Effects of different phosphorus concentrations and N/P ratios on the growth and photosynthetic characteristics of Skeletonema costatum and Prorocentrum donghaiense

The effects of different phosphorus(P) concentrations(0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen(N)/P ratios on the growth and photosynthetic characteristics of S keletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P(HP) concentration group showed the greatest algal density and highest specifi c growth rate. Changes in the maximum effi ciency of photosystem Ⅱ(F _v/F_m) were monitored under the various P and N/P conditions. The largest decrease in F _v/F_m was in the low-P(LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields(Φ _(PSⅡ)) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinofl agellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _(PSⅡ) values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang(Yangtze) River estuary from a photosynthesis perspective.     

Influence of salinity and nitrogen content on production of dimethylsulfoniopropionate (DMSP) and dimethylsulfide (DMS) by Skeletonema costatum

The effects of changing salinity and nitrogen limitation on dimethylsulfoniopropionate(DMSP) and dimethylsulfide(DMS) concentrations were investigated in batch cultures of coastal diatom Skeletonema costatum,an ecologically important species.Changes in salinity from 20-32 caused no measurable variation in cell growth or culture yield,but increased intracellular DMSP per cell by 30%.Nitrogen limitation caused up to a two-fold increase in total DMSP per cell and up to a three-fold increase in DMS per cell.These changes in DMSP and DMS per cell in the Skeletonema costatum cultures with nitrogen limitation and changing salinity were primarily attributed to the physiological functions of DMSP as an osmolyte and an antioxidant.The data obtained in this study indicated that nitrogen limitation and salinity may play an important role in climate feedback mechanisms involving biologically derived DMS.     

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide （DMS）, acrylic acid （AA）, and dimethylsulfoniopropionate （DMSP） during growth of three microalgae： Prorocentrum micans, Gephyrocapsa oceanica, and Platymonas subcordiformis. The DMSP, AA, and DMS concentrations in culture media varied significantly among algal growth stages, with the highest concentrations in the late stationary growth stage or the senescent stage. In the stationary growth stage, the average DMSP concentration per cell in P. micans （0.066 5 pmol/cell） was 1.3 times that in G. oceanica （0.049 5 pmol/ cell） and 20.2 times that in P. subcordiformis （0.003 29 pmol/cell）. The average concentrations of AA were 0.044 6, 0.026 9, and 0.003 05 pmol/cell in P. micans, G. oceanica, and P. subcordiformis, respectively, higher than the concentrations of DMS （0.272, 0.497, and 0.086 2 fmol/cell, respectively）. There were significant positive correlations between cell density and AA, DMSP, and DMS concentrations. The ratios of DMS/AA and AA/（DMSP＋AA） in the three algae differed significantly over the growth cycle. In all three microalgae, the DMS/AA ratios were less than 25% during the growth period, suggesting that the enzymatic cleavage pathway, which generates DMS, was not the main DMSP degradation pathway. The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence. In all three microalgae, the AA/（DMSP＋AA） ratio （degradation ratio of DMSP） decreased during the exponential growth phase, and then increased. The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.     

The effect of three culture methods on intensive culture system of pacific white shrimp (Litopenaeus vannamei)

Different culture methods may affect the intensive culture system of Pacific white shrimp(Litopenaeus vannamei) regarding water quality and growth and economic performance.This study evaluated the potential effects of three culture methods through cultivation of juvenile shrimps under consistent tank management conditions for 84 d.The three methods involved shrimp cultivation in different tanks,i.e.,outdoor tanks with cement bottom(mode-C),greenhouse tanks with cement bottom(mode-G) and outdoor tanks with mud-substrate(mode-M).Results showed that water temperature was significantly higher in mode-G than that in mode-C(P < 0.05).In contrast to the other two treatments,mode-M had stable pH after 50 d cultivation of shrimps.In the mid-late period,the average concentrations of TAN,NO2-N,DIP and COD were significantly lower in mode-M and mode-G compared with those in mode-C(P < 0.05).Despite lack of differences in the final shrimp weight among different treatments(P > 0.05),mode-M had significantly higher shrimp yield,survival rate and feed conversion rate(P < 0.05) than other modes.There were significant differences in revenue and net return among different treatments(P < 0.05).These demonstrated that the treatments of mode-G and mode-M were conductive to the intensive culture system of L.vannamei.     

The inhibitory degree between Skeletonema costatum and dinoflagllate Prorocentrum donghaiense at different concentrations of phosphate and nitrate/phosphate ratios

Interactions between Skeletonema costatum (S. costatum) and Prorocentrum donghaiense (P. donghaiiense) were inves-tigated using bi-algal cultures at different concentrations of phosphate (PO4-P) and nitrate/phosphate (N/P) ratios. Experiments were conducted under P-limited conditions and the Lotka-Volterra mathematical model was used to simulate the growth of S. costatum and P. donghaiense in the bi-algal cultures. Both of these two species were inhibited significantly in bi-algal culture. The results of the simulation showed that the inhibitory degree of S. costaum by P. donghaiense was high when the concentration of PO4-P was low (0.1μmolL-1/2 d), but that of P. donghaiense by S. costaum was high with increased PO4-P supply (0.6μmolL-1/2 d). At low concen-tration of PO4-P (0.1μmolL-1/2 d), or high concentration of PO4-P (0.6μmolL-1/2 d) with high N/P ratio (160), the interactions be-tween S. costatum and P. donghaiense were dependent on the initial cell densities of both species. At high concentration of PO4-P (0.6μmolL-1/2 d) with low N/P ratio (25 or 80), S. costatum exhibited a survival strategy superior to that of P. donghaiense. The de-gree of inhibition of P. donghaiense by S. costaum increased with elevated N/P ratio when the medium was supplemented with con-centration 0.1μmolL-1/2 d of PO4-P. The degree of inhibition to P. donghaiense by S. costaum increased with elevated N/P ratio at low concentration of PO4-P (0.1 μmolL-1/2 d). This trend was conversed at high concentration of PO4-P (0.6μmolL-1/2 d). However, the degree of inhibition of S. costaum by P. donghaiense increased with the increased N/P ratio at different PO4-P concentrations (0.1μmolL-1/2 d and 0.6μmolL-1/2 d). These results suggested that both phosphate concentration and N/P ratio affected the competition between S. costaum and P. donghaiense: P. donghaiense is more competitive in environments with low phosphate or high N/P ratio and the influence of N/P ratio on the competition was more significant with lower phosphate concentration.     

The Effect of Three Culture Methods on Intensive Culture System of Pacific White Shrimp （Litopenaeus vannama）

Different culture methods may affect the intensive culture system of Pacific white shrimp （Litopenaeus vannamei） re- garding water quality and growth and economic performance. This study evaluated the potential effects of three culture methods through cultivation of juvenile shrimps under consistent tank management conditions for 84 d. The three methods involved shrimp cultivation in different tanks, i.e., outdoor tanks with cement bottom （mode-C）, greenhouse tanks with cement bottom （mode-G） and outdoor tanks with mud-substrate （mode-M）. Results showed that water temperature was significantly higher in mode-G than that in mode-C （P 〈 0.05）. In contrast to the other two treatments, mode-M had stable pH after 50 d cultivation of shrimps. In the mid-late period, the average concentrations of TAN, NO2-N, DIP and COD were significantly lower in mode-M and mode-G compared with those in mode-C （P 〈0.05）. Despite lack of differences in the final shrimp weight among different treatments （P 〉0.05）, mode-M had significantly higher shrimp yield, survival rate and feed conversion rate （P 〈 0.05） than other modes. There were significant dif- ferences in revenue and net return among different treatments （P 〈 0.05）. These demonstrated that the treatments of mode-G and mode-M were conductive to the intensive culture system ofL. vannamei.     

Production of Pacific White Shrimp Polycultured with Swimming Crab at Different Densities,and Nutrient Budget in the Enclosure System

This study was to evaluate the effect of stocking densities on the nitrogen and phosphorus budgets for a polyculture of the crab Portunus trituberculatus and the Pacific white shrimp Litopenaeus vannamei.The shrimps(initial weight,0.012 g)were cultured at a density of 45 shrimp m^-2,and juvenile crabs(initial weight,0.024 g)were cultured at five densities of 0,3,6,9,and 12 crabs m^-2.The treatments were grouped as C0S45,C3S45,C6S45,C9S45,and C12S45,respectively.Water quality parameters,growth of shrimp and crabs,and nitrogen and phosphorus budget were measured.The results indicated that the shrimp performances in polyculture treatments C3S45 and C6S45 were superior to those in the crab-free treatment(C0S45).The crab in treatments C3S45 and C6S45 exhibited a significantly higher final mean weight and carapace width/length than those in treatments C9S45 and C12S45.The final size and survival of crabs had a negative correlation with the increasing crab stocking density.The contents of total phos-phorus and total nitrogen and the comprehensive contamination index values were higher in the C9S45 and C12S45 treatments than in the other treatments.The conversion ratios of nitrogen for crab and shrimp growth in treatment C3S45 were significantly higher than those in the crab-free treatment.These findings indicate that polyculturing shrimp with crabs at suitable densities can improve productivity,profitability,nutrient utilization,and the environmental quality.From the 60-day treatments,the optimal culture densi-ties were 3-6 crabs m^-2 and 45 shrimps m^-2.     

Effects of ambient DIN:DIP ratio on the nitrogen uptake of harmful dinoflagellate Prorocentrum minimum and Prorocentrum donghaiense in turbidistat

The effects of varying nitrogen (N): phosphorus (P) ratios on the growth and N-uptake and assimilation of the harmful dinoflagellates Prorocentrum minimum and Prorocentrum donghaiense were examined in turbidistat culture experiments. Algal cultures were supplied with media containing PO34? in various concentrations to obtain a wide range of N:P ratios. Experiments to determine rates of N uptake and assimilation of different N sources (NO3?, NH4+, urea and glycine by P. minimum and NO3?, NH4+ by P. donghaiense) were conducted using 15N tracer techniques at each N:P ratio. The growth rates suggested nutrient limitation at both high and low N:P ratios relative to the Redfield ratio. On a diel basis, the growth of both species was regulated by the light-dark cycle, which may be a result of regulation of both light- dependent growth and light-independent nutrient uptake. Maximum growth rates of both species always occurred at the beginning of light phase. In P-rich medium (low N:P ratio), both species had higher N assimilation rates, suggesting N limitation. Low assimilation coefficients at high N:P ratios suggested P limitation of N uptake and assimilation. NO3? and NH4+ contributed more than 90% of the total N uptake of P. minimum. Reduced N sources were more quickly assimilated than NO3?. Highest average daily growth rates were recorded near an N:P ratio of 12 for both species. The N uptake rates of cultures at N:P ratios near Redfield ratio were more balanced with growth rates. The linkage between growth rates and N uptake/assimilation rates were conceptually described by the variation of cell N quota. The N:P ratios affect the N uptake and growth of Prorocentrum spp., and may regulate their bloom progression in eutrophic ecosystems.     

Response of Microcystis aeruginosa FACHB-905 to different nutrient ratios and changes in phosphorus chemistry

Cyanobacterial blooms are a global problem, with their occurrence tightly tied to nutrient loading. We cultured Microcystis aeruginosa FACHB-905 in growth medium with either inorganic(orthophosphate) or organic(β-glycerophosphate or polyphosphate) phosphorus and at different N:P ratios with 50:1, 30:1, 16:1, 4:1 and 1:4, serving as the phosphorus source. Fluorescence parameters were measured to determine the response of cellular responses to nutrient stress. Scanning electron microscopy(SEM) and estimates of antioxidant activity were employed to examine potential mechanisms of physical change. The results demonstrate that inorganic phosphorus was more bioavailable to M. aeruginosa relative to organic phosphorus in culture. The highest cell concentration(2.21×10~6 cells/mL), chlorophyll-a(0.39 pg/cell) and phycocyanin(1.57 pg/cell) quotas and high levels of chlorophyll fluorescence parameters( rETR, E_k, α, φ_(PSⅡ) and F_v/F_m) were obtained when phosphorus was supplied as K_2 HPO_4 at a N:P ratio of 16–30. Organic sources of phosphorus(β-glycerophosphate and polyphosphate) were bioavailable to M. aeruginosa. In addition, too concentrated orthophosphate(N:P=1:4) resulted in the oxidative stress and lipid peroxidation of cell membrane(identified by the antioxidant system activity), and the photosynthetic activity declined consequently. This study has demonstrated the effects of different phosphorus chemistries and N:P ratios on the cyanobacterial growth, photosynthetic activity and cell physiology, which could be an effective tool for predicting cyanobacterial dominance or N-deficiency in natural lakes(due to the superior ability of cyanobacteria for dissolved N and fix atmospheric N in some cases).     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Removal of K^＋, Na^＋, Ca^2＋, and M^g2＋ from saline-alkaline water using the microalga Scenedesmus obliquus

The capability ofScenedesmus obliquus to remove cations （K^＋, Na^＋, Ca^2＋, Mg^2＋） from saline- alkaline water was investigated at different salinities （0, 5, 10, 15, 20, 25） and carbonate alkalinities （0, 5, 10, 15, 20, 25, 30, 35 mmol/L）. K^＋, Na^＋, Ca^2＋, and Mg^2＋ in saline-alkaline water were efficiently removed by S. obliquus. The maximum removal of the cations （29.37 mg for K^＋, 185.85 mg for Na^＋, 23.07 mg for Ca^2＋, 66.14 mg for Mg^2＋） occurred at salinity 25. The maximum removal of K^＋ （2.28 mg）, Na＋ （6.62 mg）, Ca^2＋ （1.01 mg）, and Mg2＋ （0.62 mg） occurred at carbonate alkalinities of 25 mmol/L for K＊, 35 mmol/L for Na＋, 20 mmol/L for Ca2＋, and 25 mmol/L for Mf＋, respectively. Under a salinity stress, the concentration of Na＇ in S. obliquus increased significantly, while that of K~ decreased significantly. The concentrations of Ca^2＋ and Mg2＋ decreased as well. The ratios of K＋/Na~, Ca2＋/Na^＋, and Mg^2＋/Na^＋ were significantly lower in all salinity treatments than those of the control. Under alkaline stress, the concentrations of Nan and K＋ in S. obliquus decreased significantly and the ratios of K^＋/Na^＋, Ca2＋/Na^＋, and Mg^2＋/Na^＋ were significantly higher in all treatments than in the control. Moreover, the concentrations of Ca2＋ and Mg2＋ in S. obliquus at alkalinities of 5-10 mmol/L were significantly higher than those of the other treatments. The removal of Na＋ by S. obliquus mainly occurs through biosorption, and Mg^2＋ and Ca^2 ＋ were removed through both biosorption and bioaccumulation.     

Effects of Nitrogen Sources and Concentrations on the Growth of Different Phytoplankton Taxa

Effects of different nitrogen(N) compounds and concentrations on the growth of the three different phytoplankton taxa, Skeletonema costatum(Bacillariophyceae), Prorocentrum micans(Dinophyceae), and Chattonella marina(Raphidophyceae), were investigated. The Monod equation was applied to examine effects of N concentrations on the growth of algal cells. Results showed that nitrate(NO_3~-N) and urea served as good N sources for the three phytoplankton taxa. S. costatum grew well on all of the seven N sources. C. marina can effectively use the two amino acids, glycine(Gly) and serine(Ser), however cannot utilize alanine(Ala), threonine(Thr), and asparaginic acid(Asp). P. micans cannot grow in five amino acid substrates. All of the three phytoplankton taxa grew well under different proportions of urea-N, and C. marina grew significantly better in medium with both NO_3~-and urea-N. The values of maximum growth rate(μmax) and half-saturation nutrient concentration(KS) for NO_3~-N were 0.71 divisions d~(-1) and 53.55 μmol L~(-1) for S. costatum, 0.67 divisions d~(-1) and 23.31 μmol L~(-1) for P. micans, and 0.23 divisions d~(-1) and 17.57 μmol L~(-1) for C. marina, respectively. The results suggested that S. costatum had a high N demand for growth, and was capable of using wide ranges of N compounds. The strategy of N utilization for S. costatum may make this species an advantage in N-enriched sea areas especially the dissolved organic nitrogen(DON) rich coastal waters, which might be the reason why S. costatum widely distributes in the cosmopolitan coastal and estuarine sea areas.     

Scaling of soil carbon,nitrogen, phosphorus and C:N:P ratio patterns in peatlands of China

Inspired by the importance of Redfield-type C:N:P ratios in global soils,we looked for analogous patterns in peatlands and aimed at deciphering the potential affecting factors.By analyzing a suite of peatlands soil data(n = 1031),mean soil organic carbon(SOC),total nitrogen(TN) and total phosphorous(TP) contents were 50.51%,1.45% and 0.13%,respectively,while average C:N,C:P and N:P ratios were 26.72,1186.00 and 46.58,respectively.C:N ratios showed smaller variations across different vegetation coverage and had less spatial heterogeneity than C:P and N:P ratios.No consistent C:N:P ratio,though with a general value of 1245:47:1,was found for entire peatland soils in China.The Northeast China,Tibet,Zoigê Plateau and parts of Xinjiang had high soil SOC,TN,TP,and C:P ratio.Qinghai,parts of the lower reaches of the Yangtze River,and the coast zones have low TP and N:P ratio.Significant differences for SOC,TN,TP,C:N,C:P and N:P ratios were observed across groups categorized by predominant vegetation.Moisture,temperature and precipitation all closely related to SOC,TN,TP and their pairwise ratios.The hydrothermal coefficient(RH),defined as annual average precipitation divided by temperature,positively and significantly related to C:N,C:P and N:P ratios,implying that ongoing climate change may prejudice peatlands as carbon sinks during the past 50 years in China.     

Allelopathic interactions between Prorocentrum micans and Skeletonema costatum or Karenia mikimotoi in laboratory cultures

Algal allelopathy is an ecological/physiological phenomenon that has focused attention on the interactions among algae and the production of algal toxins. We investigated the allelopathic interactions between the dinoflagellate genus Prorocentrum micans and diatom genus Skeletonema costatum and between P. micans and dinoflagellate genus Karenia mikimotoi using bi-algal cultures. Because the effects were species-specific and size-dependent, we evaluated the effect of different initial densities. At low densities of P. micans and high densities of S. costatum inoculated into the same medium, the growth of P. micans was weakly restrained, whereas the growth of S. costatum was significantly suppressed. S. costatum and K. mikimotoi were strongly inhibited by P. micans, in both the bi-algal cultures and enriched filtrates. Direct cell-to-cell contact was not necessary to gain a competitive advantage, thus, our results suggest that P. micans inhibited the growth of S. costatum and K. mikimotoi by the release of allelochemical(s). Last, a mathematical model was used to simulate growth and interactions between P. micans and S. costatum and between P. micans and K. mikimotoi in bi-algal cultures.     

Phytoplankton assemblage of Yangtze River Estuary and the adjacent east China sea in summer, 2004

A cruise was conducted from late August to early September 2004 with the intention of obtaining an interdisciplinary understanding of the Yangtze River Estuary including the biological, chemical and physical subjects. Water sample analysis indicated that total phytoplankton species richness was 137. Of them 81 were found in Bacillariophyta and 48 in Pyrrophyta, accounting for 59.1% and 35.0% respectively. The average cell abundance of surface water samples was 8.8×104 cells L-1, with the maximum, 102.9×104 cells L-1, encountered in the area (31.75°N, 122.33°E) and the minimum, 0.2×104 cells L-1, in (30.75°N, 122.17°E). The dominant species at most stations were Skeletonema costatum and Proboscia alata f. gracillima with the dominance of 0.35 and 0.27. Vertical distribution analysis indicated that obvious stratification of cell abundance and dominant species was found in the representative stations of 5, 18 and 33. Shannon-Wiener index and evenness of phytoplankton assemblage presented negative correlation with the cell abundance, with the optimum appearing in (30.75°N, 122.67°E). According to the PCA analysis of the environmental variables, elevated nutrients of nitrate, silicate and phosphate through river discharge were mainly responsible for the phytoplankton bloom in this area.     

Photosynthetic efficiency, cell volume, and elemental stoichiometric ratios in Thalassirosira weissflogii under phosphorus limitation

Nutrient limitation is known to inhibit growth and metabolism and to alter elemental stoichiometric ratios in phytoplankton. In this study, physiological changes in Thalassirosira weissflogii were measured under different dissolved inorganic phosphate (DIP) regimes in semi-continuous cultures to revisit the utility of the Redfield ratio for assessing nutrient limitation. The results showed that cell size increased with decreasing DIP availability. In the P-depleted treatment (f/2-P) the cell size was 1.48 times larger than that in the P-limited (f/100) treatment and 2.67 times larger than that in the P-saturated treatment (f/2 and f/10). The fucoxanthin to chlorophyll a ratio (Fuco/chl a) was relatively stable (about 0.3) in P-saturated cultures and was 10 times higher than that in P-limited and P-depleted cultures. During the experimental period, the photosynthetic efficiency index, ?F/Fm′, was relatively stable at ~0.50 in the P-saturated cultures, but quickly declined with decreasing DIP availability. Although cellular P content showed a significant difference between the P-saturated culture (1.6 pg/cell) and the P-limited culture (0.7 pg/cell), the N/P ratio in T. weissflogii did not show a trend with DIP availability and fluctuated slightly around 25. Our results suggest that cell division in T. weissflogii is not strictly size-gated but is probably regulated by a biochemical, and hence, an elemental stoichiometric ratio threshold, and that deviation of the cellular N/P ratio from the Redfield ratio is not a reliable indicator of algal nutrient stress.     

Responses of Maximum Photosystem H PhotochemicalEfficiency of Phytoplankton Communities to NutrientLimitation in the Coastal Sea of Qingdao,China

Nutrient enrichment experiments with nitrogen （N） and phosphorus （P） were conducted with samples from two stationsin the coastal waters of Qingdao, China, during summer to identify limiting nutrients. In late July of 2009, low P concentrations andthe maximum photochemical efficiency of photosystem II （Fv/Fm） in the initial samples together with Fv/Fm and chlorophyll a （Chl a）responses to P addition indicated P limitation at the two stations. In early August, low P levels still limited phytoplankton growth atstation A. Fv/Fm and Chl a were the highest in the NP treatments at station B, suggesting an N/P co-limitation. In mid-September,nutrient concentrations and Fv/Fm were elevated and phytoplankton communities were healthy. Greater Fv/Fm and Chl a in the treat-ments with added P than those without the addition suggested potential P limitation at station A. Lack of Fv/Fm and Chl a responsesfollowing nutrient additions indicated N and P repletion at station B. At the end of July 2010, neither N nor P was limited at station B.Additionally, Fv/Fm coupled with 24-h-long nutrient enrichment experiments can be used to detect P limitation and N/P co-limitationto natural populations. This method can be more accurate for assessing co-limitation than the use of criteria of nutrient concentrationsand ratios as indicators, and can provide more rapid results than nutrient addition bioassays using chlorophyll response as an indica-tor, when a population is potentially limited. Compared with the two conventional methods, the results based on F,/F~ can also pro-vide more detailed information about physiological states of the populations.