Optimization of inorganic carbon sources to improve the carbon fixation efficiency of the non-photosynthetic microbial community with different electron donors.

As the non-photosynthetic microbial community (NPMC) isolated from seawaters utilized inorganic carbon sources for carbon fixation, the concentrations and ratios of Na2CO3, NaHCO3, and CO2 were optimized by response surface methodology design. With H2 as the electron donor, the optimal carbon sources were 270 mg/L Na2CO3, 580 mg/L NaHCO3, and 120 mg/L CO2. The carbon fixation efficiency in response to total organic carbon (TOC) was up to 30.59 mg/L with optimal carbon sources, which was about 50% higher than that obtained with CO2 as the sole carbon source. The mixture of inorganic carbon sources developed a buffer system to prevent acidification or alkalization of the medium caused by CO2 or Na2CO3, respectively. Furthermore, CO2 and HCO3(-), the starting points of carbon fixation in the pathways of Calvin-Benson-Bassham and 3-hydroxypropionate cycles, were provided by the carbon source structure to facilitate carbon fixation by NPMC. However, in the presence of mixed electron donors composed of 1.25% Na2S, 0.50% Na2S2O3, and 0.457% NaNO2, the carbon source structure did not exhibit significant improvement in the carbon fixation efficiency, when compared with that achieved with CO2 as the sole carbon source. The positive effect of mixed electron donors on inorganic carbon fixation was much higher than that of the carbon source structure. Nevertheless, the carbon source structure could be used as an alternative to CO2 when using NPMC to fix carbon in industrial processes.

Cadmium stimulated cooperation between bacterial endophytes and plant intrinsic detoxification mechanism in Lonicera japonica thunb.

Due to the intimate association between plant physiology and metabolism, the internal colonizing microbe (endophytes) community must be adjusted to support plant productivity in response to cell damage in plants under stress. However, how endophytes coordinate their activities with plant intrinsic mechanisms such as antioxidative systems and detoxification pathways during Cd accumulation remains unknown. In this hydroponic pot study, we investigated how exposure of Lonicera japonica. thunb. to different levels of Cd (0.5, 2.5, 5, 10, and 20 mg kg-1) affected plant growth, metabolic pathways, and endophyte community structure and function. Although Cd accumulation increased at 5 mg kg-1 Cd, the biomass and height of L. japonica increased in association with elevated endophyte-involved plant detoxification activities. Endophytes, such as Sphingomonas, Klenkia, and Modestobacter, expressed major antioxidative regulators (superoxide dismutase and ascorbate acid) to detoxify Cd in L. japonica. Furthermore, L. japonica and its endophytes synergistically regulated the toxic effects of Cd accumulation via multiple plant metabolic defensive pathways to increase resistance to metal-induced stress.

Variability of soil organic carbon reservation capability between coastal salt marsh and riverside freshwater wetland in Chongming Dongtan and its microbial mechanism.

Two representative zones in Chongming Dongtan which faced the Yangtze River and East China Sea respectively were selected to study the variability of soil organic carbon (SOC) reservation capability between coastal wetland and riverside wetland in the Chongming Dongtan wetland as well as its mechanism by analyzing soil characteristics and plant biomass. The results showed the SOC content of riverside wetland was only 48.61% (P = 0.000 < 0.05) that of coastal wetland. As the organic matter inputs from plant litter of the coastal wetland and riverside wetland were approximately the same, the higher soil microbial respiration (SMR) of riverside wetland led to its lower SOC reservation capability. In the riverside wetland, the high soil microbial biomass, higher proportion of beta-Proteobacteria, which have strong carbon metabolism activity and the existence of some specific aerobic heterotrophic bacteria such as Bacilli and uncultured Lactococcus, were the important reasons for the higher SMR compared to the coastal wetland. There were additional differences in soil physical and chemical characteristics between the coastal wetland and riverside wetlands. Path analysis of predominant bacteria and microbial biomass showed that soil salinity influenced beta-Proteobacteria and microbial biomass most negatively among these physical and chemical factors. Therefore the low salinity of the riverside area was suitable for the growth of microorganisms, especially beta-Proteobacteria and some specific bacteria, which led to the high SMR and low SOC reservation capability when compared to the coastal area.

Organic carbon accumulation capability of two typical tidal wetland soils in Chongming Dongtan, China.

We measured organic carbon input and content of soil in two wetland areas of Chongming Dongtan (Yangtze River Estuary) to evaluate variability in organic carbon accumulation capability in different wetland soils. Observed differences were investigated based on the microbial activity and environmental factors of the soil at the two sites. Results showed that the organic carbon content of wetland soil vegetated with Phragmites australis (site A) was markedly lower than that with P. australis and Spartina alterniflora (site B). Sites differences were due to higher microbial activity at site A, which led to higher soil respiration intensity and greater carbon outputs. This indicated that the capability of organic carbon accumulation of the site B soils was greater than at site A. In addition, petroleum pollution and soil salinity were different in the two wetland soils. After bio-remediation, the soil petroleum pollution at site B was reduced to a similar level of site A. However, the culturable microbial biomass and enzyme activity in the remediated soils were also lower than at site A. These results indicated that greater petroleum pollution at site B did not markedly inhibit soil microbial activity. Therefore, differences in vegetation type and soil salinity were the primary factors responsible for the variation in microbial activity, organic carbon output and organic carbon accumulation capability between site A and site B.

The variability in CO2 fluxes at different time scales in natural and reclaimed wetlands in the Yangtze River estuary and their key influencing factors.

With the increasing pace of global warming, studies of the carbon cycle and carbon sink capacity of estuarine wetlands have received increasing attention. Estuarine wetlands are often located in economically developed and densely populated areas, and their reclamation has become an important way to acquire land resources. To explore the effect of reclamation on the carbon sink function of estuarine wetlands, the Chongming Dongtan reclaimed wetland and Jiuduansha natural wetland, which are located in the Yangtze River estuary, were selected to investigate their variabilities in carbon fluxes and the main influencing factors using the open path eddy covariance flux monitoring system. The CO2 uptake capacity of the Dongtan reclaimed wetland was significantly weaker than that of the Jiuduansha natural wetland (P < 0.05). The difference in carbon fluxes between the two wetlands was mainly influenced by plant growth and carbon fixation, which accounted for 70.7% of the variation, with soil respiration being the second most important factor. The soil water content and nitrogen and phosphorus concentrations in the reclaimed wetland significantly decreased due to the barrier to water flow presented by a dam (P < 0.01). Nitrogen limitation was the main reason for the poor plant growth in the reclaimed wetland. Although nitrogen inputs in the natural wetland promoted soil respiration (40.6%), they were overshadowed by the inhibitory effect of soil moisture and salinity (55.8%). It was therefore possible to improve the carbon sink function of reclaimed wetlands if plant growth and carbon fixation efficiency could be enhanced without promoting soil respiration.

Matching different inorganic compounds as mixture of electron donors to improve CO2 fixation by nonphotosynthetic microbial community without hydrogen.

The dominant bacteria in nonphotosynthetic microbial community (NPMC) isolated from the ocean were identified by PCR-DGGE. The results revealed that the dominant microorganisms in cultures of NPMC differed when Na(2)S, Na(2)S(2)O(3), and NaNO(2) were used as the electron donor to reduce CO(2). These findings implied that different microorganisms in the NPMC respond to different inorganic compound as suitable electron donor, indicating that matching of Na(2)S, Na(2)S(2)O(3), and NaNO(2) may provide mixed electron donors that increase the ability of NPMC to fix CO(2). Accordingly, the central composite response surface method (RSM) was used to predict the optimal concentration and match of Na(2)S, Na(2)S(2)O(3), and NaNO(2) as mixed electron donors to improve CO(2) fixation efficiency under aerobic and anaerobic conditions without hydrogen. The results indicated that 0.46% NaNO(2), 0.50% Na(2)S(2)O(3), and 1.25% Na(2)S were the optimal match under aerobic conditions, while 1.04% NaNO(2), 1.07% Na(2)S(2)O(3), and 0.98% Na(2)S were the optimal match under anaerobic conditions. Under these conditions, the fixed CO(2) by NPMC was determined to be 387.51 and 512.57 mg/L, respectively, which obviously exceeded those values obtained prior to optimization (5.94 and 7.14 mg/L, respectively), as well as that obtained when hydrogen was used as the electron donor (91.60 mg/L).

Evaluation of the effectiveness and safety of the thermo-treatment process to dispose of recombinant DNA waste from biological research laboratories.

The discharge of recombinant DNA waste from biological laboratories into the eco-system may be one of the pathways resulting in horizontal gene transfer or "gene pollution". Heating at 100 degrees C for 5-10 min is a common method for treating recombinant DNA waste in biological research laboratories in China. In this study, we evaluated the effectiveness and the safety of the thermo-treatment method in the disposal of recombinant DNA waste. Quantitative PCR, plasmid transformation and electrophoresis technology were used to evaluate the decay/denaturation efficiency during the thermo-treatment process of recombinant plasmid, pET-28b. Results showed that prolonging thermo-treatment time could improve decay efficiency of the plasmid, and its decay half-life was 2.7-4.0 min during the thermo-treatment at 100 degrees C. However, after 30 min of thermo-treatment some transforming activity remained. Higher ionic strength could protect recombinant plasmid from decay during the treatment process. These results indicate that thermo-treatment at 100 degrees C cannot decay and inactivate pET-28b completely. In addition, preliminary results showed that thermo-treated recombinant plasmids were not degraded completely in a short period when they were discharged into an aquatic environment. This implies that when thermo-treated recombinant DNAs are discharged into the eco-system, they may have enough time to re-nature and transform, thus resulting in gene diffusion.

Inhibitory effect of self-generated extracellular dissolved organic carbon on carbon dioxide fixation in sulfur-oxidizing bacteria during a chemoautotrophic cultivation process and its elimination.

The features of extracellular dissolved organic carbon (EDOC) generation in two typical aerobic sulfur-oxidizing bacteria (Thiobacillus thioparus DSM 505 and Halothiobacillus neapolitanus DSM 15147) and its impact on CO2 fixation during chemoautotrophic cultivation process were investigated. The results showed that EDOC accumulated in both strains during CO2 fixation process. Large molecular weight (MW) EDOC derived from cell lysis and decay was dominant during the entire process in DSM 505, whereas small MW EDOC accounted for a large proportion during initial and middle stages of DSM 15147 as its cytoskeleton synthesis rate did not keep up with CO2 assimilation rate. The self-generated EDOC feedback repressed cbb gene transcription and thus decreased total bacterial cell number and CO2 fixation yield in both strains, but DSM 505 was more sensitive to this inhibition effect. Moreover, the membrane bioreactor effectively decreased the EDOC/TOC ratio and improved carbon fixation yield of DSM 505.

Determination of cobalt at ng ml(-1) level in water using the electrophilic substiution complexation between co(II) and chlorophosphonazo-p-CL-Cu(II) complex.

The chromophore chlorophosphonazo-p-Cl (PCCPA) was used to complex Co(II) and Cu(II) at pH 9.18. The formation of Co(PCCPA)2 and Cu(PCCPA)2 complexes were characterized by the spectral correction technique. Co(II) could competitively substitute Cu(II) from the Cu(II)-PCCPA complex via electrophilic effect. With the assistance of the light-absorption ratio variation approach, the electrophilic substitution complexation showed a high selectivity and good sensitivity with 1.9 ng mL(-1) of LOD. The proposed method has been applied to the direct detection of Co(II) in surface water and wastewater with good percent of recovery.

The variability and causes of organic carbon retention ability of different agricultural straw types returned to soil.

Retaining the organic carbon (C) content of agricultural straw when returned to soil is restricted by rapid decomposition. In order to clarify the difference in returned straw decomposition and the causes, and to develop a straw returning mode with high-efficiency of organic C accumulation, the decomposition processes of corn, soybean, rice and wheat straws were systematically studied in fields. When returned in situ (the original planting area), the C in soybean straw was decomposed most quickly with a decomposition constant of 0.00542 d-1, but wheat straw showed a longer retention in soil with 0.00303 d-1. However, for ex situ return of all straw in one area away from in situ return, soybean straw was decomposed most slowly (0.00452 d-1) and wheat straw more quickly (0.00652 d-1). The sequence of C decomposition rate in 270 d was soybean > corn > rice > wheat (in situ) and corn > wheat > rice > soybean (ex situ). Both surrounding soil and straw nature were important factors influencing the decomposition rate. The farmland with rice and wheat rotation retained more C from returned straws due to its high moisture and low nitrogen (N) content, while the soybean field was a contrast. Soybean straw had a low decomposition rate after ex situ return due to its low N content and high C/N ratio. The farmland of wheat-rice rotation combined with soybean straw ex situ return may develop into a field of higher C retention ability.

Response of cbb gene transcription levels of four typical sulfur-oxidizing bacteria to the CO2 concentration and its effect on their carbon fixation efficiency during sulfur oxidation.

The variability in carbon fixation capability of four sulfur-oxidizing bacteria (Thiobacillus thioparus DSM 505, Halothiobacillus neapolitanus DSM 15147, Starkeya novella DSM 506, and Thiomonas intermedia DSM 18155) during sulfur oxidation was studied at low and high concentrations of CO2. The mechanism underlying the variability in carbon fixation was clarified by analyzing the transcription of the cbb gene, which encodes the key enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase. DSM 15147 and DSM 505 fixed carbon more efficiently during sulfur oxidation than DSM 506 and DSM 18155 at 0.5% and 10% CO2, which was mainly because their cbb gene transcription levels were much higher than those of DSM 506 and DSM 18155. A high CO2 concentration significantly stimulated the carbon fixation efficiency of DSM 505 by greatly increasing the cbb gene transcription efficiency. Moreover, the influence of the CO2 concentration on the carbon fixation efficiency of the four strains differed greatly during sulfur oxidation.

Salinity and nutrient contents of tidal water affects soil respiration and carbon sequestration of high and low tidal flats of Jiuduansha wetlands in different ways.

Soils were collected from low tidal flats and high tidal flats of Shang shoal located upstream and Xia shoal located downstream with different tidal water qualities, in the Jiuduansha wetland of the Yangtze River estuary. Soil respiration (SR) in situ and soil abiotic and microbial characteristics were studied to clarify the respective differences in the effects of tidal water salinity and nutrient levels on SR and soil carbon sequestration in low and high tidal flats. In low tidal flats, higher total nitrogen (TN) and lower salinity in the tidal water of Shang shoal resulted in higher TN and lower salinity in its soils compared with Xia shoal. These would benefit ß-Proteobacteria and Anaerolineae in Shang shoal soil, which might have higher heterotrophic microbial activities and thus soil microbial respiration and SR. In low tidal flats, where soil moisture was high and the major carbon input was active organic carbon from tidal water, increasing TN was a more important factor than salinity and obviously enhanced soil microbial heterotrophic activities, soil microbial respiration and SR. While, in high tidal flats, higher salinity in Xia shoal due to higher salinity in tidal water compared with Shang shoal benefited γ-Proteobacteria which might enhance autotrophic microbial activity, and was detrimental to ß-Proteobacteria in Xia shoal soil. These might have led to lower soil microbial respiration and thus SR in Xia shoal compared with Shang shoal. In high tidal flats, where soil moisture was relatively lower and the major carbon input was plant biomass that was difficult to degrade, soil salinity was the major factor restraining microbial activities, soil microbial respiration and SR.

[Response of Soil Respiration and Organic Carbon to Returning of Different Agricultural Straws and Its Mechanism].

Soybean, maize and rice straws were selected as raw materials to study the response of the soil respiration (SR) and soil organic carbon (SOC) to returning of different straws in the Chongming Dongtan area. The results showed that all of SR, SOC and the plant biomass of the lands with returning of different straws were higher than those of the controls. The soil with soybean straw returning possessed the lowest SR and highest SOC among the three kinds of straws, meaning its higher soil organic carbon sequestration capability than corn and maize straws returning. Straw returning significantly enhanced soil dehydrogenase, ß-glycosidase activities and microbial biomass, and soil dehydrogenase activity was significantly correlated with soil respiration. The dehydrogenase activity of the soil with soybean straw returning was the lowest, thus, the lowest SR and highest SOC. Soybean straw had the highest cellulose and lignin contents and the lowest N content among the three kinds of straws, resulting in its lowest biodegradability. Therefore, when soybean straw was returned to soil, it was difficult to degrade completely by soil microorganisms, thus the lowest soil microbial activity, eventually leading to the lowest SR and highest SOC.

[Response of Straw and Straw Biochar Returning to Soil Carbon Budget and Its Mechanism].

Direct straw returning and straw carbonization returning are the main measures of straw returning. Because of the differences in structure and nature as well as returning process between straw and straw biochar, the soil respiration and soil carbon budget after returning must have significant differences. In this study, outdoor pot experiment was carried out to study the response of soil respiration and carbon budget to straw and straw biochar returning and its possible mechanism. The results showed that soil respiration of straw biochar returning [mean value 21. 69 µmol.(m2.s)-1] was significantly lower than that of direct straw returning [mean value 65.32 µmol.(m2.s)-1], and its soil organic carbon content ( mean value 20. 40 g . kg-1) and plant biomass (mean value 138. 56 g) were higher than those of direct straw returning (mean values 17. 76 g . kg-1 and 76. 76 g). Considering the carbon loss after the biochar preparation process, its soil carbon budget was also significantly higher than that of direct straw returning, so it was a low carbon mode of straw returning. Direct straw returning significantly promoted soil dehydrogenase activity, soil ß-glycosidase activity and soil microorganism quantity, leading to higher soil respiration, but straw biochar did play an obvious role in promoting the microbial activity index. Easily oxidizable carbon (EOC) and biodegradability of straw biochar were lower than those of straw, which showed that straw biochar had higher stability, and was more difficult to degrade for soil microorganisms so its soil microbial activity was generally lower, and could be retained in the soil for a long time.

Interactions Between Autotrophic and Heterotrophic Strains Improve CO2 Fixing Efficiency of Non-photosynthetic Microbial Communities.

Five autotrophic strains isolated from non-photosynthetic microbial communities (NPMCs), which were screened from oceans with high CO2 fixing capability, were identified as Ochrobactrum sp. WH-2, Stenotrophomonas sp. WH-11, Ochrobactrum sp. WH-13, Castellaniella sp. WH-14, and Sinomicrobium oceani WH-15. The CO2 fixation pathways of all these strains were Calvin-Benson-Bassham pathway. These strains could metabolize multifarious organic compounds, which allowed switching them to autotrophic culture after enrichment in heterotrophic culture. The central composite response surface method indicated that these strains possessed many interactive effects, which increased the CO2 fixing efficiency of a combined community composed of these strains by 56 %, when compared with that of the single strain. Furthermore, another combined community composed of these autotrophic strains and NPMC had richer interactive relationships, with CO2 fixing efficiency being 894 % higher than that of the single strain and 148 % higher than the theoretical sum of the CO2 fixing efficiency of each of its microbial components. The interaction between strictly heterotrophic bacteria in NPMC and isolated autotrophic strains played a crucial role in improving the CO2 fixing efficiency, which not only eliminated self-restraint of organic compounds generated during the growth of autotrophic bacteria but also promoted its autotrophic pathway.

Feasibility of a two-step culture method to improve the CO2-fixing efficiency of nonphotosynthetic microbial community and simultaneously decrease the spontaneous oxidative precipitates from mixed electron donors.

When compared with H2, mixed electron donors (MED), comprising S(2-), S2O3 (2-), and NO2 (-), could generally improve the CO2-fixing efficiency of nonphotosynthetic microbial communities (NPMCs). However, a large amount of abiotic precipitates combined with bacteria produced during culture may be unfavorable for the recycling and reuse of bacteria. The main component of the abiotic precipitates is S(0), which influences the enrichment and reuse of bacteria but is not conducive for CO2 fixation in the subsequent step. In this study, a two-step culture method (TSCM), employing H2 and MED, respectively, was verified to be feasible for improving the CO2-fixing efficiency of NPMCs in the second step. In the TSCM, the net-fixed CO2 increased to 854 mg/L and abiotic precipitates were not produced in the medium. Sequence analysis of 16 s rDNA from NPMC indicated the presence of microbial symbioses in the NPMC, supporting the possible applications of TSCM.

Enhanced CO2 fixation by a non-photosynthetic microbial community under anaerobic conditions: optimization of electron donors.

To enhance the CO(2) fixation efficiency of the non-photosynthetic microbial community (NPMC) isolated from sea water under anaerobic conditions without hydrogen, the concentration of inorganic compounds as electron donors and their ratios were optimized by response surface methodology design (RSMD). The results indicated that the CO(2) fixation efficiency of NPMC using NaNO(2), Na(2)S(2)O(3) and Na(2)S as the electron donors was increased about 90%, 75% and 207%, respectively. Additionally, there were interactions between two electron donors and three electron donors. Central composite RSMD experimentation predicted that the optimal concentration and ratios of these inorganic compounds was 1.04% NaNO(2), 1.07% Na(2)S(2)O(3) and 0.98% Na(2)S. Under these conditions, the fixed CO(2) was 139.89 mg/L, which obviously exceeded the amount prior to optimization, as well as when H(2) was used as an electron donor. The established electron donor system can effectively enhance the CO(2) fixation efficiency of NPMC without hydrogen under anaerobic conditions.

[Establishment of microarray for detecting mutation in HBV pre-core/core and basic core promoter regions].

OBJECTIVE: To develop a sensitive and specific microarray for detecting mutations of HBV pre-core/core and basic core promoter regions in the clinic. METHODS: Site-specific oligonucleotide probes were designed and immobilized to microarray slides and hybridized to HBV gene fragments amplified with specific biotin-labeled primer using asymmetrical PCR. The specificity and sensitivity of the method were estimated. And the microarray was applied to detect 138 clinical serum samples with HBV-DNA. RESULTS: The mutations of HBV pre-core/core and basic core promoter regions can be specifically detected using the microarray, and the sensitivity was 1 x 10(1) copies/microl. Among 138 samples, 40 samples had T1762/ A1764 mutation, 11 samples had C1814 mutation, and 16 samples had A1896 mutation. The A1896 mutation rate in high HBV-DNA load group was significantly higher than that in low HBV-DNA load group (P < 0.01). CONCLUSION: An DNA microarray assay was successfully established to detect the mutations in HBV pre-core/core and basic core promoter regions. The A1896 mutation in pre-core/core region maybe involve in duplication of HBV.

Optimization of electron donors to improve CO2 fixation efficiency by a non-photosynthetic microbial community under aerobic condition using statistical experimental design.

To improve the CO(2) fixation efficiency of non-photosynthetic microbial community (NPMC) isolated from sea water under aerobic conditions without hydrogen, the concentration of inorganic compounds as electron donors and their ratios were optimized using response surface methodology design (RSMD). These results indicated that Na(2)S, followed by Na(2)S(2)O(3) and NaNO(2) enhanced the CO(2) fixation by NPMC and the efficiency was increased about 100%, 200% and 200%, respectively. Some interaction between NaNO(2) and Na(2)S(2)O(3), as well as between Na(2)S(2)O(3) and Na(2)S was observed. Central composite RSMD experimentation predicted that the optimal concentration of these inorganic compounds and their ratios was 0.457% NaNO(2), 0.50% Na(2)S(2)O(3) and 1.25% Na(2)S. Under these conditions, the fixed CO(2) was 105.76 mg/L, which obviously exceeded the amount before optimization, as well as that obtained using hydrogen as the electron donor. This indicates that the NPMC using the established electron donors system can effectively fix CO(2) without light and hydrogen gas under aerobic condition.

Waste recombinant DNA: effectiveness of thermo-treatment to manage potential gene pollution.

Heating at 100 degrees C for 5-10 min is a common method for treating wastewater containing recombinant DNA in many bio-laboratories in China. In this experiment, plasmid pET-28b was used to investigate decay efficiency of waste recombinant DNA during thermo-treatment. The results showed that the decay half-life of the plasmid was 2.7-4.0 min during the thermo-treatment, and even heating for 30 min the plasmids still retained some transforming activity. Low pH promoted the decay of recombinant DNA, but NaCl, bovine serum albumin and EDTA, which existed in the most wastewater from bio-laboratories, protected DNA from degradation. Thus, the decay half-life of plasmid DNA may be longer than 2.7-4.0 min practically. These results suggest that the effectiveness of heating at 100 degrees C for treating waste recombinant DNA is low and a gene pollution risk remains when those thermo-treated recombinant DNAs are discharged into the environment. Therefore other simple and effective methods should be developed.