基于15N示踪技术的植物-土壤系统氮循环研究进展

同位素示踪技术是指外源添加与生物体内的元素或物质完全共同运行的示踪物,用来指示生物体内某元素或物质变化过程的一种方法。利用氮稳定同位素示踪技术,能从本质上揭示生态学过程发生的机理,从而成为生态学科研工作十分重要的工具之一。对近年来15N示踪技术应用于土壤氮素固定、植物氮素营养和植物-土壤系统氮迁移的研究进展进行了综述,并对稳定氮同位素技术在解决相关生态学难题可能的前景和不足方面进行了展望。     

Cx36 expression in the AII‐mediated rod pathway is activity dependent in the developing rabbit retina

Gap junctions are composed of connexin 36 (Cx36) and play a critical role in the rod photoreceptor signaling pathways of the vertebrate retina. Despite the fact that their connection and modulation in various rod pathways have been extensively studied in adult animals, little is known about the contribution and regulation of gap junctions to the development of the AII amacrine cell (AC)‐mediated rod pathway. Using immunohistochemistry and microinjection, this study demonstrates a steady increase in relative Cx36 protein expression in both plexiform layers of the rabbit retina at around the time of eye opening. However, immediately after eye opening, most Cx36 immunoreactive AII ACs show no gap junction coupling pattern to neighboring cells and it is not until the third postnatal week that AII cells begin to exhibit an adult‐like tracer‐coupling pattern. Moreover, studies using dark‐rearing and AMPA receptor blockade during postnatal development both revealed that relative levels of Cx36 immunoreactivity in AII ACs were increased when neural activity was inhibited . Our findings suggest that Cx36 expression in the AII‐mediated rod pathway is activity dependent in the developing rabbit retina . © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 473–486, 2016     

Superfluous glutamine synthetase activity in Chinese Hamster Ovary cells selected under glutamine limitation is growth limiting in glutamine-replete conditions and can be inhibited by serine

Passaging and expansion of animal cells in lean maintenance medium could result in periods of limitation of some nutrients. Over time, such stresses could possibly result in selection of cells with metabolic changes and contribute to heterogeneity. Here, we investigate whether selection of Chinese Hamster Ovary (CHO) cells under glutamine limitation results in changes in growth under glutamine-replete conditions. In glutamine-limiting medium, compared to control cells passaged in glutamine-rich medium, the selected cells showed higher glutamine synthetase (GS) activity and attained a higher peak viable cell density (PVCD). Surprisingly, in glutamine-replete conditions, selected cells still showed a higher GS activity but a lower PVCD. We show that in glutamine-replete medium, PVCD of selected cells was restored on (a) inhibition of GS activity with methionine sulfoximine, (b) supplementation of aspartate—without affecting GS activity, and (c) supplementation of serine, which is reported to inhibit GS in vitro. Consistent with the reported effect of serine, inhibition of GS activity was observed upon serine supplementation along with reduced growth of cells under glutamine-limiting conditions. The latter observation is important for the design of glutamine-free culture medium and feed used for GS-CHO and GS-NS0. In summary, we show that CHO cells selected under glutamine limitation have superfluous GS activity in glutamine-replete medium, which negatively affects their PVCD. This may be due to its effect on availability of aspartate which was the limiting nutrient for the growth of selected cells in glutamine-replete conditions.     

Bromoform in the effluents of a nuclear power plant: a potential tracer of coastal water masses

Bromoform (tribromomethane, CHBr₃), one of the trihalomethanes, is a chlorination byproduct in cooling water of power plants and industrial complexes. We used the distribution of bromoform in seawater to monitor the movement of cooling water from the Youngkwang Nuclear Power Plant (YNPP) located on the West Coast of Korea. Bromoform concentrations were highest, 124 g l^–1, in surface water near the outlet of YNPP and decreased linearly with distance from the outlet, mimicking the dissipation of cooling water discharged from the power plant. This byproduct of chlorination is thus a potential tracer of coastal water masses, due to its conservative behavior in the cooling water, low natural background in seawater, and easy analytical detection.     

Multi-compartmental modelling and experimental design for glucose transport studies in insulin-resistant rats

Many pathologies are associated with abnormalities of glucose metabolism or with perturbations of its transport (type 2 diabetes or insulin-resistance). The pre-diabetic state is characterised by a state of insulin-resistance, in others words a defect of glucose transport in insulin-sensible tissues, such as muscles and adipose tissues. The mathematical modelling of experimental data can be an excellent method to explore the mechanisms implied in the studied biological phenomenon. Thus, starting from a symbolic formulation like the compartmental modelling, it can be possible to develop a theoretical basis for the observation and to consider the best-adapted experiments for the study. We showed with mathematical models that [123I]-6-deoxy-6-iodo-D-glucose (6-DIG), shown as a tracer of glucose transport in vitro, could point out this transport abnormality. To quantify the insulin resistance, we estimated the fractional transfer coefficients of 6-DIG from the blood to the organs. We realised many studies to lead to a satisfying model; special attention has been paid to the precision of the parameter to select the best model. The results showed that by associating experimental data obtained with 6-DIG activities and an adapted mathematical model, discriminating parameters (in and out fractional transfer coefficients) between the two groups (control and insulin-resistant rats) could be pointed out.     

Analysis of metabolic and physiological responses to gnd knockout in Escherichia coli by using C-13 tracer experiment and enzyme activity measurement

The physiological and metabolic responses to gnd knockout in Escherichia coli K-12 was quantitatively investigated by using the (13)C tracer experiment (GC-MS/NMR) together with the enzyme activity analysis. It was shown that the general response to the gene knockout was the local flux rerouting via Entner-Doudoroff pathway and the direction reversing via non-oxidative pentose phosphate pathway (PPP). The mutant was found to direct higher flux to phosphoglucose isomerase reaction as compared to the wild-type, but the respiratory metabolism was comparable in both strains. The anaplerotic pathway catalyzed by malic enzyme was identified in the mutant, which was accompanied with an up-regulation of phosphoenolpyruvate carboxylase and down-regulation of phosphoenolpyruvate carboxykinase. The presented results provide first evidence that compensatory mechanism existed in PPP and anaplerotic pathway in response to the gnd deletion.     

Marine bacterioplankton can increase evaporation and gas transfer by metabolizing insoluble surfactants from the air–seawater interface

Hydrophobic surfactants at the air–sea interface can retard evaporative and gaseous exchange between the atmosphere and the ocean. While numerous studies have examined the metabolic role of bacterioneuston at the air–sea interface, the interactions between hydrophobic surfactants and bacterioplankton are not well constrained. A novel experimental design was developed, using Vibrio natriegens and ³H-labelled hexadecanoic acid tracer, to determine how the bacterial metabolism of fatty acids affects evaporative fluxes. In abiotic systems, >92% of the added hexadecanoic acid remained at the air–water interface. In contrast, the presence of V. natriegens cells draws down insoluble hexadecanoic acid from the air–water interface as an exponential function of time. The exponents characterizing the removal of hexadecanoic acid from the interface co-vary with the concentration of V. natriegens cells in the underlying water, with the largest exponent corresponding to the highest cell abundance. Radiochemical budgets show that evaporative fluxes from the system are linearly proportional to the quantity of hexadecanoic acid at the interface. Thus, bacterioplankton could influence the rate of evaporation and gas transfer in the ocean through the metabolism of otherwise insoluble surfactants.     

Evaluating the effects of gross nitrogen mineralization,immobilization, and nitrification on nitrogen fertilizer availability in soil experimentally contaminated with diesel

Sandy clay loam soil was contaminated with 5000 mg kg⁻¹ diesel, and amended with nitrogen (15.98 atom% ¹⁵N) at 0, 250, 500, and 1000 mg kg⁻¹ to determine gross rates of nitrogen transformations during diesel biodegradation at varying soil water potentials. The observed water potential values were −0.20, −0.47, −0.85, and −1.50 MPa in the 0, 250, 500, and 1000 mg kg⁻¹ nitrogen treatments respectively. Highest microbial respiration occurred in the lowest nitrogen treatment suggesting an inhibitory osmotic effect from higher rates of nitrogen application. Microbial respiration rates of 185, 169, 131, and 116 mg O₂ kg⁻¹ soil day⁻¹ were observed in the 250, 500, control and 1000 mg kg⁻¹ nitrogen treatments, respectively. Gross nitrification was inversely related to water potential with rates of 0.2, 0.04, and 0.004 mg N kg⁻¹ soil day⁻¹ in the 250, 500, and 1000 mg kg⁻¹ nitrogen treatments, respectively. Reduction in water potential did not inhibit gross nitrogen immobilization or mineralization, with respective immobilization rates of 2.2, 1.8, and 1.8 mg N kg⁻¹ soil day⁻¹, and mineralization rates of 0.5, 0.3, and 0.3 mg N kg⁻¹ soil day⁻¹ in the 1000, 500, and 250 mg kg⁻¹ nitrogen treatments, respectively. Based on nitrogen transformation rates, the duration of fertilizer contribution to the inorganic nitrogen pool was estimated at 0.9, 1.9, and 3.2 years in the 250, 500, and 1000 mg kg⁻¹ nitrogen treatments, respectively. The estimation was conservative as ammonium fixation, gross nitrogen immobilization, and nitrification were considered losses of fertilizer with only gross mineralization of organic nitrogen contributing to the most active portion of the nitrogen pool.     

Amino Acid Availability Regulates the Effect of Hyperinsulinemia on Skin Protein Metabolism in Pigs

The effects of amino acid supply and insulin infusion on skin protein kinetics (fractional synthesis rate (FSR), fractional breakdown rate (FBR), and net balance (NB)) in pigs were investigated. Four-month-old pigs were divided into four groups as follows: control, insulin (INS), amino acid (AA), and INS + AA groups based on the nutritional and hormonal conditions. l-[ring-¹³C₆]Phenylalanine was infused. FBR was estimated from the enrichment ratio of arterial phenylalanine to intracellular free phenylalanine. Plasma INS was increased (p < 0.05) in the INS and INS + AA groups. Plasma glucose was maintained by infusion of glucose in the groups receiving INS. The interventions did not change the NB of skin protein. However, the interventions affected the FSR and FBR differently. An infusion of INS significantly increased both FSR and FBR, although AA infusion did not. When an AA infusion was added to the infusion of insulin (INS + AA group), FSR and FBR were both lower when compared with the INS group. Our data demonstrate that in anesthetized pigs INS infusion did not exert an anabolic effect, but rather it increased AA cycling into and out of skin protein. Because co-infusion of AAs with INS ameliorated this effect, it is likely that the increased AA cycling during INS infusion was related to AA supply. Although protein kinetics were affected by both INS and AAs, none of the interventions affected the skin protein deposition. Thus, skin protein content is closely regulated under normal circumstances and is not subject to transient changes in AAs or hormonal concentrations.     

Measuring tree root respiration using (13)C natural abundance: rooting medium matters

Tree root respiration utilizes a major portion of the primary production in forests and is an important process in the global carbon cycle. Because of the lack of ecologically relevant methods, tree root respiration in situ is much less studied compared with above-ground processes such as photosynthesis and leaf respiration. This study introduces a new (13)C natural tracer method for measuring tree root respiration in situ. The method partitions tree root respiration from soil respiration in buried root chambers. Rooting media substantially influenced root respiration rates. Measured in three media, the fine root respiration rates of longleaf pine were 0.78, 0.27 and 0.18 mg CO(2) carbon mg(-1) root nitrogen d(-1) at 25 degrees C in the native soil, tallgrass prairie soil, and sand-vermiculite mixture, respectively. Compared with the root excision method, the root respiration rate of longleaf pine measured by the field chamber method was 18% higher when using the native soil as rooting medium, was similar in the prairie soil, but was 42% lower if in the sand-vermiculite medium. This natural tracer method allows the use of an appropriate rooting medium and is capable of measuring root respiration nondestructively in natural forest conditions.