Utilization of glutamate and glucose for heterotrophic growth by the marine pennate diatom Nitzschia laevis

Nitzschia laevis Hustedt grew in the dark in the presence of either glutamate or glucose as substrate. Complex mixtures of yeast extract or tryptone plus lactate also supported good heterotrophic growth, while tryptone alone only supported very slow growth in the dark. The observed growth rates of N. laevis in the dark at different concentrations of glutamate or glucose could be accounted for by the measured uptake rates of these compounds. The affinity of the uptake systems for glutamate and glucose (K _s =0.03 mM for each) was quite high, and similar for dark- and light-grown cells. The lack of a lag-phase when cells were transferred from photoautotrophic to heterotrophic growth conditions can be explained by the presence of uptake systems for glutamate and glucose in ligh--grown cells, as well as in dark-grown cells. However, the uptake capacity was generally higher in the latter than the former. N. laevis also took up alanine and lactate according to Michaelis-Menten kinetics, with a K _s for alanine of 0.02 mM and for lactate of 0.4 mM. Malate and glycerol were not taken up to a significant extent by the cells. Cells grown in continous light had a doubling time of 18 h. The shortest doubling time observed in the dark on glutamate was 48 h and on glucose 24 h. Glutamate was used for heterotrophic growth with an efficiency of 43% and glucose with an efficiency of 48%.Contribution No., 945 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Primary production and heterotrophy of a pennate and a centric salt marsh diatom

Radionuclide labeled sodium bicarbonate and glucose were used to measure the rates of primary production and glucose uptake by Nitzschia obtusa var. undulata and Cymbella pusilla. The presence of glucose in the medium enhanced primary production in both diatom species. Uptake of glucose in C. pusilla was photoinhibited. A maximum of 9.3 nmol C mg dry wt^-1 h^-1 of glucose was taken up in the light without the presence of NaHCO₃ in the medium and with NaHCO₃ present the value was 18 nmol C mg dry wt^-1 h^-1. The ratio of inorganic carbon fixed to the total carbon budget (autotrophy and heterotrophy) was 0.51 for N. obtusa var. undulata and 0.62 for C. pusilla.     

Carbohydrate metabolism in the marine diatom Skeletonema costatum

This is a study on the metabolism of the algal biochemical constituents of a marine diatom, Skeletonema costatum. The carbohydrates, protein, and lipid, of the diatom grown under light and dark conditions were analyzed. The composition of the organic compounds did not vary in the alga grown under continuous light conditions throughout the experiment, although a little accumulation of carbohydrate and lipid was observed in the stationary phase of the algal growth. When the diatom, at maximum growth stage, was transferred to darkness, 44.4% of the carbohydrate, 57.8% of the protein, and 27.0% of the lipid were used by respiration within a few days. Detailed analysis of the algal carbohydrate demonstrated that low molecular weight carbohydrates, consisting of glucose, and -1,3-glucan, were readily metabolized by algal respiration, leaving cell-wall carbohydrates consisting of mannan and pentosan which are immune to algal respiration. The respiratory utilization of carbohydrates is discussed in relation to the rapid decay of glucan from marine particulate matter during the course of its sinking to further depths in the ocean, as observed previously by the present author.     

Feeding of the copepod Acartia tonsa on the diatom Nitzschia closterium and brown algae (Fucus vesiculosus) detritus

Integumental sensilla were examined in 39 species of meso- and bathypelagic shrimps and 6 species of epibenthic shrimps. A tuft organ, consisting of a group of openended, tubular setae and so probably functioning as a chemosensor, is described. A single tuft with ancillary smaller tufts occur on the dorsal median surface of the fourth abdominal segment and two pairs of tufts occur on the proximal dorsal region of the telson of all oplophorid and pandalid shrimps examined. An additional tuft is present on the fifth abdominal segment of the rare bathypelagic Physetocaris microphthalma Chace. No tuft organs were identified in Processa canaliculata Leach, suggesting that they may not occur in all families of caridean shrimps. Acanthephyrid, systellaspid and pandalid shrimps have the integument completely covered by scales. These are extremely delicate and are normally removed during the process of catching the shrimp through abrasion of the integument by the net. The scales are peculiarly orientated, pointing anteriorly in the anterior half and posteriorly in the posterior half of the body. They probably have a sensory function as distance receptors, monitoring water currents and disturbances in the environment surrounding the shrimp.     

Silicon and the ecology of marine plankton diatoms. II. Silicate-uptake kinetics in five diatom species

The variation of the rate of silicate uptake with varying silicate concentration in the medium was investigated in short-term experiments with the following marine diatom species:Skeletonema costatum, Thalassiosira pseudonana, T. decipiens, Ditylum brightwellii, andLicmophora sp. The uptake conformed to Michaelis-Menten kinetics only after a correction had been made for reactive silicate that apparently could not be utilized by the diatoms. The magnitude of this correction was in the range of 0.3 to 1.3 g-at Si/l. Mean values of the half-saturation constant of silicate uptake were calculated for the different species. The lowest value was found inS. costatum (0.80 g-at Si/l) and the highest inT. decipiens (3.37 g-at Si/l). Growth limitation by low silicate concentrations could be a cause of species succession in marine plankton-diatom blooms.     

Kinetics of light-intensity adaptation in a marine planktonic diatom

The marine planktonic diatom Thalassiosira weisflogii was grown in turbidostat culture under both continuous and 12 hL: 12 hD illumination regimes in order to study the kinetics of adaptation to growth-irradiance levels. In both illumination regimes adaptation to a higher growth-irradiance level was accompanied by an increase in cell division rates and a decrease in chlorophyll a cell^-1. The rates of adaptation for both processes, derived from first order kinetic analysis, equaled each other in each experiment. The results suggest that during the transition from low-to-high growth-irradiance levels chlorophyll a is diluted by cell division and is not actively degraded. Introduction of a light/dark cycle lowered the rate of adaptation. In transitions from high-to-low growth-irradiance levels there was a sharp drop in growth rates and a slow increase in chlorophyll a cell^-1 under both continuous and intermittent illumination. In the 12 hL:12hD cycle there was a circadian rhythm in chlorophyll a cell^-1, where cellular chlorophyll contents increased during the light cycle and decreased during the dark cycle. This circadian rhythm was distinctly different from light intensity adaptation. For kinetic analysis of light intensity adaptation in a 12 hL: 12 hD cycle, the circadian periodicity was separated from the light intensity response by subjecting the data to a Kaiser window optimization digital filter. Kinetic parameters for light-intensity adaptation were resolved from the filtered data. The kinetics of lightintensity adaptation of marine phytoplankton are discussed in relation to their spatial variations and time scales of mixing.This research was performed at Brookhaven National Laboratory under the auspices of the United States Department of Energy under Contract No. DE-AC02-76 CH00016     

岷江干旱河谷灌木幼苗的土壤养分适应性研究

岷江上游干旱河谷地区的植被恢复是目前急需解决的问题,小马鞍羊蹄甲（Bauhinia faberi var. microphylla）是干旱河谷地区的优势乡土灌木,并作为当地植被恢复过程中的重要备选物种,因而进行小马鞍羊蹄甲种群在土壤贫瘠的干旱河谷的养分限制研究十分必要。针对性地进行施肥将影响到小马鞍羊蹄甲幼苗存活、生长和定殖,而如何有效提高该物种的存活和生长速率对于干旱河谷植被恢复将具有重要的意义。采用野外调查与模拟实验相结合的方法,分别研究了干旱河谷地区3个典型群落（干旱河谷的核心区——飞虹,干旱河谷灌丛与亚高山森林的过渡区——北部的石大关、干旱河谷灌丛与温性森林的过渡区——南部的蓝新镇）中小马鞍羊蹄甲幼苗叶片的化学计量特征,和施肥试验[w（有效氮）分别为100、280、460 mg·kg–1、w（有效磷）分别为12、24、48 mg·kg–1、w（氮）分别为40、70、100 mg·kg–1、w（磷）分别为12和24 mg·kg–1]中叶片化学计量特征及幼苗生长参数（叶片数、基径和株高）和各器官（根茎叶）生物量。结果表明：在野外各演替阶段的小马鞍羊蹄甲幼苗生物量和营养元素质量分数都随着磷肥的增加而增加,表明磷素是植物生长的限制因子,同时N∶P比均大于16,也暗示其受到P养分的限制;在室内施肥试验中,施加N肥没有促进小马鞍羊蹄甲生物量的积累,反而抑制了幼苗生长;施加P肥促进了幼苗生物量的积累,表明幼苗缺乏P元素,养分限制类型为P限制。     

Membrane voltage of marine phytoplankton,measured in the diatom Coscinodiscus radiatus

In order to investigate the role of the ionic relations in buoyancy of marine phytoplankton, voltage recordings have been made on the planktonic diatom Coscinodiscus radiatus using conventional glass microelectrode techniques. The most negative Nernst equilibrium voltage in C. radiatus is E _K, the potassium equilibrium voltage of around-85 mV. Accordingly, stable voltages of-40 to-80 mV were recorded from C. radiatus which conforms to the general theory of electro-diffusion of ions through membranes (voltage range V _d). In addition, membrane voltages much more negative, e.g. up to-140 mV (voltage range V _p), have been recorded in C. radiatus; these voltages demonstrate the operation of an electrogenic pump. Within the voltage range V _d, light-on and-off (microscope illumination) caused weak hyper- and depolarizations by about 2 mV with a time constant of about 10 s. Also within V _d, spontaneous oscillations could be observed with a frequency of about 0.03 Hz and irregular amplitudes up to 30 mV. These phenomena are simulated by a model for electrocoupling of the major ion transporters in plants, as worked out for guard cells with their subtle osmoregulatory system. Equivalent mechanisms are suggested to operate in planktonic diatoms for adjustment of buoyancy by appropriate uptake and release of ions.     

Inhibition of diatom photosynthesis by germanic acid: Separation of diatom productivity from total marine primary productivity

Germanic acid was shown to inhibit photosynthetic ¹⁴CO₂ uptake in marine diatoms. Inhibition was not complete even at concentrations of 20 mg Ge/1 nor in cultures incubated for extended periods of time (up to 24 h). The decrease in photosynthesis due to Ge(OH)₄ was independent of the stage of growth of the diatom culture. At 0.5 and 1.0 mg Ge/1, the degree of inhibition was dependent on the concentration of Si(OH)₄ in the medium. At 5 and 10 mg Ge/1, inhibition was not affected by Si(OH)₄ concentrations as high as those found in the sea-120 g-at Si/1. The effect of Ge(OH)₄ on photosynthesis is specific for diatoms; other marine phytoplankton were not inhibited. In mixed cultures of diatoms and marine flagellates, the reduction in ¹⁴CO₂ fixation upon addition of Ge(OH)₄ was used to calculated the proportion of diatom photosynthesis to total photosynthesis, and calculated proportions agreed well with actual proportions. Inhibition by Ge(OH)₄ was also used to estimate the percent of diatom photosynthesis in a natural marine community, and this was compared with the diatom portion of the crop. Diatom photosynthesis was higher than one might expect from crop figures, although both diatom photosynthesis and diatom numbers in the crop were low.     

pH effect on the susceptibility to parasitoid infection in the marine diatom Coscinodiscus spp. (Bacillariophyceae)

The pH on the frustule of individual cells of the marine centric diatoms Coscinodiscus granii and Coscinodiscus wailesii (Bacillariophyceae) was measured with pH microsensors in culture media with increasing pH values of 8.04, 8.14, and 8.22, respectively. In 85–96% of the C. granii cells the pH on the frustule was up to 0.4 units higher than that of the medium, reaching a maximum pH 8.95. Only in 2–3% the surface pH exceeded that of the medium by up to 0.7 pH units. These results strongly suggest that diatoms in batch cultures differ, at least temporarily, in their individual photosynthetic activities. Infection experiments with the parasitoid nanoflagellate Pirsonia diadema (Stramenopile) showed that flagellates failed to infect when the culture pH was 8.8 and above. pH measurements on freshly infected C. granii showed that the prevalence of infection was higher in tendency on diatoms with low surface pH. Application of these results to parasitoid-diatom interactions in natural waters suggests that within phytoplankton populations a strong photosynthetic activity might prevent diatom cells temporarily from infection by pH-sensitive parasitoids.     

Artificial UV-B induced changes in pigmentation of marine diatom Coscinodiscus gigas

In vitro studies in marine diatom Coscinodiscus gigas revealed that artificial UV-B radiation (313 nm) at a dose level of 0.4W m(-2) for a continuous period of 3 hours in a UV treatment chamber caused disbursement of chromatophores from their normal loci and resulted in clumping / aggregation of chromatophores exhibiting a phenomenon called UV-B induced syntrophism. It is also understood that such clumping could cause only insignificant reduction in photosynthetic oxygen release.     

Effects of enhanced UVB radiation on a marine benthic diatom mat

In order to study the ecophysiological response of a natural diatom-dominated microbial mat to an enhanced level of ultraviolet-B radiation (UVBR), intact sediment cores from a shallow microtidal bay on the Swedish west coast were incubated in an outdoor flow-through system and exposed to either no UVBR or to an artificially enhanced level of UVBR. The microbial mat was cohesive and dominated by the large (≈ 400 μm) motile diatom Gyrosigma balticum. Functional response to UVBR was assessed by measuring carbon fixation (¹⁴C) and allocation of photosynthetic products. Sediment oxygen microprofiles were measured as indicators of the balance between the photosynthetic and respiratory activity of the community. Structural variables included biomass and composition of microalgae, pigment composition (HPLC), content of UV-absorbing compounds, and composition of fatty acids. An enhanced level of UVBR resulted in significant functional changes in the microalgal community. Significantly decreased carbon fixation and lower net oxygen production (as calculated from oxygen microprofiles) suggest that primary productivity in the type of microbial mat studied is potentially sensitive to an enhanced level of UVBR. The pattern of carbon allocation hinted a change as to the photosynthetically active part of the community when exposed to UVBR, indicating a differential sensitivity among algal cells. Despite significant functional effects, pigment or algal composition were not significantly affected by enhanced UVBR. The concentration of UV-absorbing compounds was low and did not increase at UVBR exposure. Due to the short duration of the experiment (4 d), a change in algal biomass or composition was not to be expected. Moreover, the thickness of the sampled sediment layer, may have contributed to the lack of observed structural effects of enhanced UVBR. Visual observations suggested that UVBR affected the motility of G. balticum. This study, as well as other experiments with a similar diatom mat, suggest that vertical migration is a key mechanism to be further studied in relation to UVBR exposure of diatom-dominated microbial mats. Received: 7 September 1996 / Accepted: 8 October 1996     

Effect of TiO2 nanoparticles and multiwall carbon nanotubes on the freshwater diatom Nitzschia frustulum: evaluation of growth,cellular components and morphology

Great attention should be paid to the possible impacts of various nanoparticles on aquatic organism. Freshwater diatoms are essential components of phytoplankton and play a critical role in bioassessment of nanoparticle exposure in the environment. In this study, cell growth was inhibited by TiO₂-NPs and MWCNTs, the 24?h EC₅₀ values of TiO₂-NPs and MWCNTs to Nitzschia frustulum were 20.75 and 24.64?mg?L^?1, respectively. Significant decreases of Chl a content after TiO₂-NPs exposures were detected and the Chl a content of N. frustulum was obviously increased by MWCNTs treatment at lower concentration. The ROS was detected in N. frustulum after TiO₂-NPs and MWCNTs exposures. The MDA content was significantly induced by TiO₂-NPs at lower concentrations of 24 and 48?h exposure; meanwhile, it increased at all tested concentrations at 24?h MWCNTs exposure. The SOD enzyme was induced by 72 and 96?h TiO₂-NPs exposure, and increased by MWCNTs treatment at 96?h in N. frustulum. Scanning electron microscopy results revealed that N. frustulum had obvious cell deformation after TiO₂-NPs treatment. The result showed that the physiological and biochemical response mechanisms after NPs exposure of diatom were species-specific, and in relation to the exposure concentration and time.     

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.     

Abundance and production rates of floating diatom mats (Rhizosolenia castracanei and R. imbricata var. shrubsolei) in the Eastern Pacific Ocean

Abundance and production rates were measured on freefloating mats composed of the diatoms Rhizosolenia castracanei and R. imbricata var. shrubsolei in the California Current and boundary waters of the central North Pacific during October, 1980. Mats ranged from 3.0 to 10.6 cm in maximum length and had a mean volume of 6.4 ml. Production rates of the diatom mats averaged 4.0 g C colony^-1 h^-1, more than 10³ times higher than that of an equal volume of surrounding water. However, because of their low density at these sites, about 1 mat m^-3, diatom mats contributed only about 1% of the total primary production. The two large Rhizosolenia species comprised almost all of the phytoplankton biomass within the mats. Rhizosolenia species rarely occurred in seawater between mats, where the phytoplankton community was dominated by a diverse array of nannoplankton. The Rhizosolenia species in the mats appeared to be in healthy condition and contained intracellular bacteria. The very high production rates of the colonies indicate high nutrient demand and, since these particular diatoms lack the symbiotic cyanobacterium Richelia intracellularis, which is implicated in N-fixation in other species of Rhizosolenia, our results indicate that other nutrient sources must be present. We discuss the potential role of the intracellular bacteria in nitrogen fixation.     

Comparative effects of indole derivatives as antifouling agents on the growth of two marine diatom species

Antifouling agents, used to prevent biofouling, need to be assessed for their impacts on marine organisms and environment before the application. Diatoms are one of the main components of fouling biofilms, which play important roles in the formation of biofouling. Particularly, diatoms are also the important ingredients of primary production and present interest as ecotoxicological models in marine environment. In this study, two benthic diatoms Nitzschia closterium f. minutissima and Navicula climacospheniae, widely distributed in fouling biofilm, were used as models for screening the activities of potential antifoulants. Nine indole derivatives were tested and CuSO₄ was used as a reference. Indole derivatives showed significant anti-algal activities and the EC₅₀ values of most indole derivatives were lower than that of CuSO₄. Halogen substituent enhanced the anti-algal activities of compounds, and the most efficient compounds for N. closterium f. minutissima were gramine and 7-chloroindole with the EC₅₀ values of 1.94 and 2.1?mg/L, while for N. climacospheniae, 7-chloroindole and 6-bromoindole were the most efficient and the EC₅₀ values were 3.91 and 4.25?mg/L, respectively. In conclusion, indole derivatives would be one of the promising candidates as antifoulants and our results strengthened the need to perform antifouling activity assays and environment-friendly evaluations.     

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.     

Diel variation in chlorophyll a,carbohydrate and protein content of the marine diatom Skeletonema costatum

Skeletonema costatum (Greville) Cleve isolated from Narragansett Bay, USA, was incubated at 3 light intensities (ca. 0.008, 0.040 and 0.075 ly min^-1) under a 12 h light: 12 h dark (12L:12D) photoperiod at 2°, 10° and 20°C. Cellular chlorophyll a increased at intensities less than ca. 0.040 ly min^-1; increases occured within one photoperiod at temperatures above 10°C. Cellular carbohydrate increased with light intensity at all temperatures; increases during the photophase were due to net production of the dilute acid-soluble fraction. Cellular protein increased during the photoperiod at 10° and 20°C; there was little difference in cellular protein among all cultures after one photoperiod. The rate at which cellular chlorophyll a increased in response to a decrease in light suggests that diel variation in cellular chlorophyll a is temperature-dependent in S. costatum. Protein: carbohydrate ratios ranged from ca. 0.5 to 2.0 over a diel cycle; ratios increased at lower intensities and higher temperatures. The diel range in protein:carbohydrate ratios equals that in cultures developing nitrogen deficiency; thus, use of this ratio as an index to phytoplankton physiological state must account for diel light effects.