4种海洋单胞藻生化组成的环境因子效应研究

在实验室模拟条件下,应用14C示踪法测定4种海洋单胞藻的光合作用速率,研究光、温度和营养盐等环境因子对藻类细胞生化组成的影响.结果表明,三角褐指藻、盐藻、中肋骨条藻和等鞭金藻适宜生长的光强范围为5.8×103～15×103lx.4种单胞藻光合作用速率随光强增加而增大,其中盐藻和等鞭金藻的光响应比较明显.随光强增加,4种单胞藻细胞的碳水化合物含量及其变化量呈增加趋势,而蛋白质含量及其变化量则减少,脂类含量变化很小.三角褐指藻、盐藻、中肋骨条藻和等鞭金藻最适生长温度分别为:14、26、21、26℃左右.在上述4个实验温度时,4种单胞藻光合作用速率最高,细胞内的碳水化合物、蛋白质、脂类含量及其变化量也达到最大值.三角褐指藻、盐藻、中肋骨条藻和等鞭金藻光合作用过程的表观活化能(E)分别为:23.2、38.5、22.4和61.7KJ/mol,温度系数(Q10)分别为:1.74、1.74.1.38和1.69.三角褐指藻和中肋骨条藻在氮磷比(N/P)为16时,盐藻和等鞭金藻在氮磷比为28时,光合作用速率最大.在N/P为16时,4种单胞藻细胞内的碳水化合物、蛋白质、脂类含量和变化量均达到最大值.     

三角褐指藻对铜离子长期暴露的生理响应

探讨了不同浓度Cu2+长期暴露对三角褐指藻生长、光合作用、呼吸作用和色素含量等生理特性的影响。结果表明:三角褐指藻在较低浓度下(0.25mg·L-1,0.5mg·L-1)生长受到刺激,而在高浓度下(1.5mg·L-1)则下降。叶绿素荧光参数有效光化学效率(Yield)与最大光化学效率(Fv/Fm)对Cu2+非常敏感,它们均随Cu2+的升高而显著下降;Cu2+暴露对光合作用速率也具有抑制作用;而在Cu2+长期暴露下,类胡萝卜素含量比对照组高,但提高的程度随Cu2+浓度升高而下降。这些结果说明,叶绿素荧光参数可以作为三角褐指藻受到Cu2+胁迫的灵敏指标;光合作用与生长并不耦合;类胡萝卜素含量和呼吸作用速率升高可能能够降低Cu2+对三角褐指藻的毒性。     

转基因与野生型三角褐指藻生理特性的比较研究

利用叶绿素荧光仪分别测定转基因和野生型三角褐指藻的最大光能转化率,利用流式细胞仪分别测定它们的细胞大小和叶绿素含量,通过血球计数板计数绘制其生长曲线并计算比生长速率。统计分析结果显示:在培养周期内,野生型和转基因三角褐指藻的最大细胞密度有显著差别,野生型藻株的比生长速率显著高于转基因藻株,说明转基因影响了三角褐指藻的生长;野生型与转基因藻株的F_v/F_m在第2天差别不显著,但到第8天时,转基因藻株的F_v/F_m却显著高于野生型的;特基因藻株的叶绿素含量和细胞大小均显著高于野生型藻株。研究结果表明,转基因对藻细胞的生理特性产生了一定的影响,该结果为微藻转基因研究提供一定的理论依据。     

硅限制对三角褐指藻叶绿素a、叶黄素含量和脂肪酸组成的影响

为研究硅限制对三角褐指藻脂肪酸组成的影响,用不含硅的f/2人工海水培养基对处于生长指数末期的三角褐指藻进行了0、6、12、24、48h的硅限制并测定了其叶绿素a、叶黄素含量和脂肪酸组成。结果表明,硅限制对三角褐指藻叶绿素a和叶黄素含量无影响,对脂肪酸组成影响显著。硅限制显著增加了C14∶0、C16∶0、C16∶1、C16∶2、C20∶5(EPA)百分含量,但各脂肪酸响应硅限制的时间不同。C14∶0在硅限制24、48h显著增加,分别是对照组的1.33倍、1.41倍。C16∶0在硅限制6、48h显著增加,分别是对照组的1.12倍、1.19倍;C16∶1在硅限制48h显著增加,是对照组的1.16倍;C16∶2在硅限制24h显著高于对照组,是对照组的1.27倍;EPA在硅限制24h百分含量达到最大值21.86%,分别是0h和对照组的1.11倍、1.18倍。研究结果表明,一定时间的硅限制能显著提高三角褐指藻EPA百分含量,以期为工业化生产EPA提供理论参考。     

防污剂Irgarol 1051对三角褐指藻生长及生理特性的影响

为研究防污剂Irgarol 1051对微藻生长及生理生化特性的影响,以三角褐指藻(Phaeodactylum tricornutum)为材料,分析了Irgarol 1051对P.tricornutum生长、叶绿素a含量、可溶性蛋白质含量、超氧化物歧化酶(Superoxide dismutase,SOD)活力及丙二醛(Malondialdehyde,MDA)含量等的影响。结果表明:Irgarol 1051可抑制P.tricornutum的生长,并诱导其产生耐受性;P.tricornutum中叶绿素a、可溶性蛋白质含量及SOD酶活力等均随Irgarol 1051质量浓度的增加而升高,以维持其生长和生理状态;随着Irgarol 1051质量浓度的升高,藻细胞中MDA含量显著增加,表明细胞中活性氧自由基(Reactive oxygen species,ROS)过量积累,这将破坏藻细胞的膜结构与功能。研究结果将为探讨Irgarol1051对微藻的效应机制及其环境风险评价提供数据资料和科学依据。     

汞对三角褐指藻生长影响的研究——汞与浮游植物相互作用(Ⅰ)

采用连续培养技术研究汞对三角褐指藻生长的影响。当长时间接触含HgCl_2(范围5～25μg/dm~3)的培养介质中,得到结果表明,藻的生长速率随汞浓度增加而降低,细胞所排泄的有机物增加,对细胞中Chl-a的含量有一定程度影响,并抑制对硝酸盐和磷酸盐的吸收。在最初的2—4天中,汞对生物化学过程影响最大。     

RNA干扰UGP基因表达对三角褐指藻碳流分配的影响

为深入研究RNA干扰尿苷二磷酸葡萄糖焦磷酸化酶(UGPase)基因的表达对三角褐指藻(Phaeodactylum tricornutum Bohlin)碳流分配的影响,本研究以RNA干扰UGP表达的三角褐指藻为对象,测定了野生和转基因藻株的叶绿素荧光参数和生物量,分析了它们的脂肪酸组成和金藻昆布多糖、总脂及蛋白质的含量。研究显示,转基因藻株的最大光能转化效率(Fv/Fm)从指数生长后期开始显著低于野生型,但野生和转基因藻株的终生物量无明显差别。转基因藻株(AS、IR)的C16与C18之和分别占总脂肪酸含量的66.90%和68.89%,比野生藻株分别提高了19.40%和22.97%,说明抑制UGP表达改变了三角褐指藻的脂肪酸组成。转基因藻株的金藻昆布多糖、总脂及蛋白质含量与野生型藻株差别明显:藻株AS的金藻昆布多糖含量降低25.70%,蛋白质含量上升3.80%,总脂含量提高了13.2%;藻株IR的金藻昆布多糖含量降低40.70%,蛋白质含量上升8.30%,总脂含量增加了25.20%。研究结果表明,RNA干扰UGP的表达改变了三角褐指藻的脂肪酸组成,使得在最终生物量无明显减少的情况下,光合作用固定的碳更多转向油脂合成。本研究进一步明确了RNA干扰UGP的表达对三角褐指藻碳流分配的影响机制,为通过基因工程改造藻种提高微藻的生物柴油生产潜力提供了新思路。     

海水升温与氮限制对三角褐指藻生长及光合作用的影响

以硅藻三角褐指藻为实验材料,研究升温(24℃)与氮限制(40μmol/L)对藻细胞生长及光合特性的影响。结果显示,氮限制使得电子从Q_A~-到Q_B的传递受阻,降低了其对光能的吸收利用效率,导致其对高光强的耐受能力下降,从而降低生长速率。而升温并未影响三角褐指藻细胞电子传递速率、单位反应中心耗散的能量(DI_o/RC)、捕获的用于还原Q_A的能量(TR_o/RC)以及吸收的能量(ABS/RC)等,表明升温对其PSⅡ光化学反应影响不显著。但应对高光强时,升温使得藻细胞修复速率降低,导致藻细胞受光抑制明显。本研究表明,相对于温度,氮限制对三角褐指藻细胞光合作用和生长的影响更为显著,而海水升温与氮限制耦合作用会进一步影响三角褐指藻的初级生产贡献。     

三角褐指藻与抑食金球藻的竞争及化感作用研究

为探明三角褐指藻（Phaeodactylum tricornutum）对抑食金球藻（Aureococcus anophagefferens）是否具有化感作用,本研究以三角褐指藻和抑食金球藻为实验材料,研究了抑食金球藻在单培养和共培养条件下的生长情况及三角褐指藻培养滤液对其生长和叶绿素荧光参数的影响。结果表明:三角褐指藻对抑食金球藻有明显的化感抑制作用。在单培养体系中,抑食金球藻的生长曲线可用逻辑斯谛增长模型拟合,随着起始密度的增加,环境容量（K）逐渐减小,而抑食金球藻的种群瞬间增长率（r）、进入拐点时间及稳定期细胞密度均较为接近;当三角褐指藻与抑食金球藻以不同起始密度比共同培养时,抑食金球藻的生长均受到了显著地抑制（P<0.05）,但其抑制作用并未与三角褐指藻的密度呈明显的线性关系;滤液培养实验发现,10 mL和15 mL三角褐指藻培养滤液的加入可对抑食金球藻的生长产生显著影响（P<0.05）,对其叶绿素荧光参数没有影响（P>0.05）,25 mL和35 mL三角褐指藻培养滤液的加入可对抑食金球藻的生长和叶绿素荧光参数均产生显著的抑制作用（P<0.05）,叶绿素荧光参数F_v/F_m、ΦPSⅡ、alpha的值降低,Ik的值增加,PSⅡ受到损害。     

对二甲苯和苯乙烯对海洋微藻生长和光合能力的影响

为了探讨对二甲苯和苯乙烯对海洋微藻的毒性效应,本研究以三角褐指藻和亚心形扁藻为材料,分别比较了不同浓度的对二甲苯和苯乙烯胁迫对微藻生长以及叶绿素含量的影响,并计算对二甲苯和苯乙烯对三角褐指藻和亚心形扁藻96hEC50。研究表明,对二甲苯和苯乙烯对三角褐指藻和亚心形扁藻生长的抑制呈显著的剂量-效应关系,对二甲苯和苯乙烯对三角褐指藻96hEC50分别为60.20mg/L和47.82mg/L,对亚心形扁藻的96hEC50值分别为26.49mg/L和85.62mg/L,相比而言,三角褐指藻对苯乙烯敏感,而亚心形扁藻对对二甲苯更为敏感。同时研究还发现,对二甲苯和苯乙烯胁迫下,试验微藻的叶绿素含量均随处理浓度以及处理时间而显著降低。这些结果为评估对二甲苯和苯乙烯对海洋微藻的毒性效应提供了依据。     

海水中铁的来源形态及其与浮游植物的相互关系

铁是浮游植物生长所必须的微量营养元素。铁在浮游植物对氮的吸收、固氮菌对N2的固定、叶绿素的合成、卟啉生物合成、光合作用电子的传输等生物过程发挥着非常重要的作用。浮游植物的光合作用固定了占全球近40％的碳，因此研究海洋中的铁的来源、形态及其与浮游植物的关系具有重要意义。本文论述了海水中铁的来源、形态及不同形态铁的相互转化，以及浮游植物需铁的生物过程、吸铁的方式、缺铁的生理生化指示和铁对浮游植物生长及群落组成的影响。     

兼养小球藻在不同浓度Fe3+培养下的蛋白质组学研究

为研究小球藻(Chlorella vulgaris)对不同浓度Fe~(3+)的响应,测定了自养和乙酸兼养小球藻在不同浓度Fe~(3+)条件下的生长速率和油脂含量,比较分析了兼养小球藻在不同浓度Fe~(3+)下的蛋白质组表达差异。结果表明,兼养小球藻比自养小球藻生长速率快,积累的生物量多,在缺铁条件下中性脂含量高。蛋白质组分析显示:在缺铁条件下光合作用相关蛋白含量最低;在缺铁和高浓度铁条件下,热激蛋白、蛋白合成和糖代谢等过程相关蛋白表达都下调,表明缺铁和高浓度铁条件对小球藻的生长都是逆境条件;在缺铁条件下氨基酸代谢相关蛋白表达上调,这可能是由于NO_3~–同化下降,氨基酸合成减少,需要进行氮的回收利用。上述结果表明:在兼养条件下缺铁培养的小球藻可获得较高生物量和中性脂含量;而高浓度铁能促进小球藻生长,但不能提高中性脂含量,对藻细胞也有一定的胁迫效应。     

台风对海洋叶绿素a浓度影响的延迟效应

利用MODIS、SeaWiFS 3A资料详细分析了2000-2006年间西北太平洋海域主要台风对叶绿素a浓度的影响.结果发现,台风可导致叶绿素a浓度最大增长平均值为2.385倍,个别最高达10倍以上,且增长到最大值平均延迟5.94d;同时叶绿素a浓度最大值与无台风时叶绿素a浓度具有线性相关性,相关系数达0.889;叶绿素a浓度与相应的海域平均海水深度具有负的乘幂关系,其相关系数是0.87;台风后叶绿素a浓度的最大增长量与相应海域海水平均深度也呈负乘幂关系,其相关性略低于前者,相关系数为0.75.     

The influence of increased iron concentration on survival and growth of seedlings and young plants of eelgrass Zostera marina

Iron (Fe) is an essential nutrient for plants but toxic at high concentrations. We subjected seedlings and young plants of eelgrass Zostera marina to different seawater iron concentrations (500, 600, 700, 800, 1,000 and 1,500 μg/L) for 30 days under controlled laboratory conditions. Natural seawater without added iron (500 μg/L) was used as reference seawater. No sediments were provided to avoid iron scavenging by particle surfaces in the sediment. We measured plant response in terms of survivorship, morphology, growth rate and productivity. Survival analysis combined with morphological, dynamic and productive assessment suggested that the optimum seawater iron concentration for the establishment of Z. marina seedlings and young plants is 700 μg/L. The no observed effect concentration, lowest observed effect concentration, lethal concentration that caused an increase in mortality to 10% of that of the control, and the effect concentration that caused a decrease in growth to 10% of that of the control values of young plants were significantly lower than those of seedlings, implying an increased sensitivity to high Fe concentrations (>1,000 μg/L). This study further develops our understanding of the physiological ecology of the early life stages of Z. marina and provides data that could prove helpful in the development of successful eelgrass restoration and conservation.     

Study of effects of environmental factors on biochemical compositions of four species of marine single-celled algae

INTRODUCTIONThecontentsandtheirvariationofcellularbiochemicalcomponentsinnormallygrowingmarinealgaereflectthedifferenceinecologicalenvironments,reportedbymanyresearchers(MomsandSkea,1978;SmithandMoms,1980;LiandHarrison,1982;SmithandGeider,1985;Yangetal.,1991,1992).Lightintensity,temperatureandnutrientsaremostimPOrtantenvironmentalfactorsintheocean.ThispaperemphasizestheeffectsofenvironmentalfactorsonthecompositionofPhaendactylumtricornutum,Dunaliellaspp.,SkeletonemacostatumandIsOChr…     

三角褐指藻和亚心形扁藻对太平洋纺锤水蚤繁殖与生化指标的影响

采用实验生态的方法,探讨三角褐指藻(Phaeodactylum tricornutum)、亚心形扁藻(Platymonassubcordiformis)对太平洋纺锤水蚤(Acartia pacifica)繁殖及其生化指标的影响.实验结果表明,三角褐指藻对太平洋纺锤水蚤产卵量和卵的孵化率均具有抑制作用,随着投喂时间的增加,太平洋纺锤水蚤的产卵量由15.02±3.15 ind/d下降到3.25±3.15 ind/d(p<0.05),其卵的孵化率由22.88%±2.49%下降到14.84%±1.20%(p<0.05).而投喂亚心形扁藻的太平洋纺锤水蚤其产卵量和卵的孵化率与对照组无显著差异(p>0.05).投喂三角褐指藻后,太平洋纺锤水蚤的超氧化物歧化酶(SOD)活力相对于投喂亚心形扁藻的增加了20.94±0.04μg/mg,其谷胱甘肽过氧化物酶(GPX)活力增加了15.812±1.240 U/mg.这表明相对于亚心形扁藻,三角褐指藻对太平洋纺锤水蚤的抗氧化性酶具有诱导作用.     

Effects of iron on growth, degree of pigmentization and nitrogen assimilation in Skeletonema costatum

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Evidence for organic complexation of iron in seawater

Iron occurs at very low concentrations in seawater of oceanic origin and its low abundance is thought to limit primary production in offshore waters (Martin and Fitzwater, 1988). A new electrochemical method, cathodic stripping voltammetry (CSV), is used here to determine the speciation of iron in seawater originating from the Western Mediterranean taking advantage of ligand competition of an added electroactive ligand with the natural organic complexing matter to evaluate whether iron is organically complexed. The measurements indicate that iron occurs 99% (or 99.9% depending on which value is selected for α_Fe′) complexed by organic complexing ligands throughout the water column of the Western Mediterranean and by analogy probably also in other oceanic waters. The composition of the organic complexing ligands is as yet unknown, but the data indicate a major source from microorganisms (bacteria or phytoplankton) in and immediately below the fluorescence maximum in the upper water column. The organic complexes are apparently reversible releasing iron when the competing ligand is added and binding more iron when its concentration is increased. The organic complexing ligands occur at concentrations well above those of iron ensuring full complexation of this biologically essential element, and buffer the free iron concentration at a very low level against fluctuations as a result of removal by primary producers or inputs from atmospheric sources. The new data indicate that a re-evaluation of the concept of the bioavailable fraction of iron is required.     

Control of the phytoplankton response during the SAGE experiment: A synthesis

The SOLAS Air-Sea Gas Exchange (SAGE) experiment was conducted in Sub-Antarctic waters off the east coast of the South Island of New Zealand in the late summer of 2004. This mesoscale iron enrichment experiment was unique in that chlorophyll a (chl a) and primary productivity were only 2× OUT stations values toward the end of the experiment and this enhancement was due to increased activity of non-diatomaceous species. In addition, this enhancement in activity appeared to occur without a significant build up of particulate organic carbon. Picoeukaryotes (<2 ??m) were the only members of the phytoplankton assemblage that showed a statistically significant increase, a doubling in biomass. To better understand the controls of phytoplankton growth and biomass, we present results from a series of on-deck perturbation experiments conducted during SAGE. Results suggest that the pico-dominated phytoplankton assemblage was only weakly inhibited by iron. Diatoms with high growth rates comprised a small (<1%) fraction of the phytoplankton assemblage, were likely iron limited, and potentially further limited by silicic acid and therefore did not significantly contribute to bloom dynamics. On deck experiments and comparison of SAGE with other iron addition experiments suggested that neither light availability nor deep mixed layers limited phytoplankton growth. Although no substantial increase in grazing rate or specific phytoplankton growth rate was detected, microzooplankton biomass doubled over SAGE as a result of an increase in cell size. The importance of microzooplankton grazing was highlighted by the fact that they were capable of consuming 15-49% of the total phytoplankton production per day. Removal was highest on eukaryotic picophytoplankton production with a mean value of 72% (29-143%). Patch dilution played an important role during SAGE; the mean patch net algal growth:dilution rate, 1.13 (0.4-2.2) was the lowest reported for a mesoscale iron enrichment experiment. Phytoplankton biomass, estimated by chlorophyll a, only accumulated when phytoplankton growth exceeded grazing and when net algal growth exceeded dilution rate. The SAGE results highlight the function of the smallest phytoplankton size fraction described by the ecumenical Iron Hypothesis. Thus, adding iron to HNLC-low silicic acid regions during certain times of the year may simply transfer more carbon through the microbial food web. A primary implication of this study is that any iron-mediated gain in fixed carbon with this set of environmental conditions has a high probability of being recycled in surface waters.