Indicator Of Nutrient Limitation Research Articles

Co-Limitation of Phytoplankton by N and P in a Shallow Coastal Lagoon (Ria Formosa): Implications for Eutrophication Evaluation

AbstractThe Ria Formosa coastal lagoon is a highly productive shallow ecosystem in southern Portugal, subjected to nutrient inputs from anthropogenic and natural sources. Nutrients are major abiotic drivers of phytoplankton in this system, but their effects on phytoplankton assemblages and the occurrence of nutrient limitation are still poorly understood. The main goal of this study was, thus, to evaluate the occurrence, type, and effects of nutrient limitation on phytoplankton community and specific functional groups in the Ria Formosa coastal lagoon. We conducted nutrient enrichment experiments with factorial additions of nitrogen (N) and phosphorus (P) using natural phytoplankton assemblages from distinct locations in the Ria Formosa, throughout a yearly cycle. Phytoplankton composition and abundance were evaluated using inverted and epifluorescence microscopies, and spectrophotometric methods were used for biomass. Limitation was defined as higher phytoplankton growth following enrichment with a particular nutrient in relation to the non-enriched control. The most common type of phytoplankton limitation was simultaneous co-limitation by N and P; diatoms, as r-strategists, were the most frequently limited group. Single N and P limitation, and serial P limitation were also observed, as well as negative responses to nutrient enrichment. Group-specific responses to nutrient enrichment were not reflected in the relative abundance of phytoplankton groups within the whole assemblage, due to the numerical dominance of pico-sized groups (cyanobacteria and eukaryotic picophytoplankton). Ambient nutrient ratios and concentrations did not predict phytoplankton nutrient limitation, given the different nutrient utilisation traits among phytoplankton functional groups. Therefore, nutrient ratios should not be used as indicators of nutrient limitation in eutrophication assessment.

The misuse of ratios in ecological stoichiometry.

Ecological stoichiometry is concerned with the ratios of different elements, particularly carbon, nitrogen, and phosphorus. Ratios by their nature do not respond symmetrically to changes in the numerator and denominator and do not follow normal distributions; however, researchers frequently fail to consider these properties in their analyses, which has biased reported results. Calculating means, variance, or linear slopes based on untransformed ratios results in biased results. I demonstrate the consequences of these errors for inferences from stoichiometric analyses using simple examples and several large monitoring data sets. I then review 100 studies in ecological stoichiometry and find that misuse of ratio data is common, with 93% of studies containing at least one error. These errors may be problematic, particularly in large-scale meta-analyses summarizing data over large ranges. Fortunately, most of these mistakes can be easily avoided by first log transforming elemental ratios. I therefore recommend that, to ensure robust and reproducible results, researchers in ecological stoichiometry should adopt a convention of presenting stoichiometric ratio data as the logarithm of molar ratios in the future. The widespread use of untransformed nitrogen to phosphorus ratio as an indicator of nutrient limitation has likely exaggerated the importance of phosphorus limitation, particularly in freshwater systems.

Cell death in individual freshwater phytoplankton species: relationships with population dynamics and environmental factors

ABSTRACT Understanding and predicting changes in phytoplankton populations requires knowledge of losses due not only to sedimentation and grazing, but also to intrinsic processes (here, collectively termed ‘cell death’). Cell death is poorly understood, especially in freshwater phytoplankton, but experiments in culture often suggest involvement of abiotic factors (e.g. temperature, light, nutrients). The occurrence of cell death was examined in a simple, natural environment: a small, well-mixed, temperate, urban pond during a period of phytoplankton growth, from mid-July to mid-November. Abundances of 18 phytoplankton taxa were measured weekly and fluorescence microscopy and staining was used to detect dead cells (using SYTOX which measures loss of membrane integrity) and cells undergoing cell death (using Annexin-V, which measures lipid inversions of membranes, an early signal of cell death). Dead and dying cells occurred in most phytoplankton taxa, but incidence and timing varied considerably, e.g. species like the chlorophyte Ankistrodesmus spiralis showed 20–30% of cells staining with SYTOX and Annexin in late autumn when the population was decreasing, while the dinoflagellate Peridinium sp. showed staining of up to 50% of cells with STYOX throughout the period, and the cyanobacterium Microcystis aeruginosa occasionally showed staining of 100% of cells with SYTOX. Overall, there was some association between cell death staining and growth phase with 10–15% of the total community showing SYTOX and Annexin staining in late autumn, when most populations were declining. Cell death could not be correlated with thresholds or rapid changes in abiotic conditions (e.g. temperature, irradiance) or with indicators of nutrient limitation (e.g. N:P ratios). While abiotic factors have been clearly implicated in cell death within unialgal culture experiments, in natural freshwater ecosystems interactions between biotic factors, such as pathogens or allelopathy, may play greater roles in losses related to cell death and be distinct for different taxa.

Phosphorus availability and turnover in the Chesapeake Bay: Insights from nutrient stoichiometry and phosphate oxygen isotope ratios

AbstractUnderstanding phosphorus (P) availability and its control on eutrophication in the Chesapeake Bay is complicated by variable sources and biogeochemical reactions transforming P forms. We investigated seasonal and spatial variability in P limitation and biological utilization in the Bay using nutrient stoichiometry (of both dissolved and particulate forms), phosphate oxygen isotope ratios, and alkaline phosphatase activity at three sites along the salinity gradient. We demonstrate that particulate nutrient ratios can be used as indicators of nutrient limitation in the Bay and suggest strong seasonal and spatial variability in P availability: the surface water is P limiting in spring, but this condition is alleviated in summer and in the deeper waters. Variability in P limitation is well reflected in the trends of phosphate oxygen isotope composition (δ18OP), with values approaching isotopic equilibrium under P limiting conditions, suggesting rapid biological P turnover. Furthermore δ18OP values suggest multiple phosphate sources including remobilization of terrestrial inorganic P phases and remineralization of organic P and P from both sources is sufficiently cycled by microorganisms, suggested by the extensive equilibrium oxygen isotope exchange. Our results further suggest high P utilization in the deeper euphotic zone where nutrients are abundant, raising caution on studying nutrient availability and limitation only in the surface water.

Relationship between the relative limitation and resorption efficiency of nitrogen vs phosphorus in woody plants.

Most previous studies have ascribed variations in the resorption of a certain plant nutrient to its corresponding environmental availability or level in tissues, regardless of the other nutrients’ status. However, given that plant growth relies on both sufficient and balanced nutrient supply, the nutrient resorption process should not only be related to the absolute nutrient status, but also be regulated by the relative limitation of the nutrient. Here, based on a global woody-plants dataset from literature, we test the hypothesis that plants resorb proportionately more nitrogen (or phosphorus) when they are nitrogen (or phosphorus) limited, or similar proportions of nitrogen (N) and phosphorus (P) when co-limited by both nutrients (the relative resorption hypothesis). Using the N:P ratio in green foliage as an indicator of nutrient limitation, we found an inverse relationship between the difference in the proportionate resorption of N vs P and this foliar N:P ratio, consistent across species, growth-forms, and vegetation-types globally. Moreover, according to the relative resorption hypothesis, communities with higher/lower foliar N:P (more likely P/N limited) tend to produce litter with disproportionately higher/lower N:P, causing a worsening status of P/N availability; this positive feedback may somehow be counteracted by several negative-feedback mechanisms. Compared to N, P generally shows higher variability in resorption efficiency (proportion resorbed), and higher resorption sensitivity to nutrient availability, implying that the resorption of P seems more important for plant nutrient conservation and N:P stoichiometry. Our findings elucidate the nutrient limitation effects on resorption efficiency in woody plants at the global scale, and thus can improve the understanding of nutrient resorption process in plants. This study also suggests the importance of the foliar N:P ratio as a key parameter for biogeochemical modeling, and the relative resorption hypothesis used to deduce the critical (optimal) N:P ratio for a specific plant community.

Response of two wetland graminoids to N:K supply ratios in a two-year growth experiment

Changes in nutrient availability in wetlands have been observed during the recent years, mostly due to human pressure. A shift from N limitation to P or K limitation causes changes in plant species composition, nutrient use efficiency, plant growth, interspecific competition and plant species performance. Several studies have shown that stoichiometry indices such as N:P and N:K ratios in plant biomass can be a good indicator of nutrient limitation. However, the implications of an N:K ratio for wetland vegetation have hardly been investigated. In order to estimate a critical N:K ratio that can indicate the type of nutrient limitations, a greenhouse experiment has been established. The response of two grass species: Holcus lanatus and Molinia caerulea, to the range of N and K supply was analysed for two years. The effect of six combinations of N:K supply ratios (from 0.5 to 225), combined with two levels of fertility in a factorial design, on aerial biomass production, nutrient concentrations and nutrient resorption efficiency was tested. The aerial biomass increased with an increasing N:K supply ratio during both vegetation seasons at the low level of supply. Significant differences were observed not only between species but also between the N:K ratios during the two years. In the first year, the optimal N:K supply ratio was 4.5 for Holcus lanatus and 225 for Molinia careluea at the high fertility level. In 2010, the optimal N:K supply ratio was similar for both grasses. At the high fertility level, the shoot biomass was the highest at an N:K supply ratio of 13.5; at the low level, shoot productivity reached the peak at a 225 N:K supply ratio. Moreover, both plant species showed the same pattern of aerial biomass production to N:K supply ratios at both fertility levels, but differences in the N:K biomass ratios make it impossible to determine a critical N:K ratio. The N:K nutrient supply ratio was a better indicator of plant performance than the N:K biomass ratio of the analysed species. The tested graminoids did not show a similar response to N:K supply ratios at the high and low levels of supply, indicating that nitrogen was the most important factor limiting the plant growth during the two years, and that these plant species were less sensitive to K shortage than to N deficiency.

Leaf nutrient concentration as an indicator of Populus and Tamarix response to flooding

Plants growing in infertile environments are able to produce more biomass per unit of nutrient taken up than plants of fertile habitats, and also to minimize nutrients loss by resorbing them from senescing leaves. The leaf nutrient concentration variability of two co-existing riparian tree genera (Populus and Tamarix) along a flood inundation gradient was examined to infer nutrient limitation and to compare nutrient use strategies in the two genera. To that end, seasonal and spatial variability in leaf nitrogen (N) and phosphorus (P) concentration (i.e., % dry mass of N and P) were analyzed in 720 samples of leaves (2 tree genera × 3 seasons × 12 sites × 10 tree replicates). Both Populus and Tamarix showed strong seasonal variability in leaf N and P concentrations, with values decreasing throughout the growing season. However, while N:P atomic ratio remained seasonally constant in Populus (N:P = 33), Tamarix shifted from N:P = 29 in spring to N:P = 36 and 37 in summer and fall. %N, %P and N:P atomic ratios were also spatially variable, but leaf litter N and P concentration (i.e., nutrient resorption proficiency) and leaf litter N:P generally followed the local flood inundation gradient as shown by linear mixed effects models. In particular, nutrient resorption was usually less proficient (higher terminal nutrient concentrations) at higher flood durations (in gravel bars and natural levees), whereas N:P increased in the drier sites (floodplain terrace). At floodplain level, a P-limitation that is higher than N-limitation seems to characterize the plant nutrient circulation in the riparian ecosystem studied. Tamarix was slightly more proficient in P resorption than Populus. The study shows that leaf nutrient concentration (e.g., N and P) derived from nutrient availability is partly controlled by the flood inundation regime and can be used as an indicator of nutrient limitation in forested floodplains. Subtle differences between tree genera provide an additional, novel explanation for the recent expansion of Tamarix in many arid and semi-arid rivers with altered hydrogeomorphic regimes.

Sediment microbial enzyme activity as an indicator of nutrient limitation in the great rivers of the Upper Mississippi River basin

We compared extracellular enzyme activity (EEA) of microbial assemblages in river sediments at 447 sites along the Upper Mississippi, Missouri, and Ohio Rivers with sediment and water chemistry, atmospheric deposition of nitrogen and sulfate, and catchment land uses. The sites represented five unique river reaches—impounded and unimpounded reaches of the Upper Mississippi River, the upper and lower reaches of the Missouri River, and the entire Ohio River. Land use and river chemistry varied significantly between rivers and reaches. There was more agriculture in the two Upper Mississippi River reaches, and this was reflected in higher nutrient concentrations at sites in these reaches. EEA was highest in the two Upper Mississippi River reaches, followed by the lower Missouri River reach. EEA was generally lowest in the upper Missouri River reach. Canonical correlation analysis revealed a strong correlation between EEA and the suite of water and sediment chemistry variables, and the percent of the catchment in anthropogenically dominated land uses, including agriculture and urban development. Nutrient ratios of the waters and sediments suggested carbon (C), nitrogen (N), or phosphorus (P) limitation at a large number of sites in each reach. C-limitation was most pronounced in the unimpounded Mississippi River and lower Missouri River reaches; N-limitation was prevalent in the two Missouri River reaches; and P-limitation dominated the Ohio River. Linking microbial enzyme activities to regional-scale anthropogenic stressors in these large river ecosystems suggests that microbial enzyme regulation of carbon and nutrient dynamics may be sensitive indicators of anthropogenic nutrient and carbon loading.

Plant limiting nutrients in Andean-Patagonian woody species: Effects of interannual rainfall variation, soil fertility and mycorrhizal infection

Andean-Patagonian forests are especially interesting for the study of N and P limitation because they receive minimal atmospheric pollution, have little influence of vascular N-fixing species, and grow on volcanic soils that retain P. In a previous study of 10 woody species (four broad-leaved deciduous species, three broad-leaved evergreens and three conifers) conducted during an exceptionally dry year in NW Patagonia, and on the basis of nutrient resorption efficiency and proficiency, we suggested that N was the most limiting nutrient except for the broad-leaved evergreen Lomatia hirsuta. In the present work, we compared patterns of nutrient limitation during a dry and a wet year, quantified the percentage of mycorrhizal infection, and related mycorrhizal behavior and nutrient limitation to soil fertility. We used N and P concentrations in green leaves as indicators of nutrient requirements, and N and P concentrations in senescent leaves (resorption proficiency) and the N/P ratio in green leaves as indicators of nutrient limitation. We also determined leaf mass area (LMA) and lignin concentration as indicators of structural and chemical defences. From previous works, the following soil fertility indicators were included: pH, organic C, total N, exchangeable cations, Olsen-P, potential N mineralization (pNmin) and N retained in microbial biomass (N-MB). Nitrogen, P and lignin concentrations in green and senescent leaves did not differ significantly between the dry and the wet year either by species or by functional groups. Most species behaved as N-proficient and P-non-proficient; this together with values of foliar N/P ratios lower than 14–16 confirmed N limitation in these forests. The only species limited by P but not by N was L. hirsuta (1.0–1.1% N in senescent leaves, N/P ratio = 21–23), a non-mycorrhizal species with cluster roots. The lack of P limitation in the other species was probably related to the high percentages of infection with arbuscular mycorrhizae (80–90% in Maytenus boaria and the conifers Araucaria araucana, Austrocedrus chilensis and Fitzroya cupressoides), and ectomycorrhizae (73–79% in five Nothofagus species). Nitrogen and P requirements were positively correlated among themselves and negatively with lignin and LMA. Soil fertility was positively correlated with nutrient requirements and negatively with lignin and LMA. Conifers had lower N and P requirements, higher LMA, lower foliar N/P ratio and grew on soils of lower soil N dynamics (lower pNmin and N-MB) than ectomycorrhizae-associated species.

Sediment microbial enzyme activity as an indicator of nutrient limitation in Great Lakes coastal wetlands

Summary1. We compared the extracellular enzyme activity (EEA) of sediment microbial assemblages with sediment and water chemistry, gradients in agricultural nutrient loading (derived from principal component analyses), atmospheric deposition and hydrological turnover time in coastal wetlands of the Laurentian Great Lakes.2. There were distinct increases in nutrient concentrations in the water and in atmospheric N deposition along the gradient from Lake Superior to Lake Ontario, but few differences between lakes in sediment carbon (C), nitrogen (N) or phosphorus (P). Wetland water and sediment chemistry were correlated with the agricultural stress gradient, hydrological turnover time and atmospheric deposition.3. The N : P ratio of wetland waters and sediments indicated that these coastal wetlands were N‐limited. Nutrient stoichiometry was correlated with the agricultural stress gradient, hydrological turnover time and atmospheric deposition.4. Extracellular enzyme activity was correlated with wetland sediment and water chemistry and stoichiometry, atmospheric N deposition, the agricultural stress gradient and the hydrological turnover time. The ratios of glycosidases to peptidases and phosphatases yielded estimates of nutrient limitation that agreed with those based solely on nutrient chemistry.5. This study, the first to link microbial enzyme activities to regional‐scale anthropogenic stressors, suggests that quantities and ratios of microbial enzymes are directly related to the concentrations and ratios of limiting nutrients, and may be sensitive indicators of nutrient dynamics in wetland ecosystems, but further work is needed to elucidate these relationships.

Vegetation in contrasting soil water sites of upland herbaceous grasslands and N:P ratios as indicators of nutrient limitation

Effects of differences in long-term water supply were examined on soil characteristics, primary production and species composition in a wet and a dry site of an upland herbaceous grassland. Also the responses of species to N and P enrichments were examined. N and P concentrations of non-legume species were positively related, indicating that biomass N:P ratios seem to be mainly determined by N:P supply ratios. Forbs had generally higher concentrations than graminoids. Intermittent water inundation of soil in the wet site resulted in greater soil N and P availability. The greater productivity of this site promoted the growth of forbs. A fertilizer experiment showed that biomass was limited by N only in the wet site, but by both nutrients in the dry one. The species with the higher N and P concentrations were favored more after N and P enrichment, respectively; however, species enhancement was not related to N:P ratios of species. This indicates that N and P concentrations of species, rather than N:P ratios of species, are better predictors of species responses to N and P enrichment. N:P ratios of whole communities were 8.73 for the wet and 11.36 for the dry site. These values in comparison with the responses of plant communities to N and P fertilization show that thresholds of N:P ratios indicative of N or P limitation are much lower than those found for European wetlands.

Use of nitrogen to phosphorus ratios in plant tissue as an indicator of nutrient limitation and nitrogen saturation

Summary Ratios of nitrogen to phosphorus (N:P) in plant foliage have been used to assess nutrient limitation in wetland ecosystems and to indicate nitrogen saturation. Extension of this application to ecosystems other than wetlands remains to be evaluated. We compared published N:P ratios as thresholds of nutrient limitation with published accounts from nutrient‐addition experiments and the N:P ratios of understorey vegetation (Acer spp., Dryopteris intermedia, Erythronium americanum, Lycopodium lucidulum, Oxalis acetosella and Viola macloskeyi) from the Catskill Mountains of New York State, USA. We also performed a nutrient‐addition experiment to test the response of these understorey plant species to inputs of N and P. N:P ratios of Catskill understorey species indicated they were at or near P limitation relative to N:P ratios from other upland ecosystems. Our experiment supported this finding in that none of the species responded to N addition but all increased in P concentration and one increased in biomass with added P. Collectively, these results suggest that the understorey vegetation of the Catskill Mountains is not nitrogen limited, providing further evidence that hardwood forests in this area are nitrogen‐saturated. Synthesis and applications. This study demonstrates that N:P ratios can be effective predictors of nutrient limitation in upland ecosystems. Therefore N:P ratios can be used for management and monitoring purposes in considering the nutrient status of upland ecosystems with particular relevance to the continued deposition of elevated atmospheric N and to the diagnosis of nitrogen saturation.

The nutrient stoichiometry of benthic microalgal growth: Redfield proportions are optimal

Cellular nutrient ratios are often applied as indicators of nutrient limitation in phytoplankton studies, especially the so‐called Redfield ratio. For periphyton, similar data are scarce. We investigated the changes in cellular C:N:P stoichiometry of benthic microalgae in response to different levels and types of nutrient limitation and a variety of abiotic conditions in laboratory experiments with natural inocula. C:N ratios increased with decreasing growth rate, irrespective of the limiting nutrient. At the highest growth rates, the C:N ratio ranged uniformly around 7.5. N:P ratios <13 indicated N limitation, while N:P ratios >22 indicated P limitation. Under P limitation, the C:P ratios increased at low growth rate and varied around 130 at highest growth rates. For a medium with balanced supply of N and P, an optimal stoichiometric ratio of C:N:P = 119:17:1 could be deduced for benthic microalgae, which is slightly higher than the Redfield ratio (106:16:1) considered typical for optimally growing phytoplankton. The optimal ratio was stable against changes in abiotic conditions. In conclusion, cellular nutrient ratios are proposed as an indicator for nutrient status in periphyton.

Wood Decomposition: Nitrogen and Phosphorus Dynamics in Relation to Extracellular Enzyme Activity

Because plant litter decomposition is directly mediated by extracellular enzymes (ectoenzymes), analyses of the dynamics of their activity may clarify the mechanisms that link decomposition rates to substrate quality and nutrient availability. We investigated this possibility by placing arrays of white birch sticks at eight upland, riparian, and lotic sites on a forested watershed in northern New York. For 3 yr, samples were analyzed for mass loss, protein, total Kjeldahl nitrogen (TKN), and total phosphorus (TP) accumulation, and the activity of 11 classes of extracellular enzymes involved in C, N, and P cycling. The relationship between lignocellulase activity and mass loss did not differ among sites. TKN and TP immobilization exhibited some spatial variation; rates of accumulation per 1% loss of initial mass, estimated from linear regressions, ranged from 2.2 to 4.4 mg/g OM for TP and from 43 to 139 mg/g OM for TKN, with maximum concentrations reached at °80% mass loss. The relationship between the activities of acid phosphatase (AcPase) and N—acetylglucosaminidase (NAGase), enzymes involved in the acquisition of P and N from organic sources, and mass loss displayed even greater variation among sites; the slopes of linear regressions relating mass loss and temporally integrated activity ranged from 0.019 to 0.135 activity—months per mass loss point and 0.107 to 0.775 activity—months per mass loss point, respectively, suggesting that edaphic rather than substrate quality factors were regulating activity. The extent of N limitation at each site was inferred by plotting TKN accumulation, defined as the slope of the linear regression TKN concentration vs. mass loss, in relation to NAGase activity accumulation, defined as the slope of the linear regression cumulative NAGase activity—months vs. mass loss. P limitation at each site was similarly assessed from an analogous plot of TP accumulation in relation to AcPase activity accumulation. Low N or P accumulation in conjunction with high acquisition activity was taken as an indication of nutrient limitation while the converse indicated surfeit. The diagrams suggested that decomposition at the upland hemlock and lotic sites, which displayed intermediate rates of OM loss (zero order k = 0.29 g/mo and 0.23 g/mo, respectively), was primarily N limited, while the riparian sites, which had the lowest rates of OM loss (k = 0.14 g/mo), appeared to be P limited. Relative to the others, OM loss at the upland deciduous sites (k = 0.38 g/mo) was not limited by either N or P. The concordance of field observations with predictions based on ectoenzyme regulation mechanisms suggest that enzyme activity assays in conjunction with nutrient concentration measurements may be a useful indicator of nutrient limitation. An economic model is proposed that directly links N and P availability to litter decomposition rates on the basis of microbial allocation of resources of extracellular enzyme production.

The phycocyanin to chlorophyll α ratio and other cell components as indicators of nutrient limitation in two planktonic cyanobacteria subjected to low-light exposures

The phycocyanin to chlorophyll α ratio and other cell components as indicators of nutrient limitation in two planktonic cyanobacteria subjected to low-light exposures