Overexpression of a tomato flavanone 3-hydroxylase-like protein gene improves chilling tolerance in tobacco.

Flavonoids are secondary metabolites found in plants with a wide range of biological functions, such as stress protection. This study investigated the functions of a tomato (Solanum lycopersicum) flavanone 3-hydroxylase-like protein gene SlF3HL by using transgenic tobacco. The expression of the gene was up-regulated under chilling (4 °C), heat (42 °C), salt (NaCl) and oxidative (H2O2) stresses. The transgenic plants that displayed high SlF3HL mRNA and protein levels showed higher flavonoid content than the WT plants. Moreover, the expression of three flavonoid biosynthesis-related structural genes, namely, chalcone synthase (CHS), chalcone isomerase (CHI) and flavonol synthase (FLS) was also higher in the transgenic plants than in the WT plants. Under chilling stress, the transgenic plants showed not only faster seed germination, better survival and growth, but also lower malondialdehyde (MDA) accumulation, relative electrical conductivity (REC) and H2O2 and O2(·-) levels compared with WT plants. These results suggested that SlF3HL stimulated flavonoid biosynthesis in response to chilling stress.

Occupational stress and coping strategies among emergency department nurses of China.

Emergency department(ED) nurses work in a rapidly changing environment with patients that have wide variety of conditions. Occupational stress in emergency department nurses is a common problem. The purpose of this study was to describe the relationship between coping strategies and occupational stress among ED nurses in China. A correlational, cross-sectional design was adopted. Two questionnaires were given to a random sample of 127 ED nurses registered at the Heilongjiang Nurses' Association. Data were collected from the nurses that worked in the ED of five general hospitals in Harbin China. Occupational stress and coping strategies were measured by two questionnaires. A multiple regression model was applied to analyze the relationship between stress and coping strategies. The stressors of ED nurses mainly come from the ED specialty of nursing (2.97±0.55), workload and time distribution (2.97±0.58). The mean score of positive coping strategies was 2.19±0.35, higher than the norm (1.78±0.52). The mean score of negative coping strategies was 1.20±0.61, lower than the norm (1.59±0.66), both had significant statistical difference (P<0.001). Too much documents work, criticism, instrument equipment shortage, night shift, rank of professional were the influence factors about occupational stress to positive coping styles. Too much documents work, and medical insurance for ED nurses were the influential factors on occupational stress to negative coping styles. This study identified several factors associated with occupational stress in ED nurses. These results could be used to guide nurse managers of ED nurses to reduce work stress. The managers could pay more attention to the ED nurse's coping strategies which can further influence their health state and quality of nursing care. Reducing occupational stress and enhancing coping strategies are vital not only for encouraging nurses but also for the future of nursing development.

Heterology expression of the tomato LeLhcb2 gene confers elevated tolerance to chilling stress in transgenic tobacco.

Chilling is one of the most serious environmental stresses that disrupt the metabolic balance of cells and enhance the production of reactive oxygen species (ROS). Light harvesting complex (LHC) proteins had a function in dissipating excess excitation energy and eliminating ROS to maintain the normal physiological function of cells. A tomato (Lycopersicon esculentum) LHC antenna protein gene (LeLhcb2) was isolated. The LeLhcb2-green fluorescent protein (GFP) fusion protein was targeted to the chloroplast of Arabidopsis mesophyll protoplast. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the expression of LeLhcb2 was markedly abundant in leaves and was induced by chilling (4 °C). qRT-PCR analysis and western blot confirmed that the sense gene LeLhcb2 was transferred into tobacco genome and overexpressed. Under chilling stress, the transgenic plants showed not only better growth, higher fresh weight, chlorophyll content, but also lower malondialdehyde (MDA) accumulation and relative electrical conductivity (REC), compared with the wild type (WT). The maximal photochemical efficiency of PSII (Fv/Fm), non-photochemical quenching (NPQ) and D1 protein content were also higher in the transgenic plants. Furthermore, the relatively lower hydrogen peroxide (H2O2) and superoxide radical (O2(-)) levels in the sense plants were not considered to due to the higher activity of ascorbate peroxidase (APX) and superoxide dismutase (SOD). These results suggested that the overexpression of LeLhcb2 had a key function in alleviating photo-oxidation of PSII and enhanced transgenic tobacco tolerance to chilling stress.

Overexpression of a tomato carotenoid ε-hydroxylase gene alleviates sensitivity to chilling stress in transgenic tobacco.

Chilling is one of the most serious environmental stresses that disrupt the metabolic balance of cells and enhance the production of reactive oxygen species (ROS). Lutein plays important roles in dissipating excess excitation energy and eliminating ROS to maintain the normal physiological function of cells. A tomato carotenoid epsilon-ring hydroxylase gene (LeLUT1) was isolated, and the LeLUT1-GFP fusion protein was localized in the chloroplast of Arabidopsis mesophyll protoplast. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the expression of LeLUT1 was the highest in the leaves and was down-regulated by various abiotic stresses in tomato. The transgenic tobacco plants overexpressing LeLUT1 had higher lutein content, which was decreased in cold condition. Under chilling stress, the non-photochemical quenching (NPQ) values were higher in the transgenic plants than in the wild type (WT) plants. Compared with the WT plants, the transgenic plants showed lower levels of hydrogen peroxide (H2O2), superoxide radical (O2(·-)), relative electrical conductivity, and malondialdehyde content (MDA), and relatively higher values of maximal photochemical efficiency of photosystem II (Fv/Fm), oxidizable P700 of PSI, and net photosynthetic rate (Pn). Therefore, the transgenic seedlings were less suppressed in growth and lost less cotyledon chlorophyll than the WT seedlings. These results suggested that the overexpression of LeLUT1 had a key function in alleviating photoinhibition and photooxidation, and decreased the sensitivity of photosynthesis to chilling stress.

Accelerated reduction of chlorinated nitroaromatic antibiotic chloramphenicol by biocathode.

Chlorinated nitroaromatic antibiotic chloramphenicol (CAP) is a priority pollutant in wastewaters. A fed-batch bioelectrochemical system (BES) with biocathode with applied voltage of 0.5 V (served as extracellular electron donor) and glucose as intracellular electron donor was applied to reduce CAP to amine product (AMCl2). The biocathode BES converted 87.1 ± 4.2% of 32 mg/L CAP in 4 h, and the removal efficiency reached 96.0 ± 0.9% within 24 h. Conversely, the removal efficiency of CAP in BES with an abiotic cathode was only 73.0 ± 3.2% after 24 h. When the biocathode was disconnected (no electrochemical reaction but in the presence of microbial activities), the CAP removal rate was dropped to 62.0% of that with biocathode BES. Acetylation of one hydroxyl of CAP was noted exclusive in the biocatalyzed process, while toxic intermediates, hydroxylamino (HOAM), and nitroso (NO), from CAP reduction were observed only in the abiotic cathode BES. Electrochemical hydrodechlorination and dehalogenase were responsible for dechlorination of AMCl2 to AMCl in abiotic and microbial cathode BES, respectively. The cyclic voltammetry (CV) highlighted higher peak currents and lower overpotentials for CAP reduction at the biocathode compared with abiotic cathode. With the biocathode BES, antibacterial activity of CAP was completely removed and nitro group reduction combined with dechlorination reaction enhanced detoxication efficiency of CAP. The CAP cathodic transformation pathway was proposed based on intermediates analysis. Bacterial community analysis indicated that the dominate bacteria on the biocathode were belonging to α, ß, and γ-Proteobacteria. The biocathode BES could serve as a potential treatment process for CAP-containing wastewater.

Azo dye removal in a membrane-free up-flow biocatalyzed electrolysis reactor coupled with an aerobic bio-contact oxidation reactor.

Azo dyes that consist of a large quantity of dye wastewater are toxic and persistent to biodegradation, while they should be removed before being discharged to water body. In this study, Alizarin Yellow R (AYR) as a model azo dye was decolorized in a combined bio-system of membrane-free, continuous up-flow bio-catalyzed electrolysis reactor (UBER) and subsequent aerobic bio-contact oxidation reactor (ABOR). With the supply of external power source 0.5 V in the UBER, AYR decolorization efficiency increased up to 94.8±1.5%. Products formation efficiencies of p-phenylenediamine (PPD) and 5-aminosalicylic acid (5-ASA) were above 90% and 60%, respectively. Electron recovery efficiency based on AYR removal in cathode zone was nearly 100% at HRTs longer than 6 h. Relatively high concentration of AYR accumulated at higher AYR loading rates (>780 gm(-3) d(-1)) likely inhibited acetate oxidation of anode-respiring bacteria on the anode, which decreased current density in the UBER; optimal AYR loading rate for the UBER was 680 gm(-3) d(-1) (HRT 2.5 h). The subsequent ABOR further improved effluent quality. Overall the Chroma decreased from 320 times to 80 times in the combined bio-system to meet the textile wastewater discharge standard II in China.

Antisense-mediated suppression of tomato thylakoidal ascorbate peroxidase influences anti-oxidant network during chilling stress.

Photosynthesis is a well-established source of reactive oxygen species (ROS) in plants particularly under chilling stress. Ascorbate peroxidase (APXs) plays an important role in the anti-oxidant system by utilizing AsA as specific electron donor to reduce H(2)O(2) to water. In order to investigate the possible mechanisms of ascorbate peroxidsae (APX) in photoprotection under chilling stress, a tomato (Lycopersicon esculentum Mill.) thylakoidal ascorbate peroxidase gene (LetAPX) was isolated and antisense transgenic tomato plants were produced. Under chilling stress, transgenic plants accumulated more H(2)O(2), and showed higher levels of ion leakage and malondialdehyde (MDA), lower net photosynthetic rate (Pn), lower maximal photochemical efficiency of PSII (Fv/Fm) and less content of D1 protein compared with wild type (WT) plants. On the other hand, after chilling stress, transgenic plants showed higher reduced ascorbate (AsA) and activities of catalase (CAT) and superoxide dismutase (SOD) than those in WT plants, and the expression of several known stress-responsive and antioxidative genes was also higher at the end of chilling treatment. These results suggested that the suppression of LetAPX gene induced compensatory anti-oxidant mechanisms in tomato, and inactivation of tAPX may have a regulatory role in facilitating redox signaling pathways under chilling stress. Furthermore, transient increases in ROS levels also have a vital role in stress signaling and thereby in the survival of plants under chilling conditions.

Constitutive accumulation of zeaxanthin in tomato alleviates salt stress-induced photoinhibition and photooxidation.

Zeaxanthin (Z) has a role in the dissipation of excess excitation energy by participating in non-photochemical quenching (NPQ) and is essential in protecting the chloroplast from photooxidative damage. To investigate the physiological effects and functional mechanism of constitutive accumulation of Z in the tomato at salt stress-induced photoinhibition and photooxidation, antisense-mediated suppression of zeaxanthin epoxidase transgenic plants and the wild-type (WT) tomato were used. The ratio of Z/(V + A + Z) and (Z + 0.5A)/(V + A + Z) in antisense transgenic plants were maintained at a higher level than in WT plants under salt stress, but the value of NPQ in WT and transgenic plants was not significantly different under salt stress. However, the maximal photochemical efficiency of PSII (Fv/Fm) and the net photosynthetic rate (Pn) in transgenic plants decreased more slowly under salt stress. Furthermore, transgenic plants showed lower level of hydrogen peroxide (H(2)O(2)), superoxide anion radical (O(2)(•-)) and ion leakage, lower malondialdehyde content. Compared with WT, the content of D1 protein decreased slightly in transgenic plants under salt stress. Our results suggested that the constitutive accumulation of Z in transgenic tomatoes can alleviate salt stress-induced photoinhibition because of the antioxidant role of Z in the scavenging quenching of singlet oxygen and/or free radicals in the lipid phase of the membrane.

A membrane-free, continuously feeding, single chamber up-flow biocatalyzed electrolysis reactor for nitrobenzene reduction.

A new bioelectrochemical system (BES), a membrane-free, continuous feeding up-flow biocatalyzed electrolysis reactor (UBER) was developed to reduce oxidative toxic chemicals to less- or non-toxic reduced form in cathode zone with oxidation of electron donor in anode zone. Influent was fed from the bottom of UBER and passed through cathode zone and then anode zone. External power source (0.5 V) was provided between anode and cathode to enhance electrochemical reactions. Granular graphite and carbon brush were used as cathode and anode, respectively. This system was tested for the reduction of nitrobenzene (NB) using acetate as electron donor and carbon source. The influent contained NB (50-200 mg L(-1)) and acetate (1000 mg L(-1)). NB was removed by up to 98% mainly in cathode zone. The anode potential maintained under -480 mV. The maximum NB removal rate was up to 3.5 mol m(-3) TV d(-1) (TV=total empty volume) and the maximum aniline (AN) formation rate was 3.06 mol m(-3) TV d(-1). Additional energy required was less than 0.075 kWh mol(-1)NB. The molar ratio of NB removed vs acetate consumed varied from 4.3 ± 0.4 to 2.3 ± 0.1 mol mol(-1). Higher influent phosphate or acetate concentration helped NB removal rate. NB could be efficiently reduced to AN as the power supplied of 0.3 V.

Electricity generation and brewery wastewater treatment from sequential anode-cathode microbial fuel cell.

A sequential anode-cathode double-chamber microbial fuel cell (MFC), in which the effluent of anode chamber was used as a continuous feed for an aerated cathode chamber, was constructed in this experiment to investigate the performance of brewery wastewater treatment in conjugation with electricity generation. Carbon fiber was used as anode and plain carbon felt with biofilm as cathode. When hydraulic retention time (HRT) was 14.7 h, a relatively high chemical oxygen demand (COD) removal efficiency of 91.7%-95.7% was achieved under long-term stable operation. The MFC displayed an open circuit voltage of 0.434 V and a maximum power density of 830 mW/m(3) at an external resistance of 300 Omega. To estimate the electrochemical performance of the MFC, electrochemical measurements were carried out and showed that polarization resistance of anode was the major limiting factor in the MFC. Since a high COD removal efficiency was achieved, we conclude that the sequential anode-cathode MFC constructed with bio-cathode in this experiment could provide a new approach for brewery wastewater treatment.