Biological nitrogen fixation in the upper water column in the south Taiwan Strait during summer 2011

2011年6—7月,利用15n2示踪法实测了台湾海峡南部海域的生物固氮速率,结合温度、盐度、天然颗粒物氮同位素组成的分布,分析并讨论了影响研究海域生物固氮速率的环境因素。结果表明,夏季台湾海峡南部海域的生物固氮速率介于168—1080 nMOl M-3d-1之间,平均为537 nMOl M-3d-1,较高的生物固氮速率大多出现在次表层水体中。研究站位的积分固氮速率变化范围为11—40μMOl M-2d-1,平均为23μMOl M-2d-1。积分固氮速率的空间变化与不同水团的影响有关,在受黑潮水影响的海域,生物固氮速率较高,而在上升流和受河流冲淡水影响的海域,生物固氮速率较低,说明较低的水温及较高的无机氮营养盐可能会抑制研究海域的生物固氮作用。研究海域天然颗粒物δ15n与生物固氮速率之间呈现良好的负相关关系,表明天然颗粒物氮同位素组成可定性指征研究海域生物固氮作用的强弱。Biological N2fixation in marine environments is a major component in the ocean nitrogen budget and plays an important role in global carbon cycles through the sequestration of atmospheric carbon dioxide and production of marine organic matter.N2fixation could be regulated by the abundance and chemical speciation of nutrients and many trace elements in seawater.Recent studies have revealed that N2fixation is much more widespread in marine environments than previously thought.However,little is known about the N2fixation in the Taiwan Strait,especially on N2fixation rates,and their relationship with environmental parameters.The major objectives of this study were to determine the N2fixation rates and their spatial distributions and to explore major physicochemical controlling factors in the south Taiwan Strait.During June and July 2011,seawater samples were collected from ten stations at two transects in the south Taiwan Strait for the measurements of N2fixation rate using the15N2tracer assay.Particulate nitrogen and its isotopic composition were measured with an elemental analyzer(Carlo Erba NC 2500) coupled with a Finnigan MAT DeltaplusXP isotope ratio mass spectrometer.Reproducibility of nitrogen isotope measurements(in terms of δ15N) was within 0.2‰.Our results showed that N2fixation rates in the south Taiwan Strait ranged from 168—1080 nmol m-3d-1with an average of 537 nmol m-3d-1.Most of the high rates were observed at subsurface layers.The depth-integrated N2fixation rates were 11—40 μmol m-2d-1with an average of(23±10) μmol m-2d-1.The distribution of the N2fixation rates showed regional variations with influence from water masses with distinctive temperature and salinity.Higher N2fixation rates were mostly observed in the regions influenced by the Kuroshio,with an average of 31 μmol m-2d-1,while lower rates occurred in the upwelling and river plume regions with an average of 15 μmol m-2d-1.This spatial distribution pattern indicated that biological N2fixation was largely impeded by the low temperature and the high concentration of dissolved inorganic nitrogen in the south Taiwan Strait.The contribution of N2fixation in the study area could be further quantified based on the δ15N signatures of suspended particles which could be significantly depleted during N2fixation.Indeed,a negative correlation between the δ15N signatures of suspended particles and N2fixation rates was observed regardless of using all data points or depth-averaged values within the water column.This indicated that15N-depleted particles were largely derived from the enhanced N2fixation,supporting the use of nitrogen isotopic composition(δ15N) of suspended particles as a potential indicator of N2fixation in the south Taiwan Strait.Further studies are needed to better elucidate the relationship between N2fixation rates and limiting elements and their chemical speciation,and thus the physical and biogeochemical controls on N2fixation in the south Taiwan Strait.国家自然科学基金资助项目(41125020;41076043;41206062); 国际海域资源调查与开发“十二五”项目(DY125-13-E-01

金属离子对锯缘青蟹NAGase活力的影响

本文研究了几种金属离子对锯缘青蟹(Scylla serrata)N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)活力的影响.其结果表明:Li+、Na+和K+对酶活力没有明显影响,Mg2+、Ca2+和Ba2+对该酶均有激活作用,激活程度依次为Ca2+>Ba2+>Mg2+.A l3+、Fe3+、Cd2+、Pb2+和Hg2+对该酶具有一定的抑制作用,2.0μmol/dm3的Hg2+可以使酶活力完全丧失.Co2+对酶的效应是先激活后抑制,Mn2+对酶仅有轻微的激活作用.Cd2+和Fe3+对锯缘青蟹NAGase的抑制作用都呈竞争性-反竞争性混合Ⅱ型效应,Cd2+的抑制常数KI和KIS分别为23.9、5.0 mmol/dm3,Fe3+的抑制常数KI和KIS分别为395.5、135.6μmol/dm3.KI>KIS,说明酶-底物络合物(ES)与抑制剂的亲和力比游离酶(E)与抑制剂的亲和力大

锯缘青蟹N-乙酰氨基葡萄糖苷酶在甲醛溶液中的失活动力学研究

研究甲醛对锯缘青蟹N-乙酰氨基葡萄糖苷酶(NAGase EC 3.2.1.52)活力的影响,结果表明:随着甲醛浓度的增大,酶活力呈指数下降,导致酶活力下降50%的甲醛浓度(失活半衰期,IC50)为0.60 mol/L.酶在低浓度甲醛溶液中的失活过程显示为可逆失活.用底物反应动力学方法考察酶在甲醛溶液中的失活动力学,测定游离酶(E)和酶-底物络合物(ES)在甲醛溶液中失活的微观速度常数,并比较游离酶(E)和酶-底物络合物(ES)的正向反应的微观失活速度常数k+0>k′+0,表明底物对酶被甲醛的失活作用有一定的保护作用.正向的失活速度常数k+0和k′+0随着甲醛浓度的增大而增大,而逆向的速度常数k-0随着甲醛浓度的增大而减小,表明随着甲醛浓度的增大,酶变性越来越快,而活力恢复越来越难

丙酮对锯缘青蟹N-乙酰-β-D-氨基葡萄糖苷酶活力与构象的影响

以丙酮为效应物,研究其对锯缘青蟹(Scylla serrata)N-乙酰--βD-氨基葡萄糖苷酶(NAGase)活力的影响,结果表明:该酶的剩余活力随着丙酮浓度增大而呈指数下降,当丙酮浓度达25%,酶的剩余活力仅为20%,说明丙酮对青蟹NA-Gase有明显的失活作用.导致酶活力丧失50%的丙酮浓度为7.5%.在较低丙酮浓度(<10%)的失活是可逆的反应过程.动力学研究表明,该酶的失活过程属于混合型,并进一步测定游离酶(E)和酶底物络合物(ES)与丙酮的结合常数(KI和KIS),分别为4.06%和10.49%,KI<KIS,说明底物存在对酶被丙酮的失活作用有一定的保护作用.应用荧光发射光谱研究青蟹NAGase经丙酮微扰后的分子构象变化情况,结果表明:丙酮对酶分子构象有显著的影响,酶的内源荧光强度随丙酮浓度增大而降低,说明酶分子中的生色基团Trp和Tyr残基的微环境发生了变化

Generating High-concentration Solution of Reactive Oxygen Species by Strong-field Ionization Discharge

为优化氧活性粒子(rOS)在水中的生成条件,并为rOS溶液生成装置提供优化系统参数的依据,研究了rOS质量浓度在水温、PH值、O2给气体积流量、rOS投加体积质量,以及系统气压(混溶压力)等因素作用下的变化规律。实验中,气态rOS在强电离条件下通过介质阻挡放电生成,以O3计,通过O3检测仪测定其浓度;水中rOS质量浓度采用dPd分光光度法测定,用CrS来表示。实验结果表明:水温、PH值与CrS呈极显著负相关(相关系数P0.05);CrS在水温分别为16℃与24.5℃之间、PH=6.5与PH=7.0之间、O2给气体积流量为2 l/MIn与3 l/MIn之间均为差异不显著(P>0.05),其余各水温、PH值、O2给气体积流量之间均为差异显著(PO2给气体积流量>系统气压与O2给气体积流量交互作用。To optimize the generating condition of reactive oxygen species(ROS) in solution, and to provide a reference for improving ROS preparation system, we investigated the effects of several parameters, including solution temperature, solution pH, O2 input, ROS dosages, and system pressure, on the concentration of obtained ROS solution.Gaseous ROS was generated in a strong-field ionization condition induced by under dielectric barrier discharge(DBD), and its dosage represented by O3 was measured by ozone monitor.ROS in solution was caught by DPD(N, N-diethyl-p-pHenylenediamine), which was measured by DPD spectrophotometry, and the concentration of ROS solution was denoted by CRS.According to the experiments, both water temperature and pH have significant negative correlation with CRS(relativefactor P0.05).The differences between CRS are insignificant(P>0.05) under conditions of solution temperature of 16 ℃, 20 ℃, 24.5 ℃, solution pH of 6.5 and 7.0, as well as O2 input of 2 L/min and 3 L/min(P>0.05), but they are highly significant(P<0.01) or significant(P<0.05) under other tested conditions.Moreover, CRS significantly increases with the decrease of miscibility pressure(P<0.01), and it is significantly affected by the interaction between miscibility pressure and O2 input under higher ROS dosages(P<0.01).Lower water temperature, lower pH, higher ROS dosages, and lower miscibility pressure are all beneficial to increasing the concentration of ROS(CRS), which is significantly affected by the change of several parameters including solution temperature in the lower range, pH around 7, O2 input in the range of larger amount, etc.On the condition of high ROS dosage input, CRS is influenced by miscibility pressure, O2 input, and the interaction between miscibility pressure and O2 input in a descending order.国家高技术研究发展计划(863计划)(2012AA062609); 国家科技支撑计划(2013BAC06B02); 国家杰出青年科学基金(61025001)~

MOLECULAR CLONING AND EXPRESSION ANALYSIS OF A TRANSLATIONALLY CONTROLLED TUMOR PROTEIN (TCTP) GENE FROM SOLEN GRANDIS

本研究克隆得到了一个大竹蛏(Solen grandis)翻译控制肿瘤蛋白(TCTP)基因(SgTCTP)的cDNA全长,其序列全长为1055bp,5′和3′端的非编码区(UTR)分别为54和461bp,开放阅读框(ORF)540bp,编码179个氨基酸,理论等电点为4.50,预测分子量大小19.96kDa。通过荧光定量PCR法检测了SgTCTP在健康大竹蛏各组织中和病原相关分子模式(PAMPs)刺激后的表达规律,结果表明:SgTCTP在检测组织外套膜、鳃、性腺、血细胞、肌肉和肝胰腺中都有表达,其中在肝胰腺中的表达量最高。脂多糖(LPS)、肽聚糖(PGN)和葡聚糖(β-1,3-glucan)刺激..

大竹蛏(Solen grandis)翻译控制肿瘤蛋白(TCTP)基因的分子克隆和表达特征分析

本研究克隆得到了一个大竹蛏(Solen grandis)翻译控制肿瘤蛋白(TCTP)基因(SgTCTP)的cDNA全长,其序列全长为1055bp,5′和3′端的非编码区(UTR)分别为54和461bp,开放阅读框(ORF)540bp,编码179个氨基酸,理论等电点为4.50,预测分子量大小19.96kDa。通过荧光定量PCR法检测了SgTCTP在健康大竹蛏各组织中和病原相关分子模式(PAMPs)刺激后的表达规律,结果表明:SgTCTP在检测组织外套膜、鳃、性腺、血细胞、肌肉和肝胰腺中都有表达,其中在肝胰腺中的表达量最高。脂多糖(LPS)、肽聚糖(PGN)和葡聚糖(β-1,3-glucan)刺激都能诱导SgTCTP的表达量上调,SgTCTP的表达量分别在LPS和PGN刺激后6和3h达到最高,为空白对照的3.64和3.36倍;β-1,3-glucan刺激后SgTCTP的表达量上升幅度最大,在12h达到最高,为空白对照的11.76倍。SgTCTP可能作为急性时相蛋白参与大竹蛏的免疫应答