A Monte Carlo approach to estimate the uncertainty in soil CO2 emissions caused by spatial and sample size variability

The soil CO₂ emission is recognized as one of the largest fluxes in the global carbon cycle. Small errors in its estimation can result in large uncertainties and have important consequences for climate model predictions. Monte Carlo approach is efficient for estimating and reducing spatial scale sampling errors. However, that has not been used in soil CO₂ emission studies. Here, soil respiration data from 51 PVC collars were measured within farmland cultivated by maize covering 25 km² during the growing season. Based on Monte Carlo approach, optimal sample sizes of soil temperature, soil moisture, and soil CO₂ emission were determined. And models of soil respiration can be effectively assessed: Soil temperature model is the most effective model to increasing accuracy among three models. The study demonstrated that Monte Carlo approach may improve soil respiration accuracy with limited sample size. That will be valuable for reducing uncertainties of global carbon cycle.     

Differential effects of lichens,mosses and grasses on respiration and nitrogen mineralization in soils of the New Jersey Pinelands

In the New Jersey Pinelands, severely disturbed areas often do not undergo a rapid succession to forest; rather, a patchy cover of lichens, mosses and grasses persists for decades. We hypothesized that these plant covers affect soil microbial processes in different ways, and that these effects may alter the successional dynamics of the patches. We predicted that the moss and grass covers stimulate soil microbial activity, whereas lichens inhibit it, which may in turn inhibit succession. We collected soil cores from beneath each type of cover plus bare soil within two types of highly disturbed areas—sites subjected to hot wildfires, and areas mined for sand. Organic matter (OM) content, soil respiration and potential N mineralization were measured in the cores. Soils under mosses were similar to those under grasses; they accumulated more OM and produced more mineral N, predominantly in the form of ammonium, than either the bare soils or the soils beneath lichens. Mineralization under lichens, like that of the bare soils but unlike the soils beneath mosses or grasses, was dominated by net nitrification. These patterns were reproduced in experimentally transplanted moss and lichen mats. Mosses appear to create high-nutrient microsites via high rates of OM accumulation and production of ammonium, whereas lichens maintain low-nutrient patches similar to bare soil via low OM accumulation rates and production of mineral N predominantly in the mobile nitrate form. These differences in soil properties may explain the lack of vascular plant invasion in lichen mats, in contrast to the moss-dominated areas.     

Nanoflow liquid chromatography coupled to matrix-assisted laser desorption/ionization mass spectrometry: sample preparation, data analysis, and application to the analysis of complex peptide mixtures

We report the development of a robust interface for off-line coupling of nano liquid chromatography (LC) to matrix-assisted laser desorption/ionisation-mass spectrometry (MALDI-MS) and its application to the analysis of proteolytic digests of proteins, both isolated and in mixtures. The interface makes use of prestructured MALDI sample supports to concentrate the effluent to a small sample plate area and localize the MALDI sample to a predefined array, thereby enriching the analyte molecules and facilitating automated MALDI-MS analysis. Parameters that influence the preparation of MALDI samples from the LC effluent were evaluated with regard to detection sensitivity, spectra quality, and reproducibility of the method. A procedure for data processing is described. The presented nano LC MALDI-MS system allowed the detection of several peptides from a tryptic digest of bovine serum albumin, at analyzed amounts corresponding to one femtomole of the digested protein. For the identification of native proteins isolated from mouse brain by two-dimensional gel electrophoresis, nano LC MALDI-MS increased the number of detected peptides, thereby allowing identification of proteins that could not be identified by direct MALDI-MS analysis. The ability to identify proteins in complex mixtures was evaluated for the analysis of Escherichia coli 50S ribosomal subunit. Out of the 33 expected proteins, 30 were identified by MALDI tandem time of flight fragment ion fingerprinting.     

Assessment of potential ecological disruption based on heavy metal toxicity, accumulation and distribution in media of the Lagos Lagoon

Toxicity evaluations of heavy metals against three benthic animals, Tympanotonus fuscatus, Clibanarius africanus and Sesarma huzardi of the Lagos Lagoon were carried out under laboratory conditions. On the basis of the 96hLC₅₀ values, Cd was found to be the most toxic metal tested followed by Cu, Zn and Pb, in a descending order of toxicity against T. fuscatus and S. huzardi; however, against C. africanus, Cu was the most toxic followed by Cd, Zn and Pb (least toxic). The determination of the metal concentrations in the water column and sediment of the Lagos Lagoon revealed that these media of the lagoon contained measurable concentrations of heavy metals but the levels were still several folds lower than the concentrations that will cause 50% mortality of exposed animals under laboratory conditions. The significance of the observed differences between the 96hLC₅₀ values of the test metals, the concentration of heavy metals detected in tissues of field animals and ambient levels of the metals in the Lagos lagoon were discussed in relation to the protection of aquatic lives and potential public health risks. The need to verify the possibilities of synergistic interactions between the constituent metals when acting jointly against the exposed animals was recommended.     

Extrapolation in Risk Assessment: Improving the Quantification of Uncertainty,and Improving Information to Reduce the Uncertainty

Risk assessments inevitably extrapolate from the known to the unknown. The resulting calculation of risk involves two fundamental kinds of uncertainty: uncertainty owing to intrinsically unpredictable (random) components of the future events, and uncertainty owing to imperfect prediction formulas (parameter uncertainty and error in model structure) that are used to predict the component that we think is predictable. Both types of uncertainty weigh heavily both in health and ecological risk assessments. Our first responsibility in conducting risk assessments is to ensure that the reported risks correctly reflect our actual level of uncertainty (of both types). The statistical methods that lend themselves to correct quantification of the uncertainty are also effective for combining different sources of information. One way to reduce uncertainty is to use all the available data. To further sharpen future risk assessments, it is useful to partition the uncertainty between the random component and the component due to parameter uncertainty, so that we can quantify the expected reduction in uncertainty that can be achieved by investing in a given amount of future data. An example is developed to illustrate the potential for use of comparative data, from toxicity testing on other species or other chemicals, to improve the estimates of low-effect concentration in a particular case with sparse case-specific data.     

The influence of sex and strain on trace element dysregulation in the brain due to diet-induced obesity

BackgroundThe objective of this study was to identify interaction effects between diet, sex, and strain on trace element dysregulation and gene expression alterations due to diet-induced obesity (DIO) in the hippocampus, striatum, and midbrain.MethodsMale and female C57BL/6 J (B6 J) and DBA/2 J (D2 J) mice were fed either a low fat (10 % kcal) diet (LFD) or high fat (60 % kcal) diet (HFD) for 16 weeks, then assessed for trace element concentrations and gene expression patterns in the brain.ResultsIn the hippocampus, zinc was significantly increased by 48 % in D2 J males but decreased by 44 % in D2 J females, and divalent metal transporter 1 was substantially upregulated in B6 J males due to DIO. In the striatum, iron was significantly elevated in B6 J female mice, and ceruloplasmin was significantly upregulated in D2 J female mice due to DIO. In the midbrain, D2 J males fed a HFD had a 48 % reduction in Cu compared to the LFD group, and D2 J females had a 37 % reduction in Cu compared to the control group.ConclusionsThe alteration of trace element homeostasis and gene expression due to DIO was brain-region dependent and was highly influenced by sex and strain. A significant three-way interaction between diet, sex, and strain was discovered for zinc in the hippocampus (for mice fed a HFD, zinc increased in male D2 Js, decreased in female D2 Js, and had no effect in B6 J mice). A significant diet by sex interaction was observed for iron in the striatum (iron increased only in female mice fed a HFD). A main effect of decreased copper in the midbrain was found for the D2 J strain fed a HFD. These results emphasize the importance of considering sex and genetics as biological factors when investigating potential associations between DIO and neurodegenerative disease.     

Insights into the functional role of fungi in deep-sea hydrothermal vents through the analysis of stable isotopes of carbon,nitrogen, and sulfur

The functional diversity of fungi remains poorly explored in the deep-sea, particularly in hydrothermal vents. Here, we approached this gap through the analysis of stable isotopes of carbon (δ¹³C), nitrogen (δ¹⁵N), and sulfur (δ³⁴S) of fourteen isolates obtained from three deep-sea vent systems of the southern Gulf of California. The δ¹³C results indicated that 60% of the isolates relied on mixed carbon sources fixed by the Calvin-Benson-Bassham and the reductive Tricarboxylic Acid (rTCA) cycles, whereas 40% relied exclusively on rTCA carbon. The δ¹⁵N and δ³⁴S values suggested a dependence on local and external nitrogen sources and the assimilation of chemosynthetic and photosynthetic inputs. Fungal δ¹³C and δ¹⁵N overlapped with those of primary and secondary vent macroconsumers, implying the assimilation of bacterial and invertebrate necromass and their ecological role as parasites. These findings provide insights into the unexplored trophic versatility of fungi in chemosynthetic ecosystems, highlighting their importance in deep-sea trophic dynamics.     

氮磷肥对茶树锌硒等中微量元素吸收与分配的影响

锌（Zn）和硒（Se）及其他中微量元素（铝Al,钙Ca,铁Fe,铜Cu,锰Mn）是茶叶品质的重要指标,但茶树吸收Zn、Se能力及氮（N）磷（P）肥影响中微量元素吸收与分配的过程尚不清楚。以红壤丘陵区福鼎大白茶树为研究对象,开展Zn+Se、Zn+Se+N、Zn+Se+P、Zn+Se+N+P和对照共5种处理3次重复随机化区组试验,处理第3年春季分茶叶、成熟叶、吸收根、运输根和储藏根采集植物样品,测定其元素含量。结果表明,茶树地上和地下器官Zn和Se及其他中微量元素对N、P、Zn、Se添加的响应具分异性。与对照相比,茶树地上和地下器官Zn和Se含量均显著增加,与Zn+Se相比,施N和/或P肥仅显著提高茶叶和成熟叶Se含量（P<0.05）;与对照相比,施肥处理均显著提高吸收根和运输根Al、Fe含量以及储藏根Cu含量;运输根Mn含量表现为Zn+Se+N、Zn+Se+P、Zn+Se+N+P显著高于对照,储藏根Mn含量为Zn+Se+N+P显著高于其他处理;茶树各器官Ca含量对施肥处理无显著响应。此外,茶叶和成熟叶的Zn含量与吸收根显著正相关,而Se含量则与储藏根显著正相关。茶树具有吸收和积累Zn和Se的能力,而施N、P肥有助于提高茶叶Se含量,研究结果为红壤丘陵区培育高品质锌硒茶及营建生态高值茶园提供了依据。     

Quantitative methodology for corticosteroids based on chemical oxidation to electrophilic products for electron capture-negative chemical ionization using capillary gas chromatography-mass spectrometry. I. Assessment of feasibility in the analysis of horse urine for dexamethasone

Sensitive and specific methodology based on capillary column gas chromatography-electron capture-negative chemical ionization-mass spectrometry has been developed for the quantitative analysis of corticosteroids from biological fluids. The feasibility of this method is demonstrated in the quantitative analysis of dexamethasone in horse urine following administration of the drug. A structurally similar compound, 6 alpha-methylprednisolone, is added to the urine as an internal standard. The free dexamethasone and the internal standard are extracted and oxidized to high-electron-affinity 1,4-androstadiene-3,11,17-trione structural analogs and then analyzed by capillary column gas chromatography-electron capture-negative chemical ionization-mass spectrometry.     

基于碳、氮稳定同位素研究胶州湾普氏栉虾虎鱼的摄食习性

普氏栉虾虎鱼属于小型暖温性底层鱼类,是胶州湾鱼类群落中的优势种之一,在胶州湾食物网和生态系统中发挥着重要作用.本文应用碳、氮稳定同位素技术,基于胶州湾渔业资源底拖网调查采集的样品,对普氏栉虾虎鱼的摄食习性进行了研究.结果表明: 胶州湾普氏栉虾虎鱼的δ¹⁵N值范围为11.24‰～13.99‰,平均值为(12.70±0.70)‰,δ¹³C值范围为-20.67‰～-18.46‰,平均值为(-19.08±0.36)‰;各体长组的营养级范围为3.49～3.76,平均营养级为(3.62±0.21),其δ¹⁵N值和营养级与体长呈显著负相关,δ¹³C值与体长无显著相关性.普氏栉虾虎鱼摄食的主要饵料生物类群为多毛类、虾类和软体动物,浮游动物和颗粒有机物(POM)的饵料贡献率较小.聚类分析结果表明,普氏栉虾虎鱼各体长组食物组成的相似性均在92%以上,相似性较高,说明其摄食习性随体长变化无明显差异.普氏栉虾虎鱼在胶州湾生态系统中属于中级消费者,其摄食各饵料生物类群比例的变化可能是其营养级与体长呈负相关关系的主要原因.     

Effects of nitrogen supply on the anatomy and chemical composition of leaves of four grass species belonging to the genus Poa, as determined by image-processing analysis and pyrolysis–mass spectrometry

Previous experiments have shown that the anatomy and chemical composition of leaves of inherently fast- and slow-growing grass species, grown at non-limiting nitrogen supply, differ systematically. The present experiment was carried out to investigate whether these differences persist when the plants are grown at an intermediate or a very low nitrogen supply. To this end, the inherently fast-growing Poa annua L. and Poa trivialis L., and the inherently slow-growing Poa compressa L. and Poa pratensis (L.) Schreb. were grown hydroponically at three levels of nitrate supply: at optimum (RGR_max) and at relative addition rates of 100 and 50 mmol N (mol N)^?1 d^?1 (RAR₁₀₀ and RAR₅₀), respectively. As expected, at the lowest N supply, the potentially fast-growing species grew at the same rate as the inherently slow-growing ones. Similarly, the differences in leaf area ratio (LAR, leaf area:total dry mass), specific leaf area (SLA, leaf arear:leaf dry mass) and leaf mass ratio (LMR, leaf dry mass:total dry mass) disappeared. Under optimal conditions, the fast-growing species differed from the slow-growing ones in that they had a higher N concentration. There were no significant differences in C concentration. With decreasing N supply, the total N concentration decreased and the differences between the species disappeared. The total C concentration increased for the fast-growing species and decreased for the slow-growing ones, i.e. the small, but insignificant, difference in C concentration between the species at RGR_max increased with decreasing N supply. The chemical composition of the leaves at low N supply, analysed in more detail by pyrolysis–mass spectrometry, showed an increase in the relative amounts of guaiacyl lignin, cellulose and hemicellulose, whereas those of syringyl lignin and protein decreased. The anatomy and morphology of the leaves of the four grass species differing in RGR_max were analysed by image-processing analysis. The proportion of the total volume occupied by mesophyll plus intercellular spaces and epidermis did not correlate with the amount of leaf mass per unit leaf area (specific leaf mass, SLM) at different N supply. The higher SLM at low N supply was caused partly by a high proportion of non-veinal sclerenchymatic cells per cross-section and partly by the smaller volume of epidermal cells. We conclude that the decrease in relative growth rate (and increase in SLM) at decreasing N supply is partly due to chemical and anatomical changes. The differences between the fast- and slow-growing grass species at an optimum nutrient supply diminished when plants were growing at a limiting nitrogen supply.     

Esophagus cancer and radiation exposure due to nuclear test fallout: an analysis based on the data of the Semipalatinsk historical cohort, 1960-1999

This paper describes the Semipalatinsk historical cohort study and, in particular, examines the association between combined external and internal radiation exposure and esophagus cancer. Esophagus cancer is the most frequent single cancer site in the cause of death follow-up for the Semipalatinsk cohort. Set up in the 1960s, this historical cohort included 10 exposed settlements in the vicinity of the Semipalatinsk nuclear test site in East Kazakhstan as well as 6 comparison settlements in a low exposure area of the same region. The external and internal radiation doses to the population of the settlements under study were mainly due to local fallout from atmospheric nuclear testing (1949-1962). The database includes dosimetry and health information for 19.545 inhabitants of exposed and comparison villages in the Semipalatinsk region, comprising a total of 582.750 person-years of follow-up between 1960 and 1999. Cumulative effective dose estimates in this cohort range from 20 mSv to -4 Sv, with a mean dose of 634 mSv in the exposed group. Relative risks were calculated in terms of rate ratios, using a Poisson regression model for grouped person-time data. Esophagus cancer was found substantially elevated, with a statistically significant increase of the relative risk with dose and an ERR/Sv of 2.37 (1.45; 3.28) for the total cohort. If the data set was restricted to the exposed group only, the ERR/Sv was found considerably lower (0.18 (-0.16; 0.52)), whereas the dose-response remained significant only in women. Overall, our results based on the Semipalatinsk historical cohort indicate an association between fallout exposure and the risk of esophagus cancer that should be further investigated.     

Construction of Saccharomyces cerevisiae strains that accumulate relatively low concentrations of trehalose, and their application in testing the contribution of the disaccharide to stress tolerance

Genetically related diploid strains of Saccharomyces cerevisiae that accumulate varied amounts of trehalose during starvation for nitrogen have been constructed. Strains that produced greater than 5% trehalose (dry cell weight) were more tolerant of thermal, or freeze-thaw stresses than strains that produced less than 4% trehalose. Thus trehalose appears to play a role in stress tolerance of yeast. The significance of these results is that, for the first time, a series of related, unmutated strains have been used to test the effect of trehalose on thermotolerance. Previous studies employed either heat shock treatment, or mutated strains to provide trehalose variations, and as such the contribution of the disaccharide to stress tolerance could not necessarily be separated from other factors such as heat shock proteins.     

Interactive effects of nitrogen and phosphorus on the acclimation potential of foliage photosynthetic properties of cork oak,Quercus suber,to elevated atmospheric CO2 concentrations

Leaf gas-exchange and chemical composition were investigated in seedlings of Quercus suber L. grown for 21 months either at elevated (700 μmol mol^–1) or normal (350 μmol mol^–1) ambient atmospheric CO₂ concentrations, [CO₂], in a sandy nutrient-poor soil with either ‘high’ N (0.3 mol N m^–3 in the irrigation solution) or with ‘low’ N (0.05 mol N m^–3) and with a constant suboptimal concentration of the other macro- and micronutrients. Although elevated [CO₂] yielded the greatest total plant biomass in ‘high’ nitrogen treatment, it resulted in lower leaf nutrient concentrations in all cases, independent of the nutrient addition regime, and in greater nonstructural carbohydrate concentrations. By contrast, nitrogen treatment did not affect foliar N concentrations, but resulted in lower phosphorus concentrations, suggesting that under lower N, P use-efficiency in foliar biomass production was lower. Phosphorus deficiency was evident in all treatments, as photosynthesis became CO₂ insensitive at intercellular CO₂ concentrations larger than ≈ 300 μmol mol^–1, and net assimilation rates measured at an ambient [CO₂] of 350 μmol mol^–1 or at 700 μmol mol^–1 were not significantly different. Moreover, there was a positive correlation of foliar P with maximum Rubisco (Ribulose-1,5-bisphosphate carboxylase/oxygenase) carboxylase activity (V_cmax), which potentially limits photosynthesis at low [CO₂], and the capacities of photosynthetic electron transport (J_max) and phosphate utilization (P_max), which are potentially limiting at high [CO₂]. None of these potential limits was correlated with foliar nitrogen concentration, indicating that photosynthetic N use-efficiency was directly dependent on foliar P availability. Though the tendencies were towards lower capacities of potential limitations of photosynthesis in high [CO₂] grown specimens, the effects were statistically insignificant, because of (i) large within-treatment variability related to foliar P, and (ii) small decreases in P/N ratio with increasing [CO₂], resulting in balanced changes in other foliar compounds potentially limiting carbon acquisition. The results of the current study indicate that under P-deficiency, the down-regulation of excess biochemical capacities proceeds in a similar manner in leaves grown under normal and elevated [CO₂], and also that foliar P/N ratios for optimum photosynthesis are likely to increase with increasing growth CO₂ concentrations. Symbols: A, net assimilation rate (μmol m^–2 s^–1); A_max, light-saturated A (μmol m^–2 s^–1); α, initial quantum yield at saturating [CO₂] and for an incident Q (mol mol^–1); [CO₂], atmospheric CO₂ concentration (μmol mol^–1); C_i, intercellular CO₂ concentration (μmol mol^–1); C_a, CO₂ concentration in the gas-exchange cuvette (μmol mol^–1); F_B, fraction of leaf N in ‘photoenergetics’; F_L, fraction of leaf N in light harvesting; F_R, fraction of leaf N in Rubisco; Γ*, CO₂ compensation concentration in the absence of R_d (μmol mol^–1); J_max*, capacity for photosynthetic electron transport; J_mc, capacity for photosynthetic electron transport per unit cytochrome f (mol e^–[mol cyt f]^–1 s^–1); K_c, Michaelis-Menten constant for carboxylation (μmol mol^–1); K_o, Michaelis-Menten constant for oxygenation (mmol mol^–1); M_A, leaf dry mass per area (g m^–2); O, intercellular oxygen concentration (mmol mol^–1); [P_i], concentration of inorganic phosphate (mM); P_max*, capacity for phosphate utilization; Q, photosynthetically active quantum flux density (μmol m^–2 s^–1); R_d*, day respiration (CO₂ evolution from nonphotorespiratory processes continuing in the light); Rubisco, ribulose-1,5-bisphosphate carboxylase/oxygenase; RUBP, ribulose-1,5-bisphosphate; T_l, leaf temperature (°C); U_TPU*, rate of triose phosphate utilization; V_cmax*, maximum Rubisco carboxylase activity; V_cr, specific activity of Rubisco (μmol CO₂[g Rubisco]^–1 s^–1] *given in either μmol m^–2 s^–1 or in μmol g^–1 s^–1 as described in the text.     

钾对外生菌根真菌的分泌作用及氮、磷、钾含量的影响

用无钾、低钾、中钾和高钾的液体培养基培养外生菌根真菌14～21d,随着钾离子浓度的降低,外生菌根真菌分泌H^ 和草酸的速率提高。由于H^ 能取代2：1型粘粒矿物品层和晶格内的钾,草酸能络合矿物中的Fe,Al离子,促进Al-O八面体风化,推测菌根真菌和菌根活化土壤无效钾的能力在低钾时较强,在高钾时较弱。在外生菌根真菌分泌的H^ 中,草酸电离可能不是它们的主要来源,而是一些目前尚未知道的有机酸。此外,液体培养基中的钾离子不同程度地影响外生菌根真菌体内的氮、磷、钾含量,看来环境中的钾离子供应影响了真菌菌丝对养分的吸收。     

Alterations in plasma essential trace elements selenium, manganese, zinc, copper, and iron concentrations and the possible role of these elements on oxidative status in patients with childhood asthma

The aim of the present study is to evaluate the status of plasma essential trace element selenium (Se), manganese (Mn), copper (Cu), zinc (Zn), and iron (Fe) concentrations and the effect of these elements on oxidative status in patients with childhood asthma. Plasma Se, Mn, Cu, and Zn concentrations were determined by atomic absorption spectrophotometry (AAS) and Fe concentrations, malondialdehyde (MDA), and total antioxidant capacity (TAC) were determined by the colorimetric method. The plasma MDA/TAC ratio was calculated as an index of oxidative status. Plasma albumin levels were measured to determine nutritional status. Plasma Fe concentrations, MDA levels and the MDA/TAC ratio were significantly higher (p<0.001, p<0.001, and p<0.01, respectively) and Se and Mn concentrations and TAC were lower (p<0.01, p<0.05, and p<0.01, respectively) in patients when compared to the healthy subjects. Plasma Zn, Cu, and albumin levels were not found to be significantly different in patients and controls (p>0.05). There were positive relationships between plasma MDA and Fe (r=0.545, p<0.001) and TAC and Se (r=0.485, p<0.021), and a negative correlation between TAC and MDA values (r= −0.337, p<0.031) in patients with childhood asthma. However, there was no correlation between these trace elements and albumin content in patient groups. These observations suggest that increased Fe and decreased Se concentrations in patients with childhood asthma may be responsible for the oxidant/antioxidant imbalance.     

基于SolVES模型的关中-天水经济区生态系统文化服务评估

随着人类对于生态系统服务的需求持续增加,生态系统服务的评估逐渐成为研究热点。其中文化服务因其自身的无形性常在研究中被忽视且难以衡量。选取关中-天水经济区为研究区域,应用Sol VES模型并生成5种价值指数地图和价值总和地图来评估该区域生态系统的文化服务。结果表明:审美价值在秦岭山脉和城市公园等区域较高;娱乐价值在娱乐机会较多、交通便利的城市和秦岭北麓区域更高;文化与历史价值集中在历史文化底蕴深厚的城市地区;精神价值在与城市距离较近、有山川分布的森林公园更高。Sol VES模型在大范围区域的应用取得了较好的效果同时为政府进行生态建设和规划提供了科学依据。     

Effect of lpdA gene knockout on the metabolism in Escherichia coli based on enzyme activities, intracellular metabolite concentrations and metabolic flux analysis by 13C-labeling experiments

The lipoamide dehydrogenase (LPD) encoded by lpdA gene is a component of the pyruvate dehydrogenase complex (PDHc), alpha-ketoglutarate dehydrogenase (AKGDH) and the glycine cleavage multi-enzyme (GCV) systems. In the present study, cell growth characteristics, enzyme activities and intracellular metabolite concentrations were compared between the parent strain Escherichia coli BW25113 and its lpdA knockout mutant in batch and continuous cultures. The lpdA knockout mutant produced significantly more pyruvate and L-glutamate under aerobiosis. Some D-lactate and succinate also accumulated in the culture broth. Based on the investigation of enzyme activities and intracellular metabolite concentrations, acetyl-CoA was considered to be formed by the combined reactions through pyruvate oxidase (PoxB), acetyl-CoA synthetase (Acs) and acetate kinase (Ack)-phosphoacetyltransferase (Pta) in the lpdA mutant. The effect of the lpdA gene knockout on the intracellular metabolic flux distributions was investigated based on 1H-13C NMR spectra and GC-MS signals obtained from 13C-labeling experiment using the mixture of [U-13C] glucose, [1-13C] glucose, and naturally labeled glucose. Flux analysis of the lpdA mutant indicated that the Entner-Doudoroff (ED) pathway and the glyoxylate shunt were activated. The fluxes through glycolysis and oxidative pentose phosphate (PP) pathway (except for the flux through glucose-6-phosphate dehydrogenase) were slightly downregulated. The TCA cycle was also downregulated in the mutant strain. On the other hand, the fluxes through the anaplerotic reactions of PEP carboxylase, PEP carboxykinase and malic enzyme were upregulated, which were consistent with the results of enzyme activities. Furthermore, the influence of the poxB gene knockout on the growth of E. coli was also studied because of its similar function to PDHc which connects the glycolysis to the TCA cycle. Under aerobiosis, a comparison of lpdA mutant and poxB mutant indicated that PDHc is the main enzyme which catalyzes the reaction from pyruvate to acetyl-CoA in the parent strain, while PoxB plays a very important role in the PDHc-deficient strain.     

Phloem translocation of Fe,Cu, Mn,and Zn in Ricinus seedlings in relation to the concentrations of nicotianamine,an endogenous chelator of divalent metal ions,in different seedling parts

During the first 8 days of germination the Ricinus seedling is supplied with all nutrients by the endosperm via phloem transport. In 4- to 8-days-old seedlings the concentrations and contents of Fe, Cu, Mn and Zn, and nicotianamine (NA) in the endosperm, cotyledons, hypocotyl and roots were estimated. From the data obtained translocation rates and flow profiles for the metals were established. The main sink for Fe, Mn and Zn were the cotyledons whereas Cu was mainly imported into the hypocotyl. Maximum flow rates occurred between days 5 and 7, for Zn between days 6 and 8.The time kinetics of NA and divalent metal ion concentrations and contents are interpreted as co-transport. The role of NA as transport vehicle of micronutrients in the sieve tubes is discussed.