The concept of light intensity adaptation in marine phytoplankton: Some experiments with Phaeodactylum tricornutum

The historical background on adaptation of algae to various light intensities is analysed. It is argued that there is little evidence to suggest that previous growth at low light intensities enhances the ability of an alga to utilize these low light levels. Rather, the published evidence suggests that the most general response to growth at sub-optimal light intensities is a reduced ability to utilize saturating levels. The present experiments with Phaeodactylum tricornutum Bohlin have tested this concept of light intensity adaptation. Changing photosynthetic abilities during batch growth depended on the light intensity used for growth and these changes affected interpretations of the data. When measurements were made intensities appeared to photosynthesize (at all intensities) better than did those grown at higher light levels. When the changes during batch growth were taken into account, or when the alga was grown in turbidostat cultures, a different picture was obtained. Growth at reduced light intensities was accompanied by (a) increased chlorophyll content, (b) decreased rate of light-saturated photosynthesis expressed on a chlorophyll, cell number or cell protein basis, and (c) decreased activity of RuDP carboxylase. One result suggested that growth at a suboptimal light intensity did enhance the ability to utilize lower light levels. The light-saturation curve of cells grown in batch culture at 0.7 klux showed higher slopes at the low light intensities than did those grown at 12 klux. This was most marked when photosynthesis was expressed per cell, but was also apparent when it was put on a per chlorophyll basis.     

Biosynthetically active glutamine synthetase in the marine diatom Phaeodactylum tricornutum: optimization of the forward-reaction assay

The activity of glutamine synthetase (GS) was measured in the marine diatom Phaeodactylum tricornutum Bohlin (Strain SME) by a biosynthetic assay, based on -glutamyl hydroxamate synthesis, and referred to as the forwardreaction assay. The effects of pH, temperature and different homogenizing buffer preparations on enzyme activity, linearity of reactions, and substrate-saturation kinetics were investigated. The resultant data provide the basis for establishing optimum experimental conditions for a standard assay. Affinities of P. tricornutum GS for glutamate, ATP and Mg²⁺ were similar to those recorded elsewhere for a variety of other phytoplankton species using true biosynthetic assays based on release of inorganic phosphate, whereas the affinity for hydroxylamine was two orders of magnitude lower than that for ammonium, with an apparent K _m value in the millimolar range. This, together with negative results obtained during earlier attempts to detect GS activity in P. tricornutum using the true biosynthetic assay, indicates that the GS of this alga has a lower affinity for ammonium than that of other phytoplankton species. Dual substrate kinetics demonstrated that apparent K _m and V _m values for glutamate were directly proportional to the concentration of ATP, thus giving indirect evidence of a correlation between GS activity and the adenylate energy charge. Comparisons between synthetase activities obtained with the optimized forward-reaction assay and transferase activities reported from other studies on various phytoplankton species revealed discrepancies which, to a great extent, probably arise from differences in the growth conditions of the organisms.     

Changes in the acitivy of fucoxanthin-excited photosynthesis in the marine diatom Phaeodactylum tricornutum grown under different culture conditions

Variations in the photosynthetic activity under monochromatic light was studied in Phaeodactulum tricornutum grown under various culture conditions, with special reference to the composition of photosynthetic pigments. Photosynthetic activity, under light-limiting conditions, was reduced when the cells were grown under strong light. The reduction was more extensive in activity resulting from fucoxanthin-excitation than in that from chlorophyll a-excitation. The diminution in activity for fucoxanthin-excited photosynthesis did not correlate with variations in the pigment content. A similar diminution was observed with chlorophyll a fluorescence upon excitation of fucoxanthin. The change was accelerated by lowering the culture temperature, or limiting the supply of nitrogen source. The results were interpreted in terms of a nitrogen-deficient state for algal cells induced by strong light, low temperature or a limited supply of nitrogen. This leads to a modification of the physicochemical state of in vivo fucoxanthin, so that the excitation energy of fucoxanthin is less efficiently transferred to chlorophyll a. The significance of the phenomenon in the oceanic primary production is discussed.     

海洋硅藻三角褐指藻经黑暗与UV辐射后的补偿生长(英文)

补偿生长是生物体对环境变化的一种适应性反应,主要指生物体遭受到不良环境胁迫后其生长及生理机能受到制约,但当不良环境胁迫解除后,生物体的生长能力会得到不同程度的恢复,表现出相对未受不良环境胁迫生物体超常生长的现象.迄今为止,补偿生长研究的对象主要集中在水生动物、反刍动物和高等植物上,而针对体积微小的海洋浮游植物的研究非常有限.海洋微藻是海洋生态系统最主要的初级生产者,在自然界物质和能量流动中扮演着至关重要的角色;同时,它们可以广泛应用在水产养殖、食品加工、医药保健、环境保护和生物制能等各种行业,具有非常好地开发利用前景.本文利用海洋硅藻三角褐指藻为试验材料,设置黑暗胁迫和UV辐射胁迫处理,着重测定微藻在恢复生长时期的藻生物量,叶绿素a含量和蛋白质含鼍等指标,探讨经黑暗与UV辐射胁迫后三角褐指藻的补偿生长现象.结果显示,无论是先经正常光照处理还是2 d黑暗处理,UV辐射0.1 rain和0.8 rain的微藻生物量都明显高于不经UV辐射胁迫的微藻生物置;此外,经过不同程度的黑暗与UV辐射处理后,微藻在恢复生长期的叶绿素a和蛋白质含量在某特定生长时期中明显高于对照组的叶绿素a和蛋白质含最.结果表明,三角褐指藻对外界环境变化具有适应能力,一旦不良环境胁迫被解除,微藻会表现出一定的补偿生长效应.我们的研究结果不但拓展和丰富了目前补偿生长的研究对象范围,而且还能为当前赤潮的形成、预测和防治提供一个新的研究思路,为有益经济微藻的高效率培养与利用提供理论指导.     

Effects of 9–10 dihydroanthracene and its biodegradation products on the marine diatom Phaeodactylum tricornutum

Growth, photosynthetic capacity and chlorophyll a content of the marine diatom Phaeodactylum tricornutum Bohlin were observed after exposure to the aromatic hydrocarbon 9–10 dihydroanthracene and its biodegradation products. Growth was inhibited after exposure to the aromatic hydrocarbon, whereas no inhibition occurred in the presence of the biodegradation products alone. The degradation products were found to enhance the chlorophyll a cellular content. Synergistic effects between dihydroanthracene and its biodegradation products increased the toxicity of this aromatic hydrocarbon.     

邻苯二甲酸二丁酯对三角褐指藻的生态毒性效应

为了研究邻苯二甲酸二丁酯（DBP）对海洋微藻的生态毒性效应,选择三角褐指藻（Phaeodactylum tricornutum）作为受试对象,设置7个DBP质量浓度处理,即丙酮对照组、2.5、3、3.5、4、5和7.5mg·L-1,测定了三角褐指藻的生长、光合色素质量浓度和叶绿素荧光特性等指标。结果表明：DBP各暴露组对三角褐指藻的生长均有显著抑制作用,三角褐指藻不能耐受7.5mg·L-1的DBP,当DBP质量浓度为5mg·L-1时,三角褐指藻在前2天几乎没有生长,从第3天开始恢复生长,并很快进入指数生长期;叶绿素a、c及类胡萝卜素质量浓度也随着DBP暴露浓度的增大而逐渐降低;随着DBP暴露浓度的升高,三角褐指藻光系统Ⅱ（PSⅡ）的最大光能转化效率（Fv/Fm）、PSⅡ的潜在活性（Fv/F0）、实际光能转化效率（Yield）和光合电子传递效率（ETR）均降低,但与对照组相比,低质量浓度处理组（≤4mg·L-1）,各指标下降不明显,而5mg·L-1的实验组在胁迫12h后,Fv/Fm、Fv/F0、Yield、ETR逐渐上升,其参数值甚至超过其它处理组。     

Variations in pigment contents of the diatom Phaeodactylum tricornutum during growth

Changes in the amounts of chlorophyll a, chlorophyll c, fucoxanthin, diadinoxanthin and -carotene were determined during Phaeodactylum tricornutum growth. The transition of the culture from the logarithmic to the stationary phase is accompanied by an increase in the carotenoids: chlorophyll a ratio, associated with variations in the percentage of individual carotenoids. While fucoxanthin content decreases with the age of the culture, diadinoxanthin content increases and -carotene remains almost constant. The furanoid isomer of diadinoxanthin, absent during logarithmic growth, appears increasingly during the nutrient-deficient period. Changes in the amounts of chlorophyll a, chlorophyll c and fucoxanthin are quite similar.     

Growth and competition of the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. II. Light limitation

The marine diatoms Phaeodactylum tricornutum (Bohlin) and Thalassiosira pseudonana (Hasle and Heimdal) were grown under both continous illumination and a 14 h light: 10 h dark cycle at light intensities ranging from 1.53×10^-4 to 2.95×10^-1 ly min^-1. Under both photoperiods, T. pseudonana exhibited higher division rates than P. tricornutum at high light intensities, but the reverse was true at all light intensities <3×10^-3 ly min^-1. Comparison of these results with available data on light-limited growth of other planktonic algae suggests that P. tricornutum may be unusually efficient at maintaining its cell division rate at low light intensity. This efficiency may contribute substantially to its success in turbid, nutrient-enriched mass algal culture systems, the only environments in which it is known to attain great numbers.Contribution No. 4086 from the Woods Hole Oceanographic Institution.     

Growth and competition of the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. I. Nutrient effects

Two marine diatoms, Phaeodactylum tricornutum (Bohlin) and Thalassiosira pseudonana (Hasle and Heimdal), were grown both separately and together in batch cultures on a mixture of waste water and seawater enriched with different components of f medium. At 17°C, the maximum division rates of the two species were statistically indistinguishable. The waste water-seawater mixture used proved to have insufficient Si, relative to N and P, for the growth of T. pseudonana, which requires approximately 5x10^-14 g-at Si cell^-1 to divide at a maximum rate. P. tricornutum, on the other hand, although capable of taking up nearly 9x10^-15 g-at Si cell^-1, could sustain maximum rates of division with 4.3x10^-18 g-at Si cell^-1 or less. No allelopathic interaction between the two species could be detected. We conclude that P. tricornutum enjoys a considerable competitive advantage over T. pseudonana in a waste water-seawater-based mariculture system that is not supplemented with Si. Although Si proved necessary for T. pseudonana to complete more successfully with the other diatom, the presence of excess amounts of Si is not necessarily sufficient for the maintenance of T. pseudonana in mixed continuous culture with P. tricornutum: other factors, such as light-related or photoperiod-related growth response, are believed to determine the ultimate outcome of competition between these algae in light-limited continuous culture.Contribution No. 3999, from the Woods Hole Oceanographic Institution.Communicated by M.R. Tripp, Newark     

An indirect method for the rapid estimation of carotenoid contents in Phaeodactylum tricornutum: Possible application to other marine algae

The equations of Parsons and Strickland (1963) have been applied to calculate the concentration of total carotenoids in Phaeodactylum tricornutum, and the same pigments have simultaneously been determined on the individual fractions after TLC (thinlayer chromatography) separation. Comparison of both sets of data shows that for diverse carotenoids: chlorophyll a ratios, the direct spectrophotometric values range from 80 to 130% of the chromatographic values; wide variations in the proportions of different carotenoids account for such difference. On the basis of correlations between the proportion of each carotenoid and the absorbance ratio 480:663 nm, an indirect method for the rapid estimation of individual carotenoids of P. tricornutum is proposed. Its eventual application to other Chrysophyta species and to sea water is discussed.Contribución del Instituto de Biología Marina No. 320.     

Factors affecting the photosynthetic capacity of laboratory cultures of the diatom Phaeodactylum tricornutum

The light-saturated photosynthetic capacity of cultures of Phaeodactylum tricornutum Bohlin grown under different conditions has been measured. In batch cultures grown in a regime of alternating light and dark periods, the photosynthetic capacity reaches a maximum before the end of the exponential phase of growth, and declines thereafter. In cultures illuminated at 0.7 mW (milliwatt)/cm², there is a 75% falloff in photosynthetic capacity per cell over an 8 day period following the time of maximum photosynthetic capacity. At 1.75 mW/cm², the corresponding fall-off is 85% over a 4 day period. Cultures exposed to a prolonged period of darkness (up to 16 days at 18°C) maintain a high photosynthetic capacity. Incubation in darkness also protects the cells from the deleterious effects of high temperature (28°C) upon photosynthetic capacity. The various fluctuations of photosynthetic capacity occur without any accompanying major changes in the concentration of chlorophyll a. Evidence from estimations of total protein and of the gross pattern of photosynthetic assimilation under different conditions suggests that the changes in photosynthetic capacity are largely controlled by the enzymic component of the photosynthetic machinery. By carefully controlling the conditions of dark incubation, the photosynthetic capacity can be reduced to a very low level without significantly affecting chlorophyll a concentration. Since the effect on photosynthetic capacity is reversible, it is possible to study aspects of chloroplast development without the complication of an associated synthesis of chlorophyll.     

Feeding deterrence properties of apo-fucoxanthinoids from marine diatoms. I. Chemical structures of apo-fucoxanthinoids produced by Phaeodactylum tricornutum

Methanol extracts of freshly harvested cells of the marine diatom Phaeodactylum tricornutum were found to produce a feeding deterrent effect in the copepod Tigriopus californicus. Bioassay guided fractionation of the methanol extracts led to the isolation of four compounds possessing feeding deterrent activity. The compounds were identified as apo-10-fucoxanthinal (1), apo-12-fucoxanthinal (2), apo-12-fucoxanthinal (3), and apo-13-fucoxanthinone (4) by detailed spectroscopic analysis and comparison with authentic compounds produced semi-synthetically from fucoxanthin. Compounds 1 to 4 exhibited feeding deterrent responses in T. californicus at concentrations of less than 20 ppm.     

Photosynthetic carboxylating enzymes in Phaeodactylum tricornutum: Assay methods and properties

Rapid freezing (in liquid nitrogen) of the marine diatom Phaeodactylum tricornutum Bohlin followed by thawing permits a convenient and sensitive measurement of the activities of carboxylating enzymes without the need to prepare a cell-free extract. Using this method, the properties of RuDP and PEP carboxylases have been compared with those assayed in cell-free extracts. The most significant difference was in the Michaelis' constants (K _m 's); the values being lower in the freeze/thaw assay. The absolute rate of carbon-dioxide fixation by the enzymes was less than the rate of photosynthesis by the intact alga. Significantly, the activity of PEP carboxylase was comparable (in some experiments, greater) to that of RuDP carboxylase. The significance of this and the possibility of an enzymatic approach to measurements of marine primary productivity are discussed.     

Development of the ability to take up L-lysine by the diatom Phaeodactylum tricornutum

Freshly harvested cells of Phaeodactylum tricornutum Bohlin grown with nitrate, ammonium or lysine as a sole nitrogen source had a low ability to take up lysine or arginine, but this ability increased when cells were deprived, over 48 h, of either nitrogen or carbon. The effects of nitrogen and carbon deprivation were additive, and the uptake ability was greatest in cells incubated in darkness in nitrogen-free medium. Uptake ability increased in cells illuminated in the presence of 10^-5 M 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) an inhibitor of photosynthetic electron transport. An inverse relationship between rate of development of the uptake system and rate of photosynthesis was also established. Development of the uptake system was prevented by cycloheximide or by anaerobiosis. Following transfer to a normal nitrate medium, illuminated cells lost the lysineuptake system by dilution as the cells grew. There was a linear and positive correlation between the initial rate of uptake of lysine and the maximum concentration which was maintained in the cells when equilibrium was reached, indicating that transinhibition of lysine uptake may occur and that the extent of this inhibition is related to the size of the internal amino acid pool. The relevance of the findings to the growth of phytoplankton in natural waters is discussed.     

The effect of heavy metals on photosynthesis and loss of cell potassium in two species of marine algae,Dunaliella tertiolecta and Phaeodactylum tricornutum

Observations on sixth-stage nauplius (pre-cyprid) of the barnacle Balanus eberneus Gould treated with L-proline-3,4-H³ suggest that the cement gland in this stage is a functional organ with considerable biosynthetic activity; the activity of the cement glands in the cyprid similarly treated is relatively low, ceasing gradually during, and completely after, metamorphosis. In the early phases of this stage, the glands become highly vacuolated (degenerate), and disappear completely in 3 days. The origin of the cement apparatus in the newly metamorphosed barnacle is discussed.     

不同氮浓度下三角褐指藻生长特性和化学组成

三角褐指藻是一类海洋单细胞硅藻,富含多不饱和脂肪酸,可以作为鱼、虾、贝等理想的饵料。而近年该藻曾多次在我国沿海海域发生暴发性增殖,给当地生态环境带来了一定的影响。为了探讨不同氮浓度对三角褐指藻生长特性和化学组成的影响,设置了低氮(44 μ mol.L-1)、中氮(880 μmol.L-1)和高氮(4 400 μmol.L-1)浓度三种处理,着重测定三角褐指藻的细胞密度、比生长率、生物量、可溶性糖、蛋白质含量和叶绿素含量等指标。结果表明,高氮浓度明显地促进了藻细胞的生长繁殖。高氮浓度下的藻细胞密度、比生长率和生物量分别比低氮浓度下的提高了 5.38 倍、0.81 倍和 2.86 倍。藻生长前期,高氮浓度和中氮浓度下的生长曲线相似,呈现一个"S"型的曲线。另外,高氮浓度下的藻细胞可溶性糖、蛋白质和叶绿素a含量分别是低氮浓度下的 2.5 倍、1.5 倍和 15 倍,说明高氮浓度促进了藻细胞化学组成的转化和积累。结果揭示,氮浓度可能是导致三角褐指藻近年在我国沿海海域发生暴发性增殖的重要因素。     

鼠尾藻和鸭毛藻水提液对三角褐指藻的抑制作用

调查了鼠尾藻Sargassum thunbergii和鸭毛藻Symphyocladia latiuscula水提液对三角褐指藻Phaeodactylum tricornutum 生长的抑制作用.结果表明,海藻水提液明显抑制三角褐指藻生长,并且这种抑制作用来自藻体活性物质(化感物质);鼠尾藻和鸭毛藻的化感物质既溶于水也溶于乙醇,随时间可能被三角褐指藻消耗尽或发生降解,因而抑制作用减弱以至消失,最大抑制发生在接种4 d.鼠尾藻和鸭毛藻水提液抑制三角褐指藻生长的半效应质量浓度(EC_(50,96h))分别为4.37g·L~(-1)和3.59g·L~(-1),暗示三角褐指藻对鸭毛藻水提液抑制的反应更敏感.     

铁浓度诱导的三角褐指藻生长和生化组分变化

采用试验生态学方法,以海洋硅藻三角褐指藻(Phaeodactylum tricornutum)为研究材料,设置一系列铁浓度处理(3.15mg·L-1、6.30 mg·L-1、9.45 mg·L-1、18.90 mg·L-1和34.65 mg·L-1),着重测定藻液光密度(OD450)、比生长率、藻细胞密度、藻生物量、叶绿素含量和蛋白质含量等生理生化指标,探讨铁浓度对海洋微藻生长特性和生化组分的影响.结果表明,铁浓度对三角褐指藻的生长状况产生了显著的影响.三角褐指藻经培养48 h后,不同铁浓度下的藻液光密度(OD450)存在显著的差异,6.30 mg·L-1铁浓度下的藻液光密度值(OD450)最高,而随着铁浓度的进一步升高,藻液光密度值(OD450)却明显降低;比生长率和藻细胞密度在3.15 mg·L-1到9.45 mg·L-1铁浓度范围内随着铁浓度升高而增大(最大值分别约为0.609 d-1和1200×104 cell·mL-1),但高于9.45 mg·L-1的铁浓度显著降低了比生长率和藻细胞密度;在试验所设置的铁浓度范围内,藻生物最表现出随铁浓度的升高而增大的趋势,34.65 mg·L-1铁浓度下的藻生物量高达0.460 mg·ML-1.同样地,微藻叶绿素a含量和蛋白质含量也明显地受到铁浓度的影响.在3.15 mg·L-1到18.90 mg·L-1铁浓度范围内,叶绿素a含量逐渐增高(最大值为2.41 mg·L-1);同样地,蛋白质含量在9.45 mg·L-1.铁浓度下达到最大值(0.153 mg·mL-1),而随着铁浓度的逐渐升高,叶绿素和蛋白质含量却明显降低.研究结果表明.铁浓度诱导海洋微藻的生长及代谢发生变化.一定较高浓度的铁显著地促进了藻细胞的生长繁殖和藻细胞化学组分的转化和积累.这些发现将有利于加深认识铁浓度在海洋生态系统中扮演的角色,进一步明确赤潮爆发的生理生态机制,从而有助于人们采取有效的预测、预防和管理措施以降低赤潮的危害.     

Effects of ethyl 2-methyl acetoacetate (EMA) on the growth of Phaeodactylum tricornutum and Skeletonema costatum

We investigated the effects of ethyl 2-methyl acetoacetate (EMA) on growth of the marine diatom algae Phaeodactylum tricornutum (P. tricornutum) and Skeletonema costatum (S. costatum). Growth of P. tricornutum was significantly inhibited by the minimum concentration (3.5 mmol·L ^?1) of EMA at lower initial algal densities (IADs) (3.6×10⁴ and 3.3×10⁵ cells·mL ^?1). However, at the highest IAD, significant growth inhibition was found at above 7 mmol·L ^?1 of EMA exposure. In S. costatum, EMA concentrations of 10.5 mmol·L ^?1 or more significantly inhibited growth at lower IAD (3×10⁴ and 1.8×10⁵ cells·mL ^?1); at the highest IAD, only EMA concentrations above 14 mmol·L ^?1 obviously inhibited the growth of S. costatum. Changes in specific growth rates and pigment were consistent with algal growth, but only at higher EMA concentrations or lower IAD values was the ratio of chlorophyll a (Chla) to carotenoid significantly lower than the control. Medium effective concentration (EC ₅₀) values were in the order 4.07, 8.03 and 12.27 mmol·L ^?1 for P. tricornutum and 7.48, 11.92 and 17.22 mmol·L ^?1 for S. costatum. All these results show that the effect of EMA on the growth of algae was species specific and mainly depended on IAD, which might be an important factor to influence algal growth.     

Nitrate uptake in marine phytoplankton: Energy sources and the interaction with carbon fixation

Field studies of whole natural phytoplankton communities from Knight Inlet, B. C., Canada and laboratory cultures of the diatom Skeletonema costatum indicate inorganic carbon fixation may be temporarily suppressed following 10 to 15% enrichment with NO ₃ ^- or NH ₄ ⁺ . (This effect is suggested to be due to competition between inorganic carbon and nitrogen for adenosine triphosphate (ATP), and is reduced when chlorophyll a is increased intracellularly after 6 to 8 h.) Results imply that the source of ATP for nitrate uptake is primarily from Photosystem I (cyclic photophosphorylation) in the presence of light. It would appear that a transient nutrient-adaptive response occurs upon addition of extracellular nitrogen.