Effect of light regime upon growth rate and chemical composition of a clone of Skeletonema costatum from the Trondheimsfjord, Norway

Nutrient-saturated cultures of Skeletonema costatum were grownat 15C and 42 combinations of photon flux density (PFD) anddaylength. The growth rate increased with the daylength andPFD up to 460–630 µE m^–2 s^–1 (maximum2.5 doublings day At 2000 µE m^–2 s^–1 the growthrate was reduced by 45%. The chlorophyll (chl) content of thecells and the rate of production of carbon per unit chlorophylland ambient light increased for declining light regimes as didcellular nitrogen and carbon. The N/C ratio, cellular phosphorusand ratios between in vivo fluorescence, with and without DCMU,and chlorophyll varied negligibly. The ATP/C ratio was linearlyrelated to the growth rate. The results were described mathematically.The chl/C ratio was low both in strong light and in marginallylow light, corresponding to low cellular chlorophyll and highcellular carbon, respectively. The observed increase in cellularnitrogen and carbon at shade adaptation probably represent anincrease in the size of internal stores of organic nitrogenand may imply that Skeletonema cells become enriched with organicnitrogen when staying in nitrate-rich subsurface layers, e.g.in or below a nutricline. However, close to zero growth in marginallight the cells become greatly enriched with respect to everymeasured factor. Such cells may be physiologically resting stageswhich may ensure survival during dark periods and promote rapiddevelopment during the initial phase of blooms. Cultures andnatural blooms of Skeleronema in the Trondheimsfjord exhibitvery similar patterns of variation.     

Differential effects of changes in spectral irradiance on photoacclimation,primary productivity and growth in Rhodomonas salina (Cryptophyceae) and Skeletonema costatum (Bacillariophyceae) in simulated blackwater environments

The underwater light field in blackwater environments is strongly skewed toward the red end of the electromagnetic spectrum due to blue light absorption by colored dissolved organic matter (CDOM). Exposure of phytoplankton to full spectrum irradiance occurs only when cells are mixed up to the surface. We studied the potential effects of mixing‐induced changes in spectral irradiance on photoacclimation, primary productivity and growth in cultures of the cryptophyte Rhodomonas salina and the diatom Skeletonema costatum. We found that these taxa have very different photoacclimation strategies. While S. costatum showed classical complementary chromatic adaption, R. salina showed inverse chromatic adaptation, a strategy previously unknown in the cryptophytes. Transfer of R. salina to periodic full spectrum light (PFSL) significantly enhanced growth rate (μ) by 1.8 times and primary productivity from 0.88 to 1.35 mg C · (mg Chl^?1) · h^?1. Overall, R. salina was less dependent on PFSL than was S. costatum, showing higher μ and net primary productivity rates. In the high‐CDOM simulation, carbon metabolism of the diatom was impaired, leading to suppression of growth rate, short‐term ¹⁴C uptake and net primary production. Upon transfer to PFSL, μ of the diatom increased by up to 3‐fold and carbon fixation from 2.4 to 6.0 mg C · (mg Chl^?1) · h^?1. Thus, a lack of PFSL differentially impairs primarily CO₂‐fixation and/or carbon metabolism, which, in turn, may determine which phytoplankton dominate the community in blackwater habitats and may therefore influence the structure and function of these ecosystems.     

Responses of Two Coastal Algae (Skeletonema costatum and Chlorella vulgaris) to Changes in Light and Iron Levels1

Iron (Fe) is essential for phytoplankton growth and photosynthesis, and is proposed to be an important factor regulating algal blooms under replete major nutrients in coastal environments. Here, Skeletonema costatum, a typical red-tide diatom species, and Chlorella vulgaris, a widely distributed Chlorella, were chosen to examine carbon fixation and Fe uptake by coastal algae under dark and light conditions with different Fe levels. The cellular carbon fixation and intracellular Fe uptake were measured via ¹⁴C and ⁵⁵Fe tracer assay, respectively. Cell growth, cell size, and chlorophyll-α concentration were measured to investigate the algal physiological variation in different treatments. Our results showed that cellular Fe uptake proceeds under dark and the uptake rates were comparable to or even higher than those in the light for both algal species. Fe requirements per unit carbon fixation were also higher in the dark resulting in higher Fe: C ratios. During the experimental period, high Fe addition significantly enhanced cellular carbon fixation and Fe uptake. Compared to C. vulgaris, S. costatum was the common dominant bloom species because of its lower Fe demand but higher Fe uptake rate. This study provides some of the first measurements of Fe quotas in coastal phytoplankton cells, and implies that light and Fe concentrations may influence the phytoplankton community succession when blooms occur in coastal ecosystems.     

Seasonal Development and Morphological Variability of Cyclotella ocellata (Bacillariophyceae) in the Eutrophic Lake Dagow (Germany)

The development of Cyclotella ocellata Pantocsek was studied systematically in the eutrophic, dimictic hardwater Lake Dagow from March to October, 1994. Cyclotella ocellata was the most important centric diatom in the lake with a maximum cell density of 6 × 10⁶ cells 1⁻¹. The seasonal development, characterized by a spring and a summer maximum, is considered in relation to environmental factors and the succession of the phytoplankton community. The amount of Cyclotella ocellata biomass as a proportion of the total phytoplankton biomass varied from 0.5 to 35%. In addition, seasonal changes in cell size and feature associated with sexual reproduction of Cyclotella ocellata were documented. Light and electron microscopic investigations demonstrate an extremely wide range of morphological variability of this natural population.     

Physiological reactions to a change in light regime in cultured Skeletonema costatum (Bacillariophyta): implications for estimation of phytoplankton biomass

The marine planktonic diatom Skeletonema costatum (Cleve) was grown in batch culture under a 12 h light: 12 h dark (LD) regime for several generations before the experiment. At Time 0, half of the culture was transferred to continuous light (CL). Particulate organic carbon (POC), nitrogen (PON), and photosynthetic pigments (measured by spectrophotometry and high performance liquid chromatography) were monitored for 7 to 9 days in the two cultures. Under CL, POC and PON production were always lower. In addition, we measured a much higher proportion of chlorophyll degradation products (as chlorophyllid a, phaeophytin a and phaeophorbid a (PrD)). These indicate different physiological conditions between the two cultures, as reflected also be the lower POC/PON and chlorophyll a/POC ratios. Under CL illumination cells appear stressed, probably due to the total quantity of light to which cells are exposed. In fact, a higher proportion of the two xanthophylls diadinoxanthin and diatoxanthin in the CL culture indicates a photoprotective reaction of the cells. In contrast with the LD culture, parameters measured were not significantly inter-correlated in the CL culture. The lack of correlation between chlorophyll a and POC in CL does not encourage the use of the chlorophyll a/POC ratio as a biomass estimator. On the opposite, the ratio of total pigment content to POC did not vary much as a function of the light condition of the culture, remaining near 0.040 in both light regimes. Although further observations are needed, this index seems to be a reliable indicator of phytoplankton carbon biomass.     

The toxicity of crude oil for the marine alga Skeletonema costatum (Greville) Cleve in relation to nutrient limitation

The toxicity of crude oil in relation to nutrient limitation was studied in Skeletonema costatum cultures. The addition of 100 mg/l of crude oil, although slightly toxic for the algae grown in complete media, was eventually lethal for the algae growing in phosphorus and nitrogen limited media, indicating the importance of those two nutrients for the algal resistance to oil pollution problems. Less severe effects of crude oil were observed in the silicon limited media, suggesting that the adsorptive properties of silica play an important role in the uptake and intracellular distribution of hydrocarbons. Chl a and carbon uptake were found to be more sensitive parameters for assessing hydrocarbon toxicity than cell counting.     

Photosynthetic parameters, pigment composition and respiration rates of the marine diatom Skeletonema costatum grown in continuous light and a 12:12 h light-dark cycle

Nutrient-sufficient cultures of a Trondheimsfjord (Norway) cloneof the marine centric diatom Skeleionema costatum (Grev.) Clevewere grown at 75 µmol m^–2 s^–1 and 15C at24 and 12 h daylength to study diurnal variations and the effectof daylength on pigment and chemical composition, photosyntheticparameters, dark respiration rates and scaled fluorescence excitationspectra (F), the latter used as estimates for the absorptionof energy available to Photosystem II. Specific growth rateswere 1.06 and 0.56 day^– in 24 and 12 h daylength, respectively,while dark respiration rates were generally 85% of the net growthrate. The Chla-normalized photosynthetic coefficients P^B_m anda^B were {small tilde}20–25% higher in continuous lightthan at 12 h daylength, while the Chla:C ratio was {small tilde}15%lower (0.051 versus 0.061 w:w). Thus, the carbon-normalizedcoefficients P^c_m and a^c were <11% lower at 24 h than at 12h daylength. The maximum quantum yield _max, the Chla:C ratioand F differed negligibly, as did the light saturation indexl_k, the N:C ratio and the ratios Chlc:Chla and Fucoxanthin:Chla. P^B_m and l_k did not exhibit diurnal variations at 24 hdaylength, and varied within 23% of the daily mean at 12 h daylength.Predictions of the daily gross photosynthetic rate based ondata for a given time of the day should thus not be >10%in error relative to an integrated value based on several datasets collected through 24 h. _max was 0.084–0.117 mol O₂(mol photons)^– for gross oxygen evolution. However, ifused in mathematical models for predicting the gross and netgrowth rates (i.e. the gross and net carbon turnover rates),‘practical’ values of 0.076 and 0.040 g-at C (molphotons)^–, respectively, should be employed. Correspondingly,values for a^B and P^B_m should be adjusted pro rata. ¹Present address: College of Marine Studies, Sjmannsveien 27,N-6008 lesund, Norway     

Micro-quantitation of lipids in serum-free cell culture media: a critical aspect is the minimization of interference from medium components and chemical reagents

Lipids (fatty acids) at a concentration range of 10-100 microg/L are essential components included in most serum-free cell culture medium formulations. A gas chromatography/mass spectrometry (GC/MS) method for the micro-quantitation of lipids, determined as fatty acid methyl esters (FAMEs), in complex serum-free cell culture media was developed. The interference of derivatizing reagents, extraction solvents and medium additives in the micro-quantitation of lipids was also examined. The results show that the concentration of fatty acids such as palmitic and stearic acids detected in derivatizing reagents or extraction solvents was in the range of 10-230 microg/L. Tween-80, a surfactant and medium additive, produced nearly 20 FAMEs alone when methylated using a derivatizing agent. Moreover, the surfactant Pluronic F-68, a medium additive, interfered in the FAME recovery. Procedures, which include use of low volumetric ratio of reagent to medium and precipitation of the above surfactants, were developed to minimize background FAMEs to levels which do not significantly affect the quantitation of medium lipids and to diminish the interference caused by Pluronic F-68. Fatty acid concentrations in several complex serum-free culture media were quantitated by this method and were very close to the values indicated in their formulations.     

Morphological variability of Aulacoseira granulata (Ehr.) Simonsen (Bacillariophyceae) in the Lower Paraná River (Argentina)

In this paper we consider the morphological variation within a natural population of Aulacoseira granulata (Ehr.) Simonsen in relation to environmental factors. This species was dominant in the phytoplankton of the Lower Paraná River (Argentina) and exhibited seasonal fluctuations of cell dimensions. The mean cell diameter was directly correlated with the river water level and inversely with pH and nitrate concentrations, whereas cell length was directly correlated with transparency and nitrate concentration and inversely with suspended solids. This pattern was similar to that observed for filament length. The cell length : diameter ratio was inversely related to water level and discharge and directly related to pH, transparency, and nitrate concentration. Maximum diameters did not coincide with maximum lengths. A tendency to maintain cell volume throughout the annual cycle was observed, which probably relates to both buoyancy and photosynthetic capacity. These results associate the water ascendancy and the size recovery phases to discharge. Cells become smaller on the ebbing of the flood phase, and the decreasing depth increases the probability that the alga will be disentrained from the turbulent field. The loss during low water would act as a stimulus for auxosporulation, contributing to the production of large cells to start off the next population. Received: August 12, 2000 / Accepted: November 14, 2000     

Qualitative and quantitative composition of pigments in Phaeodactylum tricornutum (Bacillariophyceae) stressed by iron

The effect of Fe(III) deficiency on qualitative and quantitative changes in pigment composition in Phaeodactylum tricornutum Bohlin was demonstrated by HPLC and AAS. Maximum content of pigments showed the diatom cells incubated at the optimum iron concentration, i.e., 10 M. The contents of chlorophyll a, chlorophyll c ₁+c ₂, fucoxanthin, diadinoxanthin and ,-carotene were 109.99, 20.16, 40.39, 1.29 and 1.48 fg per cell, respectively. The results obtained showed that Fe(III) affected qualitative and quantitative pigment composition in P. tricornutum. The content of individual pigments, proportions between accompanying pigments and their ratios to chlorophyll a were important indicators of phytoplankton response to iron stress. The strong reduction in ,-carotene content, several times (2–5) increase in diadinoxanthin level as compared to ,-carotene, and high amount of diadinoxanthin in relation to chlorophyll a were observed in algae growing at very low Fe(III) concentrations, 0.001 and 0.01 M. The data suggested that phytoplankton pigments could be a potential physiological marker.     

High CO2 increases lipid and polyunsaturated fatty acid productivity of the marine diatom Skeletonema costatum in a two-stage model

Journal of Applied Phycology - Lipid and polyunsaturated fatty acids (PUFA) from microalgae can be used as biodiesel and health care products. How to enhance their productivity is crucial for...     

Effect of media composition on yield values of bacteria growing on binary and ternary substrate mixtures in continuous culture.

Pseudomonas aeruginosa 142 and a presumed variant were grown axenically in chemostats on salicylate/benzoate or salicylate/glucose binary feeds. Each substrate was supplied at 2, 10, 50, 90, 98, or 100% of the total energy flux. Two experiments were also run with ternary mixtures using the same substrates. Aliquots were transferred to fed-batch reactors receiving the same substrates at the same specific rates as the chemostat, but with one substrate radiolabeled with 14C. Radiolabel incorporated into biomass, 14CO2, and soluble microbial products over a period of 8 minutes was used to establish the biomass yield, CO2 yield, and product yield, respectively, associated with a given substrate. The effect of the percent substrate in the feed on the yields depended on the pair of substrates supplied. When benzoate comprised 50% or more of the applied substrate in salicylate/benzoate feeds, the fraction of benzoate in the feed had a small effect on the yield values associated with benzoate. However, when benzoate constituted 2% or 10% of the feed, CO2 yields were lower, biomass yields were slightly lower, and product yields were higher. In contrast, the percent of salicylate in the feed had little effect on any of the salicylate yields for cells growing on the salicylate/benzoate feeds. When salicylate was mixed with glucose, the yields associated with salicylate behaved quite differently. Biomass and CO2 yields were lower and product yields higher when salicylate was 2% or 10% of the feed than when it was higher. In the same substrate mixtures, glucose-based biomass yields were higher and CO2 yields were lower when glucose constituted 2% or 10% of the feed but were constant for higher percentages. The results suggest that the fate of a substrate is relatively independent of the feed composition as long as the substrate in question constitutes a significant percentage of the mixture. Thus, in those situations the assumption of a constant biomass yield in multicomponent substrate modeling is justified. However, when a given substrate constitutes a small percentage of the feed, significant changes in yield may occur.     

Morphological investigations of the frustule, perizonium and initial valves of the freshwater diatom Achnanthes crenulata Grunow (Bacillariophyceae)

The present study clarifies the fine structure of the vegetative frustules, initial valves and perizonium of Achnanthes crenulata Grunow. The valves of the vegetative cell are distinctly linear‐lanceolate with an undulate margin. The valve face is quite flat and in girdle view is smoothly curved as in species of Gephyria (Bacillariophyceae). However, the valve face of the initial cells is slightly rounded and does not have an undulate margin. Furthermore, the rapheless sternum is centrally positioned along the apical axis of the araphid initial valve. As this taxon develops from auxospore to initial valve, it forms only longitudinal perizonial bands; no transverse bands arise. The perizonium consists of three silicified bands: one large, central longitudinal plate and two bands that underlie this plate; these two bands are either open or closed. This taxon has several conspicuous structures compared to other marine species of Achnanthes, but the structure of the perizonium supports the position of A. crenulata within Achnanthes sensu stricto.     

Salt adaptation of the cyanobacterium synechococcus 6311 growing in a continuous culture (turbidostat)

Physiological aspects of salt-adaptation in the cyanobacterium Synechococcus 6311 growing in a continuous culture (turbidostat) were studied. The process of salt-adaptation was completed within 3 days, as expressed by the specific growth rate of cells grown in the presence of 0.2 and 0.4 molar NaCl. An increase in photosynthetic activity during the adaptation period leads to the accumulation of soluble sugars, essential for osmoregulation in the salt-grown cells. Cells grown in the presence of 0.4 molar NaCl showed an initial inhibition in the rate of protein synthesis which was enhanced after the 1st day of salt stress. After adaptation, salt-grown cells showed an increase in phycobiliprotein synthesis and a higher phycobiliprotein to protein ratio.     

Morphological study of the atrioventricular conduction system and Purkinje fibers in yak

We studied the morphology of the atrioventricular conduction system (AVCS) and Purkinje fibers of the yak. Light and transmission electron microscopy were used to study the histological features of AVCS. The distributional characteristics of the His‐bundle, the left bundle branch (LBB), right bundle branch (RBB), and Purkinje fiber network of yak hearts were examined using gross dissection, ink injection, and ABS casting. The results showed that the atrioventricular node (AVN) of yak located in the right side of interatrial septum and had a flattened ovoid shape. The AVN of yak is composed of the slender, interweaving cells formed almost entirely of the transitional cells (T‐cells). The His‐bundle extended from the AVN, and split into left LBB and RBB at the crest of the interventricular septum. The LBB descended along the left side of interventricular septum. At approximately the upper 1/3 of the interventricular septum, the LBB typically divided into three branches. The RBB ran under the endocardium of the right side of interventricular septum, and extended to the base of septal papillary muscle, passed into the moderator band, crossed the right ventricular cavity to reach the base of anterior papillary muscle, and divided into four fascicles under the subendocardial layer. The Purkinje fibers in the ventricle formed a complex spatial network. The distributional and cellular component characteristics of the AVCS and Purkinje fibers ensured normal cardiac function.     

Morphological and chemical variations in natural populations of Pituranthos triradiatus in the Negev Desert

Morphochemical variations among six populations of Pituranthos triradiatus have been studied in the Negev Desert, in areas located within an air distance of 80 km, with an aridity gradient from 150 to 50 mm of average annual rainfall. Based on morphological florisitic characters, as well as on the content of furanocoumarins in the shoots of the shrub, five different populations have been observed. Some of the morphological characters which are commonly used for classification in the species level were found useful for the subspecies level, e.g. number of rays per umbell or per umbellet. Others, such as the shape of petals or stylopodium, or petals color, were not efficient. In addition to the commonly used characters, average fruit weight was found useful. Seven different furanocoumarins were found in the examined populations of P. triradiatus. Four of them (isoimperatorin, imperatorin, bergapten and isopimpinellin) were in each of the populations studied. Three other furanocoumarins were found to be specific: cnidilin for the Revivim population; pimpinellin for Zin and isobergapten for the Marzeva and Neqarot populations. In addition to these specific furanocoumarins, the amount of bergapten in the shoots seems to be specific for each population and is suggested to be used as a chemical marker for further screening of chemotype differentiation in R. triradiatus. It is suggested that the differences in the floristic characters, as well as in the content of furanocoumarins are of genetic nature and not an artifact. To better establish this view and to evaluate the efficiency of furanocoumarins content as a chemotaxanomic marker, further work is needed. It is also suggested that levels of isolation among the investigated population account for the differences encountered. No association could be found between different levels of aridity and different levels of furanocoumarin content.     

Ozone exposure over two growing seasons alters root-to-shoot ratio and chemical composition of birch (Betula pendula Roth)

Physiological and chemical responses of 17 birch (Betula pendula Roth) clones to 1.5–1.7 × ambient ozone were studied in an open‐field experiment over two growing seasons. The saplings were studied for growth, foliar visible injuries, net photosynthesis, stomatal conductance, and chlorophyll, carotenoid, Rubisco, total soluble protein, macronutrient and phenolic concentrations in leaves. Elevated ozone resulted in growth enhancement, changes in shoot‐to‐root (s/r) ratio, visible foliar injuries, reduced stomatal conductance, lower late‐season net photosynthesis, foliar nutrient imbalance, changes in phenolic composition, and reductions in pigment, Rubisco and soluble protein contents indicating accelerated leaf senescence. Majority of clones responded to ozone by changing C allocation towards roots, by stomatal closure (reduced ozone uptake), and by investment in low‐cost foliar antioxidants to avoid and tolerate ozone stress. A third of clones, showing increased s/r ratio, relied on inducible efficient high‐cost antioxidants, and enhanced leaf production to compensate ozone‐caused decline in leaf‐level net photosynthesis. However, the best ozone tolerance was found in two s/r ratio‐unaffected clones showing a high constitutive amount of total phenolics, investment in low‐cost antioxidants and N distribution to leaves, and lower stomatal conductance under ozone stress. The results highlight the importance of phenolic compounds in ozone defence mechanisms in the birch population. Depending on the genotype, ozone detoxification was improved by an increase in either efficient high‐cost or less efficient low‐cost antioxidative phenolics, with close connections to whole‐plant physiology.