorphae CCNWGS0123 under differing levels of pH, agitation speed, temperature, initial copper concentration, biosorbent dose and contact time using flame atomic absorption spectroscopy for metal estimation. The maximum copper removal rate was achieved at pH 5.0, agitation speed 150×g, temperature 28°C and initial Cu (II) concentration of 100 mg L(-1). Maximum biosorption capacity was at 0.5 g L⁻¹ and equilibrium was attained within 30 min. Langmuir and Freundlich isotherms showed correlation coefficients of 0.958 and 0.934, respectively. Fourier transform-infrared spectroscopy (FT-IR) analysis indicated that many functional groups, such as O-H, N-H, C-H, C=O, -NH, -CN, C-N, C-O, amide -I, -II, -III and unsaturated alkenes, alkyls and aromatic groups on the cell surface were involved in the interaction between CCNWGS0123 and Cu. Scanning electron microscope and energy dispersive X-ray scanning results showed deformation, aggregation, and cell-surface damage due to the precipitation of copper on the cell surface. Dead cells of CCNWGS0123 showed potential as an efficient biosorbent for the removal of Cu²⁺ from aqueous solutions.

The appearance of chromium in the aqueous effluent is a major concern for the modern industry. In this work, Mesorhizobium amorphae strain CCNWGS0123 was investigated as a biosorbent to remove chromium from aqueous solutions. The optimum pH for Cr(III) and Cr(VI) biosorption were 4 and 2, respectively. This isolate showed an experimental maximum Cr(III) adsorption capacity of 53.52 mg L(-1), while the result was 47.67 mg L(-1) for Cr(VI), with an initial 100 mg L(-1) Cr ions and 1.0 g L(-1) biomass. In terms of time equilibrium, Cr(III) ion was more readily adsorbed than Cr(VI) by this isolate. The biosorption data of both ions fit the Langmuir isotherm better than that of Freundlich model. Meanwhile, this organism exhibited a good capability to release Cr ions, with desorption efficiency of 70 % for Cr(III) and 76 % for Cr(VI). Fourier transform infrared spectroscopy analysis showed that -OH, -COO, -NH, amide I, and C=O were involved in Cr(III) and Cr(VI) binding. The biosorbent was further characterized by scanning electron microscopy and energy-dispersive X-ray spectrometry, which indicated an accumulation of chromium on the cellular level. In the binary mixtures, the removal ratio of total Cr and Cr(III) increased from pH 2 to 4. The highest removal ratio of the total Cr was observed in the 25/25 mg L(-1) mixture at pH 4. In addition, the removal efficiency of Cr(VI) was closely influenced by Cr(III) in the mixture, decreasing to 23.57 mg g(-1) in the 100/100 mg L(-1) mixture system, due to the competition of Cr(III). The potential usage of the chromium-resistant rhizobium for the remediation of chromium-contaminated effluents has been demonstrated based on the above results.

Mesorhizobium alhagi strain CCNWXJ12-2(T) is a novel species of soil-dwelling, nitrogen-fixing bacteria that can form symbiotic root nodules with Alhagi sparsifolia. Moreover, the strain has high resistance to salt and alkali. Here we report the draft genome sequence of Mesorhizobium alhagi strain CCNWXJ12-2(T). A large number of osmotic regulation-related genes have been identified.

Here, we describe the draft genome sequence of Mesorhizobium amorphae strain CCNWGS0123, isolated from nodules of Robinia pseudoacacia growing on zinc-lead mine tailings. A large number of metal(loid) resistance genes, as well as genes reported to promote plant growth, were identified, presenting a great future potential for aiding phytoremediation in metal(loid)-contaminated soil.

The bioaccumulation characteristics of Zn(2+) and Cd(2+) by a novel species, Streptomyces zinciresistens CCNWNQ0016(T), were investigated. S. zinciresistens accumulated Zn(2+) and Cd(2+) mainly on the cell wall followed by intracellular accumulation. The mycelium was deformed, aggregated and formed precipitate of zinc and cadmium on the cell surface. Electron dense granules were detected on the cell wall as well as within the cytoplasm. The amino, carboxyl, hydroxyl and carbonyl groups were responsible for the biosorption of Zn(2+) and Cd(2+). The Langmuir isotherm model fitted the experimental data of metals adsorption processes better than Freundlich isotherm model. Cu(2+) and Cr(3+) competed for adsorption sites on the cell surface with Zn(2+) and Cd(2+). 87.33% and 98.11% recovery of Zn(2+) and Cd(2+), respectively, could be obtained at pH≤2 from metal-loaded biomass of S. zinciresistens desorption.

Copper is a highly utilized metal for electrical, automotive, household objects, and more recently as an effective antimicrobial surface. Copper-containing solutions applied to fruits and vegetables can prevent bacterial and fungal infections. Bacteria, such as Salmonellae and Cronobacter sakazakii, often found in food contamination, are rapidly killed on contact with copper alloys. The antimicrobial effectiveness of copper alloys in the healthcare environment against bacteria causing hospital-acquired infections such as methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli O157:H7, and Clostridium difficile has been described recently. The use of copper and copper-containing materials will continue to expand and may lead to an increase in copper mining and production. However, the copper mining and manufacturing industry and the consumer do not necessarily enjoy a favorable relationship. Open pit mining, copper mine tailings, leaching products, and deposits of toxic metals in the environment often raises concerns and sometimes public outrage. In addition, consumers may fear that copper alloys utilized as antimicrobial surfaces in food production will lead to copper toxicity in humans. Therefore, there is a need to mitigate some of the negative effects of increased copper use and copper mining. More thermo-tolerant, copper ion-resistant microorganisms could improve copper leaching and lessen copper groundwater contamination. Copper ion-resistant bacteria associated with plants might be useful in biostabilization and phytoremediation of copper-contaminated environments. In this review, recent progress in microbiological and biotechnological aspects of microorganisms in contact with copper will be presented and discussed, exploring their role in the improvement for the industries involved as well as providing better environmental outcomes.

Studying rhizobia in the root nodules of Sphaerophysa salsula (Pall.) DC in the northwest of China, we obtained five strains classified as genus Rhizobium on the basis of their 16S rRNA gene sequences. The sequence similarity of strain CCNWQTX14(T) with the most related species was 99.0%. Further phylogenetic analysis of housekeeping genes (recA and atpD) suggested the five strains comprised a novel lineage within Rhizobium. The nifH and nodD gene sequences of CCNWQTX14(T) were phylogenetically closely related with those of Sinorhizobium kummerowiae and R. sphaerophysae, respectively. The five strains isolated from different places were also distinct from related Rhizobium species using ERIC fingerprint profiles. The DNA-DNA hybridization value was 41.8% between CCNWQTX14(T) and Rhizobium sphaerophysae CCNWGS0238(T). Our novel strains were only able to form effective nodules on its original host Sphaerophysa salsula. Our data showed that the five Rhizobium strains formed a unique genomic species, for which a novel species Rhizobium helanshanense sp. nov. is proposed. The type strain is CCNWQTX14(T) (=ACCC 16237(T) =HAMBI 3083(T)).

In the title compound, C10H11ClO3, obtained by the reaction of 2,4-dihydroxyacetophenone, potassium carbonate and 1-bromo-2-chloroethane, an intramolecular O—H...O hydrogen bond occurs.

Heavy-metal pollution in soil is a major threat to human health and the entire ecosystem. Legumes and their associated rhizobial microorganisms are important for the biogeochemical cycles in agriculture and natural ecosystems. Legume-rhizobia symbiosis is very important in restoration of heavy-metal contaminated soils because the fixation of atmospheric nitrogen by legume-rhizobium symbiosis can increase the accumulation of nitrogen and organic matter. We reviewed the importance of remediation of heavy-metal contamination in soil and the current situation of remediation techniques, analysed the advantages and disadvantages of each remedial techniques. We address especially the superiority, recent advances and potential application of the legume-rhizobium symbiosis in remediation of heavy-metal contaminated soils.

A total of seventy-five symbiotic bacterial strains isolated from root nodules of wild Sophora alopecuroides grown in different regions of China's Loess Plateau were characterized. Based on the combined RFLP patterns, thirty-five genotypes were defined among the rhizobia and they were classified into nine genomic species, including Mesorhizobium alhagi and M. gobiense as the main groups, as well as Agrobacterium tumefaciens, M. amorphae, Phyllobacterium trifolii, Rhizobium giardinii, R. indigoferae, Sinorhizobium fredii and S. meliloti as the minor groups according to the 16S rRNA and recA gene analyses. Five and three lineages of nodA and nifH were found, respectively, in these strains, implying that the symbiotic genes of the S. alopecuroides rhizobia had different origins or had divergently evolved. Results of correspondence analysis showed that there was a correlation between rhizobial genotypes and the geographic origins. Possible lateral transfer of the recA and 16S rRNA genes between the P. trifolii and A. tumefaciens strains, and that of symbiotic genes (nodA, nifH) between different genera, was shown by discrepancies of the phylogenetic relationships of the four gene loci. These results revealed diverse rhizobia associated with wild S. alopecuroides grown in different regions of China's Loess Plateau, and demonstrated for the first time the existence of symbiotic A. tumefaciens strains in root nodules of S. alopecuroides.

An aerobic, brown-pigmented, non-spore-forming, endophytic bacterium, designated strain Zy-3T, was isolated from root nodules of Sphaerophysa salsula, a native leguminous herb belonging to the family Leguminosae growing in north-western China. Cells of strain Zy-3T were non-motile, Gram-negative rods. Strain Zy-3T produced siderophores and showed antifungal activity. Phylogenetic analysis of the 16S rRNA gene sequence indicated that the closest relative of this organism was Paracoccus halophilus HN-182T (96.6 % sequence similarity). On the basis of genotype, fatty acid patterns and physiological characteristics, a novel species Paracoccus sphaerophysae sp. nov. is proposed, with Zy-3T (=ACCC 05413T =HAMBI 3106T) as the type strain.

A filamentous actinomycete, designated strain CCNWNQ 0016T, was isolated from soil from a zinc and copper mine in Shaanxi province, north-western China. A polyphasic taxonomic study was carried out to establish the status of strain CCNWNQ 0016T. Chemical and morphological properties of the isolate were similar to those of species of the genus Streptomyces. Analysis of the almost complete 16S rRNA gene sequence placed strain CCNWNQ 0016T in the genus Streptomyces where it formed a distinct phyletic line with recognized species of this genus. The strain exhibited highest sequence similarities to Streptomyces niveoruber NBRC 15428T (98.8 %), Streptomyces griseoviridis NBRC 12874T (98.7 %) and Streptomyces aurantiogriseus NBRC 12842T (98.7 %). Furthermore, DNA–DNA hybridization studies between the novel isolate and these three strains showed relatedness values of 49.6±0.6, 47.2±0.9 and 46.9±1.1 %, respectively. A number of phenotypic properties also enabled the isolate to be differentiated from related species of the genus Streptomyces. Therefore, it is proposed that strain CCNWNQ 0016T (=K42T =ACCC 41871T =HAMBI 3107T) should be classified as the type strain of a novel species in the genus Streptomyces, Streptomyces zinciresistens sp. nov. The maximum level of resistance to Zn2+ for strain CCNWNQ 0016T is 35 mM.

Eleven strains that formed symbiotic root nodules on Alhagi sparsifolia, designated previously as genospecies II, were identified as a new lineage of Mesorhizobium (Alphaproteobacteria) that could be differentiated from all previously recognized species of the genus Mesorhizobium by using 16S rRNA gene sequences (dnaJ, dnaK, recA, glnA, nifH, nodA and nodC gene sequences, fatty acid profiles (C18 : 1 ω7c, 35 %;11-methyl C18 : 1 ω7c, 30 %) and numerical taxonomy. These strains are therefore considered to represent a novel species, for which the name Mesorhizobium alhagi sp. nov. is proposed, with isolate CCNWXJ12-2T (=ACCC 15461T=HAMBI 3019T) as the type strain.

The biosorption characteristics of Zn(II) using live and dead cells of Streptomyces ciscaucasicus strain CCNWHX 72-14 as biosorbents have been investigated in the present research. Optimum conditions for biosorption were determined to be: pH adjusted to 5.0, agitated at 90 rpm and at a dose of 2 g/L. For initial zinc concentrations of 1-150 mg/L, batch biosorption data of live biomass preferred to be simulated with Freundlich model while those of dead strain fit Langmuir isotherm well. Experimental maximum biosorption capacity turned out to be 42.75 mg/g (0.654 mmol/g) for living material and 54 mg/g (0.826 mmol/g) for dead sorbents, respectively. The pseudo-second-order equation, instead of the pseudo-first-order one, was chosen to describe the time course biosorption process. In contrast to live biosorbents, dead biomass seemed to have lower binding strength with higher desorption efficiency at pH 1.0. Competitive biosorption revealed the order of competing metal ion to be: Cu(2+)>Cd(2+)>Ni(+). FT-IR analysis indicated that more functional groups were involved in the biosorption process of dead adsorbents, compared with those linked to live biomass. Taken together, it can be concluded that dead cells of CCNWHX 72-14 were better and cheaper biosorbents than live ones.

The taxonomic position of an actinomycete isolated from a lead-polluted soil in Gansu province, north-west China, was determined by using a polyphasic approach. Chemical and morphological properties of the isolate, designated strain CCNWHX 13-160(T), were similar to those of streptomycetes. Analysis of the almost complete 16S rRNA gene sequence placed strain CCNWHX 13-160(T) in the genus Streptomyces where it formed a distinct phyletic line with recognized Streptomyces species. The strain was most similar to Streptomyces pseudovenezuelae NBRC 12904(T) (98.9 %) and Streptomyces resistomycificus NBRC 12814(T) (98.8 %). Furthermore, DNA-DNA hybridization studies between the novel isolate and these two strains showed relatedness values of 49.7+/-0.8 and 43.2+/-1.1 %, respectively. It is proposed that strain CCNWHX 13-160(T) (=ACCC 41207(T)=HAMBI 2991(T)) be classified as the type strain of a novel species in the genus Streptomyces, Streptomyces plumbiresistens sp. nov. The MIC of Pb(2+) for growth of strain CCNWHX 13-160(T) was 4.0 mM.

The aim of this work is to describe the diversity of potentially symbiotic bacteria associated with the invasive introduced legume Robinia pseudoacacia in China. Thirty-three isolates from 33 separate trees and nodules were characterized using restriction length fragment polymorphism and sequencing of 16S rRNA, nodA, nodC and nifH genes. Their 16S rRNA gene patterns and sequences placed them in three clades: 85% of isolates were related to the Mesorhizobium mediterraneum/temperatum group, whereas the remaining were similar either to Mesorhizobium amorphae or to Sinorhizobium meliloti. However, despite their diverse taxonomic positions, the nodA, nodC and nifH genes' phylogenies indicated that these R. pseudoacacia symbionts share similar symbiosis genes, implying gene transfers and a degree of host specificity. Comparison of R. pseudoacacia symbiotic diversity in native and other invaded areas suggests that most Chinese symbionts may not have arrived with the seed but were local bacteria that acquired specific symbiotic genes from native American rhizobia.

International audience; We isolated 33 nodule bacteria from the legume Alhagi sparsifolia growing in the desert of northwest China. They fell into three groups by restriction analysis of their rrs (small subunit ribosomal RNA) genes, and these, together with dnaK and dnaJ genes, were sequenced from representative isolates to assess their taxonomic position by phylogenetic analysis. The bacteria in each group belonged to different lineages that might represent three different new Mesorhizobium species, two of which form a novel clade very distinct from other species in the genus. Most A. sparsifolia symbionts harboured closely related nodA and nodC genes forming new lineages. The presence of these closely related symbiosis genes in various genomic backgrounds and the incongruence observed between the different gene phylogenies indicate a history of horizontal gene transfer of symbiosis genes between the A. sparsifolia symbionts.

A copper-resistant bacteria CCNWSX2332 was isolated from root nodules of Lespedeza cuneata growing in a gold mining tailing region in northwest of China. The specific growth rate of the strain was 0.62 microh(-1) in the presence of 2.0 mM Cu(2+) in TY liquid media, and the maximum copper accumulation of whole cell reached 147.03 microM Cu(2+) per gram (dry weight) after 4 h incubation. A partial sequence of the copper resistance gene copA was amplified from the strain, and the phylogenetic analysis based on 16S rDNA sequence showed that CCNWSX2332 belonged to Agrobacterium, and it had 100% similarity with Agrobacterium tumefaciens type strain IAM13129(T).

A total of 108 strains of bacteria were isolated from root nodules of wild legumes growing in gold mine tailings in northwest of China and were tested for heavy metal resistance. The results showed that the bacterial strain CCNWRS33-2 isolated from Lespedeza cuneata was highly resistant to copper, cadmium, lead and zinc. The strain had a relatively high mean specific growth rate under each heavy metal stress test and exhibited a high degree of bioaccumulation ability. The partial sequence of the copper resistance gene copA was amplified from the strain and a sequence comparison with our Cu-resistant PCR fragment showed a high homology with Cu-resistant genes from other bacteria. Phylogenetic analysis based on the 16S rRNA gene sequence showed that CCNWRS33-2 belongs to the Rhizobium–Agrobacterium branch and it had 98.9% similarity to Agrobactrium tumefaciens LMG196.

To screen high strength phenol degrading bacteria, we selected 108 rhizobial strains isolated from nodules of eight wild legumes species in the mining tailing region of Shaanxi province, northwest of China, and cultivated them in a basal salt (BS) medium supplemented with different phenol concentrations as a sole carbon source. The results showed that some of the strains could use phenol as sole carbon source. In order to study the characteristics of phenol degradation, the strain CCNWTB701 isolated from Astragalus chrysopteru was used as well, due to the fact that it was very efficient in phenol degradation. The phenol degradation was around 99.5 and 78.3%, with an initial concentration of 900 and 1000 mg/l phenol in 62 and 66 h, respectively. Kinetic studies indicated that the strain had a high KS (743.1 microM) and an extremely high KSI (10,469 microM) in Haldane's model. The phylogenetic analysis based on 16S rRNA gene sequences showed that CCNWTB701 belonged to the Rhizobium genus, and it was closely related to Rhizobium mongolense and Rhizobium gallicum.

Type VI secretion systems (T6SSs) are widespread multi-component machineries that translocate effectors into either eukaryotic or prokaryotic cells, for virulence or for interbacterial competition. Herein, we report that the T6SS-4 from Yersinia pseudotuberculosis displays an unexpected function in the transportation of Zn2+ to combat diverse stresses and host immunity. Environmental insults such as oxidative stress induce the expression of T6SS-4 via OxyR, the transcriptional factor that also regulates many oxidative response genes. Zinc transportation is achieved by T6SS-4-mediated translocation of a novel Zn2+-binding protein substrate YezP (YPK_3549), which has the capacity to rescue the sensitivity to oxidative stress exhibited by T6SS-4 mutants when added to extracellular milieu. Disruption of the classic zinc transporter ZnuABC together with T6SS-4 or yezP results in mutants that almost completely lost virulence against mice, further highlighting the importance of T6SS-4 in resistance to host immunity. These results assigned an unconventional role to T6SSs, which will lay the foundation for studying novel mechanisms of metal ion uptake by bacteria and the role of this process in their resistance to host immunity and survival in harmful environments., Author Summary One unique feature of type VI secretion system is the presence of multiple distinct systems in certain bacterial species. It is well established that some of these systems function to compete for their living niches among diverse bacterial species, whilst the activity of many such transporters remains unknown. Because metal ions are essential components to virtually all forms of life including bacteria, eukaryotic hosts have evolved complicated strategies to sequester metal ions, which constitute a major branch of their nutritional immunity. Therefore the ability to acquire metal ions is critical for bacterial virulence. This study reveals that the T6SS-4 of Yersinia pseudotuberculosis (Yptb) functions to import Zn2+ from the environment to mitigate the detrimental effects such as hydroxyl radicals induced by diverse stresses. Expression of the transporter is activated by multiple regulatory proteins, including OxyR and OmpR that sense diverse environmental cues. Zinc ion acquisition is achieved by translocating a Zn2+-binding substrate YezP, which is co-regulated with T6SS-4 by OxyR. Our results reveal a novel role for type VI secretion system, which is important in the study of the mechanism of metal ion acquisition by bacteria and the role of this process in bacterial pathogenesis and survival in detrimental environments.

We report the draft genome sequence of Pseudomonas psychrotolerans strain L19, isolated from a European 50-cent copper alloy coin. Multiple genes potentially involved in copper resistance were identified; however, it is unknown if these copper ion resistance determinants contribute to prolonged survival of this strain on dry metallic copper.

Sinorhizobium meliloti CCNWSX0020 was isolated from Medicago lupulina plants growing in lead-zinc mine tailings, which can establish a symbiotic relationship with Medicago species. Also, the genome of this bacterium contains a number of protein-coding sequences related to metal tolerance. We anticipate that the genomic sequence provides valuable information to explore environmental bioremediation.

We report the draft genome sequence of arsenite-oxidizing Halomonas sp. strain HAL1, isolated from the soil of a gold mine. Genes encoding proteins involved in arsenic resistance and transformation, phosphate utilization and uptake, and betaine biosynthesis were identified. Their identification might help in understanding how arsenic and phosphate metabolism are intertwined.

Message RNA (mRNA) carries a large number of local secondary structures, with structural stability to participate in the regulations of gene expression. A worthy question is how the local structural stability is maintained under the constraint that multiple selective pressures are imposed on mRNA local regions. Here, we performed the first genome-wide study of natural selection operating on high structural stability regions (HSRs) of mRNAs in Escherichia coli. We found that HSR tends to adjust the folded conformation to reduce the harm of mutations, showing a high level of mutational robustness. Moreover, guanine preference in HSR was observed, supporting the hypothesis that the selective constraint for high structural stability may partly account for the high percentage of G content in Escherichia coli genome. Notably, we found a substantially reduced synonymous substitution rate in HSRs compared with that in their adjacent regions. Surprisingly and interestingly, the non-key sites in HSRs, which have slight effect on structural stability, have synonymous substitution rate equivalent to background regions. To explain this result, we identified compensatory mutations in HSRs based on structural stability, and found that a considerable number of synonymous mutations occur to restore the structural stability decreased heavily by the mutations on key sites. Overall, these results suggest a significant role of local structural stability as a selective force operating on mRNA, which furthers our understanding of the constraints imposed on protein-coding RNAs.

Microbial transformations of arsenic influence its mobility and toxicity. We report the draft genome sequence of the arsenite-oxidizing strainAgrobacterium tumefaciens5A isolated from an As-contaminated soil in the Madison River Valley, MT. A large number of metal (or metalloid) resistance genes, especially contributing to arsenite oxidation, were identified.

In the title compound, C12H15ClO3, the ethoxy group is nearly coplanar with the benzene ring, making a dihedral angle of 9.03 (4)°, and is involved in an intramolecular O—H...O hydrogen bond to the neighbouring hydroxy group.