Metabolic adaptation of Zostera marina (eelgrass) to diurnal periods of root anoxia

The temperate seagrass Zostera marina L. is common in coastal marine habitats characterized by the presence of reducing sediments. The roots of this seagrass grow in these anoxic sediments, yet eelgrass is highly productive. Through photosynthesis-dependent oxygen transport from leaves to roots, aerobic respiration is supported in eelgrass roots only during daylight; consequently, roots are subjected to diurnal periods of anoxia. Under anoxic root conditions, the amino acids alanine and -amino butyric acid accumulate within a few hours to account for 70% of the total amino acid pool, while glutamate and glutamine decline. Little ethanol is produced, and the pool size of the organic acid malate changes little or declines slowly. Upon the resumption of shoot photosynthesis and oxygen transport to the roots, the accumulated -amino butyric acid declines rapidly, glutamate and glutamine pools increase, and alanine declines over a 16-h period. These adaptive metabolic responses by eelgrass to diurnal root anoxia must contribute to the successful exploitation of shallow-water marine sediments that have excluded nearly all vascular plant groups. A metabolic scheme is presented that accounts for the observed changes in organic and amino acid pool sizes in response to anoxia.     

Predictive modelling of eelgrass (Zostera marina) depth limits

Empirical models relating secchi depths to maximum depth limits of eelgrass (Zostera marina L.) can describe basic differences in depth limits between areas or time periods exhibiting large differences in secchi depth. However, these models cannot predict the precise depth limit at a particular site at any specific time. In this study we aim to improve the ability of regression models to predict maximum depth limits by: (1) assuming that eelgrass depth limits respond to changes in secchi depth with a temporal delay of 1–2 years, (2) including other water-quality variables in addition to secchi depth, and (3) taking into account that factors regulating depth limits may vary between years and between sites. We were not able to improve the models by introducing a systematic delay in the response of depth limits to changes in secchi depths. The reason for this failure is likely to have been the systematic nature of our approach, since some sites responded with a delay, while others did not. The explanatory power of the models increased when additional water-quality variables were added in a multiple regression model. Where secchi depth alone explained 58% of the variations in depth limits, addition of winter [NH₄⁺] and maximum water depth as independent variables increased the explanatory power to 71%. These models applied to data from one specific year, but when data from several years (1989–1998) were included, only 35% of the variation in depth limits could be explained by the three factors. More detailed analyses showed that the regulation of eelgrass depth limits varied considerably between years and between sites, and the models were further improved by taking this information into account. Our results confirmed previous studies by showing light to be the most important parameter in the regulation of eelgrass depth limits, but also revealed a complexity in the regulation of depth limits not expressed in earlier studies. Limited colonisation potentials may delay the response to improved light conditions, and hypoxia/anoxia and indirect effects of nutrients may prevent eelgrass from attaining the depth limit that light levels would allow. The power to predict depth limits on the basis of secchi depths can therefore be improved by taking site-specific information on eelgrass growth conditions into account.Communicated by M. Kühl, Helsingør     

Uptake and translocation of phosphorus in eelgrass (Zostera marina)

The uptake and translocation of phosphorus in eelgrass (Zostera marina L.) was studied in two-compartment chambers using ³²P and autoradiography. Eelgrass was able to take up phosphorus both with leaves and rootrhizomes. Only a small amount (less than 4%) of the total amount of phosphorus taken up from either source was translocated within the plants during an incubation time of 120 h. Release of translocated phosphorus from the plant tissues to the surrounding water was not detected. The autoradiographs showed that the translocated phosphorus was localized mainly in the actively growing plant tissues, i.e. the stem and young leaves, and nodes of the rhizomes and young roots. The uptake of phosphorus in leaves and the bidirectional translocation in the plants was significantly lower in the dark. The uptake of phosphorus in rootrhizomes was unaffected by light-conditions. The concentration of phosphate in the water of one compartment did not affect the phosphorus uptake by the plant parts in the opposite compartment. However, the translocation was significantly reduced when both compartments were supplied with identical phosphate concentrations. The results of the present investigation indicate that the quantitative significance of eelgrass in the cycling of phosphorus between sediment and water may have been greatly overestimated in earlier studies. The amount of phosphorus taken up by root-rhizomes and subsequently translocated to leaves is probably insignificant during most of the growing season in comparison to the amount taken up by the leaves from the surrounding water. However the uptake of phosphorus by root-rhizomes may be significant in the nutrition of eelgrass during periods with low phosphate levels in the water.     

Sediment ammonium availability and eelgrass (Zostera marina) growth

The interaction of sediment ammonium (NH ₄ ⁺ ) availability and eelgrass (Zostera marina L.) growth, biomass and photosynthesis was investigated using controlled environment and in-situ manipulations of pore water ammonium concentrations. Sediment diffusers were used to create pore water diffusion gradients to fertilize and deplete ammonium levels in sediments with intact eelgrass rhizospheres. Between October, 1982 and September, 1983 controlled environment experiments using plants from shallow (1.3 m) and deep (5.5 m) stations in a Great Harbor, Woods Hole, Massachusetts, USA eelgrass meadow along with in-situ experiments at these stations provided a range of sediment ammonium concentrations between 0.1 and 10 mM (adsorbed+interstitial NH ₄ ⁺ ). The results of the in-situ experiments indicate that nitrogen limitation of eelgrass growth does not occur in the Great Harbor eelgrass meadow. A comparison of NH ₄ ⁺ regeneration rates and eelgrass nitrogen requirements indicates an excess of nitrogen supply over demand and provides an explanation for the lack of response to the manipulations. Results of controlled environment experiments combined with in-situ results suggest that sediment ammonium pool concentrations above approximately 100 mol NH ₄ ⁺ per liter of sediment (interstitial only) saturate the growth response of Zostera marina.     

The nitrogen supply to intertidal eelgrass (Zostera marina)

The uptake of ammonium and nitrate by eelgrass (Zostera marina L.) was studied in two-compartment chambers. The plants were collected in 1992 from a population growing on a tidal flat in the S.W. Netherlands. They were incubated under conditions which reflected field conditions; this implied the use of natural seawater and sediment porewater as incubation media. In all six experiments, carried out over the course of a major part of the growing period (from July to the end of September), ammonium appeared to be much more important as a source of nitrogen than nitrate. The largest part was taken up by the leaves: uptake of ammonium by the leaves accounted for 68 to 92% of total plant nitrogen uptake. The uptake of nitrogen compounds by the root-rhizome system represented only 4 to 30% of total plant uptake. Thus, at least during flood tide, the leaves play the major role in nitrogen uptake in this intertidal population. During ebb tide, most of the plants are submerged in very shallow tidepools. It is suggested that during this phase of the tidal cycle, influx of porewater ammonium into the tide-pool water may enable the leaves to exploit local sediment resources.     

Ammonium regeneration and assimilation in eelgrass (Zostera marina) beds

Regeneration and assimilation of ammonium in the water column and in sediments of eelgrass (Zostera marina L.) beds of Izembek Lagoon and Crane Cove, Alaska, USA and Mangoku-Ura, northeastern Japan, were investigated by using a ¹⁵N isotope dilution technique. In the water column of Mangoku-Ura, ammonium was regenerated at a rate of 12 nmol l^-1 h^-1 and assimilated at a rate of 74 nmol l^-1 h^-1. The ammonium regeneration rate in sediments ranged from 2 to 150 nmol g^-1 h^-1, and with one exception, exceeded ammonium assimilation in sediments (0.3 to 77 nmol g^-1 h^-1). The ammonium regeneration in the water column was of little significance for the nitrogen supply to the eelgrass bed ecosystem. Net ammonium production (regeneration minus assimilation) in the sediment of Izembek Laggon met nitrogen demand for eelgrass growth, suggesting that ammonium regeneration in the sediments was very important for the nitrogen cycle in the eelgrass bed ecosystem.     

Irradiance reduction: Effects on standing crops of the eelgrass Zostera marina in a coastal lagoon

Abundance of the eelgrass Zostera marina L. was studied in a coastal lagoon in southern California (USA), and was found to correlate with the level of irradiance at depths greater than 0.5 m below tidal datum. Results of controlled field experiments, using canopies to reduce downwelling illuminance by 63%, confirmed that turion density is a function of the irradiance the plants receive. By Day 18 of the experiment, turion density in the shaded experimental areas had decreased compared to the density of adjacent unshaded controls. Turion densities were continually lower throughout the 9-month study in the experimental areas, which at the end of the study had a turion density only 5% that of the control areas. Flowering in the experimental areas was also inhibited by shading. The biological implications of these findings are discussed with respect to seasonal changes in incident solar radiation, water transparency, and changes in water quality due to man's increased intervention in the natural processes of coastal lagoons.Contribution No. 4 from the Center for Marine Studies, San Diego State University, San Diego, California 92182, USA.     

13C label identifies eelgrass (Zostera marina) Carbon in an Alaskan estuarine food web

The food web of Izembek Lagoon, Alaska draws most of its carbon from eelgrass (Zostera marina) and phytoplankton. The¹³C:¹²C ratios of these primary producers are sufficiently different to enable their contributions to consumers to be estimated from consumer¹³C:¹²C ratios. Although the technique is conceptually simple, carbon inputs from other sources and isotope fractionations occurring in the food web limit its precision. Isotopic data nevertheless helps to establish the major carbon fluxes through the community and to assess the importance of eelgrass carbon to individual animals. It is particularly useful when dealing with detritus food chains, where direct observations of animal feeding habits are difficult to make. The Izembek community draws much of its carbon from eelgrass. Detritus food chains provide the major pathway for assimilation of eelgrass carbon by the community, but grazers are also important. Eelgrass carbon is more important to benthic animals than to the eelgrass epibiota and the fishes, which depend mainly on phytoplankton carbon.Publication No. 381 of the Institute of Marine Science, University of Alaska.     

Development of epiphytic communities on eelgrass (Zostera marina) along a nutrient gradient in a Danish estuary

The effect of nutrient enrichment on epiphyte development was examined by following the seasonal development of epiphyte biomass on eelgrass (Zostera marina L.) at four localities along a nutrient gradient in Roskilde Fjord, Denmark between March and December 1982. In the most nutrient-poor area, epiphyte biomass followed a distinct bimodal seasonal pattern with maxima in spring and early fall. Low nutrient availability and a high rate of eelgrass leaf renewal kept epiphyte biomass at a low level throughout the summer period. Unlike phytoplankton, the epiphytic community was not stimulated by nutrient enrichment during spring, however, from May through August, the biomass of both components increased exponentially with increasing concentrations of total N in the water. Along the nutrient gradient, phytoplankton biomass increased 5- to 10-fold, while epiphyte biomass increased 50- to 100-fold. Thus differences in nutrient conditions among study sites were more clearly reflected by epiphytes than phytoplankton.Contribution No. 419 from the Freshwater-Biological Laboratory, University of Copenhagen     

Inhibition of the growth of micro-algae and bacteria by extracts of eelgrass (Zostera marina) leaves

The effects of the water-soluble fraction of dead leaves of the eelgrass Zostera marina L. on the growth of 8 species of micro-algae (pennate and centric diatoms, dinoflagellates, and a green flagellate) and a bacterium were studied on agar plates and in liquid culture. The extracts of leaves which had been dead from a few days to 2 wk inhibited growth and often killed cells in all test organisms. Extracts were lethal even at concentrations equivalent to as little as 0.25 mg dry leaf ml^-1, but inhibition decreased when extracts were prepared from leaves aged in the laboratory for 35 d (loss of anti-bacterial activity) or 90 d (loss of anti-algal activity). Extracts of leaves which had aged and dried several months in the field had no effect, except at very high concentrations (13 mg dry leaf ml^-1) when the lag phase of growth was prolonged several days in a culture of the chlorophyte Platymonas sp. The active fraction in eelgrass leaves may be important in controlling initial growth of micro-organisms on eelgrass detritus, and it could determine the composition and activity of the epiphytic community on living leaves.     

Characterization of disjunct populations of Zostera marina (eelgrass) from California: genetic differences resolved by restriction-fragment length polymorphisms

Comparative restriction-fragment analysis was used to analyze the nuclear ribosomal DNA, and alcohol dehydrogenase-1 loci of Zostera marina L., for variation within and among populations. Eelgrass is a perennial marine flowering plant that is widespread and ecologically significant throughout the temperate northern hemisphere. A chemical method was developed to obtain restriction-quality DNA without CsCl fractionation from experimentally relevant quantities of seagrass tissues (0.5 to 1.0 g). The yield was 25 g g^-1 fresh weight. The three morphologically distinct forms of Z. marina from disjunct populations examined in this study were found to be genetically distinct; morphologically similar populations were indistinguishable genetically. Genetic distinction also correlated with habitat depth, as subtidal and intertidal populations were clearly divergent. Homologous probes for the 17S and 28S ribosomal DNA genes were used to map 24 restriction sites on the rDNA repeat of Z. marina, which was determined to be about 14 kb in length. At least 1 length mutation and 5 restriction-site changes were identified that distinguished Z. marina populations from San Diego and Monterey Bay (Del Monte Beach) from Z. marina populations from Elkhorn Slough and Tomales Bay. Estimated sequence variation (100×p) between eelgrass populations ranged from 0.00 to 0.69. Individual plants were observed to contain as many as four different rDNA-repeat length variants. The mean number of rDNA-repeat length variants per individual in Z. marina was about two. Intrapopulation variation in rDNA-repeat type was observed in only one individual from the Tomales Bay population.     

Reintroduction of eelgrass (Zostera marina) in the Dutch Wadden Sea; review of research and suggestions for management measures

Eelgrass (Zostera marina L.) in the Dutch Wadden Sea historically covered an area varying from 65–150 km² in the eulittoral as well as the sublittoral zones. At present, this area comprises less than 1 km² eulittoral eelgrass stands, with an associated decrease in habitat diversity. The causes for this decline are presumably connected with the ‘wasting disease’ and the closure of the former Zuiderzee in the early 1930s resulting in increased tidal range and increased currents. After a slight recovery of the eelgrass populations on the intertidal flats a definite decline started in the early 1970s, possibly connected to increased turbidity. The present water quality and turbidity do not negatively influence eelgrass growth up to a depth of at least 0.6m below Mean Sea Level. Based on mesocosm experiments and field experiments it is concluded that re-establishment of eelgrass should be possible in sheltered bays and on unexposed tidal flats. The most suitable depths for a reintroduction are those between 0 and 20–40 cm below mean sea level.     

Significance of salinity and silicon levels for growth of a formerly estuarine eelgrass (Zostera marina) population (Lake Grevelingen, The Netherlands)

Since the early 1980s, the eelgrass, Zostera marina L., population in the saline Lake Gevelingen, The Netherlands, is rapidly declining. An earlier study, in which long-term data on eelgrass coverage in this former estuary were correlated with several environmental variables, showed only one significant correlation: coverage was positively related to water column silicon levels. In addition, a negative correlation with salinity was observed, but this was not significant. In the present study, the effect of silicon and the effect of salinity on the development of Z. marina were investigated experimentally. Enhancement of dissolved silicon concentrations in the water did not stimulate Z. marina above-ground production or an increase in final above- and below-ground biomass. The highly significant correlation between eelgrass coverage and water column silicon levels, thus, remains to be explained. The results of the growth experiments did, however, demonstrate a clear effect of salinity on Z. marina growth. Plants cultured at 22 psu showed a higher production of shoots and leaves, resulting in more above-ground biomass, than plants grown at 32 psu. In addition, below-ground biomass was also higher at 22 psu. Measurements of chlorophyll a fluorescence, performed with a PAM-fluorometer, indicated a reduction of photosynthesis in the high-salinity treatments. Thus, low salinity stimulates development of Z. marina from Lake Grevelingen. Eelgrass from such a historically estuarine area may be more sensitive to high salinities than other, more marine populations. Recovery of the autochthonous eelgrass population is expected to be favoured when the estuarine conditions of the seagrass area are re-established, or when restoration programmes are carried out with allochthonous ecotypes that are less sensitive to high salinities. Received: 23 June 1998 / Accepted: 19 November 1998     

Size and estimated age of genets in eelgrass, Zostera marina, assessed with microsatellite markers

We examined the spatial distribution of genotypes in a perennial population of eelgrass, Zostera marina L., at two spatial scales. We mapped and sampled 80 ramets in a subtidal area of 20 × 80 m, and an additional 15 ramets in two 1-m² sub-quadrats. Ramets were genotyped for seven polymorphic microsatellite loci. Among a total number of 54 genotypes detected, 12 occurred more than once. The ramets of ten of these genotypes occurred in clusters and represented genets based on their expected multi-locus genotype frequencies. The size distribution of genets was uneven with estimated ramet numbers ranging from 2 to 5000. Whereas some areas displayed a high genet diversity, which may indicate past disturbances, large genets (≥10 m²) predominated in other locations of the sampled plot. Spatial heterogeneity in clone distribution was also obvious at the smaller sampling scale (15 ramets sampled within 1 m²) where the clonal diversity (genets identified/ramets sampled) was 0.24 in one quadrat, and 0.077 in the other. Ramets belonging to the largest clone were maximally 17 m apart, which corresponds to a genet age of 67 yr based on annual rhizome growth rates. We conclude that the spatial arrangement of clones in seagrasses allows inferences about the past demography and the disturbance regime at a given site which may prove useful for coastal zone management of ecologically valuable seagrass meadows. Received: 15 September 1998 / Accepted: 14 November 1998     

Utilization of growth parameters of eelgrass,Zostera marina,for productivity estimation under laboratory and in situ conditions

Allometry was used for monitoring aboveground growth of the marine angiosperm Zostera marina L. Dry weight was regressed with leaf length and width, allowing estimation of aboveground net productivity and biomass of individual plants. At the termination of the experiment, rhizome productivity of the same plants was determined by harvesting. Plants in shaded and unshaded seawater tanks were monitored from June until September, 1976; in situ plants were also monitored at Point Judith Pond, Rhode Island, USA. Unshaded plants had shorter leaves, a lower net productivity, lower biomass, and a lower aboveground-torhizome productivity ratio than shaded plants. Unshaded plants had a higher rate of rhizome branching and the resulting new shoot formation than in situ plants.     

Effect of nutrient enrichment on growth of the eelgrass Zostera marina in the Chesapeake Bay,Virginia, USA

The addition of two commerical fertilizers, one 5% NH₄NO₃, 10% P₂O₅, 10% K₂O, and the other 10% NH₄NO₃, 10% P₂O₅, 10% K₂O, ahd a dramatic effect on the growth of Zostera marina in the Chesapeake Bay. There was a significant increase in the length, biomass and total number of turions of fertilized plots compared with controls during a 2 to 3 month period. Data from this short-term field experiment suggest that Z. marina beds in the Chesapeake Bay are nutrient-limited, that the grwoth form of Z. marina may be related to the sediment nutrient supply, and that Z. marina may competitively exclude Ruppia maritima by light-shading.     

Labyrinthula sp., a marine slime mold producing the symptoms of wasting disease in eelgrass,Zostera marina

Coastal ecosystems along the eastern United States are presently threatened by a recurrence of the wasting disease of eelgrass, Zostera marina L. Using Koch's postulates, a species of the marine slime mold, Labyrinthula, is identified as the causal microorganism of this disease. Our disease tests for pathogenicity performed on eelgrass, using four Labyrinthula spp., indicate only one species produces the disease symptoms identical to those found associated with the wasting disease. The pathogenic Labyrinthula sp. has morphological characteristics that distinguish it from the other three species. Identification of Labyrinthula spp. is difficult because species described in the literature are not clearly characterized or identifiable. Tests at various salinities demonstrate that disease symptoms appear infrequently at salinities of 10%. or less.     

Cadmium and manganese flux in eelgrass Zostera marina I. Modelling dynamics of metal release from labelled tissues

Release of cadmium and manganese radionuclides from eelgrass leaves and root-rhizomes into seawater and seawater plus 1×10^-4 disodium ethylenediaminetetraacetate (EDTA) was monitored over periods of 6 h following incubations of 1 to 98 h. Flux of both isotopes from tissues is initially rapid and enhanced by the addition of EDTA. The initial release rate is independent of incubation time, indicating desorption of metals from exterior tissue surfaces. High initial release rates become rapidly attenuated. Fitting a power curve to the data proved valuable in inferring uptake capacity of tissues from observed metal release characteristics. In addition to desorption, and diffusion from intercellular spaces, diffusion from cells and biologically controlled release could be described with the aid of curve fits. Manganese apparently is released more slowly during later phases of release than is cadmium, pointing to greater biological accumulation potential.This research was supported by grants to B. H. Brinkhuis from the New York Sea Grand Institute and the Nassau-Suffolk Regional Planning Board, and by a Jessie Smith-Noyes Fellowship and Sigma Xi grant to W. F. PenelloMarine Sciences Research Center Contribution No. 247     

Nitrate metabolism in sediments from seagrass (Zostera capricorni) beds of Moreton Bay,Australia

The fate of nitrate in sediments from seagrass (Zostera capricorni Aschers.) beds of Moreton Bay on the subtropical eastern coast of Queensland, Australia, was investigated. Added nitrate was metabolised at rates of 0.4 to 3.4 g N cm^-3 d^-1 when sediments were incubated under anaerobic conditions with a large excess of nitrate. The potential rate of nitrate utilization was as rapid in sediments from subtidal bare areas as from adjacent seagrass beds. Ammonium was produced rapidly from¹⁵N-nitrate by microbial action in all the subtidal sediments examined. After 12 h of incubation, 13 to 28% of the¹⁵N initially added as labelled nitrate was detected as labelled ammonium in the sediments. Denitrification, although not measured directly, appeared to be a relatively minor fate of nitrate. Benthic microbes took up large amounts of¹⁵N but only after a delay of 6 h; this pattern could have been due to induction and synthesis of the enzymes necessary for nitrate uptake, and the assimilation of labelled ammonium. Under field conditions, assimilation by seagrasses and denitrification by bacteria were probably not significant sinks for nitrate in comparison with uptake by benthic microbes and dissimilatory reduction to ammonium.     

Interactions between two introduced species: Zostera japonica (dwarf eelgrass) facilitates itself and reduces condition of Ruditapes philippinarum (Manila clam) on intertidal flats

Dwarf eelgrass (duckgrass; Zostera japonica) and Manila clams (Ruditapes philippinarum) are two introduced species that co-occur on intertidal flats of the northeast Pacific. Through factorial manipulation of clam (0, 62.5, 125 clams m⁻²) and eelgrass density (present, removed by hand, harrowed), we examined intra- and interspecific effects on performance, as well as modification of the physical environment. The presence of eelgrass reduced water flow by up to 40% and was also observed to retain water at low tide, which may ameliorate desiccation and explain why eelgrass grew faster in the presence of conspecifics (positive feedback). Although shell growth of small (20–50 mm) clams was not consistently affected by either treatment in this 2-month experiment, clam condition improved when eelgrass was removed. Reciprocally, clams at aquaculture densities had no effect on eelgrass growth, clam growth and condition, or porewater nutrients. Overall, only Z. japonica demonstrated strong population-level interactions. Interspecific results support an emerging paradigm that invasive marine ecosystem engineers often negatively affect infauna. Positive feedbacks for Z. japonica may characterize its intraspecific effects particularly at the stressful intertidal elevation of this study (+1 m above mean lower low water).