氮磷营养盐对中肋骨条藻生长及硝酸还原酶活性的影响

通过实验室培养,在不同氮磷浓度及氮磷比率的营养条件下,对中肋骨条藻（Skeletonema costatum）的生长及藻细胞硝酸还原酶的活性进行研究。实验结果表明,中肋骨条藻属于营养型藻类,氮磷营养盐的添加,极大地促进了藻细胞的增殖。在接种后的第4～5天,各培养组藻密度达到最大值并与对照组形成极显著性差异（P〈0．01）。实验进一步发现,环境中的氮、磷浓度及氮磷比率都会影响中肋骨条藻的生长及藻细胞硝酸还原酶活性（NRA）。此外,在各培养组中,中肋骨条藻硝酸还原酶活性的最大值（NRAmax）均出现在指数生长期（接藻后第1,2天）,早于最大藻密度的出现时间（第4,5天）,这表明藻对营养盐的同化速率与生长速率并不一致,后者存在一定的滞后效应。在本实验条件下,中肋骨条藻的硝酸还原酶活性存在一定的阈值。     

Nitrogen transformations as inferred from the activities of key enzymes in the Arabian Sea oxygen minimum zone

Vertical distributions of the potential activities of some key enzymes mediating nitrification and denitrification were investigated within the oxygen (O₂) minimum zone of the Arabian Sea at a number of locations between latitudes 17°N and 21°N and longitudes 63°E and 68°E so as to get an insight into the predominant biochemical mode(s) of production and consumption of nitrous oxide (N₂O). Results revealed that the dissimilatory nitrate (NO⁻₃) reduction activity was generally very low or absent within the σ_θ range 26.6–26.8, which corresponds to the Persian Gulf Watermass (PGW). Depth profiles of nitrate reductase (NaR), nitrite reductase (NiR) and ammonia monooxygenase (AMO) activities were compared with those of O₂, NO⁻₃, nitrite (NO⁻₂) and N₂O, and it is concluded that nitrifier denitrification rather than heterotrophic denitrification is active within the core of PGW. The presence of multiple peaks of AMO activity coinciding with distinct maxima in the O₂ profile and with a trend opposite to that of NaR activity indicates that the two processes, viz., classical and nitrifier denitrification, occur in discrete layers, probably determined by the variations in the ambient O₂ concentrations at various depths surrounding the PGW core. Further, it appears that at the depths where nitrifier denitrification is active in the absence of heterotrophic denitrification, N₂O builds up as its consumption may be inhibited by O₂. Possible reasons for the occurrence of appreciable nitrate deficit within the core of PGW, where dissimilatory NO⁻₃ reduction is lacking, are discussed.     

Inorganic nitrogen uptake and related enzymatic activity in the seagrass Zostera noltii

The preferential inorganic nitrogen source for the seagrass Zostera noltii was investigated in plants from Ria Formosa, South Portugal. Rates of ammonium and nitrate uptake were determined at different concentrations of these nutrients (5, 25 and 50 μm ), supplied simultaneously (NH₄NO₃) or separately (KNO₃ and NH₄Cl). The activity of the enzymes nitrate reductase (NR) and glutamine synthetase (GS) was also assessed. The results showed that ammonium is the preferential inorganic nitrogen source for Z. noltii, but, in the absence of ammonium, the species also has a high nitrate uptake capacity. The simultaneous availability of both inorganic nitrogen forms enhanced the uptake rate of ammonium and decreased the uptake rate of nitrate compared to when only one of the nitrogen forms was supplied. The activity of both enzymes was much higher in the leaves than in the roots, highlighting the importance of the leaves as primary reducing sites in the nitrogen assimilation process.     

丙溴磷对蜈蚣藻的毒性效应 及致毒机理的研究

实验研究了丙溴磷对蜈蚣藻的生 长、光合和呼吸作用以及叶绿素、类胡萝卜素、可溶性糖含量、电解质外渗率、过氧化物酶 、过氧化氢酶和硝酸还原酶活性的影响。结果表明：(1)低浓度(50)为3.7mg/L,对光合作用的 48hEC50为6.19mg/L,对呼吸作用的48hEC50为8.47mg/L。此外,高浓度的丙溴 磷还可引起藻体可溶性糖含量增加,叶绿素、类胡萝卜素含量降低,过氧化氢酶、过氧化物 酶活性降低,细胞膜损伤,电解质外渗率升高。(3)研究还发现藻体在受到丙溴磷伤害时其 硝酸还原酶活性急剧升高,最高可达到对照的几十倍,硝酸还原酶活性有可能成为指示藻体 受伤害程度的一项生化指标。     

Nitrate assimilation by zooxanthellae maintained in laboratory culture

The ability of cultured zooxanthellae to use ammonium and nitrate was tested. The zooxanthellae were initially isolated from the anemone Aiptasia pulchella. The zooxanthellae were able to grow on media supplemented with either form of dissolved inorganic nitrogen (DIN) although ammonium was inhibitory above 300 μM. In accord with the intact symbiosis and freshly isolated algae, the cultured zooxanthellae took up ammonium. In contrast, the cultured algae demonstrated nitrate uptake and utilization. This was enhanced with DIN deprivation. Nitrate reductase was found in the algae and appeared to be inducible. Within the animal host, excretory ammonium may repress this induction.     

基于转录组测序对两种氮素营养条件下多形微眼藻氮代谢途径的解析

探究微藻的氮代谢通路对了解其对不同氮源利用的分子机制具有重要意义。本研究利用Illumina高通量测序技术对两种氮素营养条件下(硝酸氮和尿素)多形微眼藻的转录组进行分析,通过基因功能注释及数字基因表达谱分析,研究了多形微眼藻细胞内氮代谢的调控机制。结果检测出15种参与氮代谢的酶,对应76个编码基因,构建了多形微眼藻的氮代谢通路图。其中10个酶编码基因在两种不同氮素营养条件下存在差异表达,最显著的是谷氨酸合成酶、谷氨酸脱氢酶和谷氨酰胺合成酶相关基因。有机氮源(尿素)实验组中,多形微眼藻细胞内的硝酸盐还原酶、亚硝酸盐还原酶等基因的差异表达明显高于无机氮源(硝酸钠)实验组,表明当环境中的氮源为尿素时,会对多形微眼藻细胞内硝酸盐的转化和利用有一定影响。本研究初步阐述了硝酸盐、尿素的吸收转运对多形微眼藻细胞内氮代谢的影响机制,可为硅藻在不同氮素营养条件下的吸收利用机制及氮代谢响应研究提供依据。     

纳米氧化锌和菲对长江口附近海域沉积物反硝化作用及功能菌群落结构的影响

本研究在长江口附近海域采集表层沉积物,采用实验室模拟培养与分子生物学手段相结合的方法,通过测定纳米氧化锌(ZnO NPs)和菲(Phe)胁迫下沉积物中NO^-₃-N和NO^-₂-N浓度和反硝化还原酶活性及反硝化细菌基因丰度和群落多样性变化,目的是比较研究ZnO NPs和Phe对河口区沉积物反硝化作用及功能菌群落结构的影响,并探讨其作用过程和可能的作用机制。结果表明:ZnO NPs和Phe对沉积物硝酸盐还原能力和亚硝酸还原能力均产生抑制作用,浓度越高,抑制作用越强,其中亚硝酸盐还原过程受到2种污染物抑制更强烈,加重了沉积物亚硝酸盐的累积。ZnO NPs对沉积物硝酸盐还原能力、硝酸还原酶活性、narG基因丰度的抑制程度大于Phe,Phe对沉积物亚硝酸盐还原能力、亚硝酸还原酶和nirS基因丰度的抑制程度大于ZnO NPs,表明对反硝化还原酶和反硝化功能基因的抑制是外源污染物胁迫影响反硝化过程的主要机制。ZnO NPs和Phe降低了沉积物反硝化菌群落多样性水平,增加沉积物中Halomonas的优势度,降低了Bacillus的优势度,但Phe对沉积物群落多样性和组成的影响更加明显,说明Phe对长江口海区的生态影响大于ZnO-NPs。     

钝顶螺旋藻的硝酸还原酶与氢酶之间的电子竞争

于1994年6－11月，运用生物化学方法以钝项螺旋藻为材料制备整体细胞和无细胞提取物。测定表明，整体细胞硝酸还原酶与氢酶活性的相互关系和无细胞提取物氢酶与硝酸还原酶的（对还原甲基紫精）Km值，分别为2．299×10－7mol／L和1．098×10－7mol／L；证实在硝酸还原酶与氢酶之间存在着电子竞争。     

贻贝富硒提取物中硒生物活性的初步探讨

于１９９５年１１月对贻贝富硒提取物中硒生物活性进行了初步探讨。大鼠按体重随机分为三组，分别饲喂基础饲料、添加亚硒酸钠饲料和贻贝提取物饲料，实验４周后测每组鼠体重并进行代谢实验，收集７２ｈ尿及粪，测定硒含量，实验８周后处死大鼠，分析血及肝中谷胱甘肽过氧化物酶活性硒含量。     

Common catabolic enzyme patterns in a microplankton community of the Humboldt Current System off northern and central-south Chile: Malate dehydrogenase activity as an index of water-column metabolism in an oxygen minimum zone

An extensive subsurface oxygen minimum zone off northern and central-south Chile, associated with the Peru–Chile undercurrent, has important effects on the metabolism of the organisms inhabiting therein. Planktonic species deal with the hypoxic and anoxic environments by relying on biochemical as well as physiological processes related to their anaerobic metabolisms. Here we characterize, for the first time, the potential enzymatic activities involved in the aerobic and anaerobic energy production pathways of microplanktonic organisms (<100 μm), their relationship, and this relationship's association with the oxygen concentration and microplanktonic biomass in the oxygen minimum zone and adjacent areas of the Humboldt Current System water column. Our results demonstrate significant potential enzymatic activity of catabolic pathways in the oxygen minimum zone. Malate dehydrogenase had the highest oxidizing activity of nicotinamide adenine dinucleotide (reduced form) in the batch of catabolic enzymatic activities assayed, including potential pyruvate oxidoreductases activity, the electron transport system, and dissimilatory nitrate reductase. Malate dehydrogenase correlated significantly with almost all the enzymes analyzed within and above the oxygen minimum zone, and also with the oxygen concentration and microplankton biomass in the water column of the Humboldt Current System, especially in the oxygen minimum zone off Iquique. These results suggest a possible specific pattern for the catabolic activity of the microplanktonic realm associated with the oxygen minimum zone spread along the Humboldt Current System off Chile. We hypothesize that malate dehydrogenase activity could be an appropriate indicator of microplankton catabolism in the oxygen minimum zone and adjacent areas.     

海藻提取物对菠菜硝酸盐积累的影响及机理

采用对比试验的方法,对海藻提取物对菠菜硝酸盐积累的影响及机理进行了研究。结果表明:在300 kg/hm2的氮肥施用量下,用不同浓度的海藻提取物喷施菠菜叶片,均显著降低硝酸盐含量,且浓度与硝酸盐含量呈极显著的负相关;海藻提取物对菠菜硝酸盐含量不存在稀释作用,并且对土壤硝化细菌数量无明显影响;浓度与硝酸还原酶活性呈极显著的正相关,说明海藻提取物降低菠菜硝酸盐含量的机理是通过增强硝酸还原酶活性来实现的。     

Effects of tributyltin at environmental levels on monooxygenase system of digestive gland in hard clam Meretrix meretrix

The effects on ethoxyresorufin O-deethylase （EROD）, NADPH-cytochrome c reductase and NADH cytochrorne b5 reductase activities of digestive gland in Meretrix meretrix exposed to tributyltin （TBT） at environmental levels （0.1,1.0,10.0 ng/dm as stannum concentration）,in experimental condition, were evaluated. The EROD, NADH cytochrorne b5 reductase activities were significantly inhibited after exposure to 10.0 ng/dm^3 TBT for 8 and 20 d, the NADPH cytochrorne c reductase activities were significantly inhibited after exposure to 0.1,1.0 and 10.0 ng/dm TBT for 8 d and to 1.0 and 10.0 ng/dm for 20 d, as compared with the matched control, while NADH cytochrorne b5 reductases and NADPH cytochrome c reductase activities were induced after exposure to 10.0 ng/dm^3 TBT for 2 d. The EROD activity in the 10 ng/dm^3 group,and the NADH cytochrome b5 reductases activities in 1.0 and 10.0 ng/dm groups, were significantly induced when transferred to clean recovery tanks for 7 d. The three enzymic activities in the clams exposed to TBT were recovered to the level corresponding to that of the control group after transfer to clean recovery tanks for 20 d. NADPH cytochrome c reductase activity in Meretrix meretrix seems to be more sensitive to exposure of TBT than that of the EROD and NADH cytochrome b5 reductases. The results suggest that induction and inhibition by TBT to the monooxygenase system enzymic activity in Meretrix meretrix would simultaneously exist. The enzymic activities were inhibited by exposure for a long time. The results suggest that inhibition of the monooxygenase system should be an indication of the exposure to environmentally relevant concentrations of TBT for a long time.     

Selenium distribution in the Rhône delta and the Gulf of Lions

Total dissolved selenium and selenium species have been measured in the Rhône river delta and the Gulf of Lions within the framework of the EROS-2000 project. The Rhône river concentration of total selenium averages 2.30 nM with important variations related to river discharge. During estuar-ine mixing, the concentrations of total dissolved selenium, selenite and selenate (calculated as the difference between total dissolved and selenite) decrease linearly with increasing salinity, without significant interconversion between selenium species. In the open Mediterranean waters of the Gulf of Lions the total dissolved selenium increases from 0.5 nM in the surface waters to 0.9 nM in the deep waters. Organic selenium has been observed in Mediterranean deep water, an observation which is different from those from the Atlantic and the Pacific Oceans. The distributions of total inorganic selenium (Σ5e), selenite and selenate are strongly related to phosphate and silicate concentrations as observed previously for the major oceans.     

硒化物在文蛤响应镉胁迫过程中作用的转录组学分析

研究两种硒化物在文蛤富集、排出镉过程中的作用,对文蛤进行转录组测序,并进行生物信息学分析,以探讨硒化物对Cd2+代谢相关机制。硒化卡拉胶组和亚硒酸钠组共产出587 855条高质量reads。基因本体分析功能注释到15 380条unigenes, KEGG通路注释到18 866条unigenes。差异基因主要富集到肌球蛋白复合物(myosin complex)、离子通道抑制(ion channel suppression)等基因本体分析功能中。差异基因KEGG通路富集显示,在嘌呤代谢(purinemetabolism)、ECM受体相互作用(extracellularmatrix receptor interaction)、硒化合物代谢(metabolism of selenium compounds)等通路显著富集。从结果分析Cd2+可以使文蛤体内蛋白转录修饰出现异常;有机硒可以通过调控金属离子通道活性来排出细胞内的Cd2+;无机硒主要作用于文蛤细胞表面,增强其表面活性抵御Cd2+进入细胞;差异基因KEGG注释结果表明有机硒与无机硒在文蛤重金属代谢中作用机制以及途径不相同。     

海水人工湿地系统脱氮效果与基质酶活性的相关性

利用海水人工湿地系统处理海水养殖外排水,分析了人工湿地对不同形态氮的净化效果,探讨了人工湿地表层基质酶活性变化及其对系统脱氮效果的影响。选取互花米草作为人工湿地植物,煤渣、珊瑚石和细砂作为人工湿地基质,实验期间连续进水,系统运行稳定。研究结果表明:海水人工湿地系统对氨氮(NH4-N)、亚硝态氮(NO2-N)、硝态氮(NO3-N)、总氮(TN)和可溶性有机氮(DON)去除效果显著,去除率分别为(99.6±0.7)%、(99.9±0.0)%、(98.2±2.0)%、(92.6±1.5)%和(86.1±4.8)%。人工湿地表层基质下行池脱氢酶、硝酸还原酶和脲酶的酶活性均高于上行池,下行池对污染物的去除效果更好。脱氢酶活性与海水人工湿地系统氨氮的去除有关;硝酸还原酶活性影响着海水人工湿地硝态氮的去除;脲酶活性与人工湿地总氮和硝态氮的去除存在明显相关趋势。下行池硝酸还原酶和脲酶的酶活性间具有显著相关性(r=0.76, P0.05)。人工湿地微生物种类丰富,下行池微生物多样性高于上行池,植物根部微生物多样性最高,提高了系统脱氮的效率。上述研究结果将有助于阐明海水人工湿地系统中不同形态氮的迁移转化机理。     

Effects of tributyltin at environmental levels on monooxygenase system of digestive gland in hard clam Meretrix meretrix

The effects on ethoxyresorufin O-deethylase (EROD), NADPH-cytochrome c reductase and NADH cytochrome b5 reductase activities of digestive gland in Meretrix meretrix exposed to tributyltin (TBT) at environmental levels (0.1,1.0, 10.0 ng/dm3 as stannum concentration), in experimental condition, were evaluated. The EROD, NADH cytochrome b5 reductase activities were significantly inhibited after exposure to 10.0 ng/dm3 TBT for 8 and 20 d, the NADPH cytochrome c reductase activities were significantly inhibited after exposure to 0.1,1.0 and 10.0 ng/dm3 TBT for 8 d and to 1.0 and 10.0 ng/dm3 for 20 d, as compared with the matched control, while NADH cytochrome b5 reductases and NADPH cytochrome c reductase activities were induced after exposure to 10.0 ng/dm3 TBT for 2 d. The EROD activity in the 10 ng/dm3 group, and the NADH cytochrome b5 reductases activities in 1.0 and 10.0 ng/dm3 groups, were significantly induced when transferred to clean recovery tanks for 7 d. The three enzymic activities in the clams exposed to TBT were recovered to the level corresponding to that of the control group after transfer to clean recovery tanks for 20 d. NADPH cytochrome c reductase activity in Meretrix meretrix seems to be more sensitive to exposure of TBT than that of the EROD and NADH cytochrome b5 reductases. The results suggest that induction and inhibition by TBT to the monooxygenase system enzymic activity in Meretrix meretrix would simultaneously exist. The enzymic activities were inhibited by exposure for a long time. The results suggest that inhibition of the monooxygenase system should be an indication of the exposure to environmentally relevant concentrations of TBT for a long time.     

东海北部海水中硝酸盐(NO_3-N)的分布及其成因

本文根据“中日黑潮合作调查”在东海的调查资料,探讨了硝酸盐的分布特征,以及水文条件对其分布的影响,并比较了不同年份夏季硝酸盐的分布差异及其原因。分析表明,表层海水中硝酸盐分布明显受长江冲淡水影响,陆架区测值高于外海,冬季测值高于夏季; ５０ｍ ,１００ｍ 层上陆架、黑潮锋区存在着硝酸盐锋面,黑潮主干的摆动可能是影响锋面位置的主要因素     

The dual isotopes of deep nitrate as a constraint on the cycle and budget of oceanic fixed nitrogen

We compare the output of an 18-box geochemical model of the ocean with measurements to investigate the controls on both the mean values and variation of nitrate δ¹⁵N and δ¹⁸O in the ocean interior. The δ¹⁸O of nitrate is our focus because it has been explored less in previous work. Denitrification raises the δ¹⁵N and δ¹⁸O of mean ocean nitrate by equal amounts above their input values for N₂ fixation (for δ¹⁵N) and nitrification (for δ¹⁸O), generating parallel gradients in the δ¹⁵N and δ¹⁸O of deep ocean nitrate. Partial nitrate assimilation in the photic zone also causes equivalent increases in the δ¹⁵N and δ¹⁸O of the residual nitrate that can be transported into the interior. However, the regeneration and nitrification of sinking N can be said to decouple the N and O isotopes of deep ocean nitrate, especially when the sinking N is produced in a low latitude region, where nitrate consumption is effectively complete. The δ¹⁵N of the regenerated nitrate is equivalent to that originally consumed, whereas the regeneration replaces nitrate previously elevated in δ¹⁸O due to denitrification or nitrate assimilation with nitrate having the δ¹⁸O of nitrification. This lowers the δ¹⁸O of mean ocean nitrate and weakens nitrate δ¹⁸O gradients in the interior relative to those in δ¹⁵N. This decoupling is characterized and quantified in the box model, and agreement with data shows its clear importance in the real ocean. At the same time, the model appears to generate overly strong gradients in both δ¹⁸O and δ¹⁵N within the ocean interior and a mean ocean nitrate δ¹⁸O that is higher than measured. This may be due to, in the model, too strong an impact of partial nitrate assimilation in the Southern Ocean on the δ¹⁵N and δ¹⁸O of preformed nitrate and/or too little cycling of intermediate-depth nitrate through the low latitude photic zone.     

Effects of the inhibitor ellipticine on cytochrome P450-reductase and cytochrome P450 (1A) function in hepatic microsomes of flounder (Platichthys flesus)

The effects of the mammalian inhibitor ellipticine (5,11-dimethyl-[6H]-pyrido[4,3b] carbazole) were examined in a mechanistic study of the cytochrome P450 monooxygenase system of control and β-naphthoflavone (βNF)-induced hepatic microsomes of Platichthys flesus. Ellipticine was indicated to bind to the haem moiety of cytochrome P450s (gave type II binding spectra) and to inhibit the transfer of electrons from both the hydrophobic binding site of cytochrome P450 reductase (P450R) to P450 (inhibited P450R reductase activity) and the hydrophilic binding site of P450R to soluble electron acceptors (inhibited NAD(P)H-cytochrome c reductase activity). No effect was seen on cytochrome b₅ reductase activity. Ellipticine inhibition indicated the involvement of (i) P450R (possibly also P450s) in NADPH- but not NADH- dependent hydroxyl radical production, and (ii) electron transfer and P450/P450R interaction in NADPH-dependent cytochrome P450 1A-catalysed monooxygenation (7-ethoxyresorufin O-deethylase activity and benzo(a)pyrene (BaP) metabolism). Differential effects of ellipticine on cumene hydroperoxide (CHP)-dependent BaP metabolism (P450 peroxidase activity) with CHP concentration indicated the existence of at least two forms of P450 with different substrate affinities for CHP, and different mechanisms of formation for protein adducts and free metabolites. Overall, the studies indicate the primary site of action of ellipticine in P. flesus is binding between Fe³⁺-P450 and P450R.     

Nutrient distributions and new production in polar regions: parallels and contrasts between the Arctic and Antarctic

Polar regions are poorly understood components of global biogeochemical cycles. This paper discusses the influences of nitrate and ammonium concentrations on nitrate uptake (and hence new production), particularly with regard to data collected within marginal ice zones in the Arctic and Antarctic. Subsurface ammonium maxima in waters over 150 m are frequently encountered in the Arctic and occasionally in the Antarctic. Such maxima result from the heterotrophic remineralization of organic matter, and because stratified environments occur more frequently in the Arctic, significant concentrations of ammonium accumulate as a result of lower diffusive losses. Causal agents (bacteria vs. Zooplankton) may also be different in the Arctic from those in the Antarctic. Elevated ammonium concentrations significantly reduce nitrate uptake, and it is suggested that this nutrient interaction may play a significant role in controlling new production, particularly in open water regions. The new production of the Southern Ocean is estimated, and it is suggested that the production and flux from the surface layer is significant relative to the world's oceans.