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Soil priming is a microbial-driven process, which determines key soil–climate feedbacks in response to fresh carbon inputs. Despite its importance, the microbial traits behind this process are largely undetermined. Knowledge of the role of these traits is integral to advance our understanding of how soil microbes regulate carbon (C) emissions in forests, which support the largest soil carbon stocks globally. Using metagenomic sequencing and C-glucose, we provide unprecedented evidence that microbial traits explain a unique portion of the variation in soil priming across forest biomes from tropical to cold temperature regions. We show that microbial functional profiles associated with the degradation of labile C, especially rapid simple sugar metabolism, drive soil priming in different forests. Genes involved in the degradation of lignin and aromatic compounds were negatively associated with priming effects in temperate forests, whereas the highest level of soil priming was associated with β-glucosidase genes in tropical/subtropical forests. Moreover, we reconstructed, for the first time, 42 whole bacterial genomes associated with the soil priming effect and found that these organisms support important gene machinery involved in priming effect. Collectively, our work demonstrates the importance of microbial traits to explain soil priming across forest biomes and suggests that rapid carbon metabolism is responsible for priming effects in forests. This knowledge is important because it advances our understanding on the microbial mechanisms mediating soil–climate feedbacks at a continental scale., This work were financially supported by the National Natural Science Foundation of China (41907031), the Chinese Academy of Sciences “Light of West China” Program for Introduced Talent in the West, the National Natural Science Foundation of China (31570440, 31270484), the Key International Scientific and Technological Cooperation and Exchange Project of Shaanxi Province, China (2020KWZ-010), the 2021 First Funds for Central Government to Guide Local Science and Technology Development in Qinghai Province (2021ZY002), the i-LINK +2018 (LINKA20069) from CSIC, and a Ramón y Cajal grant from the Spanish Ministry of Science and Innovation (RYC2018-025483-I)

Outer membrane vesicles (OMVs) can function as nanoscale vectors that mediate bacterial interactions in microbial communities. How bacteria recognize and recruit OMVs inter-specifically remains largely unknown, thus limiting our understanding of the complex physiological and ecological roles of OMVs. Here, we report a ligand-receptor interaction-based OMV recruitment mechanism, consisting of a type VI secretion system (T6SS)-secreted lipopolysaccharide (LPS)-binding effector TeoL and the outer membrane receptors CubA and CstR. We demonstrated that Cupriavidus necator T6SS1 secretes TeoL to preferentially associate with OMVs in the extracellular milieu through interactions with LPS, one of the most abundant components of OMVs. TeoL associated with OMVs can further bind outer membrane receptors CubA and CstR, which tethers OMVs to the recipient cells and allows cargo to be delivered. The LPS-mediated mechanism enables bacterial cells to recruit OMVs derived from different species, and confers advantages to bacterial cells in iron acquisition, interbacterial competition, and horizontal gene transfer (HGT). Moreover, our findings provide multiple new perspectives on T6SS functionality in the context of bacterial competition and HGT, through the recruitment of OMVs.

Oenococcus oeni plays a key role in inducing malolactic fermentation in wine. Acid stress is often encountered under wine conditions. However, the lack of systematic studies of acid resistance mechanisms limits the downstream fermentation applications. In this study, the acid responses of O. oeni were investigated by combining transcriptome, metabolome, and genome-scale metabolic modeling approaches. Metabolite profiling highlighted the decreased abundance of nucleotides under acid stress. The gene-metabolite bipartite network showed negative correlations between nucleotides and genes involved in ribosome assembly, translation, and post-translational processes, suggesting that stringent response could be activated under acid stress. Genome-scale metabolic modeling revealed marked flux rerouting, including reallocation of pyruvate, attenuation of glycolysis, utilization of carbon sources other than glucose, and enhancement of nucleotide salvage and the arginine deiminase pathway. This study provided novel insights into the acid responses of O. oeni, which will be useful for designing strategies to address acid stress in wine malolactic fermentation.

Aims Licorice (Glycyrrhiza uralensis Fisch.) is a crucial medicinal herb as it accumulates glycyrrhizin and liquiritin in roots. Licorice root-associated bacterial communities shaped by soil characteristics are supposed to regulate the accumulation of root secondary metabolites. Methods The soil characteristics, root secondary metabolites, and root-associated bacterial communities were analyzed in licorice plants of different ages to explore their temporal dynamics and interaction mechanisms. Results Temporal variation in soil characteristics and root secondary metabolites was distinct. The alpha-diversity of root-associated bacterial communities decreased with root proximity, and the community composition was clustered in the rhizosphere. Different taxa that were core-enriched from the dominant taxa in the bulk soil, rhizosphere soil, and root endosphere displayed varied time–decay relationships. Soil total potassium (TK) as a key factor regulated the temporal variation in some individual taxa in the bulk and rhizosphere soils; these taxa were associated with the adjustment of root secondary metabolites across different TK levels. Conclusions Licorice specifically selects root-associated core bacteria over the course of plant development, and TK is correlated with root secondary metabolites and individual core-enriched taxa in the bulk and rhizosphere soils, which may have implications for practical licorice cultivation. Electronic supplementary material The online version of this article (10.1007/s11104-020-04692-0) contains supplementary material, which is available to authorized users.

Background Hydrogen sulphide (H2S) is involved in regulating physiological processes in plants. We investigated how H2S ameliorates iron (Fe) deficiency in soybean (Glycine max L.) seedlings. Multidisciplinary approaches including physiological, biochemical and molecular, and transcriptome methods were used to investigate the H2S role in regulating Fe availability in soybean seedlings. Results Our results showed that H2S completely prevented leaf interveinal chlorosis and caused an increase in soybean seedling biomass under Fe deficiency conditions. Moreover, H2S decreased the amount of root-bound apoplastic Fe and increased the Fe content in leaves and roots by regulating the ferric-chelate reductase (FCR) activities and Fe homeostasis- and sulphur metabolism-related gene expression levels, thereby promoting photosynthesis in soybean seedlings. In addition, H2S changed the plant hormone concentrations by modulating plant hormone-related gene expression abundances in soybean seedlings grown in Fe-deficient solution. Furthermore, organic acid biosynthesis and related genes expression also played a vital role in modulating the H2S-mediated alleviation of Fe deficiency in soybean seedlings. Conclusion Our results indicated that Fe deficiency was alleviated by H2S through enhancement of Fe acquisition and assimilation, thereby regulating plant hormones and organic acid synthesis in plants.

H2S is emerging as a signaling molecule involved in the regulation of many physiological processes in plants. Here, we investigated the potential antioxidant role of H2S in soybean (Glycine max)-rhizobia (Sinorhizobium fredii) symbiotic root nodules. An endogenous H2S production deficit rhizobia mutant ∆CSE was constructed to study the effect of decreased content of H2S in soybean nodules. Fluorescent probes and confocal microscope were used to observe the production and accumulation of H2S and reactive oxygen species. Transmission electronic microscopy was conducted to study the structural changes in ∆CSE soybean nodules. Finally, qRT-PCR, enzymatic activity, and oxidative damage parameters were measured. The results demonstrated that abundant H2S was generated in the nitrogen-fixing zone of soybean nodules. The deletion of the cystathionine γ-lyase (CSE) gene in S. fredii (∆CSE) caused a sharp decrease in H2S production in both free-living rhizobia and soybean nodules. We found that decrease in the H2S level in nodule cells inhibited nitrogenase activity. In addition, to elevated H2O2 and malondialdehyde accumulation, increased protein carbonyl content and decreased O2− scavenging ability was observed in ∆CSE root nodules. Transmission electron microscopy revealed that an H2S deficit caused the deformation of bacteroids and damage of peribacteroid membranes in nodule cells. Moreover, the expression of some rhizobial and soybean genes related to antioxidant defense was up-regulated in ∆CSE nodules. H2S is crucial for the nitrogen-fixation ability of soybean nodules by acting as an antioxidant element that protects nodule cells and bacteroids from oxidative damage.

Legume plants are pioneer species in desert ecosystem that can be used to reestablish degraded ecosystem functions and soil properties, and reshape underground microbial communities. However, the distribution in soil nutrients and bacterial communities after introducing leguminous shrubs have not been well described. Here, we quantified the variations in soil nutrients and bacterial communities in soil profiles (0–100 cm) across stand ages (5-year, 8-year, 15-year) and severe desertification (SD), and further investigated the bacterial contributions to soil nutrients during stand development. The total phosphorus (TP), total nitrogen (TN) and organic carbon (OC) contents significantly increased, whereas the available nitrogen (AN) and available phosphorus (AP) contents showed a downward trend across stand ages. Bacterial diversity increased after introducing leguminous shrubs. Vertical spatial variation of bacterial communities attenuated during stand development. Bacterial beta-diversity, specific microbial phyla (i.e., Proteobacteria and Firmicutes) and nitrification process were strongly associated with vertical spatial variation in soil nutrients. Combined analyses revealed that leguminous shrubs can ameliorate soil OC, TN and TP with increasing legumes growth time, but could still exhibit no facilitating effect on the improvement of soil available N and P even for legumes aged 15 years. Improving the bioavailability of nutrient elements and microbial function diversity could be targeted for further studies in arid barren sandy.

Determining the mechanisms underlying complex plant-microbe interactions and feedbacks is crucial for effective rhizoremediation of contaminated soils. Here, rhizosphere bacterial communities of 10 common legumes planted in arable and aged oil-contaminated soils for 90 days were used to degrade phenanthrene (PHE). Bacterial community structures were analyzed to reveal the mechanisms underlying the varying PHE degradation capacities. We found that bacterial β-diversity within the rhizospheres played a major role in predicting PHE degradation rates, and more complex bacterial communities exhibited higher PHE degradation capacities. The highest PHE degradation rates were obtained with bacterial communities of Coronilla varia and Vigna unguiculata planted in oil-contaminated soils (almost 80%). There were significant differences in bacterial community composition between arable and contaminated soils, and among different legumes rhizospheres and the unplanted soils, although minimal differences were observed in bacterial α-diversity. The potential degradation-related taxa were more abundant in bacterial communities from contaminated soils, indicating more effective PHE degradation than in arable soils. Network analysis revealed the vital ecological roles of the potential degradation-related taxa in the maintenance of complex associations among bacterial taxa. Oil contamination reduced the differences in bacterial community composition between rhizosphere soils with high and low PHE degradation rates. These results suggest that the formulation of effective rhizoremediation strategies ought to focus on more diverse and complex interactions between rhizosphere-inhabiting organisms from aged oil-contaminated soils through the selection of optimal plant-microbiota association.

The rhizobium–legume symbiotic system is crucial for nitrogen cycle balance in agriculture. Hydrogen sulfide (H2S), a gaseous signaling molecule, may regulate various physiological processes in plants. However, whether H2S has regulatory effect in this symbiotic system remains unknown. Herein, we investigated the possible role of H2S in the symbiosis between soybean (Glycine max) and rhizobium (Sinorhizobium fredii). Our results demonstrated that an exogenous H2S donor (sodium hydrosulfide [NaHS]) treatment promoted soybean growth, nodulation, and nitrogenase (Nase) activity. Western blotting analysis revealed that the abundance of Nase component nifH was increased by NaHS treatment in nodules. Quantitative real-time polymerase chain reaction data showed that NaHS treatment upregulated the expressions of symbiosis-related genes nodA, nodC, and nodD of S. fredii. In addition, expression of soybean nodulation marker genes, including early nodulin 40 (GmENOD40), ERF required for nodulation (GmERN), nodulation signaling pathway 2b (GmNSP2b), and nodulation inception genes (GmNIN1a, GmNIN2a, and GmNIN2b), were upregulated. Moreover, the expressions of glutamate synthase (GmGOGAT), asparagine synthase (GmAS), nitrite reductase (GmNiR), ammonia transporter (GmSAT1), leghemoglobin (GmLb), and nifH involved in nitrogen metabolism were upregulated in NaHS-treated soybean roots and nodules. Together, our results suggested that H2S may act as a positive signaling molecule in the soybean–rhizobia symbiotic system and enhance the system’s nitrogen fixation ability.

Astragalus membranaceus (Fisch.) Bge. var. mongholicus, which is used in traditional Chinese medicine, contains several bioactive ingredients. The root-associated microbial communities play a crucial role in the production of secondary metabolites in plants. However, the correlation of root-associated bacteria and fungi with the bioactive ingredients production in A. mongholicus has not been elucidated. This study aimed to examine the changes in soil properties, root bioactive ingredients, and microbial communities in different cultivation years. The root-associated bacterial and fungal composition was analyzed using high-throughput sequencing. The correlation between root-associated bacteria and fungi, soil properties, and six major bioactive ingredients were examined using multivariate correlation analysis. Results showed that soil properties and bioactive ingredients were distinct across different cultivation years. The composition of the rhizosphere microbiome was different from that of the root endosphere microbiome. The bacterial community structure was affected by the cultivation year and exhibited a time-decay pattern. Soil properties affected the fungal community composition. It was found that 18 root-associated bacterial operational taxonomic units (OTUs) and four fungal OTUs were positively and negatively correlated with bioactive ingredient content, respectively. The abundance of Stenotrophomonas in the rhizosphere was positively correlated with astragaloside content. Phyllobacterium and Inquilinus in the endosphere were positively correlated with the calycosin content. In summary, this study provided a new opportunity and theoretical reference for improving the production and quality of in A. mongholicus, which thus increase the pharmacological value of A. mongholicus.

Nodulation outer proteins secreted via type 3 secretion systems are involved in the process of symbiosis between legume plants and rhizobia. To study the function of NopT in symbiosis, we mutated nopT in Mesorhizobium amphore CCNWGS0123 (GS0123), which can nodulate black locust (Robinia pseudoacacia). The nopT mutant induced higher levels of jasmonic acid, salicylic acid, and hydrogen peroxide accumulation in the roots of R. pseudoacacia compared with wild-type GS0123. The ΔnopT mutant induced higher disease-resistant gene expression 72 hours post-inoculation (hpi), whereas GS0123 induced higher disease-resistant gene expression earlier, at 36 hpi. Compared with the nopT mutant, GS0123 induced the up-regulation of most genes at 36 hpi and the down-regulation of most genes at 72 hpi. Proteolytically active NopT_GS0123 induced hypersensitive responses when expressed transiently in tobacco leaves (Nicotiana benthamiana). Two NopT_GS0123 targets in R. pseudoacacia were identified, ATP-citrate synthase alpha chain protein 2 and hypersensitive-induced response protein. Their interactions with NopT_GS0123 triggered resistance by the plant immune system. In conclusion, NopT_GS0123 inhibited the host plant immune system and had minimal effect on nodulation in R. pseudoacacia. Our results reveal the underlying molecular mechanism of NopT function in plant–symbiont interactions.

Background Excessive application of chemical fertilizer has exerted a great threat to soil quality and the environment. The inoculation of plants with plant-growth-promoting rhizobacteria (PGPR) has emerged as a great prospect for ecosystem recovery. The aim of this work to isolate PGPRs and highlights the effect of bacterial inoculants on available N/P/K content in soil and on the growth of wheat under conditions of reduced fertilizer application. Results Thirty-nine PGPRs were isolated and tested for their growth-promoting potential. Thirteen isolates had nitrogen fixation ability, of which N9 (Azotobacter chroococcum) had the highest acetylene reduction activity of 156.26 nmol/gh. Eleven isolates had efficient phosphate solubilizing ability, of which P5 (Klebsiella variicola) released the most available phosphorus in liquid medium (231.68 mg/L). Fifteen isolates had efficient potassium solubilizing ability, of which K13 (Rhizobium larrymoorei) released the most available potassium in liquid medium (224.66 mg/L). In culture medium supplemented with tryptophan, P9 (Klebsiella pneumoniae) produced the greatest amount of IAA. Inoculation with the bacterial combination K14 + 176 + P9 + N8 + P5 increased the alkali-hydrolysed nitrogen, available phosphorus and available potassium in the soil by 49.46, 99.51 and 19.38%, respectively, and enhanced the N, P, and K content of wheat by 97.7, 96.4 and 42.1%, respectively. Moreover, reducing fertilizer application by 25% did not decrease the available nitrogen, phosphorus, and potassium in the soil and N/P/K content, plant height, and dry weight of wheat. Conclusions The bacterial combination K14 + 176 + P9 + N8 + P5 is superior candidates for biofertilizers that may reduce chemical fertilizer application without influencing the normal growth of wheat.

Hydrogen sulfide (H2 S) is emerging as an important signalling molecule that regulates plant growth and abiotic stress responses. However, the roles of H2 S in symbiotic nitrogen (N) assimilation and remobilization have not been characterized. Therefore, we examined how H2 S influences the soybean (Glycine max)/rhizobia interaction in terms of symbiotic N fixation and mobilization during N deficiency-induced senescence. H2 S enhanced biomass accumulation and delayed leaf senescence through effects on nodule numbers, leaf chlorophyll contents, leaf N resorption efficiency, and the N contents in different tissues. Moreover, grain numbers and yield were regulated by H2 S and rhizobia, together with N accumulation in the organs, and N use efficiency. The synergistic effects of H2 S and rhizobia were also demonstrated by effects on the enzyme activities, protein abundances, and gene expressions associated with N metabolism, and senescence-associated genes (SAGs) expression in soybeans grown under conditions of N deficiency. Taken together, these results show that H2 S and rhizobia accelerate N assimilation and remobilization by regulation of the expression of SAGs during N deficiency-induced senescence. Thus, H2 S enhances the vegetative and reproductive growth of soybean, presumably through interactions with rhizobia under conditions of N deficiency.

Zinc oxide nanoparticles (ZnO NPs) are attracting much interest due to their potential toxicity and ubiquity in consumer products. However, understanding of pristine and transformed ZnO NPs impact on soil microbial communities is still limited. Here, we explored changes in the microbial communities of soils treated with pristine and sulfidized ZnO NPs (s-ZnO NPs), and their corresponding Zn ions (ZnSO4) for 30 and 90 days exposures at 100 and 500 mg Zn kg−1. The similarity in bacterial community responses was observed between ZnO NPs and s-ZnO NPs, and these Zn treatments significantly affected the bacterial communities at 90 days, which exhibited distinct patterns compared to ZnSO4. The single-time tested DTPA and H2O extractable Zn ions could not fully explain the observed ZnO NPs and s-ZnO NPs impact on bacterial communities. The two most dominant phylum Nitrospirae and Actinobacteria, associated with the reduction of NH4+–N and dissolved organic carbon, demonstrated significant changes in soils exposed to ZnO NPs and s-ZnO NPs. This suggests the potential long-term impact of transformed ZnO NPs on soil carbon and nitrogen cycling. For fungal communities, we did not find the distinct response patterns of fungal communities between nanoparticulate and ionic Zn exposures.

To improve the knowledge about the biogeographic patterns of bacteria in soils contaminated with crude oil, we studied the effects of local geochemical properties and geographic distance on bacterial community structure in oil contaminated soil in five oil refineries (46–360 km apart). The microbial structure was significantly affected by soil environmental factors such as pH, total petroleum hydrocarbons (TPH), total nitrogen, and cadmium level. Microbial alpha-diversity was positively correlated with pH but negatively correlated with TPH. Among sampling sites, the community dissimilarities increased with spatial distance. Variation of bacterial community was mostly attributed to simultaneous effects of spatial distance and environmental factors, and purely spatial distance contributed more to the variation. Microbial generalist OTUs were broadly distributed and dominant in contaminated soils. Their populations were in low proportion (15.75%), but they had high relative abundance (65.05%), and some were associated with TPH-degradation. Network analysis indicated that microbial communities had non-random co-occurrence patterns. Keystone taxa were Rubrivivax, Nitrospira, Methylotenera, Methyloversatilis and Acidaminobacter. Microbial taxa from the same module had strong ecological linkages and were involved in biological electron-transfer, C and N-cycles, and organic contaminant degradation. Our results indicate that the same microbial groups with TPH-degradation ability can be assembled from indigenous microorganisms in separate regions through long term exposure to contamination.

An actinomycete, strain D34T, was isolated from a soil sample collected from the rhizosphere of Stipa grandis at Yunwu Mountain in Ningxia, north-west China. Strain D34T showed highest 16S rRNA gene sequence similarity to Saccharothrix espanaensis DSM 44229T (99.0 %), Saccharothrix texasensis NRRL B-16107T (98.7 %) and Saccharothrix variisporea NRRL B-16296T (98.6 %). The strain contained meso-diaminopimelic acid, alanine, glycine and glutamic acid as major cell-wall amino acids. Mannose, rhamnose and galactose were the characteristic whole-cell sugars. The fatty acid profile consisted predominantly of iso-C15 : 0, iso-C16 : 0, iso-C16 : 1, C17 : 1ω6c, anteiso-C17 : 0 and anteiso-C15 : 0. The phospholipid profile included phosphatidylethanolamine (typical of phospholipid pattern type II). Furthermore, a combination of some physiological and biochemical properties and low DNA–DNA relatedness values indicated that strain D34T was differentiated from members of closely related species. On the basis of these phenotypic, genotypic and chemotaxonomic data, strain D34T represents a novel species of the genus Saccharothrix, for which the name Saccharothrix stipae sp. nov. is proposed. The type strain is D34T ( = JCM 30560T = ACCC19714T).

With ongoing global climate change and human activities, increasing desertification plays a predominant role in increasing soil nutrient losses. Soil nitrogen (N) is the essential limiting nutrient supporting plant growth and evaluating soil nutrient content, especially in desert ecosystems. N microbial processes will ultimately restore and maintain the balance in the soil N cycle, but the damage caused by desertification to soil N functional microorganisms associated with N supply, transformation, and loss is poorly understood. We examined changes in soil N and N functional gene (NFG) abundances within vertical profiles (i.e., soil depths of 0–100 cm) throughout the desertification process. We found that the abundance of N functional genes (NFGs) (except the N-fixation gene) decreased during the desertification process, almost all NFGs were attenuated along the vertical profiles, and available phosphorus (AP), soil water content (SWC) and pH were the best explanatory variables. The sums of (AOA + AOB) (associated with available N transformation) and (nirK + nirS + qnorB + nosZ) (associated with N loss) significantly decreased as desertification progressed, leading to decreased N transformation and loss potential. The (nifH + chiA) associated with available N supply increased along the desertification process gradient, suggesting enhanced potential for the acquisition available N. The increased ratio of (nifH + chiA + AOA + AOB)/(nirK + nirS + qnorB + nosZ) (associated with available N storage) collaboratively contributed to enhanced available N storage potential throughout the desertification process. Our results lay the foundation for better understanding the distinct responses of NFGs to desertification and the process through which they ultimately regulate available N storage in the degraded soils of desert ecosystems.

The rhizobium-legume symbiotic system is crucial for nitrogen cycle balance in agriculture. Hydrogen sulfide (H2S), a gaseous signaling molecule, may regulate various physiological processes in plants. However, whether H2S has regulatory effect in this symbiotic system remains unknown. Herein, we investigated the possible role of H2S in the symbiosis between soybean (Glycine max) and rhizobium (Sinorhizobium fredii). Our results demonstrated that exogenous H2S donor (sodium hydrosulfide, NaHS) treatment promoted soybean growth, nodulation and nitrogenase (Nase) activity. Western blotting analysis revealed that the abundance of nitrogenase component nifH was increased by NaHS treatment in nodules. Quantitative real-time PCR data showed that NaHS treatment up-regulated the expressions of symbiosis-related genesnodCandnodDofS. fredii. Besides, expression of soybean nodulation marker genes including early nodulin 40 (GmENOD40), ERF required for nodulation (GmERN), nodulation signaling pathway2b (GmNSP2b) and nodulation inception genes (GmNIN1a, GmNIN2aandGmNIN2b) were up-regulated. Moreover, the expressions of glutamate synthase (GmGS), nitrite reductase (GmNiR), ammonia transporter (GmSAT1), andnifHinvolved in nitrogen metabolism were up-regulated in NaHS-treated soybean roots and nodules. Together, our results suggested that H2S may act as a positive signaling molecule in soybean-rhizobia symbiotic system and enhance their nitrogen fixation ability.HighlightWe demonstrated for the first time that H2S as a signaling molecule may promote the establishment of symbiotic relationship and nitrogen fixation ability in the soybean-rhizobia symbiotic system.

Despite numerous reports that legume-rhizobium symbiosis alleviates Cu stress in plants, the possible roles of legume-rhizobium symbiosis and the regulatory mechanisms in counteracting Cu toxicity remain unclear. Here, Sinorhizobium meliloti CCNWSX0020 was used for analyzing the effects of rhizobium inoculation on plant growth in Medicago sativa seedlings under Cu stress. Our results showed that rhizobium inoculation alleviated Cu-induced growth inhibition, and increased nitrogen concentration in M. sativa seedlings. Moreover, the total amount of Cu uptake in inoculated plants was significantly increased compared with non-inoculated plants, and the increase in the roots was much higher than that in the shoots, thus decreasing the transfer coefficient and promoting Cu phytostabilization. Cu stress induced lipid peroxidation and reactive oxygen species production, but rhizobium inoculation reduced these components' accumulation through altering antioxidant enzyme activities and regulating ascorbate-glutathione cycles. Furthermore, legume-rhizobium symbiosis regulated the gene expression involved in antioxidant responses, phytochelatin (PC) biosynthesis, and metallothionein biosynthesis in M. sativa seedlings under Cu stress. Our results demonstrate that rhizobium inoculation enhanced Cu tolerance by affecting Cu uptake, regulating antioxidant enzyme activities and the ascorbate-glutathione cycle, and influencing PC biosynthesis-related gene expression in M. sativa. The results provide an efficient strategy for phytoremediation of Cu-contaminated soils.

Mesorhizobium alhagi CCNWXJ12-2T, isolated from root nodules of the desert plant Alhagi sparsifolia, contains two type III secretion systems (T3SSs). T3SSs are specialized machinery with wide distribution in bacteria that inject effector proteins into target cells. Our previous study showed that the expression of M. alhagi T3SS1 is upregulated in high-salt conditions. Here, phylogenetic analysis of T3SS1 using the core protein RhcU suggested that T3SS1 belongs to the α-Rhc II subgroup of the Rhc T3SS family. To elaborate the function of M. alhagi CCNWXJ12-2T T3SS1 in stress adaption, two T3SS1 mutants (ΔrhcQ and ΔMA29250) were constructed and analyzed. β-galactosidase transcriptional fusion assays showed that activity of the promoter of T3SS1 was induced by salts. Mutant ΔrhcQ was more sensitive to NaCl and LiCl than the wild-type, but ΔMA29250 was not. Both mutants were more sensitive to KCl than the wild-type. The intracellular Na+ concentration in ΔrhcQ in high-NaCl conditions (0.4 M) increased by 37% compared to that of the wild-type strain, while the Na+ concentration in ΔMA29250 increased by 13%. The K+ concentration in both mutants increased by 16% compared to the wild-type in high-KCl conditions (0.3 M). Strain ΔrhcQ showed decreased survival compared to the wild-type after treatment with H2O2, while the survival rate of ΔMA29250 was almost the same as that of the wild-type. Antioxidant enzyme activities in ΔrhcQ were lower than those in the wild-type strain, but this was not the case for ΔMA29250. Our data elucidate the beneficial effects of T3SS1 in the adaption of M. alhagi CCNWXJ12-2T to stress.

The objective of this study was to evaluate the effects of cowpea with root mucilage on phenanthrene (PHE) dissipation and microbial community composition. The experiment was conducted using three cowpea breeding lines (Vigna unguiculata) that differed in mucilage production. The highest PHE dissipation rate was achieved in soils planted with high root mucilage producing cowpea C1 inoculated with Bradyrhizobium, leading to 91.6% PHE disappearance in 5 weeks. Relative changes in microbial populations of different treatments were investigated by monitoring PLFA signatures. Data was analyzed by principle component analysis (PCA) and by construction of artificial neural network (ANN). Based on the PCA profiles, the ten treatments were separated into four groups. The ANN results demonstrated that fungal biomass were significantly influenced by root mucilage produced by cowpea (C1), and gram positive to gram negative bacteria ratios were shown to have a positive correlation with the presence of Phenanthrene (PHE) via a Kohonen self-organizing map (KSOM). Total PLFA biomass was significantly higher in the soil planted with cowpea line C1 inoculated with Bradyrhizobium than other treatments. Results suggest that root mucilage may contribute to the changes of the microbial biomass and composition in the rhizosphere, which in turn may positively impact PAH bioremediation.

Rhizobia typically produce a lower level of ACC deaminase compared with free-living plant growth-promoting bacteria. While the endogenous rhizobial ACC deaminase is important in legume nodulation, it is not sufficient to protect host plants against environmental stresses. The main goal of this study was to assess the effects of a genetically engineered Sinorhizobium meliloti strain overproducing ACC deaminase, and its symbiotic performance in Medicago lupulina under copper stress conditions. The engineered strain was transformed with an exogenous acdS gene by triparental conjugation. A plant growth assay was conducted to assess its plant growth promotion ability under copper stress conditions. The expressions of antioxidant genes in these plants were analyzed using quantitative real-time PCR. Plants nodulated with the engineered strain showed a greater dry weight, a decreased ethylene level in roots, a higher total copper uptake but a lower level of copper translocation to aerial parts, as compared with the plants nodulated with the wild-type strain under copper stress conditions. These results were positively correlated with higher expression of antioxidant genes in the roots of these plants exposed to severe copper stress. The engineered strain could improve plant growth as well as copper tolerance of M. lupulina, and enhance the antioxidant defense system.

The legume-rhizobium symbiosis has been proposed as an important system for phytoremediation of heavy metal contaminated soils due to its beneficial activity of symbiotic nitrogen fixation. However, little is known about metal resistant mechanism of rhizobia and the role of metal resistance determinants in phytoremediation. In this study, copper resistance mechanisms were investigated for a multiple metal resistant plant growth promoting rhizobium, Mesorhizobium amorphae 186. Three categories of determinants involved in copper resistance were identified through transposon mutagenesis, including genes encoding a P-type ATPase (CopA), hypothetical proteins, and other proteins (a GTP-binding protein and a ribosomal protein). Among these determinants, copA played the dominant role in copper homeostasis of M. amorphae 186. Mutagenesis of a hypothetical gene lipA in mutant MlipA exhibited pleiotropic phenotypes including sensitivity to copper, blocked symbiotic capacity and inhibited growth. In addition, the expression of cusB encoding part of an RND-type efflux system was induced by copper. To explore the possible role of copper resistance mechanism in phytoremediation of copper contaminated soil, the symbiotic nodulation and nitrogen fixation abilities were compared using a wild-type strain, a copA-defective mutant, and a lipA-defective mutant. Results showed that a copA deletion did not affect the symbiotic capacity of rhizobia under uncontaminated condition, but the protective role of copA in symbiotic processes at high copper concentration is likely concentration-dependent. In contrast, inoculation of a lipA-defective strain led to significant decreases in the functional nodule numbers, total N content, plant biomass and leghemoglobin expression level of Robinia pseudoacacia even under conditions of uncontaminated soil. Moreover, plants inoculated with lipA-defective strain accumulated much less copper than both the wild-type strain and the copA-defective strain, suggesting an important role of a healthy symbiotic relationship between legume and rhizobia in phytostabilization.

The symbiosis of the highly metal-resistant Sinorhizobium meliloti CCNWSX0020 and Medicago lupulina has been considered an efficient tool for bioremediation of heavy metal-polluted soils. However, the metal resistance mechanisms of S. meliloti CCNWSX00200 have not been elucidated in detail. Here we employed a comparative transcriptome approach to analyze the defense mechanisms of S. meliloti CCNWSX00200 against Cu or Zn exposure. Six highly upregulated transcripts involved in Cu and Zn resistance were identified through deletion mutagenesis, including genes encoding a multicopper oxidase (CueO), an outer membrane protein (Omp), sulfite oxidoreductases (YedYZ), and three hypothetical proteins (a CusA-like protein, a FixH-like protein, and an unknown protein), and the corresponding mutant strains showed various degrees of sensitivity to multiple metals. The Cu-sensitive mutant (Δ cueO ) and three mutants that were both Cu and Zn sensitive (Δ yedYZ , Δ cusA -like, and Δ fixH -like) were selected for further study of the effects of these metal resistance determinants on bioremediation. The results showed that inoculation with the Δ cueO mutant severely inhibited infection establishment and nodulation of M. lupulina under Cu stress, while inoculation with the Δ yedYZ and Δ fixH -like mutants decreased just the early infection frequency and nodulation under Cu and Zn stresses. In contrast, inoculation with the Δ cusA -like mutant almost led to loss of the symbiotic capacity of M. lupulina to even grow in uncontaminated soil. Moreover, the antioxidant enzyme activity and metal accumulation in roots of M. lupulina inoculated with all mutants were lower than those with the wild-type strain. These results suggest that heavy metal resistance determinants may promote bioremediation by directly or indirectly influencing formation of the rhizobium-legume symbiosis. IMPORTANCE Rhizobium-legume symbiosis has been promoted as an appropriate tool for bioremediation of heavy metal-contaminated soils. Considering the plant-growth-promoting traits and survival advantage of metal-resistant rhizobia in contaminated environments, more heavy metal-resistant rhizobia and genetically manipulated strains were investigated. In view of the genetic diversity of metal resistance determinants in rhizobia, their effects on phytoremediation by the rhizobium-legume symbiosis must be different and depend on their specific assigned functions. Our work provides a better understanding of the mechanism of heavy metal resistance determinants involved in the rhizobium-legume symbiosis, and in further studies, genetically modified rhizobia harboring effective heavy metal resistance determinants may be engineered for the practical application of rhizobium-legume symbiosis for bioremediation in metal-contaminated soils.

A comparative study was conducted on the toxicity of Cd to alkaline phosphatase activity (ALP) and dehydrogenase activity (DHA) in 18 top soils with contrasting soil properties representative of 14 major soil types in China. Soil pH and carbonate content, soil organic matter, and cation exchange capacity (CEC) largely affected the Cd toxicity on two enzyme activities; with the soil pH having only minor effect on the median ecological dose values based on total Cd concentrations (ED50 T). The values of ED50 T/ED50 W (based on water-soluble Cd content) of alkaline phosphatase and dehydrogenase were strongly influenced by pH and CEC contents, which explained up to 71% of the variation for alkaline phosphatase, 82% of the variation for dehydrogenase, and also were significantly correlated with the parameter KF derived from Freundlich adsorption isotherms. This study suggests that the values of ED50 T/ED50 W could be useful to evaluate the buffer capacity of soils which protects soil enzymes from harmful effec...

Biosorption of zinc by living biomasses of metal resistant symbiotic bacterium Mesorhizobium amorphae CCNWGS0123 was investigated under optimal conditions at pH 5.0, initial metal concentrations of 100 mg L−1, and a dose of 1.0 g L−1. M. amorphae exhibited an efficient removal of Zn2+ from aqueous solution with maximum biosorption capacity of 120.85 mg g−1. Moreover, more than 70% Zn2+ could be recovered from Zn-loaded biomass at pH 1.0. Both the Langmuir and Freundlich isotherms provided a better fit to experimental data for Zn2+ sorption with correlation coefficients of 0.9885. Kinetics models suggested there was more than one step involved in the Zn2+ sorption process, while a pseudo-second-order model was more suitable to describe the kinetic behavior accurately, indicating a chemisorption process. Carbonyl, amino, carboxyl, and aromatic groups were responsible for the biosorption of Zn2+ by M. amorphae. Cellular deformation, precipitate, and damage were found after Zn2+ treatment. Competitive sorptio...

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 1.4 mM 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 (N H), primarily involved in metal ion binding.

The main aim of this work was to study molecular characterization of a DNA fragment conferring resistance to Cu(II) in Sinorhizobium meliloti CCNWSX0020. The strain CCNWSX0020, resistant to 1.4 mmol l(-1) Cu(II) in tryptone-yeast extract medium was isolated from Medicago lupulina growing in mine tailings of Fengxian County, China. The availability of the complete genome sequence of S. meliloti CCNWSX0020 provides an opportunity for investigating genes that play significant roles in Cu(II) resistance. A copper resistance gene, with a length of 1,445 bp, encoding 481 amino acids, designated omp, was identified by cDNA-amplified fragment length polymorphism from S. meliloti CCNWSX0020. The expression of omp gene strongly increased in the presence of Cu(II). The omp-defective mutants display sensitivities to Cu(II) compared with their wild types. The Cu(II)-sensitive phenotype of the mutant was complemented by a 1.5-kb DNA fragment containing omp gene. BLAST analysis revealed that this gene encoded a hypothetical outer membrane protein with 75 % similarity to outer membrane efflux protein in Rhizobium leguminosarum bv. viciae 3841. These studies suggested that the omp product was involved in the Cu(II) tolerance of S. meliloti CCNWSX0020.

Soil dehydrogenase plays a role in the biological oxidation of soil organic matter and can be considered a good measure of the change of microbial oxidative activity under environmental pollutions. However, the kinetic characteristic of soil dehydrogenase under heavy metal stresses has not been investigated thoroughly. In this study, we characterized the kinetic characteristic of soil dehydrogenase in 14 soil types, and investigated how kinetic parameters changed under spiked with different concentrations of cadmium (Cd). The results showed that the Km and Vmax values of soil dehydrogenase was among 1.4-7.3mM and 15.9-235.2μMh-1 in uncontaminated soils, respectively. In latosolic red soil and brown soil, the inhibitory kinetic mechanism of Cd to soil dehydrogenase was anticompetitive inhibition with inhibition constants (Ki) of 12 and 4.7mM, respectively; in other soils belonged to linear mixed inhibition, the values of Ki were between 0.7-4.2mM. Soil total organic carbon and Ki were the major factors affecting the toxicity of Cd to dehydrogenase activity. In addition, the velocity constant (k) was more sensitive to Cd contamination compared to Vmax and Km, which was established as an early indicator of gross changes in soil microbial oxidative activity caused by Cd contamination.

Background Serine/threonine protein kinases are highly conserved kinases with a wide distribution in microbes and with multiple functions. Mesorhizobium alhagi CCNWXJ12-2, a α-proteobacterium which could be able to form symbiosis with Alhagi sparsifolia in northwest of China, contains a putative PrkA-family serine protein kinase, PrkA. In our previous study, the expression of prkA was found to be downregulated in high-salt conditions. To elucidate the function of M. alhagi PrkA, a prkA deletion mutant was constructed and the phenotypes of the mutant were analyzed. Results The salt and alkaline tolerance and antioxidant capacity of the wild-type strain and the prkA deletion mutant was measured. Our results showed that the deletion mutant had higher salt and alkaline tolerance than the wild-type strain. The total cellular Na+ content was measured and showed no significant difference between the wild-type strain and the mutant. The prkA deletion mutant also showed a higher H2O2 tolerance than the wild-type strain. Therefore the activities of antioxidant enzymes were measured. Catalase activity was similar in the wild-type and the deletion mutant, while the superoxide dismutase activity in the mutant was higher than that in the wild-type. Conclusions We firstly analyze the function of a serine protein kinase, PrkA, in M. alhagi. Our data indicate that PrkA could reduce the survival of M. alhagi under environmental stress and deletion of prkA dramatically improved the salt and alkaline tolerance and antioxidant capacity of M. alhagi.

Elucidating the mechanisms underlying microbial succession is a major goal of microbial ecology research. Given the increasing human pressure on the environment and natural resources, responses to the repeated introduction of organic and inorganic pollutants are of particular interest. To investigate the temporal dynamics of microbial communities in response to pollutants, we analyzed the microbial community structure in batch microcosms that were inoculated with soil bacteria following exposure to individual or combined pollutants (phenanthrene, n-octadecane, phenanthrene + n-octadecane, and phenanthrene + n-octadecane + CdCl2). Subculturing was performed at 10-day intervals, followed by high-throughput sequencing of 16S rRNA genes. The dynamics of microbial communities in response to different pollutants alone and in combination displayed similar patterns during enrichment. Specifically, the repression and induction of microbial taxa were dominant, and the fluctuation was not significant. The rate of appearance for new taxa and the temporal turnover within microbial communities were higher than the rates reported in other studies of microbial communities in air, water, and soil samples. In addition, conditionally rare taxa that were specific to the treatments exhibited higher betweenness centrality values in the co-occurrence network, indicating a strong influence on other interactions in the community. These results suggest that the repeated introduction of pollutants could accelerate microbial succession in microcosms, resulting in the rapid re-equilibration of microbial communities. This article is protected by copyright. All rights reserved.

The Sinorhizobium meliloti (S. meliloti) strain CCNWSX0020 displayed tolerance to high levels exposures of multiple metals and growth promotion of legume plants grown in metal-contaminated soil. However, the mechanism of metal-resistant strain remains unknown. We used five P1B-ATPases deletions by designating as ∆copA1b, ∆fixI1, ∆copA3, ∆zntA and ∆nia, respectively to investigate the role of P1B-ATPases in heavy metal resistance of S. meliloti. The ∆copA1b and ∆zntA mutants were sensitive to zinc (Zn), cadmium (Cd) and lead (Pb) in different degree, whereas the other mutants had no significant influence on the metal resistance. Moreover, the expression of zntA was induced by Zn, Cd and Pb whereas copA1b was induced by copper (Cu) and silver (Ag). This two deletions could led to the increased intracellular concentrations of Zn, Pb and Cd, but not of Cu. Complementation of ∆copA1b and ∆zntA mutants showed a restoration of tolerance to Zn, Cd and Pb to a certain extent. Taken together, the results suggest an important role of copA1b and zntA in Zn homeostasis and Cd and Pb detoxification in S. meliloti CCNWSX0020.

Aims: Eukaryotic typical 2-cysteine (Cys) peroxiredoxins (Prxs) are multifunctional proteins subjected to complex regulation and play important roles in oxidative stress resistance, hydrogen peroxide (H2O2) signaling modulation, aging, and cancer, but the information on the biochemical functions and regulation mechanisms of prokaryotic atypical 2-Cys Prxs is largely lacking. Results: In this study, we show that at low peroxide concentrations, the atypical 2-Cys Prx in Corynebacterium glutamicum (CgPrx) mainly exists as monomers and displays thioredoxin (Trx)-dependent peroxidase activity. Moderate oxidative stress causes reversible S-mycothiolation of the H2O2-sensing Cys63 residue, which keeps CgPrx exclusively in dimer form with neither peroxidase nor chaperone activity. Then, the increased levels of H2O2 could act as a messenger to oxidize the redox-sensitive regulator hydrogen peroxide-inducible gene activator, leading to activation of expression of the more efficient mycothiol peroxidase and...

Mining generates large amounts of wastewater, which may contain elevated levels of potentially toxic metals. We evaluated the use of live cells of Mesorhizobium amorphae CCNWGS0123 as a new, relatively inexpensive and environment-friendly adsorbent material for removal of toxic metals from aqueous solutions, such as mine tailings leachate. Maximum copper uptake and removal efficiency was attained at a dosage of 0.5 g L−1, a pH of 5.0, an agitation speed of 150 rpm, and at a temperature of 28 °C for an initial Cu2+ concentration of 100 mg L−1; equilibrium was achieved within 30 min. The adsorption isothermal data matched Langmuir and Freundlich isotherms well, with correlation coefficients of 0.931 and 0.932, respectively. FT-IR analysis indicated that many functional groups with a negative charge on the cell surface were involved in the interaction between live CCNWGS0123 and copper ions. Scanning electron microscope results confirmed that there was some copper sediment on the cell surface, causing deformation, aggregation, and cell surface damage. Cu2+ accumulation was related to cell growth, and copper uptake was not only due to cell-surface binding, but also to intracellular accumulation. Live cells of CCNWGS0123 are a new biosorbent that have the potential to be economical when small batches of biomass are used to remove Cu2+ from an aqueous solution.

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.

Mine drainage leads to serious contamination of soil. To assess the effects of mine drainage on microbial communities in riparian soils, we used an Illumina MiSeq platform to explore the soil microbial composition and diversity along a settling pond used for mine drainage treatment. Non-metric multidimensional scaling analysis showed that the microbial communities differed significantly among the four sampling zones (influent, upstream, downstream and effluent), but not seasonally. Constrained analysis of principal coordinates indicated heavy metals (zinc, lead and copper), total sulphur, pH and available potassium significantly influenced the microbial community compositions. Heavy metals were the key determinants separating the influent zone from the other three zones. Lower diversity indices were observed in the influent zone. However, more potential indicator species, related to sulphur and organic matter metabolism were found there, such as the sulphur-oxidizing genera Acidiferrobacter, Thermithiobacillus, Limnobacter, Thioprofundum and Thiovirga, and the sulphur-reducing genera Desulfotomaculum and Desulfobulbus; the organic matter degrading genera, Porphyrobacter and Paucimonas, were also identified. The results indicated that more microorganisms related to sulphur- and carbon-cycles may exist in soils heavily contaminated by mine drainage.

An actinomycete, strain K55T, was isolated from a composite soil sample from a nickel mine, collected from Yueyang, Shaanxi Province, PR China. Strain K55T showed 16S rRNA gene sequence similarities of 98.73 %–98.51 % to species of the genus Micromonospora, including Micromonospora haikouensis 232617T, Micromonospora coxensis 2-30-b(28)T, Micromonospora wenchangensis 2602GPT1-05T, Micromonospora matsumotoense IMSNU 22003T, Micromonospora maoerensis NEAU-MES19T, and Micromonospora humi P0402T. This strain harboured meso-diaminopimelic acid, alanine and glycine as the major cell-wall amino acids, xylose and glucose as the characteristic whole-cell sugars, and iso-C15 : 0 (20.53 %),iso-C17 : 0 (12.74 %), iso-C16 : 0 (12.15 %), anteiso-C17 : 0 (7.97 %), C17 : 1ω8c (7.49 %) and C17 : 0 (6.63 %) as the dominant fatty acids. The major menaquinones were MK-10(H4) and MK-10(H6). The phospholipid profile comprised phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol and unknown phosphoglycolipids. The DNA G+C content was 71.4 mol%. A comprehensive analysis of several physiological and biochemical traits and DNA–DNA relatedness indicated that strain K55T was different from closely related species. These phenotypic, genotypic and chemotaxonomic data suggest that strain K55T represents a novel species of the genus Micromonospora, for which the name Micromonospora nickelidurans sp. nov., is proposed. The type strain is K55T ( = JCM 30559T = ACCC19713T).

Improving the microbial tolerance to stresses is very important for bioprocesses. Our previous study showed that IrrE, a global regulator from the extremely radioresistant bacterium Deinococcus radiodurans, dramatically enhanced the multi-stress tolerance of Escherichia coli when expressed exogenously. However, the function of IrrE is still unclear. In this study, we used whole-genome microarray assays to profile the global gene expression of the IrrE-expressing E. coli strain MGE and the control strain MGT with or without salt shock. The analysis showed that IrrE expression led to many differentially expressed genes in E. coli, which were responsible for the transport and metabolism of trehalose and glycerol, nucleotide biosynthesis, carbon source utilization, amino acid utilization, and acid resistance, including many RpoS-dependent genes, e.g., the trehalose biosynthesis genes otsAB, the acid-resistance genes gadABC and uspB, the osmotic and oxidative stress response genes katE (response to DNA damage stimulus and stress) and osmBC (response to stress), and gadWX (which controls the transcription of pH-inducible genes). The intracellular content of trehalose and glycerol increased significantly in the IrrE-expressing strain after NaCl treatment for 0 and 60 min as determined by HPLC. These results indicated the possibility that IrrE regulates the global regulator RpoS. Interestingly, we found that although IrrE did not affect the level of the rpoS transcript, it enhanced the accumulation of the RpoS protein by increasing the expression of the antiadaptors, AppY, IraM and IraD, which inhibit RpoS degradation, suggesting that the accumulation of RpoS due to IrrE regulation is an important way to improve tolerance to salt and other stresses in E. coli.

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.

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.

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.

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.

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.

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.

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.

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.