Leaf vs. epiphyte nitrogen uptake in a nutrient enriched Mediterranean seagrass (Posidonia oceanica) meadow

In situ nitrogen uptake by leaves and epiphytes was studied in a Mediterranean seagrass (Posidonia oceanica) meadow impacted from a fish farm and a pristine meadow, using ¹⁵NH₄ and ¹⁵NO₃ as tracers. In the impacted meadow both leaves and epiphytes yielded higher N concentrations and showed higher specific N uptake, suggesting a linkage between N uptake and its accumulation. Epiphytes took up N faster than leaves in relation to their corresponding biomass, but when assessed per unit area, N uptake was higher in leaves. Leaf N uptake was negatively correlated with epiphyte N uptake. With increasing epiphyte load on leaves, N leaf uptake decreased while N epiphyte uptake increased, indicating that epiphyte overgrowth hinders N uptake by P. oceanica leaves. Epiphyte contribution to total N uptake increased, while that of leaves decreased at the impacted meadow. However, 2-3 times less N was transferred daily from the water column to the benthic compartment, through seagrass and epiphyte uptake on total, at the impacted meadow. Therefore, it is probably still the loss of the key species - the seagrass - which plays the most important role in N cycling in seagrass ecosystems.     

Effects of nitrogen addition on nitrogen metabolism and carbon reserves in the temperate seagrass Posidonia oceanica

The effect of repeated N additions on a dense, shallow meadow of Posidonia oceanica (L.) Delile in the NW Mediterranean was studied over a year. N was added biweekly both to the sediment and to the water column as ammonium and nitrate. The most obvious result of these additions was an overall increase in N content (% DW) in all tissues of fertilized plants; this increase was maximum in rhizomes, with values of 5% N reached, which confirmed the storage capacities of these organs.Fertilization affected the different N fractions in distinct ways. The free amino acid (FAA) concentration increased the most, particularly in rhizomes and roots, suggesting the function of these compounds for N storage and, probably, translocation. The non-soluble N fraction also increased greatly. The total soluble protein (TSP) and the inorganic N forms concentrations were less sensitive to fertilization, and only increased moderately in a few cases. N assimilation, assessed through in vivo glutamine synthetase (GS) activity, was maximum in leaves after the peak of growth, which coincided with the lowest N values in both control and fertilized plants. Thus assimilation was probably greatest at the period of highest N deficiency. Growth rates did not respond to N enrichment. Another clear effect of N addition was to decrease carbon reserves. In effect, the concentration of total non-structural carbohydrate (TNC) greatly decreased in rhizomes of fertilized plants, coinciding with the increase in FAA. We conclude that increased nitrogen availability can affect plant survival through the decrease in their carbon reserves, crucial for P. oceanica overwintering. This interaction between N and C metabolism helps to explain changes in benthic vegetation after steadily increasing coastal water eutrophication.     

Posidonia oceanica in the Marmara Sea

The seagrass Posidonia oceanica is a stenohaline species endemic to the Mediterranean Sea, where it normally lives at a salinity of between 36.5 and 39.5 ppt. Surveys carried out at the North-eastern distribution limits revealed large beds in the Dardanelles Strait and isolated beds in the Marmara Sea, where the salinity ranges between 21.5 and 28 ppt. Microsatellite analysis of these low-salinity tolerant P. oceanica beds, show different signs of genetic isolation: excess of heterozygosity and a presence of fixed alleles. These particularities are rarely found in the whole distributional range of the species. Moreover, all the populations considered in the analysis have a very low genetic diversity in comparison with most of the meadows sampled throughout the Mediterranean Sea. Taking into consideration the genetic data, rhizome expansion rate and the actual extent of the isolated beds in the Marmara Sea and knowing the reproductive rate and dissemination characteristics of P. oceanica, we hypothesize that the isolated population of the Marmara Sea has been established since the Middle Holocene, before the catastrophic intrusion of brackish water into the Marmara Sea and the strong and persistent flow coming from the Black Sea.     

Secondary metabolites of Posidonia oceanica (Posidoniaceae)

Posidonia oceanica Delile is an endemic of the Mediterranean Sea. P. oceanica is one of the few marine higher plants of the Mediterranean and plays a significant role in coastal ecosystems of the Mediterranean Sea. In the past, a number of phytochemical investigations with different focus were carried out. The results of these investigations are summarized and appraised here. A total of 51 natural products was reported from P. oceanica, including phenols, phenylmethane derivatives, phenylethane derivatives, phenylpropane derivatives and their esters, chalkones, flavonols, 5α-cholestanes, and cholest-5-enes. Many of the compounds reported for P. oceanica were, however, not detected by appropriate phytochemical methods and some most probably represent artifacts and are no genuine natural products.     

Posidonia oceanica seedling root structure and development

The seedling root system of the seagrass Posidonia oceanica consists of a primary root and up to four adventitious roots. Under culture, germination and early growth began with the emergence of the primary root in the first week. Then the two adventitious root primordia originally present in the seed emerged at 3 and 5 weeks respectively, followed successively by further adventitious roots. Primary roots reached 17 mm at 4 weeks, but then their growth decreased markedly. In contrast the adventitious roots showed a pattern of continued elongation. Anatomical observations of both primary and adventitious roots revealed a multilayered hypodermis of thick-walled cells enclosing a wide cortex (99% of the root transverse area) and narrow stele. A well-distinguished endodermis was only observed in the primary roots, while differentiated xylem elements were found solely in the adventitious roots, but it is unclear to what degree differences between the two root types are due to different root maturity or to their role in water and nutrient uptake. Overall, the P. oceanica seedling root system is composed of multiple, rapidly formed roots which are strong yet flexible due to a large proportion of cortical tissue and further strengthened by a multilayered hypodermis, characteristics which could potentially facilitate initial anchorage and establishment.     

Comparing shoot population dynamics methods on Posidonia oceanica meadows

Prediction capacity of three main shoot population dynamics methods (age structure, net shoot recruitment per plagiotropic rhizome and shoot census) have been tested for a period of four years (2002-2006) on a Posidonia oceanica meadow. Accuracy of each method was checked by comparing measured and predicted densities at the end of the study period. Predicted densities came from the evolution of initial densities (measured in 2002) by a basic exponential model of population growth. The exponential model used the different net shoot recruitment rate estimates by each population dynamics method on three depths (upper, medium and lower limit) and three localities at each depth. Predictions performed by shoot census and net shoot recruitment per plagiotropic rhizome methods matched with measured densities at the end of the study period. Conversely, age structure method underestimated shoot densities at each depth, indicating an unreal decrease of shoot population in the meadow.     

Organic and inorganic human-induced contamination of Posidonia oceanica meadows

In coastal environments, plants are used for phytoremediation of contamination. Organic and inorganic contaminants may be due to natural and/or anthropogenic sources. The aim of this study is to compare inorganic (trace metal) and organic (PAH) contamination in Posidonia oceanica and to analyse the relationship between these types of pollutants indeed very few studies have been interested in their correlations and common sources. P. oceanica leaves were collected in two sites exhibiting different levels of human-induced pressure. Higher values were recorded in the more polluted site (Toulon) for trace metals (Ag, Hg, Pb) as well as for PAHs (Medium Molecular Weight and High Molecular Weight) due to the presence of the city and/or harbour in proximity. For the first time in a coastal environment, correlations were observed between metals and PAHs.     

Recovery of deep Posidonia oceanica meadows degraded by trawling

The recovery capacity of meadows of the Mediterranean seagrass Posidonia oceanica (L.) Delile in an area affected by illegal trawling were assessed after protection by anti-trawling reefs. The differences in vegetative growth between two impacted and two undisturbed localities were tested using growth, shoot balance, aborted branches, and leaf and rhizome production of both plagiotropic and orthotropic rhizomes. The organic matter in sediments, silt clay fraction and light intensity incident on the bottom were also measured in order to evaluate the physical conditions. Environmental and plant variables were measured in three sites placed inside each locality. The vegetative growth was positive in both impacted and control meadows but growth rates were lower in impacted than in control meadows. Average growth, production and shoot balance were greater in plagiotropic rhizomes from undisturbed localities (40.7±1.75 vs. 28.4±1.34 mm/year, 1133±0.06 vs. 708±0.04 mg DW/shoot/year, 1.36±0.08 vs. 0.96±0.06 shoots/year, respectively). Significantly greater values were also found in undisturbed localities for orthotropic rhizomes in terms of shoot balance and rhizome production (0.07±0.01 vs. 0.01±0.003 shoots/shoot/year and 155 vs. 124 mg DW/shoot/year, respectively). Of the physical parameters measured, only light intensity differed significantly between impacted and undisturbed localities. This parameter was 15.5% to 67.6% lower in impacted localities than in undisturbed localities, and this is the factor that causes the retardation of vegetative growth. The results show that recovery of P. oceanica meadows is possible after eliminating the cause of the impact. However, the very low rates of vegetative growth may prolong the time to total recuperation to almost 100 years. Therefore, effective management of P. oceanica meadows should aim to prevent meadow loss.     

Seasonal nitrogen speciation in temperate seagrass Posidonia oceanica (L.) Delile

To better understand some basic aspects of the nitrogen economy in Posidonia oceanica and, specifically, the seasonality of the processes of storage, translocation and assimilation, we examined nitrogen speciation into soluble compounds, both inorganic (nitrates, nitrites and ammonium) and organic (free amino acids, FAA, and total soluble protein, TSP), and the nitrogen assimilation potential (through the glutamine synthetase activity measurement) in the leaves, rhizomes and roots of P. oceanica over a 1-year cycle. Only a limited amount of inorganic nitrogen was found, accounting for less than 3.3% of the total nitrogen content, and it was mostly in the form of ammonium. Nitrate and nitrite concentrations were very low, always below 7.2 μmol g⁻¹ dw in annual average. Among the organic soluble fractions, FAAs were the most abundant, accounting for up to 50% of N pools. Rhizomes were the organs in which FAA concentrations reached their maximum value. The leaves showed higher nitrogen assimilation potential than the roots and this assimilation potential was highest during and after the period of maximum leaf growth, probably corresponding to the assimilation of both new and recycled nitrogen. Our results suggest that 5% of the total nitrogen assimilation occurs in roots and 79% in leaves on an annual average. In addition, rhizomes contributed to the total shoot nitrogen assimilation by 32-54% between autumn and spring. Rhizomes appear as key organs in the nitrogen economy of the plant, not only as a major site for nitrogen assimilation but also as an organ for nitrogen storage. This storage, mostly in the form of FAA, occurs during periods of high availability and low demand (winter). This stored nitrogen can supply up to 33% of plant demands during the moment of maximum leaf growth (i.e. late spring).     

Patch dynamics of the Mediterranean seagrass Posidonia oceanica: Implications for recolonisation process

Patch dynamics of the Mediterranean slow-growing seagrass Posidonia oceanica was studied in two shallow sites (3–10 m) of the Balearic Archipelago (Spain) through repeated censuses (1–2 year⁻¹). In the sheltered site of Es Port Bay (Cabrera Island), initial patch density (October 2001) was low: 0.05 patches m⁻², and the patch size (number of shoots) distribution was bimodal: most of the patches had less than 6 shoots or between 20 and 50 shoots. Mean patch recruitment in Es Port Bay (0.006 ± 0.002 patches m⁻² year⁻¹) exceeded mean patch loss (0.001 ± 0.001 patches m⁻² year⁻¹), yielding positive net patch recruitment (0.004 ± 0.003 patches m⁻² year⁻¹) and a slightly increased patch density 3 years later (July 2004, 0.06 patches m⁻²). In the exposed site of S’Estanyol, the initial patch density was higher (1.38 patches m⁻², August 2003), and patch size frequency decreased exponentially with size. Patch recruitment (0.26 patches m⁻² year⁻¹) and loss (0.24 patches m⁻² year⁻¹) were high, yielding a slightly increased patch density in the area 1 year later (October 2004, 1.40 patches m⁻²). Most recruited patches consisted of rooting vegetative fragments of 1–2 shoots. Seedling recruitment was observed in Summer 2004 at both sites. Episodic, seedling recruitment comprised 30% and 25% of total patch recruitment in Es Port Bay and S’Estanyol, respectively. Patch survival increased with patch size and no direct removal was observed among patches of 5 shoots or more. Most patches grew along the study, shifting patch distribution towards larger sizes. Within the size range studied (1–150 shoots), absolute shoot recruitment (shoots year⁻¹) increased linearly with patch size (R² = 0.64, p < 4 × 10⁻⁵, N = 125), while specific shoot recruitment was constant (about 0.25 ± 0.05 year⁻¹), although its variance was large for small patches. Given the slow growth rate and the high survival of patches with 5 or more shoots, even the low patch recruitment rates reported here could play a significant role in the colonisation process of P. oceanica.     

Configuration at C-24 of sterols from the marine phanerogames Posidonia oceanica and Cymodocea nodosa

Sterols were extracted from two marine phanerogames, Posidonia oceanica and Cymodocea nodosa. The two plants contain 24α-ethyl sterols, while the 24α-methyl sterols are accompanied by 24β-epimers. The most abundant components are sitosterol, cholesterol and stigmasterol.     

Effect of nutrient enrichment on growth and photosynthesis of the zooxanthellate coral Stylophora pistillata

The effect of prolonged (9 week) nutrient enrichment on the growth and photosynthetic rates of the zooxanthellate coral Stylophora pistillata was investigated. The main questions were: (1) what is the exposure time needed to induce measurable change in growth rate? (2) which are the concentrations of nitrogen and phosphorus required to cause changes in these rates? (3) what is the recovery potential of the corals after the nutrient stress? For this purpose, three tanks (N, P, NP) were enriched with ammonium (N), phosphorus (P) or both nutrients (NP), respectively. A fourth tank (C) served as a control. The growth of 40 nubbins (10 in each tank) was monitored during four periods: period 1 (nutrient-poor conditions), period 2 (10?μm NH₄ and/or 2?μm PO₄ enrichment), period 3 (20?μm NH₄ and/or 2?μm PO₄) and period 4 (nutrient-poor conditions). Period 4 was performed to study the recovery potential of corals after a nutrient stress. During period 1, growth rates remained constant in all tanks. In the P tank, growth rates declined during the two enrichment periods, with a total decrease of 60% by the end of period 3. In the N tank, growth rates remained nearly constant during period 2 but decreased in period 3 (60% decrease). In the NP tank, 50% and 25% decreases were observed during periods 2 and 3. At the end of the recovery period, a regain in growth rate was observed in the N and NP tanks (35 and 30% increase, respectively, compared with the rates measured at the end of period 3) and growth rates returned to 60% of the initial rates. By contrast, in the P tank, there was no regain in growth and a further decrease of 5% was observed. Rates of photosynthesis were often higher during the enriched than the nutrient-poor period (up to 150% increase). Corals with the highest percent increases in maximal gross photosynthetic rate (P ^g _max ) had the smallest decreases in growth rate due to nutrient enrichment. In conclusion, high ammonium (20?μm) and relatively low phosphorus concentrations (2?μm) are required to induce a significant decrease in coral growth rate. The largest reduction was observed with both ammonium and phosphorus enrichment. The decrease in growth rate was rapid following nutrient enrichment, since a 10% decrease or more could be observed after the first week of treatment.     

Effects of external and internal nutrient supplies on decomposition of wild rice, Zizania palustris

We examined effects of external supplies of nitrogen (N) and phosphorus (P) from the environment and internal supplies of N and P from within litter tissue on wild rice shoot and root litter decomposition and N and P dynamics. To investigate the effects of external supplies, wild rice shoot and root litterbags were decayed in mesocosms in the field over 115 days with either added N or P or a control in ambient conditions. To investigate the effects of the internal nutrient supply, wild rice plants were grown with added N, P, both N and P, or no supplemental nutrient, to produce enriched litters, which were then decayed for 168 days under controlled temperature in the laboratory. Both external and internal N and P supplies affected shoot litter decay more than decay of root litter. Increased external P supply significantly increased the rate of wild rice shoot decay and P mineralization but adding N had no effect on decay rates through time. Neither adding N nor P influenced root decay. Enrichment of P internally in the litter through fertilization increased the concentration of P (0.16%) and water-soluble compounds (28.7% WS) in shoot litter compared to control shoot litter (0.11% P, 19.8% WS), which likely caused the significant increase in shoot decay rates, particularly in the labile pool. In contrast, N enrichment not only increased plant growth but also increased lignin concentrations (7.5%) compared to control shoot litter (2.7% lignin) for added structural support. This significantly inhibited decay and nearly doubled the amount of mass remaining after 168 days (42.1% OM) when compared to control shoots (22.4% OM). Increased lignin likely overrides a concomitant increase in nitrogen concentration in shoot litter and appears to control wild rice decomposition. Lignin and phosphorus appear to play a key role in driving wild rice decay through the effects on litter quality.     

Effects of salinity on leaf growth and survival of the Mediterranean seagrass Posidonia oceanica (L.) Delile

The main aim of this study was to estimate the effects of salinity variation on the Mediterranean seagrass Posidonia oceanica (L.) Delile and its attached epiphytes. Leaf growth and survival of this plant were tested in several short-term (15 days) mesocosms experiments under controlled conditions between February 2001 and November 2001. Plants collected from shallow meadows at Alicante (SE Spain), with an ambient salinity of 36.8-38.0 psu, were placed in tanks of 300 L with an additional overhead light and exposed to different salinity treatments (ranging from 25 to 57 psu) during 15 days. To estimate the mortality and growth recuperation, in some experiments shoots were returned to control salinity (38 psu). Leaf growth was measured in the laboratory where epiphytic fauna and flora were removed from leaves, with a razor blade, to determine their biomass.P. oceanica was negatively influenced by increased salinity. Shoots showed a significant decrease in growth and survival, whereas epiphyte biomass did not show a clear response because of their high variability. Maximum leaf growth occurred between 25 and 39 psu. In addition, plants suffered considerable mortality at salinities above 42 psu and below 29 psu, with 100% mortality at 50 psu. In salinities between 39 and 46 psu, surviving plants were able to regain their original growth rate when returned to normal seawater salinity (38 psu). These results suggest that P. oceanica is one of the most sensitive seagrasses to salinity increments it is more tolerant to salinity reductions (25.0-36.4 psu), perhaps due to the terrestrial origin of seagrasses.     

Grazing history effects on above- and below-ground litter decomposition and nutrient cycling in two co-occurring grasses

Large herbivores may alter carbon and nutrient cycling in soil by changing above- and below-ground litter decomposition dynamics. Grazing effects may reflect changes in plant allocation patterns, and thus litter quality, or the site conditions for decomposition, but the relative roles of these broad mechanisms have rarely been tested. We examined plant and soil mediated effects of grazing history on litter mass loss and nutrient release in two grazing-tolerant grasses, Lolium multiflorum and Paspalum dilatatum, in a humid pampa grassland, Argentina. Shoot and root litters produced in a common garden by conspecific plants collected from grazed and ungrazed sites were incubated under both grazing conditions. We found that grazing history effects on litter decomposition were stronger for shoot than for root material. Root mass loss was neither affected by litter origin nor incubation site, although roots from the grazed origin immobilised more nutrients. Plants from the grazed site produced shoots with higher cell soluble contents and lower lignin:N ratios. Grazing effects mediated by shoot litter origin depended on the species, and were less apparent than incubation site effects. Lolium shoots from the grazed site decomposed and released nutrients faster, whereas Paspalum shoots from the grazed site retained more nutrient than their respective counterparts from the ungrazed site. Such divergent, species-specific dynamics did not translate into consistent differences in soil mineral N beneath decomposing litters. Indeed, shoot mass loss and nutrient release were generally faster in the grazed grassland, where soil N availability was higher. Our results show that grazing influenced nutrient cycling by modifying litter breakdown within species as well as the soil environment for decomposition. They also indicate that grazing effects on decomposition are likely to involve aerial litter pools rather than the more recalcitrant root compartment.     

Carex sempervirens tussocks induce spatial heterogeneity in litter decomposition, but not in soil properties, in a subalpine grassland in the Central Alps

Tussocks of graminoids can induce spatial heterogeneity in soil properties in dry areas with discontinuous vegetation cover, but little is known about the situation in areas with continuous vegetation and no study has tested whether tussocks can induce spatial heterogeneity in litter decomposition. In a subalpine grassland in the Central Alps where vegetation cover is continuous, we measured soil properties [concentration of N, C, organic matter (OM) and pH] and monitored litter decomposition traits (dry mass loss, loss of C, N, P and K) inside and outside tussocks of Carex sempervirens. Soil C, N, OM concentrations or pH inside tussocks did not differ significantly from those outside tussocks. After 1 year of decomposition, litter dry mass loss, C and K loss were significantly smaller inside than outside tussocks. The slower litter decomposition inside tussocks was likely caused by the elevated tussock base, which made environmental conditions inside tussocks much dryer than those outside in early spring when snow melts. Our results suggest that in areas with continuous vegetation cover tussocks induce spatial heterogeneity in litter decomposition but not in soil properties.     

Shoot age as a confounding factor on detecting the effect of human-induced disturbance on Posidonia oceanica growth performance

The response of orthotropic rhizome elongation and primary production of Posidonia oceanica to anthropogenic perturbations and potential confounding effects of shoot age were assessed using a Linear Multilevel Model (LMM). This model examined the confounding effect of age by comparing the estimates of impact and variance components obtained by excluding and including Age as an explanatory variable. Age had a negative effect on rhizome elongation and primary production with an annual decrease of 0.6 mm y^− 1 and 7 mg dw y^− 1 respectively. According to the LMM when age effect was omitted, the differences between disturbed and control locations in rhizome elongation and primary production were 2.62 mm y^− 1 and 0.044 g dw y^− 1 respectively. These effects were statistically not significant. On the contrary, when age effect was included in the statistical model, impacts became evident for both variables, with significant differences between disturbed and control locations of 5.85 mm y^− 1 and 0.081 g dw y^− 1 for rhizome elongation and primary production, respectively. Thus, particular attention should be paid to the potential confounding effect of shoots age in analyses of impacts on growth performance of P. oceanica.