LIGHT/DARK PERIODICITY IN NITROGEN ASSIMILATION OF THE MARINE PHYTOPLANKTERS SKELETONEMA COSTATUM AND COCCOLITHUS HUXLEYI IN N-LIMITED CHEMOSTAT CULTURE1,2

Skeletonema costatum and Coccolithus huxleyi were grown in nitrogen-limited chemostat cultures with illumination provided in light/dark cycles. S. costatum assimilated nitrate and ammonium primarily during the day and less so at night. Conversely, the concentration of nitrate and ammonium in the culture medium varied periodically, increasing at night and decreasing in the light. C. huxleyi assimilated both N sources at a rate sufficient to keep them at very low levels both day and night. However, the activity of N-assimilating enzymes, measured in cell-free extracts, were higher in the light than in the dark periods, implying light/dark differences in the capacity to assimilate nitrogen. Such periodicity in the rate of uptake and enzymatic activity appears to complicate the mathematical expression of nutrient-limited growth of phytoplankton exposed to natural light/dark cycles. Three aspects of dial periodicity in N assimilation have been observed in natural phytoplankton communities in the sea: (1) in assimilation rate, (2) in activity of enzymes of N-assimilation, and (3) in the ammonium concentration of the seawater. The cultures also showed periodicity in these parameters and appear to be useful model systems for study.     

Changes in Leaf Nitrate Reductase Activity In Vivo and In Vitro During Light-Dark Transitions

Nitrate reductase activity in the leaves of a number of plants after transfer from light to dark was assayed both by in vivo and in vitro methods. The initial activity persisted during the dark phase for a considerable length of time and declined gradually. After exposure to light again, the NR activity increased rapidly. The possibility of nitrate assimilation in complete darkness is discussed.     

In situ microcosm experiments on the influence of nitrate and light on phytoplankton community composition

Phytoplankton blooms are fundamental features of coastal ecosystems, but the processes that select for blooms of certain species are not well understood. The aim of this work was to investigate experimentally the interaction of light and nutrients (nitrate) in structuring phytoplankton community composition in Pelorus Sound, New Zealand. Microcosm experiments were conducted in situ nine times throughout the year, providing controls and treatments for increased nutrients and decreased light. Nitrate availability was found to be limiting to phytoplankton growth during spring and summer. Small- to medium-sized, chain-forming diatom taxa such as Chaetoceros sp., Skeletonema sp., Pseudonitzschia sp. and Thalassiosira sp. responded most rapidly to nitrate enrichment, increasing their biovolume up to 32-fold during the 5-day experiments. A long-term phytoplankton monitoring database showed that these taxa have historically dominated the phytoplankton assemblage, suggesting that intense competition for nitrate is a key component in structuring the phytoplankton community. Many of the taxa that were able to withstand light reduction in the shaded treatments were rare historically in Pelorus Sound, suggesting that light is secondary to nitrate availability in structuring the phytoplankton community composition in this coastal embayment.     

Light and dark assimilation of nitrate in plants

Abstract. Heterotrophic assimilation of nitrate in roots and leaves in darkness is closely linked with the oxidative pentose phosphate pathway. The supply of glucose-6-phosphate to roots and chloroplasts in leaves in darkness is essential for assimilation of nitrite into amino acids. When green leaves are exposed to light, the key enzyme, glucoses-phosphate dehydrogenase, is inhibited by reduction with thioredoxin. Hence the dark nitrate assimilatory pathway is inhibited under photoautotrophic conditions and replaced by regulatory reactions functioning in light. On account of direct photo-synthetic reduction of nitrite in chloroplasts and availability of excess NADH for nitrate reduclase, the rate of nitrate assimilation is extremely rapid in light. Under dark anaerobic conditions also nitrate is equally rapidly reduced to nitrite on account of abolition of competition for NADH between nitrate reductase and mitochondrial oxidation.     

Germination characteristics of some plant species from calcareous fens in southern Germany and their implications for the seed bank

Seeds of 25 plant species from calcareous fen hay meadows were exposed to different experimental conditions and their germination was characterised. Constant temperature inhibited germination especially in Cyperaceae . Both gibberellic acid and potassium nitrate failed to terminate dormancy. Increased germination rates were found in dicot species after treatment with gibberellic acid. Temperature fluctuations increased germination of Cyperaceae as well as dicotyledons. Treatment with gibberellic acid removed the chilling requirement in some of the species. Dormancy of small seeds with thin seed coats was broken by the application of gibberellic acid or fluctuating temperature; large thick-coated seeds were unaffected by gibberellic acid. No obligatory darkness requirement was found in any species; three species germinated irrespective of light treatments. All other species achieved higher percentage germination in daylight or in red (670 nm) light. Permanent darkness and far-red light (730 nm) reduced germination drastically. The results indicate that germination characteristics of the species investigated can be related to their seed bank types.     

台湾海峡上升流区浮游植物对营养盐添加的响应

2006年6月在台湾海峡近岸上升流区通过表层水体营养盐添加的现场培养实验,研究该海区营养盐限制情况及其浮游植物水华产生的主要影响因素.对营养盐,叶绿素a浓度和浮游植物细胞丰度进行了测定,结果表明,实验中不存在明显的硅限制;氮磷营养盐均存在明显的限制,且氮限制情况更为严重.营养盐添加后,冰河拟星杆藻(Asterionellopsis glacialis)等硅藻迅速生长成为优势藻种,其对氮磷的利用机制有所不同.对氮营养盐采取吸收后迅速同化利用,相较于硝酸盐的补充,氨氮补充条件下优势硅藻更易迅速生长并迅速死亡;对磷营养盐的利用则由于体内磷库的存在,采用迅速吸收后贮存在体内慢慢消耗的利用机制.氮营养盐的补充是上升流期间浮游植物水华产生的主要因素.     

Dependence of phytoplankton assimilation quotients on light and nitrogen source: implications for oceanic primary productivity

Photosynthetic production of oxygen by phytoplankton assemblagedominated by Peridinium in Lake Kinneret, Israel, generallyexceeds the molar equivalent rate of carbon assimilation. Carbonassimilation occurs only if oxygenic photosynthesis exceedsa light-dependent threshold. Assimilation quotients (mol C molO₂^–1) are a variable function of irradiance, and typicallyonly about one-half of the photoreductant produced during oxygenicphotosynthesis is used for reduction of carbon dioxide. Mostof the residual oxygenic photoreductant probably is used forlight-dependent reduction of nitrate, which competes with carbondioxide for oxygenic photoreductant. Nitrate is an importantsource of nitrogen for this algal assemblage, and light-dependentnitrate reduction probably is much larger than carbon dioxidereduction at lowest irradiances in the euphotic zone. Oxygenproduction also may be much larger than carbon assimilationat low light levels in other environments where oxidized formsof nitrogen are important nitrogenous nutrients for phytoplankton,as in the lower euphotic zone of the sea, where low rates ofcarbon assimilation by phytoplankton have been thought to beinconsistent with the amount of oxygen that accumulates duringsummer.     

Utilisation of organic compounds by osmotrophic algae in an acidic lake of Patagonia (Argentina)

We investigated whether algal osmotrophy in naturally acidic Lake Caviahue is an important process for acquisition of organic carbon and organic nitrogen. To accomplish this, we quantified algal assimilation of organic compounds, measured the specific growth rate and biomass yield, and documented incorporation of organic compounds by phytoplankton in situ using microautoradiography. Substrate uptake quantification and microautoradiographic investigations were performed using ³H-leucine, ³H-glucose, ³H-thymidine, ¹⁴C-aspartic acid, ¹⁴C-acetic acid and ¹⁴C-bicarbonates. The results showed that the most important species of the phytoplankton community, Keratococcus rhaphidioides and Watanabea sp., took up various sources of organic carbon and nitrogen under both light and dark conditions. They were also able to assimilate leucine, thymidine, aspartic acid and acetate under high levels of inorganic nitrogen and phosphorus, while they could use leucine, arginine, glutamine and glucose under low levels of nitrogen and phosphorus. The assimilation rates were higher in light than in darkness, and the algal specific growth rates increased when organic sources were added. We proposed that osmotrophy complements the main photosynthetic process of the phytoplankton in Lake Caviahue, which helps to overcome the scarcity of light and inorganic nitrogen and carbon in the water column.     

Seasonal variation in the diel vertical distribution of the migratory alga Cryptomonas marssonii (Cryptophyceae) in a small,highly humic lake

The diel vertical distribution patterns of a migratory alga Cryptomonas marssonii in a small, steeply stratified humic lake were investigated during a summer season (10 diurnal experiments between May and September) using a close-interval Blakar-type sampler. The results indicate that the cells were phototactic; they were typically concentrated at the surface or subsurface during daylight, while in darkness the highest densities were recorded in deeper water, usually near the upper limit of anoxia. During a dense blue-green bloom in August the cells of C. marssonii were also concentrated by day into the same water layer, where oxygen was depleted. However, the cells seemed to avoid totally anoxic water. Because the vertical distribution pattern of C. marssonii had special diurnal and seasonal characteristics, care is needed when designing a sampling programme for a phytoplankton population dominated by this species.     

Modelling the interactions between ammonium and nitrate uptake in marine phytoplankton

An empirically based mathematical model is presented which can simulate the major features of the interactions between ammonium and nitrate transport and assimilation in phytoplankton. The model (ammonium-nitrate interaction model), which is configured to simulate a generic microalga rather than a specified species, is constructed on simplified biochemical bases. A major requirement for parametrization is that the N:C ratio of the algae must be known and that transport and internal pool sizes need to be expressed per unit of cell C. The model uses the size of an internal pool of an early organic product of N assimilation (glutamine) to regulate rapid responses in ammonium-nitrate interactions. The synthesis of enzymes for the reduction of nitrate through to ammonium is induced by the size of the internal nitrate pool and repressed by the size of the glutamine pool. The assimilation of intracellular ammonium (into glutamine) is considered to be a constitutive process subjected to regulation by the size of the glutamine pool. Longer term responses have been linked to the nutrient history of the cell using the N:C cell quota. N assimilation in darkness is made a function of the amount of surplus C present and thus only occurs at low values of N:C. The model can simulate both qualitative and quantitative temporal shifts in the ammonium-nitrate interaction, while inclusion of a derivation of the standard quota model enables a concurrent simulation of cell growth and changes in nutrient status. <br>     

闽南-台湾浅滩近岸上升流区浮游植物碳同化速率的研究

本文采用~(14)C同位素示踪法,测定了浮游植物光合作用速率,结合浮游植物细胞含碳量,计算了不同季节、不同深度下浮游植物碳同化速率常数,讨论了不同环境条件对浮游植物碳同化速率常数的影响。结果表明,闽南-台湾浅滩近岸上升流的形成,是该海域高生产力的主要原因,上升流期间浮游植物复制时间要比非上升流期间浮游植物复制时间缩短1.8倍。同时还表明,温度,光照强度、营养盐是控制浮游植物生长的主要因子。上升流期间营养盐始终保持在较高的水平。是上升流区具有较大的浮游植物碳同化速率常数(3.3d~(-1))的原因之一。适宜该海域浮游植物生长的光照强度在3 000—15 000lx之间,温度的影响可用Goldman和Carpenter模式近似表示。     

Dark Leaf Respiration in Light and Darkness of an Evergreen and a Deciduous Plant Species

Dark respiration in light as well as in dark was estimated for attached leaves of an evergreen (Heteromeles arbutifolia Ait.) and a deciduous (Lepechinia fragans Greene) shrub species using an open gas-exchange system. Dark respiration in light was estimated by the Laisk method. Respiration rates in the dark were always higher than in the light, indicating that light inhibited respiration in both species. The rates of respiration in the dark were higher in the leaves of the deciduous species than in the evergreen species. However, there were no significant differences in respiration rates in light between the species. Thus, the degree of inhibition of respiration by light was greater in the deciduous species (62%) than in the evergreen species (51%). Respiration in both the light and darkness decreased with increasing leaf age. However, because respiration in the light decreased faster with leaf age than respiration in darkness, the degree of inhibition of respiration by light increased with leaf age (from 36% in the youngest leaves to 81% in the mature leaves). This suggests that the rate of dark respiration in the light is related to the rate of biosynthetic processes. Dark respiration in the light decreased with increasing light intensity. Respiration both in the light and in the dark was dependent on leaf temperature. We concluded that respiration in light and respiration in darkness are tightly coupled, with variation in respiration in darkness accounting for more than 60% of the variation in respiration in light. Care must be taken when the relation between respiration in light and respiration in darkness is studied, because the relation varies with species, leaf age, and light intensity.     

Energy balance of a plant under normal and unfavourable conditions

Two main components of plant energy balance are analyzed--photosynthesis and dark respiration. Different plant species, growing in optimal conditions and differing in photosynthetic metabolism, productivity and potential resistance to stress, have constant ratio between total dark respiration and grossphotosynthesis. The ratio about 38-40% at the phase of active growth. This value reflect plant state, when income (assimilation) is maximized, and expense (total oxidation) is minimized. Intensities of dark respiration of plants in darkness and light are similar despite the fact that the plants have considerable differences: 1) different carbon sources--young assimilates in light and reserve ones in the dark; 2) biochemical modifications (or inhibition) of some stages of dark respiration in light; 3) intensification of alternative respiration in unfavourable conditions differing in dark and light variants. Ratio between intensity of dark respiration and photosynthesis increases in plants growing in unfavourable conditions. This increase is more considerable in plants that are less resistant to the given stress. Characters of energy balance can be used for estimation of physiological status of plants, prediction of resistance and potential productivity at seedling stage.     

Asynchronous vertical migration and bimodal distribution of motile phytoplankton

Some motile phytoplankton have the capability to exploit deepsources of nutrients in a vertical migration cycle: photosynthesisin the near-surface layer, transit to depth, uptake of the limitingnutrient and transit back to the surface layer. If all foursteps can be completed within 24 h, then migrations can be synchronizedto the day/night cycle to maximize photosynthetic efficiency.Alternatively, if physiological, behavioral or environmentalfactors make it impossible for the cycle to be completed in24 h, then migration may be asynchronous. Many observationsof phytoplankton reveal bimodal vertical distributions of organisms,with maxima near the surface and the nutricline. We demonstratehow bimodal vertical distributions of phytoplankton may be symptomaticof asynchronous vertical migration using a Lagrangian Ensemblenumerical model. We simulate vertical migration of the dinoflagellateAlexandrium fundyense in conditions similar to those in theGulf of Maine, where bimodal distributions of A. fundyense havebeen observed. Migration is regulated by internal nutritionalstate—organisms swim down toward the nitracline when depletedof nitrogen, and return to the surface after nutrient uptake.We test the sensitivity of the results to growth rate, nitrogenuptake rate and swimming speed, and find that organism distributionscan be bimodal or unimodal depending on conditions. Finally,we develop an analytical estimate for population distributionbased on organism characteristics and nutricline depth.     

N‐ASSIMILATION IN THE NOXIOUS FLAGELLATE HETEROSIGMA CARTERAE (RAPHIDOPHYCEAE): DEPENDENCE ON LIGHT,N‐SOURCE,AND PHYSIOLOGICAL STATE1

The capabilities of the diel vertically migrating flagellate Heterosigma carterae Hulburt for assimilating ammonium and nitrate into cell‐N in light and in darkness were studied using cells of different N‐status. Ammonium utilization in darkness, except by N‐replete cells, attained>50% of use in the light. However, the capacity to use nitrate was poor in darkness, and less than 20% of nitrate‐N that was taken up in darkness was then actually incorporated into cell‐N. The ability to assimilate N in darkness improved as N‐status (N:C) declined, concurrent with an increasing content of water‐soluble carbohydrate. This carbohydrate was used to support dark N‐assimilation. Cells held in darkness for over a day and that had halted nitrate‐uptake were still capable of taking up ammonium. Furthermore, the act of taking up ammonium appeared to make available a source of C to support nitrate uptake that was previously unavailable. The implications of these results for the ecophysiology of this organism and for the construction of mathematical models of algal growth are considered.     

Prediction of proton exchange and bacterial growth on various substrates using constraint-based modeling approach

Proton exchange between cells and medium is one of the most important factors affecting culture pH, and hence its prediction is advantageous. In this research, proton exchange flux across the cell membrane was predicted using a genome-scale model. Calculated proton exchange flux was then exploited as a criterion to predict the trends and intensities of pH changes in cultures of Bacillus subtilis containing various C-sources, i.e. glucose, sucrose, glycerol, lactate, and citrate, as well as N-sources, i.e. ammonium chloride, sodium nitrate, urea, and histidine. The results showed that glucose, sucrose, and glycerol lowered culture pH as compared to citrate and lactate, which raised it. With regard to N-sources, the model predicted that ammonium chloride lowered culture pH while other sources raised pH. Furthermore, maximum theoretical biomass yield using the various C&N-sources was calculated, and sensitivity of microbial growth to proton exchange flux was investigated using robustness analysis to identify the effect of pH on growth of B. subtilis using different substrates. Finally, the effect of ammonium nitrate, a widely used nitrogen source, on growth of B. subtilis was studied. Experimental data obtained by cultivation of B. subtilis DSM 3256 on mineral salt media containing various C&N-sources were used to confirm model predictions. Model predictions were in good agreement with the experimental results for all of the examined C-sources as well as ammonium chloride and sodium nitrate as N-sources. However, the predictions for the N-sources urea and histidine showed deviations, possibly because these two compounds serve as both C&N-sources.     

PHYSICAL AND CHEMICAL CONDITIONS SURROUNDING THE DIURNAL VERTICAL MIGRATION OF CRYPTOMONAS SPP. (CRYPTOPHYCEAE) IN A SEASONALLY STRATIFIED MIDWESTERN RESERVIOR (USA)

Cross Reservoir, a small mesotrophic reservoir located at the University of Kansas Ecological Reserves (Kansas, USA), contained a dense metalimnetic community of algae and photosynthetic bacteria between early July and late October 1997–1999. Within this community, various Cryptomonas species, primarily C. erosa (Ehrenberg), C. erosa var. reflexa (Marsson), and C. rostratisformis (Skuja), diurnally migrated as indicated by in situ fluorescence monitoring and direct phytoplankton enumeration. The Cryptomonas spp. typically resided near the oxic–anoxic boundary of the water column; however, they actively migrated upward during the day and descended to lower anoxic locations at night, apparently responding to diurnal changes in their local habitat. Their nocturnal environment had moderate levels of sulfide, elevated secondary nutrients, and a community of anoxygenic phototrophic bacteria, whereas their daytime environment had higher light, lower nutrients, and no local photosynthetic bacteria. Monitoring indicated that the Cryptomonas spp. migration was generally linked to daily variations in absolute light intensity (e.g. sunny vs. cloudy days) and the level of other potentially growth-limiting resources, particularly nitrogen and phosphorus. However, further analyses showed that the primary factor that determined whether the Cryptomonas spp. migrated or not on a given day was the slope of the light gradient immediately above the Cryptomonas spp. peak.