Sodium-dependent amino acid transport in the chlorophyte Platymonas subcordiformis

Sodium-dependent transport of alanine and serine by Platymonas subcordiformis was demonstrated by evaluating the kinetics of entry of these substrates over a range of concentrations of ambient sodium. The reciprocal of the concentration at which entry rate was half maximal (K _t ) was linearly related to ambient sodium concentration; maximum entry (V _max) was not affected. Entry of labeled amino acids as measured by determining radioactivity in the medium was close to but slightly less than rates of net entry as determined by high-performance liquid chromatography. This difference presumably reflects extrusion of labeled carbon by the cells in a form not detectable by the chromatographic techniques employed. The coupling coefficient (sodium ions per amino acid molecule) for entry of alanine and serine was evaluated and determined to be 2. Methylaminoisobutyric acid was not taken up by P. subcordiformis and had no inhibitory effect on uptake of alanine or serine. B-2-aminobicycloheptane-2-carboxylic acid competitively inhibited uptake of both substrates. The internal sodium concentration of P. subcordiformis was measured, and the maximum gradient energetically favorable for amino acid transport was calculated from data in the present work and drawn from the literature. Assuming that uptake of amino acids is strictly sodium-dependent, an amino acid concentration gradient of the order of 10⁶ (cell:medium) can be achieved. This concentration differential permits net uptake of amino acids by P. subcordiformis from ambient amino acid levels in the nanomolar range.     

大气CO2体积分数升高对植物N素吸收的影响

从影响植物N素吸收的因素来看，大气CO2体积分数升高条件下植物净光合作用增强，碳同化产物增多，利于改善N素吸收的能量和物质基础：植物根系生长增强，生物量增多且空间分布加大，有利于N素吸收；但土壤有效N供应能力的变化存在增强和减弱两种观点。从植物N素吸收的实际情况来看，大气CO2体积分数升高条件下植物N吸收总量并末增加，植物体内N质量分数普遍降低，某些种类植物N吸收形态也发生了改变。因此要阐明大气CO2体积分数升高对植物N素吸收的影响机制，必须探明土壤有效N供应能力的变化：CO2体积分数升高条件下N矿化作用是否增强，微生物和植物间是否存在对有效N的竞争，此外，CO2体积分数升高条件下植物根系形态特征变化和N素吸收(包括主动和被动吸收)的生理机制及其与环境因素的关系也值得进一步研究。     

Effect of nitrogen source on short-term light and dark CO2 uptake by a marine diatom

Based on a series of short-term incubations involving the marine diatom Chaetoceros simplex (Bbsm), precultured in NH ₄ ⁺ -, NO ₃ ^- -and urea-limited continuous cultures at several dilution rates, we found that both the short-term specific rate of ¹⁴CO₂ uptake and the amount of CO₂ fixed after 8- and 16-min incubations were unaffected by enrichment with NH ₄ ⁺ , urea, or NO ₃ ^- when NH ₄ ⁺ or urea were the preconditioning forms of N, but were slightly suppressed when the cells were first grown on NO ₃ ^- . Similar enrichments in the dark, however, led to significant CO₂ uptake under all conditions of NH ₄ ⁺ enrichment and to similarly enhanced CO₂ uptake, but only at high growth rates, when urea was the source of enrichment nitrogen. Our light results are contrary to some contemporary findings, but there does seem to be agreement that photosynthetic rates of rapidly growing phytoplankton will not be affected by exposure to pulses of nitrogen. Enhanced dark uptake, in contrast, appears to be characteristic of phytoplankton under all degrees of N limitation, and, as such, may be useful as an “all or nothing” index of the nitrogen status of natural waters. There is some indication that the index may be useful in determining both the form of and the degree of N limitation as well.     

Kinetics of glucose and amino acid uptake by attached and free-living marine bacteria in oligotrophic waters

Kinetics of glucose and amino acid uptake by attached and free-living bacteria were compared in the upper 70 m of the oligotrophic north-western Mediterranean Sea. Potential uptake rates of amino acids were higher than those of glucose in all the samples analysed. Cell-specific potential uptake rates of attached bacteria were up to two orders of magnitude higher than those of total bacteria, both for amino acids and glucose (0.72–153 amol amino acids cell⁻¹ h⁻¹ and 0.05–58.42 amol glucose cell⁻¹ h⁻¹ for attached bacteria and 0.34–1.37 amol amino acids cell⁻¹ h⁻¹ and 0.07–0.22 amol glucose cell⁻¹ h⁻¹ for total bacteria). The apparent K _m values were also higher in attached bacteria than in total bacteria, both for amino acids and glucose. These results would reflect the presence of different uptake systems in attached and free-living bacteria, which is in accordance with the different nutrient characteristics of their microenvironments, ambient water and particles. Attached bacteria show transport systems with low affinity, which characterise a bacterial community adapted to high concentration of substrates. Received: 13 June 2000 / Accepted: 6 December 2000     

Effect of salinity adaptation on nitrogen metabolism in the freshwater fish Tilapia mossambica. I. Tissue protein and amino acid levels

Water, proteins and total free amino acids were estimated in different tissues of the euryhaline fish Tilapia mossambica after adaptation to various strenghts of sea water. The water content did not vary significantly in any tissue on salinity adaptation. The soluble and insoluble proteins displayed a general and considerable decrease in muscle, liver and heart; the decrease in the soluble fraction in the heart and the proteins of the muscle in 75% sea water (100% sea water=32.5 S) were significant. The gill proteins did not alter with salinity; the kidney proteins tended to increase slightly in 100% sea water (SW). The total free amino-acid content decreased insignificantly in all tissues on adaptation to 25% SW; in higher salinities, however, the content increased significantly. This increase was sudden and steep in 50% SW, and gradual and less steep in 75 and 100% SW. It is suggested that constancy in water content may contribute to the great adaptability of T. mossambica to heterosmotic media, and that the total free amino acids may be involved in isosmotic intracellular regulation. The possibility of amino acid increase as a result of protein breakdown is also indicated.     

Effect of nitrogen supply on nitrogen uptake,accumulation and assimilation in Porphyra perforata (Rhodophyta)

Porphyra perforata J. Ag. was collected from a rocky land-fill site near Kitsilano Beach, Vancouver, British Columbia, Canada and was grown for 4 d in media with one of the following forms of inorganic nitrogen: NO ₃ ^- , NH ₄ ⁺ and NO ₃ ^- plus NH ₄ ⁺ and for 10 d in nitrogen-free media. Internal nitrogen accumulation (nitrate, ammonium, amino acids and soluble protein), nitrate and ammonium uptake rates, and nitrate reductase activity were measured daily. Short initial periods (10 to 20 min) of rapid ammonium uptake were common in nitrogen-deficient plants. In the case of nitrate uptake, initial uptake rates were low, increasing after 10 to 20 min. Ammonium inhibited nitrate uptake for only the first 10 to 20 min and then nitrate uptake rates were independent of ammonium concentration. Nitrogen starvation for 8 d overcame this initial suppression of nitrate uptake by ammonium. Nitrogen starvation also resulted in a decrease in soluble internal nitrate content and a transient increase in nitrate reductase activity. Little or no decrease was observed in internal ammonium, total amino acids and soluble protein. The cultures grown on nitrate only, maintained high ammonium uptake rates also. The rate of nitrate reduction may have limited the supply of nitrogen available for further assimilation. Internal nitrate concentrations were inversely correlated with nitrate uptake rates. Except for ammonium-grown cultures, internal total amino acids and soluble protein showed no correlation with uptake rates. Both internal pool concentrations and enzyme activities are required to interpret changes in uptake rate during growth.     

不同氮肥对油菜B与Mo吸收的影响

利用盆栽试验研究了潮土中不同氮肥对油菜硼钼吸收的影响，结果表明，前期(5周)施氮并没有促进油菜的生长，而且大部分氮肥对油菜生长表现抑制作用，氯化铵处理的作用最大，硫酸铵和尿素处理次之，硝酸铵处理再次之；不同氮肥均有抑制油菜生长前期(5周)硼吸收和累积的作用，其中以氯化铵、硝酸铵和尿素的作用为甚；不同氮肥同样抑制油菜生长前期(5周)对钼的吸收和累积，以硫酸铵处理作用最突出，氯化铵处理次之。但随着时间的推移，10、15周不同氮肥大多促进油菜的生长，增加油菜对硼钼的吸收和累积，且各氮肥处理间的差异逐步减小。     

Effect of nitrogen deposition on China's terrestrial carbon uptake in the context of multifactor environmental changes

The amount of atmospheric nitrogen (N) deposited on the land surface has increased globally and by nearly five times in China from 1901 to 2005. Little is known about how elevated reactive N input has affected the carbon (C) sequestration capability of China's terrestrial ecosystems, largely due to the lack of reliable data on N deposition. Here we have used a newly developed data set of historical N deposition at a spatial resolution of 10 km x 10 km in combination with other gridded historical information on climate, atmospheric composition, land use, and land management practices to drive a process-based ecosystem model, the dynamic land ecosystem model (DLEM) for examining how increasing N deposition and its interactions with other environmental changes have affected C fluxes and storage in China's terrestrial ecosystems during 1901-2005. Our model simulations indicate that increased N deposition has resulted in a net C sink of 62 Tg C/yr (1 Tg = 1012 g) in China's terrestrial ecosystems, totaling up to 6.51 Pg C (1 Pg = 10(15) g) in the past 105 years. During the study period, the N-induced C sequestration can compensate for more than 25% of fossil-fuel CO2 emission from China. The largest C sink was found in southeast China, a region that experienced the most significant increase of N deposition in the period 1901-2005. However, the net primary productivity induced by per-unit N deposition (referred to as ecosystem N use efficiency, ENUE, in this paper) has leveled off or declined since the 1980s. This indicates that part of the deposited N may not be invested to stimulate plant growth, but instead leave the ecosystem by various pathways. Except shrubland and northwest/southwest China, signs of N saturation are apparent in the rest major biome types and regions, with ENUE peaking in the 1980s and leveling off or declining thereafter. Therefore, to minimize the excessive N pollution while keeping the N-stimulated C uptake in China's terrestrial ecosystems, optimized management practices should be taken to increase N use efficiency rather than to keep raising N input level in the near future.     

Interactions of inorganic nitrogen in the uptake and assimilation by marine phytoplankton

The effect of ambient ammonium concentration on the nitrate uptake rate of marine phytoplankton was investigated. These studies consisted of laboratory experiments using unialgal species and field experiments using natural phytoplankton communities. In laboratory experiments, ammonium suppressed the uptake rates of nitrate and nitrite. Approximately 30 min were required for ammonium to exhibit its fully inhibitory effect on nitrate uptake. At high ammonium concentration (>3 g-at/l), a residual nitrate uptake rate of approximately 0.006 h^-1 was observed. When the ambient ammonium concentration was reduced to a value less than 1 g-at/l, the suppressed nitrate uptake rate subsequently attained a value comparable to that observed before the addition of ammonium. A range of 25 to 60% reduction in the nitrate uptake rate of natural phytoplankton communities was observed at ambient ammonium concentrations of 1.0 g-at/l. A mechanism is proposed for the suppression of nitrate uptake rate by ammonium through feedback control of the nitrate permease system and/or the nitrate reductase enzyme system. The feedback control is postulated to be regulated by the level of total amino acids in the cell.Contribution No. 936 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA. This paper represents a portion of a dissertation submitted to the Department of Oceanography, University of Washington, Seattle, in partial fulfillment of the requirements for the Ph.D. degree.     

氮输入对沼泽湿地植物生长和氮吸收的影响

氮是湿地植物生长必不可少的营养元素之一,但当外源氮输入超出植物生长需要时,氮素将抑制植物生长。不同植物对氮输入的响应不同,同一植物不同器官对氮输入的响应也不一致。为了探讨氮输入对湿地植物生长和氮吸收的影响机制,本文选取滇西北典型湖泊湿地纳帕海湖滨挺水植物茭草（Zizania caduciflora）和水葱（Scirpus validus）为对象,通过控制实验,研究了3个不同氮输入水平[0 g·m-2·a-1（对照,CK）、20 g·m-2·a-1（N20）、40 g·m-2·a-1（N40）]对茭草和水葱生物量积累、根冠比、氮吸收的影响。结果表明：培养期内,茭草地上生物量始终表现为N40〉N20〉CK,即氮输入促进茭草地上生物量积累;而水葱地上生物量随培养时间不同而发生变化,培养早期N20处理促进水葱地上生物量积累,N40处理抑制水葱地上生物量积累。茭草地下生物量表现为N40〉CK〉N20,即氮输入不足抑制茭草地下生物量积累,足够氮输入促进茭草地下生物量积累;水葱地下生物量表现为CK〉N20〉N40,即氮输入抑制水葱地下生物量积累。植物地上部分和地下部分生长对氮输入的响应也不一致,导致植物根冠比发生变化,茭草根冠比表现为N20     

Amino acid uptake by the toxic dinoflagellate Alexandrium fundyense

A toxic axenic strain of Alexandrium fundyense is shown to be capable of removing dissolved free amino acids (DFAAs) until concentrations are similar (low nM) to those found in natural waters. Uptake is greatest during exponential growth, rather than during C and/or N-stress as is usual in diatoms and other flagellates. A wide range of amino acids can be taken up, their concentration being decreased within a few hours to the levels observed prior to DFAA addition. The maximum rate of DFAA-N uptake, during early exponential phase, was 0.8 pmol-N cell⁻¹ h⁻¹, equivalent to ≃20% of the total N requirement. More typically, the contribution of DFAA-N was only ≃5%. However, these uptake rates are not sustainable. It is apparent that this organism cannot use amino-N to support significant growth, even though it can take up DFAAs. This, and the fact that the composition of the internal amino acid pool differed from that externally, is further evidence that the N-physiology of this genus is abnormal (differences to other dinoflagellates include an abnormally high concentration of glutamine and arginine, an effective absence of amine X, and release of nitrite during the␣concurrent assimilation of nitrate and ammonium in␣darkness). There is no evidence that the use of DFAAs enhance toxin content, except when cells are supplied with very high (unnatural) concentrations of arginine. Received: 8 May 1989 / Accepted: 14 September 1998     

Effect of seawater velocity on inorganic nitrogen uptake by morphologically distinct forms of Macrocystis integrifolia from wave-sheltered and exposed sites

In conditions of low water motion (<0.06 ms^–1), the availability of essential nutrients to macroalgae, and thus their potential productivity, may be limited by thick diffusion boundary-layers at the thallus surface. The ability of macroalgae to take up nutrients in slow moving water may be related to how their blade morphology affects diffusion boundarylayer thickness. For the giant kelp, Macrocystis integrifolia Bory, morphological measurements indicate that blades of plants from a site exposed to wave action are thick, narrow and have a heavily corrugated surface. In contrast, blades from a site with a low degree of water motion are relatively thin, with few surface corrugations and large undulations along their edges. The aim of our work was to test the hypothesis that morphological features of M. integrifolia blades from a sheltered site allow enhanced inorganic nitrogen uptake at low seawater velocities compared to blades with a wave-exposed morphology. The rate of nitrate and ammonium uptake by morphologically distinct blades of M. integrifolia, from sites that were sheltered from and exposed to wave action, were measured in the laboratory at a range of seawater velocities (0.01 to 0.16 ms^–1), between March and May 1993. For both sheltered and exposed blade morphologies, nitrate and ammonium uptake rates increased with increasing seawater velocity, reaching a maximum rate at 0.04 to 0.06 ms^–1. Uptake parameters V _max (maximum uptake rate) and U _0.37 (the velocity at which the uptake rate is 37% of the maximum rate) were estimated using an exponential decay formula. These parameters were similar for both blade morphologies, at all seawater velocities tested. Additional measurements suggest that the nitrogen status of M. integrifolia blades from wavesheltered and exposed sites were similar throughout the experimental period, and thus nitrogen status did not affect the rate of nitrogen uptake in these experiments. on the basis of these results, we conclude that blade morphology does not enhance nitrogen uptake by M. integrifolia in conditions of low water motion. Potential effects of diffusion boundary-layers on kelp productivity are discussed.     

The effect of nitrogen deficiency,methionine and inhibitors of methionine metabolism on the DMSP contents of Tetraselmis subcordiformis (Stein)

The unicellular alga Tetraselmis (Platymonas) subcordiformis (Prasinophyceae) displayed a two-phase increase in DMSP (dimethylsulfoniopropionate) contents in response to nitrogen deficiency. In the first phase, DMSP pools increased by 75% within 24 h. The second increase in DMSP was observed after 14 d and was accompanied by cyst formation. Methionine metabolism was involved in DMSP accumulation. A 2.6-fold increase of the DMSP pool could be induced by supplementing a complete ASP (artifical seawater Provasoli) medium with 100 M L-methionine. Inhibition of methionine utilization by T. subcordiformis caused by the addition of S-adenosylhomocystein, a competitive inhibitor of S-adenosylmethionine-dependent transmethylases, resulted in a 24% increase of DMSP contents after 7 h. DMSP accumulation after hyperosmotic shock was significantly delayed when methionine liberation from proteins was inhibited by a set of protease inhibitors. A model for the mechanism of DMSP accumulation is presented on the basis of these data and additional information from the literature. It is proposed that methionine availability in T. subcordiformis determines the rate of DMSP synthesis.     

接种微生物对油菜吸收Cd效果的影响研究

采用盆栽土壤模拟试验方法,以油菜（Brassica campestris）为供试植物,通过向土壤中接种微生物,研究了四种菌株JA1,JA2,JA3,JA4对油菜吸收Cd的效果影响。结果表明：菌株JA1,JA2,JA4不仅可以促进植株的生长,分别使油菜的生物量提高17%,28%和32%,而且增强了植株对土壤Cd的吸收,使油菜地上部富集量较对照提高了26.8%,48.8%和65.9%,植物提取量较对照增加了48.9%,76.1%和119%,并且可有效促进土壤Cd的活化,分别使土壤Cd的有效态含量提高了25.9%,59.2%和41.9%;菌株JA3则使其植株地上部富集量、植物提取量和土壤Cd有效态含量降低,较对照降低了14.6%,11.8%和13.6%。各处理的叶绿素和可溶性蛋白含量均略高于对照。     

Amino acid uptake and respiration by marine heterotrophs

The concentration and turnover of dissolved free amino acids were measured in samples from 25 and 100 m on three occasions at a station 6 miles off the California (USA) coast. Individual amino acid concentrations varied from undetectable (<0.05 g/l) to 3 g/l, the total amino acid concentration from 1.8 to 8.5 g/l. The greater concentration of total amino acids was always found at 25 m. The predominant amino acids were serine, lysine, aspartate, glutamate and alanine; reliable analyses could not be made for glycine because of a high blank. For the 10 individual amino acids studied, the rate of heterotrophic turnover ranged from undetectable to 1.2 g/l day^-1; serine, aspartate, alanine and glutamate showed the highest rates. In samples from 25 m, the rates were 15 to 20 times higher than those taken from 100 m. The total calculated flux of the amino acids studied varied from 0.015 to 3.2 g/l day^-1 and amounted to 1–10% of photosynthetic carbon dioxide fixation.     

硅酸盐、腐殖酸对Cr污染土壤中小白菜生理及吸铬量的影响

针对在铬污染区土壤中的叶菜类吸铬量是果菜类2~3倍,以及硅酸盐或腐植酸在重金属污染土壤上的最佳施用量研究较少的问题,为保障蔬菜的品质安全,文章通过土壤盆栽试验,研究硅酸盐、腐殖酸不同用量对Cr污染土壤中小白菜(Brassica Chinensis)生理指标和吸铬量的影响.结果表明,施低质量分数的硅酸盐(1.0 g·kg-1)效果最佳,既能提高受Cr污染小白菜的生物量、叶绿素相对含量,又能减轻Cr毒害对抗氧化酶系统中SOD酶活性的抑制,缓解小白菜受到的过氧化胁迫,有效降低小白菜吸铬量;高质量分数硅酸盐(2.0 g·kg-1)对生长产生抑制.腐殖酸在设计的3个质量分数水平上均不能有效降低小白菜吸铬量,对生长的促进作用也不明显.     

Ammonium and free amino acid uptake by a deep-sea mussel (Bathymodiolus sp., undescribed) containing methanotrophic bacterial symbionts

The morphology of the lophophore and mouth was measured for freshly collected colonies of seven common bryozoan reef species at three sites along the central Caribbean coast of Panamá during the dry season, from the end of December 1988 through May 1989. Characters measured or scored include the number of tentacles, circumference of the lophophore, abanal and anal tentacle length, abanal and anal tentacle spacing, and perimeter of the mouth. Taxa studied include three species each of Stylophoma Levinsen, and Steginoporella (Smitt), and one species of Trematooecia Osburn. Individuals were collected from both continental and offshore fringing reefs from depths of 1 to>30 m. Terrestrial runoff from heavy tropical rains, sedimentation, and sea exposure vary greatly between sites. Congeneric species showed small but consistent differences in lophophore size, but little or no difference in spacing of tentacles or perimeter of the mouth. This suggests that closely related species feed on similarly sized particles despite their overall differences in zooidal size. Lopophore morphology also varied intraspecifically between reefs, particularly in the length of the tentacles and spacing between tentacles. Patterns of variation were similar for all species, which implies that species shift their diets in similar directions between the different reefs. The combination of these patterns suggests that niche diversification in feeding does not occur among congeneric species in the cheilostomes examined.     

长期定位施肥对冬小麦吸收利用氮素的影响

研究不同施肥措施对冬小麦（Triticum aestivum L.）吸收利用氮素的影响,为新形势下制定合理培肥施肥技术,减少氮素流失,提高氮素利用效率提供借鉴。以黄淮海平原国家潮土土壤肥力与肥料效益监测站1990年以来的长期定位施肥土壤为基础,研究长期不施肥（CK）、氮钾配施（NK）、氮磷钾配施（NPK）、氮磷钾配施有机肥（MNPK）、氮磷钾配合秸秆还田（SNPK）5种施肥措施对冬小麦吸收利用氮素的影响。研究结果表明,与CK、NK施肥措施相比,NPK、MNPK、SNPK施肥措施有利于小麦各生育时期各器官对土壤氮素的吸收利用,能显著提高小麦各生育阶段的田间群体数、干物质积累量以及有效穗数,有利于灌浆期、成熟期氮素在茎、叶、鞘、穗等器官的累积。小麦对氮素吸收累积量、氮肥利用率以NPK最大,MNPK/SNPK次之,均极显著（P≤0.01）高于CK和NK;氮素吸收累积量大小依次为NPK〉MNPK〉SNPK〉NK〉CK。施NPK、NPK配施有机肥或秸秆还田的施肥措施均有利于冬小麦在整个生育时期对土壤氮素养分的吸收,提高籽粒产量。     

Impact of water on CO2 capture by amino acid ionic liquids

The reversible capture of CO₂ from fossil-fueled industries and the absorption of CO₂ for natural-gas-sweetening purposes are industrial issues closely related to very important environmental, economical, and technological concerns. Biological amino acids can be used for task-specific ionic liquids for reversible CO₂ capture. Several groups have reported efficient and reversible CO₂ capture by such ionic liquids under rigorously dry conditions. However, we have observed that CO₂ capture by amino acid ionic liquids is hugely impacted by the presence of water. In addition, the amino acid anions appear to play only a transitory role in the CO₂ capture in the first minutes of exposure to a wet CO₂ stream. Here, we studied the interaction of two ionic liquids—tetramethylammonium glycinate ([N₁₁₁₁][Gly]) and tetraethylammonium prolinate ([N₂₂₂₂][Pro])—with CO₂ under wet conditions, by ¹³C-NMR. Results show that CO₂ is initially captured in a carbamate form by the amine-functionalized anions of these salts. This capture mode is unambiguously confirmed by a single-crystal X-ray study of the CO₂-ionic liquid complex. However, in solution, as additional CO₂ is added, the carbamate releases the covalently bound CO₂, and the CO₂ remaining in solution shifts in form to an equilibrium mixture of carbonate and bicarbonate. Indeed, when the amount of CO₂ present in the system exceeds about one-half mole per mole of ionic liquid present, the ionic liquid–carbamate complex is detected in only trace amounts, and the neutralized amino acids are readily identifiable by NMR.     

Dark CO2 uptake by the diatom Chaetoceros simplex in response to nitrogen pulsing

In a continuing investigation of dark CO₂ uptake by nitrogen-limited cultures of the marine diatom Chaetoceros simplex (Bbsm), we expanded on several of our earlier conclusions regarding the potential application of this physiological response for measuring the degree and type of nitrogen limitation in phytoplankton populations. First, the duration over which the maximal enhancement of dark ¹⁴CO₂ uptake was sustained after NH ₄ ⁺ enrichment was a function both of the concentration of added NH ₄ ⁺ and the standing crop of phytoplankton nitrogen — in effect, the total N demand. Second, pulsing with NH ₄ ⁺ for a given degree of N-limitation always produced the same level of enhanced dark CO₂ uptake regardless of whether the cultures were preconditioned with oxidized or reduced nitrogen. In contrast, urea pulsing led to reduced dark CO₂ uptake, but the effect was most pronounced in cells grown on NO ₃ ^– . And third, the assay could be used to distinguish readily between no, moderate, and severe N limitation. The degree of severe N limitation was quantitatively correlated with the degree of enhanced dark CO₂ uptake, but this relationship was not so clear in the region of moderate N limitation. The main advantage of the assay is that it is a relatively simple and effective alternative to more complicated techniques for gauging the degree and form of N limitation in phytoplankton. Further evaluation will be required, both in the laboratory and field, before the assay can be calibrated for quantitative use.Contribution No. 5982 from the Woods Hole Oceanographic Institution