有机氮比例和光强对赤潮藻球形棕囊藻生长和光合作用的影响

采用实验室一次性培养的方法,研究了有机氮比率和光照强度相互作用对赤潮藻球形棕囊藻（Phaeocystis globosa）生长和光合作用的影响。结果表明,在高光条件下（120μmol.m-2.s-1）,有机氮含量为20%时,其细胞密度、比生长率均显著高于对照组和其他处理组,球形棕囊藻生长最好;在低光条件下（10μmol.m-2.s-1）,有机氮含量大于50%时,其细胞密度要高于硝态氮和有机氮为20%的实验组。无论是高光还是低光条件下,有机氮含量变化对球形棕囊藻Fv/Fm影响不大。但在高光条件下,不同氮源对其Fv/Fm有显著的影响,硝氮组显著高于尿素组而在低光照条件下,以尿素为唯一氮源的对照组达到最大光能转化效率0.752,但与硝氮组差别不显著。对于既能利用无机氮又能利用有机氮的海洋微藻来说,有机氮的存在可能显著促进其增殖,从而改变浮游植物群落结构,并可能在适宜的环境条件下形成赤潮。     

2,2′,4,4′-四溴联苯醚（BDE-47）对4种海洋微藻的急性毒性

多溴联苯醚(PBDEs)是一类具有生态风险性的新型持久性有机污染物,其中BDE-47是对生物和人体毒性最强的PBDEs同系物之一.选择4种海洋微藻(海水小球藻、牟氏角毛藻、中肋骨条藻和赤潮异弯藻),采用概率单位-浓度对数法研究了BDE-47对海洋微藻的急性毒性效应(BDE-47浓度梯度设置为0、0.1、1、5、10、50μg·L-1).结果显示,BDE-47对海水小球藻、牟氏角毛藻、中肋骨条藻和赤潮异弯藻的96h半效应浓度(96hEC50)分别为0.79、1.52、1.99和2.25μg·L-1,表明BDE-47对海洋微藻属于极高毒性物质.     

2,2'',4,4''-四溴联苯醚(BDE-47)对4种海洋微藻的急性毒性

多溴联苯醚(PBDEs)是一类具有生态风险性的新型持久性有机污染物,其中BDE-47是对生物和人体毒性最强的PBDEs同系物之一.选择4种海洋微藻(海水小球藻、牟氏角毛藻、中肋骨条藻和赤潮异弯藻),采用概率单位一浓度对数法研究了BDE-47对海洋微藻的急性毒性效应(BDE-47浓度梯度设置为0、0.1、1、5、10、50μg·L-1).结果显示,BDE-47对海水小球藻、牟氏角毛藻、中肋骨条藻和赤潮异弯藻的96h半效应浓度(96h EC50)分别为0.79、1.52、1.99和2.25μg·L-1,表明BDE-47对海洋微藻属于极高毒性物质.     

BDE-47对4种海洋微藻抗氧化酶活性的影响

运用实验生态学和生物化学的方法,研究了不同质量浓度的2,2',4,4'-四溴联苯醚(BDE-47)对海水小球藻Chlorellaautotropica,牟氏角毛藻Chaetoceros muelleri,中肋骨条藻Skeletonema costatum和赤潮异弯藻Heterosigam akashiwo的抗氧化酶活性的影响.结果表明:在实验设定质量浓度范围内(0.1～2.5 μg·L~(-1)),4种微藻的超氧化物歧化酶(SOD)和过氧化氢酶(CAT)均能出现一定的应激活性,以减少BDE-47胁迫对藻细胞自身的危害,但是酶活增加的幅度却有很大不同.与赤潮异弯藻和中肋骨条藻相比,海水小球藻和牟氏角毛藻的SOD和CAT对BDE-47具有较高的敏感性.4种海洋微藻的SOD和CAT对BDE-47的敏感性顺序依次为:海水小球藻>牟氏角毛藻>中肋骨条藻>赤潮异弯藻.因此,海水小球藻的SOD和CAT可以作为指示海洋环境中BDE-47污染水平的生物标志物.     

米氏凯伦藻在三种无机氮源的生长情况

米氏凯伦藻引发的赤潮危害鱼类和无脊椎动物,给养殖业带来严重的经济损失,研究其营养生理生态,对掌握该藻形成赤潮的规律有着重要的意义。采用一次培养的实验方法研究了米氏凯伦藻(Karenia mikimotoi Hansen)在三种无机氮源不同氮磷浓度比(cN/cP)下的生长情况。结果表明:在分别以氯化铵、硝酸钠和亚硝酸钠为氮源时,米氏凯伦藻生长的最适cN/cP比分别为32、32和100。米氏凯伦藻对氯化铵的利用效率相对低于硝酸钠和亚硝酸钠,以氯化铵为氮源该藻生长缓慢,细胞密度低;而分别以硝酸钠和亚硝酸钠为氮源时该藻生长情况较好,可较长时间维持高的细胞密度。     

Hg(Ⅱ)和Pb(Ⅱ)对海洋单细胞藻的急性毒性效应

近年来,海洋重金属污染日益严重,为考察重金属对海洋藻类的生物毒性效应,进而评估其对海洋生态环境的影响,本研究采用一次性培养的实验方法,测定了2种重金属Hg(Ⅱ)和Pb(Ⅱ)对8种海洋单细胞藻的急性毒性效应,包括赤潮异弯藻(Heterosigma akashiwo Hada)、旋链角毛藻(Chaetoceros curv...     

邻苯二甲酸二丁酯对海洋微藻生长的影响

近年来,内陆和近海水体中的一种典型环境激素邻苯二甲酸二丁酯(DBP)含量显著上升,但其对海洋生态系统的影响并不明确。本研究选取7种海洋微藻为实验材料,包括1种甲藻(东海原甲藻Prorocentrum donghaiense),2种定鞭藻(小普林藻Prymesium parvum和球形棕囊藻Phaeocystis globosa),2种硅藻(中肋骨条藻Skeletonema costatum和三角褐指藻Phaeodactylum tricornutum),以及1种隐藻(红胞藻Rhodomonus salina)和1种绿藻(海洋小球藻Chlorella sp.),设置5个DBP暴露浓度(5、10、20、50、100μg·L-1),研究其对海洋微藻生长的影响,并探索DBP暴露对微藻抗氧化系统和光合系统的影响。结果表明:在所有暴露组中,球形棕囊藻、小普林藻、东海原甲藻、红胞藻、海洋小球藻生长速率显著增加;5~20μg·L-1的DBP暴露下中肋骨条藻不受影响,50、100μg·L-1下受到抑制;所有DBP暴露组对三角褐指藻没有显著影响。在50μg·L-1DBP暴露下,随着时间的延长,球形棕囊藻的SOD、CAT、MDA均表现出先升高后下降的趋势,第7天藻的叶绿素a、叶绿素b,类胡萝卜素含量较对照组分别增加了23%、10%、48%,球形棕囊藻光系统II(PSII)的最大光能转化效率(Fv/Fm)、PSII的潜在活性(Fv/F0)、光合性能指数PI分别增加了4.8%、16%、69%。研究发现DBP对海洋微藻的影响具有种间差异性,能够显著促进有害赤潮藻球形棕囊藻的生长,近岸水体DBP含量的增加可能改变浮游植物群落组成,进而增加有害藻华暴发的风险。     

Cu（Ⅱ）对海洋浮游植物生长影响的初步研究

选择赤潮异弯藻(Heterosigma akashiwo Hada)、三角褐指藻(Pheodactylum tricornutum Bohlin)、海洋原甲藻(Prorocentrum micans Ehrenber)、裸甲藻(Gymnodinim sp.)、亚心型扁藻(Platymonas subcordiforus)、旋链角毛藻(Chaetoceros curvisetus Cleve)、中肋骨条藻(Skeletonema costatum(Greville)Cleve)、青岛大扁藻(Platymonashelgolanidica)8种浮游植物,采用一次培养实验方法,研究了重金属Cu(Ⅱ)对海洋浮游植物生长的影响,并在Logistic生长模型的基础上结合Lorentz方程和GaussAmp方程,引入Cu(Ⅱ)浓度项,建立了Cu(Ⅱ)胁迫下海洋浮游植物生长动力学模型——Logistic-W模型,描述了Cu(Ⅱ)存在条件下海洋浮游植物的生长过程.结果表明,较高浓度Cu(Ⅱ)对8种浮游植物的生长普遍具有抑制作用,而较低浓度Cu(Ⅱ)则可促进旋链角毛藻、中肋骨条藻、青岛大扁藻的生长;Lorentz方程可以描述Cu(Ⅱ)浓度对浮游植物生长速率参数的影响,而GaussAmp方程可以描述Cu(Ⅱ)浓度对浮游植物生物量的影响;Cu(Ⅱ)胁迫下浮游植物的生长可用动力学方程Logistic-W描述,实验验证该模型合理,其拟合相关系数R2为0.817～0.993,平均为0.916.论文提出的生长模型可以预测不同浓度Cu(Ⅱ)胁迫下海洋浮游植物的生长情况,也可根据浮游植物的生长情况推测相应海区的Cu(Ⅱ)浓度.     

海洋赤潮藻球形棕囊藻在氮磷富营养下的细胞增殖

利用常见海洋赤潮微藻球形棕囊藻(Phaeocystis globosa)为试验研究材料,以舵海洋微藻营养液为对照(1P1N:磷质量浓度为5×10-3g·L-1.氮质量浓度为75×10-3g·L-1),设置3组富磷和富氮营养处理(3P1N:磷质量浓度为15×10-3 g·L-1,氮质量浓度为75×10-3 g·L-1;1P3N:磷质量浓度为5××10-3 g·L-1,氮质量浓度为225×10-3 g·L-1;3P3N:磷质量浓度为15××10-3 g·L-1,氮质量浓度为225×10-3 g·L-1),利用细胞记数和叶绿素荧光测定等方法研究了藻细胞在不同富磷和富氮条件的增殖情况.结果显示,不同浓度磷和氮营养下的藻体荧光值变化在试验周期内均呈现"S"型曲线,表明藻细胞的生长经历缓慢期,快速期和平缓期3个阶段;同时,不同的富磷和富氮营养条件对球形棕囊藻的叶绿素荧光值有一定的影响,其中在对照1P1N下的藻体荧光值最低,在试验结束时(第10天)只有850 μg·L-1,而在3P1N,1P3N和3P3N条件下的藻体荧光值均达到900 μg·L-1以上,显著高于1P1N下的藻体荧光值,表明富磷和富氮营养可以促进藻细胞的生长增殖,但在试验设置的不同富磷和富氮营养下的藻体荧光值之间没有显著的差异.就不同磷和氮营养条件下的藻最大比生长速率而言,3P3N和3P1N条件下的最大,均达到0.77 d-1,明显高于1P1N和1P3N条件下的藻最大比生长速率(分别只有0.70 d-1和0.69 d-1).此外,试验结束时细胞密度的变化趋势与藻体荧光值相似,富磷和富氮营养条件下的细胞密度显著高于1P1N下的细胞密度,而富磷和富氮营养条件下的细胞密度间也不存在显著的差异.研究结果揭示,水体中的高磷和高氮营养浓度是导致藻细胞大量快速增殖的一个主要因素,而利用叶绿素荧光来测定藻细胞增殖是一种快速、简便,灵敏和可靠的方法,可在今后赤潮监测过程中多加利用,以能及时、准确地预测预报赤潮爆发.从而减少其对环境和经济的影响.     

Cu和Pb对赤潮异弯藻(Heterosigma akashiwo)生长的影响

以添加了不同浓度的CuSO_4和Pb(NO_3)_2混合液的f/2培养基培养赤潮异弯藻,通过分析藻的最大生长密度、生长周期及比生长率这3个参数与Cu和Pb之间的相关关系,讨论2种重金属对赤潮异弯藻生长的影响以及2种重金属之间的互作效应。结果表明,Cu为0～0.02 mg·L~(-1)或Pb为0～0.32 mg·L~(-1)时,对赤潮异弯藻生长有促进作用;当Pb浓度不同的条件下,Cu浓度达到0.5～2.5 mg·L~(-1)时,赤潮异弯藻生长受到明显抑制,甚至无法生长;在Cu浓度不同的条件下,Pb在1～9 mg·L~(-1)范围内,随着浓度的增加,对赤潮异弯藻生长抑制作用逐步增强。在Cu浓度为0～2.5 mg·L~(-1)或Pb浓度为1～9 mg·L~(-1)及两者互作条件下,赤潮异弯藻最大生长密度和生长周期都受到显著影响(P0.01)。另外,Cu和Pb对赤潮异弯藻比生长率无显著影响,两者相互作用不明显。结合湛江海域已报道的这2种重金属实际含量,进一步评估了海区中重金属的潜在生态效应。     

Growth of marine planktonic diatoms on inorganic and organic nitrogen

A study was conducted to determine if coastal diatoms from eutrophic waters (Werribee, Port Phillip Bay, Australia) are able to grow better than diatoms from oligotrophic waters (Bass Strait, Australia) on organic nitrogen compounds as their principal nitrogen sources. Eight clones of marine planktonic diatoms, belonging to 5 species (Skeletonema costatum, Asterionella japonica, Nitzschia closterium, Coscinodiscus sp., and Fragilaria sp.), were incubated with inorganic (either nitrate or ammonia) or organic (either urea, uric acid, alanine, aspartic acid, glutamic acid, glycine, serine, threonine, or valine) nitrogen sources and growth response was measured under high and low light intensities. All clones grew well on the organic as well as thorganic nitrogen compounds under both light regimes. Intraspecific differences were not great, as no appreciable difference was noted between clones from oligotrophic and eutrophic waters. The two negatively-charged amino acids, aspartic and glutamic acids, were somewhat less effective in supporting growth of some clones than were the other amino acids. Virtually all of the dissolved organic nitrogen (DON) compounds tested were utilizable for algal growth. Further, all clones appeared able to utilize at least some natural DON (uncharacterized) for cell division; in 1980, DON represented 97% of total nitrogen in Bass Strait and 83% of total nitrogen in Werribee waters. The results are consistent with previous findings on algal utilization of certain DON compounds and indicate comparable abilities of cells from oligotrophic and eutrophic coastal waters to assimilate these nutrients.Publication No. 307 in the Ministry for Conservation, Victoria, Environmental Studies Series     

Interactions between nitrate and ammonium uptake: variation with growth rate,nitrogen source and species

The interaction between nitrate and ammonium uptake was examined as a function of preconditioning growth rate and nitrogen source by adding nitrate, ammonium, or both to nitrogen-sufficient,-deficient, and-starvedSkeletonema costatum (Grev.) Cleve and nitrogen-deficientChaetoceros debilis Cleve. By simultaneously measuring the internal accumulation of intermediates of nitrogen assimilation and the rates of nitrogen assimilation, the metabolic control of nitrogen uptake could be assessed. After the simultaneous addition of nitrate and ammonium to culture, both nitrate and ammonium uptake rates were decreased in comparison with the rates observed when each was added alone, although nitrate uptake was usually decreased more than ammonium uptake. Since both nitrate and ammonium uptake rates vary with time, preconditioning growth conditions, nitrogen sources present, and species, it was necessary to use several different indices to quantify inhibition. In general, ammonium inhibition of nitrate uptake inS. costatum was greatest in cultures preconditioned to ammonium and those at low growth rates, whereas ammonium uptake was inhibited most in cultures preconditioned to nitrate. In nitrogen-deficientC. debilis, nitrate uptake was more inhibited by ammonium, but uptake returned to normal rates more quickly than inS. costatum, whereas inhibition of ammonium uptake was similar. These results explain why the interaction between nitrate and ammonium uptake in the field can be so variable. Inhibition of uptake is not controlled by internal ammonium or total amino acids, nor is it related to the inability to reduce nitrate. Instead, inhibition must be determined in part by the external concentration of nitrogen compounds and in part by some intermediate(s) of nitrogen assimilation present inside the cell.Bigelow Laboratory Contribution No. 82022     

Partitioning of nitrogen and carbon in cultures of the marine diatom Thalassiosira fluviatilis supplied with nitrate,ammonium, or urea

Small or negligible differences in growth rates, average cell size, yields in cell numbers and total cell volumes were found in cultures of Thalassiosira fluviatilis inriched with nitrate, ammonium, or urea. Intracellular pools of unassimilated nitrate, nitrate, and ammonium were found in nutrient-rich conditions, but urea was not accumlated internally. Nitrogen assimilation into organic combination rather than nitrogen nutrient uptake was a critical rate-limiting step in nitrogen utilization. The free amino acid pool, protein, lipid-associated nitrogen, pigments, and total cell nitrogen were all highest in young or mature phase cells and decreased with age in senescent cells, whereas chitan, lipid, carbohydrate, and total cellular carbon all continued to increase during senescence. Dissolved organic nitrogen compounds accumulated in the medium only during senescence. C:N and lipid:protein were sensitive indicators of nitrogen depletion and age in T. fluviatilis.     

大亚湾裸甲藻种群动态及其关键调控因子

2008年1—12月对大亚湾养殖海域裸甲藻种群动态和主要环境因子进行了周年调查。结果表明,大亚湾海域裸甲藻类群以直径约为16～22μm的小型裸甲藻（Gymnodinium sp.）为主,另外米氏凯伦藻（Karenia mikimotoi）、链状裸甲藻（Gymnodnium catenatum）和血红哈卡藻（Akashiwo sanguinea）也有少量出现。裸甲藻种群密度呈现出明显的季节性变化特征：5月出现裸甲藻密度高峰,全年最大密度达到903 cells.mL-1,秋冬季节密度最小。不同站位裸甲藻密度也具有明显的空间分布差异,养殖及近岸海域密度普遍高于外海对照区。相关性分析结果表明,裸甲藻密度的关键调控因子包括温度、化学需氧量（COD）、可溶性有机氮（DON）和尿素浓度。裸甲藻高密度、高频率出现的温度范围在24～26℃,DON和尿素的质量浓度范围分别为N 156.38～187μg·L-1和N 17.4～38.9μg·L-1。在温度适宜的条件下,尿素等有机氮含量的增加可能成为裸甲藻赤潮的触发因子。     

红球菌Chr-9降解吡啶和苯酚的特性

研究了红球菌(Rhodococcus)Chr-9菌株在基础盐培养基中降解吡啶和苯酚的特性,分析了菌株降解苯酚和吡啶间的差异.结果表明,菌株Chr-9能够在72 h内将基础盐培养基中的吡啶(200 mg L-1)和苯酚(200 mg L-1)完全降解,同时利用吡啶和苯酚进行生长.菌株降解吡啶的最适温度为35℃,最适pH为8.0.菌株降解吡啶和苯酚的速度与底物的初始浓度呈负相关;在无其它氮源的基础盐培养基中,菌株能够利用吡啶和苯酚协同生长.图7参12     

不同氮源对2种微藻生长及总脂含量的影响

采用一次培养的方式研究硝态氮（NaNO3）、氨态氮（NH4Cl）、有机氮（尿素）及硝铵混合氮（NH4NO3）（氮浓度17.6 mmol.L-1）对新分离的两株热带淡水微藻——网状空星藻Coelastrum reticulatum及栅藻Scenedesums sp.生长情况及总脂含量的影响。结果表明,2株微藻在不同氮源影响下生长状况不同,并且造成最终的干质量、总脂含量也有较为明显的差异。2种藻添加NaNO3和NH4Cl情况下分别获得了最高的生物量（（0.72±0.08）g.L-1和（0.80±0.03）g.L-1）和最高总脂含量（（38.35±1.32）%和（30.24±3.13）%）。最终二者在添加NaNO3的情况下单位体积总脂产量最高,分别为199.26 mg.L-1和190.76 mg.L-1,均可作为生物柴油的良好来源。     

Utilization of L-lysine and L-arginine by the diatomPhaeodactylum tricornutum

The rate of growth ofPhaeodactylum tricornutum Bohlin with L-lysine as sole nitrogen source is about half that with ammonium or nitrate, and only one of the two nitrogen atoms in the lysine molecule appears to be used for growth. The organism cannot grow heterotrophically with lysine as carbon source. The rate of lysine metabolism is slow and most of that utilized is incorporated into protein, this process being faster in light than in darkness. The dark incorporation of lysine-carbon into protein is stimulated by addition of ammonium, whereas incorporation in light is unaffected. Arginine-carbon is also mainly incorporated into protein. Light has little effect on this incorporation, and addition of ammonium decreases it both in light and darkness. There is no appreciable conversion of the carbon of either amino acid to carbon dioxide. Although, under anaerobiosis, lysine is taken up and accumulated, negligible metabolism of lysine occurs.     

Nitrogen utilization during spring blooms of marine phytoplankton in Bedford Basin,Nova Scotia,Canada

Spring diatom flowerings in Bedford Basin, Nova Scotia, almost exhausted the available nitrogen supplies. The supply of nitrate appeared to regulate the magnitude of the bloom. Other nutrient supplies may also have been limiting. Certain alterations in quantities of major metabolites could be interpreted in terms of nitrogen limitation. Nitrate, (nitrite), and ammonium were generally utilized simultaneously by Thalassiosira nordenskioldii, the dominant bloom organism. This species did not effectively utilize urea in either laboratory culture or in the natural environment. T. fluviatilis simultaneously utilized all nitrogen sources provided in laboratory experiments simulating natural nutrient conditions.     

费氏中华根瘤菌ntrBC基因突变株的构建及其特性

从费氏中华根瘤菌(Sinorhizobium fredii)HN01基因文库的重组质粒pGXHN300上获得ntrB、ntrC基因完整序列.通过自杀质粒pK18mob同源单交换的方法,分别获得了HN01菌株的ntrB和ntrC基因的突变株GXHNB、GXHNC.功能检测证明GXHNB、GXHNC能以硝酸盐为唯一氮源进行正常生长,但不能以铵源作为唯一氮源进行正常生长.图4表2参15