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Mupirocin, a polyketide antibiotic produced by Pseudomonas fluorescens, is used as a topical antimicrobial treatment to cure various skin infections. Quorum sensing system plays an important role in regulation of mupirocin biosynthesis in P. fluorescens NCIMB 10586. In Pseudomonas, the RpeA/RpeB two-component signal transduction (TCST) system regulates quorum sensing system. However, the influences of the RpeA/RpeB TCST system on mupirocin production or other cell activities have not been studied. In this work, the homologous genes of rpeA and rpeB in P. fluorescens NCIMB 10586 were identified and inactivated in the chromosome, respectively. The deletion of rpeA reduced the mupirocin production from 160 in the wild-type to 21.3 mg/L along with slightly decreased cell growth, while no significant effected on mupirocin production in the rpeB mutant. Next, it was found that the RpeA/RpeB TCST system regulated the biosynthesis of mupirocin by modulating the quorum sensing system. Furthermore, untargeted metabolomics analysis was employed to detect the influences of RpeA on other cell activities modulated by quorum sensing system. Combined with quantitative real-time PCR, the results demonstrated that RpeA also regulated other cell activities including central carbon, amino acids, fatty acids, and purine metabolism. Overall, this study expands the current understanding of the RpeA/RpeB TCST system and provides several targets for increasing yields of mupirocin. Key points: • In P. fluorescens, the RpeA/RpeB TCST system regulates the biosynthesis of mupirocin. • RpeA modulates the cell activities through effecting the central carbon metabolism. [ABSTRACT FROM AUTHOR]

Glycerol is a by-product of biodiesel, and it has a great application prospect to be transformed to synthesize high value-added compounds. Pseudomonas chlororaphis GP72 isolated from the green pepper rhizosphere is a plant growth promoting rhizobacteria that can utilize amount of glycerol to synthesize phenazine-1-carboxylic acid (PCA). PCA has been commercially registered as "Shenqinmycin" in China due to its characteristics of preventing pepper blight and rice sheath blight. The aim of this study was to engineer glycerol utilization pathway in P. chlororaphis GP72. First, the two genes glpF and glpK from the glycerol metabolism pathway were overexpressed in GP72ANO separately. Then, the two genes were co-expressed in GP72ANO, improving PCA production from 729.4 mg/L to 993.4 mg/L at 36 h. Moreover, the shunt pathway was blocked to enhance glycerol utilization, resulting in 1493.3 mg/L PCA production. Additionally, we confirmed the inhibition of glpR on glycerol metabolism pathway in P. chlororaphis GP72. This study provides a good example for improving the utilization of glycerol to synthesize high value-added compounds in Pseudomonas. [ABSTRACT FROM AUTHOR]

The effects of P supplementation on chromium(VI) uptake by Leersia hexandra Swartz were studied using pot-culture experiment. P-deficiency and zero-P addition controls were included. The Cr(VI) uptake followed Michaelis-Menten kinetics. Compare with the control, the P-supply decreased the Michaelis constant (Km) by 16.9% and the P-deficiency decreased the maximum uptake velocity (Vmax) by 18%, which indicated no inhibition and competition between P and Cr(VI) uptake by L. hexandra. Moreover, there were a synergistic action between P and Cr(VI) suggests that Cr(VI) uptake by the roots of L. hexandra may be an active process. The bioconcentration factor (BCF) and the transport factor (TF′) increased with the increase in P supply. The highest BCF was 3.6-folds higher than the control, indicating that the additional P contribute to a higher ability of L. hexandra transporting Cr from root to the aboveground parts. [ABSTRACT FROM AUTHOR]

Involvement of genes ( CYS-A1, CYS-C1 and NIT4) encoded with cysteine synthase, β-cyanoalanine synthase, nitrilase and cyanide metabolisms are evident in Arabidopsis. In the present study, identifications of CYS-A1, CYS-C1 and NIT4, predictions of conserved motifs, and constructions of phylogenetic relationships, based on their amino acid sequences in rice, were conducted. In order to elucidate the transcriptional responses of these cyanide-degrading genes, two candidate homologues were selected for each gene to test their expression changes upon exposure to exogenous KCN in rice seedlings using RT-PCR. Results showed that all selected candidate homologous genes were differentially expressed at different exposure points in roots and shoots of rice seedlings, suggesting their distinct roles during cyanide assimilation. Both candidate homologues for CYS-A1 constantly exhibited more abundant transcripts in comparison to control. However, only one candidate homologue for CYS-C1 and NIT4 showed a remarkable up-regulation during KCN exposure. Analysis of both tissue and solution cyanide indicated that rice seedlings were quickly able to metabolize exogenous KCN with minor accumulation in plant tissues. In conclusion, significant up-regulation of CYS-A1 suggested that the endogenous pool of cysteine catalyzed by cysteine synthase does not restrict the conversion of exogenous KCN into cyanoalanine through the β-cyanoalanine pathway. However, insufficient responses of the transcription level of NIT4 suggested that NIT enzyme may be a limiting factor for cyanoalanine assimilation by rice seedlings. [ABSTRACT FROM AUTHOR]

Hydroponic experiments were conducted with different species of plants (rice, maize, soybean and willow) exposed to ferri-cyanide to investigate the half-saturation constant ( K) and the maximal metabolic capacity ( v) involved in phyto-assimilation. Three varieties for each testing species were collected from different origins. Measured concentrations show that the uptake rates responded biphasically to ferri-cyanide treatments by showing increases linearly at low and almost constant at high concentrations from all treatments, indicating that phyto-assimilation of ferri-cyanide followed the Michaelis-Menten kinetics. Using non-linear regression, the highest v was by rice, followed by willows. The lowest v was found for soybean. All plants, except maize (DY26) and rice (XJ12), had a similar K value, suggesting the same enzyme was active in phyto-assimilation of ferri-cyanide. Transcript level, by real-time quantitative PCR, of enzymes involved in degradation of cyanides showed that the analyzed genes were differently expressed during different cyanides exposure. The expression of CAS and ST genes responded positively to KCN exposure, suggesting that β-CAS and ST pathways were two possible pathways for cyanide detoxification in rice. The transcript level of NIT and ASPNASE genes also showed a remarkable up-regulation to KCN, implying the contribution to the pool of amino acid aspartate, which is an end product of CN metabolism. Up-regulation of GS genes suggests that acquisition of ammonium released from cyanide degradation may be an additional nitrogen source for plant nutrition. Results also revealed that the expressions of these genes, except for GS, were relatively constant during iron cyanide exposure, suggesting that they are likely metabolized by plants through a non-defined pathway rather than the β-CAS pathway. [ABSTRACT FROM AUTHOR]

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Hydroponic experiments were conducted with rice seedlings ( Oryza sativa L. cv. XZX45) exposed to gallium nitrate (Ga) to investigate the accumulation of Ga in plant tissues and phytotoxic responses. Results showed that phyto-transport of Ga was apparent, and roots were the dominant site for Ga accumulation. The total accumulation rates of Ga responded biphasically to Ga treatments by showing increases at low (1.06-8.52 mg Ga/L) and constants at high (8.52-15.63 mg Ga/L) concentrations, suggesting that accumulation kinetics of Ga followed a typical saturation curve. Higher amount of Ga accumulation in plant tissues led to significant inhibition in relative growth rate and water use efficiency in a dose-dependent manner. DNA-protein cross-links (DPCs) analysis revealed that overaccumulation of Ga in plant tissues positively stimulated formation of DPCs in roots. Likewise, the measure of root cell viability evaluated by Evan blue uptake showed a similar trend. These results suggested that Ga can be absorbed, transported, and accumulated in plant materials of rice seedlings. Overaccumulation of Ga in plant tissues provoked the formation of DPCs in roots, which resulted in cell death and growth inhibition of rice seedlings. [ABSTRACT FROM AUTHOR]

Hydroponic experiments were conducted with rice seedlings ( Oryza sativa L. cv. ND1) exposed to methylene blue (MB) to investigate the accumulation of hydrogen peroxide (HO) and activities and gene expression of antioxidant enzymes. Results showed that a linear decrease in relative growth rate and water use efficiency was observed with rice seedlings with increasing MB concentrations. MB-induced accumulation of HO was evident in both roots and shoots. The activities of antioxidative enzymes were analyzed and found to be different at different MB treatment concentrations. The activities of enzymes related to the ascorbate-glutathione cycle were more sensitive to MB treatments than other antioxidative enzymes. Transcript level, by real-time quantitative PCR, of antioxidative enzymes showed that the analyzed genes were differently expressed during different MB concentrations in both roots and shoots. The isoform analysis of superoxide dismutase (SOD) gene showed that the expressions of Cu/ ZnSOD and MnSOD were relatively constant, where significant upregulation of FeSOD was observed with rice seedlings exposed to high MB concentrations. Furthermore, the expression of CAT, POD, and MDHAR genes responded biphasically to MB treatments by showing negligible changes at 1.56-15.63 μM MB and significant induction at 31.36-62.52 μM MB. The expression of GR, APX, and DHAR genes showed a remarkable induction to MB. Our results suggest that on transcription level, and in accordance with enzymatic responses, enzymes of GR, APX, and DHAR play central role in the HO detoxification in rice seedlings under MB exposure. [ABSTRACT FROM AUTHOR]

Solution culture was carried to investigate capacity of synthetic aminopolycarboxylic acids (ethylenediamine tetraacetate, N-hydroxyethylenediaminetriacetic acid, and diethylenetriamine-pentaacetate) for enhancing botanical removal and transport of heavy metals (Cu and Zn) by plants. Biodegradable organic acids (citric acid, malic acid, and oxalic acid) were also selected as alternatives to compare them with synthesized chelating agents for effectiveness. Young rice seedlings ( Oryza sativa L. cv. XZX 45) were grown in nutrient solutions treated with single or combined metal solutions in presence or absence of chelating compounds. Calculation by chemical equilibrium program VISUAL MINTEQ showed that different chelating compounds had various complex potential with Cu and Zn ions, in which synthetic chelators exhibited higher complexed capability than biodegradable organic acids. All applied synthetic aminopolycarboxylic acids significantly decreased removal of metal from nutrient solution ( p < 0.01), while more or less effects of organic acids supplied on biosorptive potential were observed with all treatments ( p > 0.05), compared with the treatment without metal ligands. Synthetic aminopolycarboxylic acids significantly decreased metal concentrations in plant materials in all treatments ( p < 0.01). However, biodegradable organic acids decreased metal concentrations in roots ( p < 0.01), but enhanced them in shoots ( p < 0.01). Results obtained indicated that synthetic aminopolycarboxylic acids decreased uptake of metals by rice seedlings, but translocation of metals complexed within plant materials was evident. Although exogenous biodegradable organic acids showed negligible effect on botanical removal of metals, metals complexed with organic acids was more mobile than those complexed with other chelating agents. These information collected here had important implication for the use of biodegradable metal chelators in transport of essential micronutrients in plant nutrition. [ABSTRACT FROM AUTHOR]

Adsorption behaviors of chromate(VI) on two different iron oxides were investigated through batch experiments under different operational conditions. The operational variables studied included sorbent doses, sorbent species, solution pH, contact time, and initial metal ion concentrations. Both ferri hydroxide and ferri oxohydroxides were capable of removing Cr(VI) from aqueous solution effectively. The extent of metal ion removed from aqueous solution by both sorbents increased with increasing contact time and initial metal ion concentrations, while decreased with elevating solution pH initially. Sorption equilibrium time was observed in 150 min. Equilibrium isotherm data were analyzed using the Langmuir and Freundlich isotherms. The Freundlich model yields a better fit than the Langmuir model. Although kinetic measurements with both sorbents had a similar pattern, ferri hydroxide always showed higher sorptive potential for Cr(VI) than ferri oxohydroxides. Results also indicated that sorptive removal of Cr(VI) by both sorbents was observed to be negative in response to solution pH, due to changes in speciation of Cr(VI) in aqueous solution. At pH lower than 3.0, both iron oxides exhibited much greater capacities of removing Cr(VI) from aqueous solution than other treatments. Therefore, it is to conclude that adsorptive removal of Cr(VI) by both iron oxides is effective. Adsorption capacity is mainly determined by speciation of Cr(VI) in solutions, which is controlled and driven by solution pH. [ABSTRACT FROM AUTHOR]

Revegetation and ecological restoration of a Mn mineland are important concerns in southern China. To determine the major constraints for revegetation and select suitable plants for phytorestoration, pedological and botanical characteristics of a Mn mine in Guangxi, southern China were investigated. All the soils were characterized by low pH and low nitrogen and phosphorus levels except for the control soil, suggesting that soil acidity and poor nutrition were disadvantageous to plant growth. In general, the studied mine soils had normal organic matter (OM) and cation exchange capacity (CEC). However, OM (8.9 g/kg) and CEC (7.15 cmol/kg) were very low in the soils from tailing dumps. The sandy texture and nutrient deficiency made it difficult to establish vegetation on tailing dumps. Mn and Cd concentrations in all soils and Cr and Zn concentrations in three soils exceeded the pollution threshold. Soil Mn and Cd were above phytotoxic levels, indicating that they were considered to be the major constraints for phytorestoration. A botanical survey of the mineland showed that 13 plant species grew on the mineland without obvious toxicity symptoms. High Mn and Cd concentrations have been found in the aerial parts of Polygonum pubescens, Celosia argentea, Camellia oleifera, and Solanum nigrum, which would be interesting for soil phytoremediation. Miscanthus floridulus, Erigeron acer, Eleusina indica, and Kummerowia striata showed high resistance to the heavy metal and harsh condition of the soils. These species could be well suited to restore local degraded land in a phytostabilization strategy. [ABSTRACT FROM AUTHOR]

Responses of free amino acids to botanical assimilation of free cyanide were investigated. Young rice seedlings ( Oryza sativa L. cv. XZX 45) were grown in nutrient solution amended with free cyanide (KCN). Cyanide was analyzed in solution as well as in plant materials to estimate the phyto-assimilation potential. Free amino acids in different parts of plants were also measured to determine metabolic responses to KCN exposure. Phyto-assimilation of KCN was obvious, and the rates were positively correlated to the concentration supplied. Although changes in total amino acid content in plant materials were negligible during KCN metabolism ( p > 0.05), responses of different amino acids to KCN treatments were quite different. All treatments with KCN increased the content of proline (Pro) and isoleucine (Ile) in roots significantly compared with control ( p < 0.05), while changes of aspartic acid, lysine, and histidine in roots were more evident at higher KCN treatments ( p < 0.05). Results indicate that the content of Pro, Ile, and tyrosine showed pronounced increase in shoots of rice seedlings exposed to KCN at 1.44 mg CN/L or higher ( p < 0.05). Other amino acids slightly changed in all plant materials exposed to KCN ( p > 0.05). Results indicate that specific amino acids in rice seedlings showed positive response to non-toxic concentrations of exogenous KCN. These findings could provide additional insights into the inducible mechanisms underlying the involvement of amino acids in KCN metabolism. [ABSTRACT FROM AUTHOR]

High-throughput Illumina RNA-seq was used for deep sequencing analysis of the transcriptome of poly(A)+ RNA from mycelium grown under three different conditions: 30 days darkness (sample 118), 80 days darkness (313W), and 30 days darkness followed by 50 days in the light (313C), in order to gain insight into the molecular mechanisms underlying the process of light-induced brown film (BF) formation in the edible mushroom, Lentinula edodes. Of the three growth conditions, BF formation occurred in 313C samples only. Approximately 159.23 million reads were obtained, trimmed, and de novo assembled into 31,511 contigs with an average length of 1,746 bp and an N of 2,480 bp. Based on sequence orientations determined by a BLASTX search against the NR, Swiss-Prot, COG, and KEGG databases, 24,246 (76.9 %) contigs were assigned putative descriptions. Comparison of 313C/118 and 313C/313W expression profiles revealed 3,958 and 5,651 significantly differentially expressed contigs (DECs), respectively. Annotation using the COG database revealed that candidate genes for light-induced BF formation encoded proteins linked to light reception (e.g., WC-1, WC-2, phytochrome), light signal transduction pathways (e.g., two-component phosphorelay system, mitogen-activated protein kinase pathway), and pigment formation (e.g., polyketide synthase, O-methyltransferase, laccase, P monooxygenase, oxidoreductase). Several DECs were validated using quantitative real-time polymerase chain reaction. Our report is the first to identify genes associated with light-induced BF formation in L. edodes and represents a valuable resource for future genomic studies on this commercially important mushroom. [ABSTRACT FROM AUTHOR]

Compound of 1-hydroxyphenazine (1-OH-PHZ) is an important fungicide in agricultural production. The ranges of 1-OH-PHZ solubilities in dichloromethane at temperature of 278.5-308.5 K and in ethyl acetate, methanol and water at temperature of 278.5-333.5 K were measured using the analytical stirred flask method at barometric pressure. The color of the liquid remained unchanged throughout. The solubilities of 1-OH-PHZ in water, methanol, ethyl acetate and dichloromethane increased orderly and these solubilities also increased with increasing the temperature. The experimental data was correlated by the modified Apelblat equation. The root mean square deviations were all within 0.5% and the maximum relative average deviation was 5.2%. The calculated solubility shows a good relationship with the experimental solubility. [ABSTRACT FROM AUTHOR]

In this work, molecularly imprinted solid-phase extraction (MISPE) has been used to selectively enrich, purify, or remove synephrine from Aurantii Fructus Immaturus. To this end, a molecularly imprinted polymer (MIP) was prepared by self-assembly from the template synephrine, the functional monomer methacrylic acid, and the crosslinker ethylene glycol dimethacrylate in 1:4:20 molar ratio. Subsequent molecular interrogation of the MIP binding sites revealed preferential structural selectivity for synephrine relative to other structurally related naturally occurring compounds (i.e. octopamine and tyramine ). This selectivity was subsequently exploited to achieve substantial sample clean-up of extracts of crude Aurantii Fructus Immaturus and Aurantii Fructus Immaturus stir-baked with bran. The purity of synephrine in the extracts after MISPE represented approximately 24.21-fold enrichment of the synephrine in the untreated extracts of Aurantii Fructus Immaturus stir-baked with bran. High recoveries (85-90%) from the samples proved that the method was valid for selective enrichment, purification, or removal of synephrine from Aurantii Fructus Immaturus. [ABSTRACT FROM AUTHOR]

Purpose: The objective of this paper is to assess the impact of long-term electroplating industrial activities on heavy metal contamination in agricultural soils and potential health risks for local residents. Methods: Water, soil, and rice samples were collected from sites upstream (control) and downstream of the electroplating wastewater outlet. The concentrations of heavy metals were determined by an atomic absorption spectrophotometer. Fractionation and risk assessment code (RAC) were used to evaluate the environmental risks of heavy metals in soils. The health risk index (HRI) and hazard index (HI) were calculated to assess potential health risks to local populations through rice consumption. Results: Hazardous levels of Cu, Cr, and Ni were observed in water and paddy soils at sites near the plant. According to the RAC analysis, the soils showed a high risk for Ni and a medium risk for Cu and Cr at certain sites. The rice samples were primarily contaminated with Ni, followed by Cr and Cu. HRI values >1 were not found for any heavy metal. However, HI values for adults and children were 2.075 and 1.808, respectively. Conclusion: Water, paddy soil, and rice from the studied area have been contaminated by Cu, Cr, and Ni. The contamination of these elements is related to the electroplating wastewater. Although no single metal poses health risks for local residents through rice consumption, the combination of several metals may threaten the health of local residents. Cu and Ni are the key components contributing to the potential health risks. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the relationship between different hCG priming-to-oocyte retrieval intervals and assisted reproductive technology (ART) outcome. Methods: We systematically searched PubMed, EMBASE, the Cochrane Library, Science Citation Index, Chinese biomedicine (CBM) literature database, and Chinese Journal Full-text Database for randomized controlled trials (RCTs) published up to November 2010. Data was extracted from the studies by two independent reviewers. Statistical analysis was performed with Cochrane Collaboration's Review Manager (RevMan) 5.0.2. From extracted data, Risk Ratio (RR) with 95% confidence interval (CI) was calculated. Results: 5 RCTs totaling 895 participants were included. Oocyte maturation rate was higher in the long interval group compared with short interval group (RR, 0.67; 95% CI, 0.62-0.73). There were no significant difference between the two groups with regard to fertilization rate (RR, 0.99; 95% CI, 0.94-1.04), implantation rate (RR, 0.91; 95% CI, 0.40-2.04), and pregnancy rate (RR, 0.79; 95% CI, 0.58-1.08). Conclusion: The percentage of mature (MII) oocytes can be increased by prolonging the interval between hCG priming and oocyte retrieval. The prolonged interval could not increase the fertilization rate, implantation rate, and pregnancy rate. Although there was evidence to confirm the results, they still need to be confirmed by large-sample, multicenter, randomized controlled trials. The time interval dependent mechanisms responsible for ART performance need to be elucidated. [ABSTRACT FROM AUTHOR]

Effect of chemical fertilizers (urea, NH4Cl, Ca(NO3)2, KCl and KH2PO4) on the fractionation of Cu, Cr and Ni was studied by a 4-month incubation experiment. Using sequential extraction procedure, it was found that the application of fertilizers could change the distribution of Cu, Cr and Ni in the fractions of soil. Applying urea (CO(NH2)2) significantly decreased the concentrations of Cu, Cr and Ni in water soluble plus exchangeable (WE) fraction, but increased those in Fe–Mn oxides bound (FM) fraction ( p < 0.01). However, application of NH4Cl caused an increase in the WE fraction by 27.7% for Cu, 111.5% for Cr and 20.4% for Ni. The CO(NH2)2 raised the soil pH from 4.51 to 4.96, whereas NH4Cl lowered the pH of soil by 0.44 units. The WE fraction of the three heavy metals was significantly increased, while the FM fraction was significantly decreased by adding KCl ( p < 0.01). Moreover, the supply of KH2PO4 reduced the WE and carbonate bound (CB) fractions of Cu, Cr and Ni in the soil, however, it raised Cu and Ni in the residual (RS) fraction and Cr in the FM fraction. In addition, the mobility index indicated that KCl and NH4Cl increased the mobility of Cu, Cr and Ni in the soil, whereas urea and KH2PO4 decreased the mobility of the three metals in the soil. These results suggest that applying chemical fertilizers does not only provide plant nutrients, but may also change the speciation and mobility of heavy metals in the soil. [ABSTRACT FROM AUTHOR]