Temperature and emergence effects on the net photosynthesis of two Zostera noltii Hornem. morphotypes

Apparent photosynthetic rates (APS) of two Zostera noltii Hornem. morphotypes were measured in air and in water at different temperatures with a closed infra-red gas analysis system (IRGA).Hyperbolic functions accurately described the photosynthesis-CO₂ relationships when the leaves were exposed to air. The photosynthetic behaviour in water, on the contrary, could not be described by Michaelis type kinetics, due to the existence of a rapid transition from the initial slope to the saturation phase. Both morphotypes (narrow-leaved, NLM and large-leaved, LLM) showed higher APS rates in water than in air, although the highest APS rates, in air as well in water, were recorded for the NLM.Temperature had a significant influence on the photosynthetic parameters: APS_max (maximum photosynthetic rate) decreased (in air and in water) with increased temperature in both morphytypes; compensation points (CP) in air increased at high temperature, especially in the LLM. NLM specimens showed enhanced affinity (lower Km) with increasing temperature in air. On the contrary, Km values in water were not significantly affected by temperature.The results suggest that NLM specimens are better adapted than the LLM to occur exposed to air. The distributional pattern of the two morphotypes in the Palmones Estuary is discussed on the basis of their photosynthetic behaviour.     

Morphological and physiological differences between two morphotypes of Zostera noltii Hornem. from the south-western Iberian Peninsula

The morphological and physiological differences between two morphotypes of Z. noltii Hornem. were studied in the intertidal meadows on the south-western Iberian Peninsula (Palmones river estuary and Ria Formosa). A small-leaved morphotype (SM) grows mainly at high intertidal sites, meadow edges or in recently deposited sandbanks, whereas a large-leaved morphotype (LM) generally thrives in well-structured beds or in deeper places. This study deals with the morphological, biochemical and physiological differences between these morphotypes as well as the ecological implications of the occurrence of different morphotypes in the same meadow. Shoot length, leaf width, rhizome internode length, roots per node, root length, leaf nutrient and pigment contents, and photosynthetic rates of both morphotypes were compared. The below-ground architecture (root and rhizome complex) of both morphotypes was more developed in sites characterized by higher hydrodynamics and/or a lower nitrogen content in sediments. Both morphotypes showed similar values for photosynthetic efficiency, dark respiration rate and compensation irradiance. On the other hand, the net photosynthetic capacity was much greater (5-fold) for the SM. This difference could explain the greater growth rate and faster leaf turnover rate of the SM compared with the LM. The occurrence of the SM in newly settled areas (and in the meadow edges) could be explained on the basis of its higher growth rate, which would allow a faster spreading of the meadow and/or better recovery after burial resulting from stormy weathers. Received: 20 December 1998 / Received in revised form: 21 April 1999 / Accepted: 23 April 1999     

Short-term phosphate uptake kinetics in Zostera noltii Hornem: a comparison between excised leaves and sediment-rooted plants

Short-term phosphate uptake by excised leaves of Zostera noltii Hornem. as well as by leaves of sediment-rooted plants were characterized and compared in a kinetic framework. Time courses of phosphate disappearance were measured over a wide range of initial substrate concentrations. Phosphate uptake determined by this perturbation method did not follow Michaelis-Menten kinetics. Both excised leaves and sediment-rooted plants exhibited a biphasic uptake pattern as a function of phosphate concentration. However, rooted plants showed higher uptake rates and accumulated higher amounts of phosphate than excised leaves. The results point out the importance of the structural and functional coupling between shoots and underground parts during the nutrient foliar uptake processes. Our study also indicates that Zostera noltii leaves function as a phosphate sink in the water column.A second objective of this work is to compare the perturbation and the multiple flask methods in determining the uptake kinetic parameters. The obtained results support that both methods provide valuable and complementary information in determining the uptake rates.     

Factors influencing primary production of seagrass beds (Zostera noltii Hornem.) in the Thau lagoon (French Mediterranean coast)

The primary production and the respiration of Zostera noltii beds in the Thau lagoon were studied by means of the benthic bell jar technique. Concurrently, environmental data (temperature, light and nutrients) as well as morphological data of seagrass meadows (leaf width and height, density of shoots, above/below-ground biomass ratio) were collected with the purpose of explaining most of the observed variations in metabolism. Seagrass plus epiphyte respiration rates were influenced mainly by the water temperature, showing a typical exponential response to an increase in temperature. Surprisingly, measurements of production rates were not related to incoming light intensities recorded at the seagrass canopy level. An equation frequently used for terrestrial standing crops, involving the leaf area index (LAI) and the characteristics of the canopy architecture (parameter K, depending on leaves optical and geometrical properties), was applied to the seagrass ecosystem in order to estimate the light energy actually available for the plants, i.e. the light intercepted by the seagrass canopy (Q(abs)). Linear relationships were then validated between gross production rates and calculated Q(abs) for Z. noltii beds, and the best fits were obtained with K values nearing 0.6, confirming the similarities between terrestrial graminaceae and seagrasses. A linear regression model for primary production is proposed, involving the calculated Q(abs), the water temperature and the leaf nutrient content.     

Leaf dynamics and standing stocks of intertidal Zostera noltii Hornem. and Cymodocea nodosa (Ucria) Ascherson on the Banc d'Arguin (Mauritania)

Leaf dynamics and standing stocks of intertidal seagrasses were studied in the Baie d'Aouatif (Parc National du Banc d'Arguin, Mauritania) in April and September 1988. Standing stocks of Zostera noltii Hornem. suggest a unimodal seasonal curve similar to what is found for populations at higher latitudes. Also, leaf growth rates (0.03 cm² cm^–2 day^–1 on average) were similar to those found at higher latitudes in these months. Variation in leaf loss over tidal depth, time and different locations in the Baie d'Aouatif was larger and more often significant than variation in leaf growth. In general, Z. noltii beds in the Baie d'Aouatif had comparable leaf growth rates and standing stocks. In both months losses were almost always higher than or equal to growth.Variation in leaf loss over time was much higher in the plots that were situated high in the intertidal than in lower plots. This is explained by differences in susceptibility to sloughing, which is presumably higher in periods with low tide around noon for shallow depths.In an experiment using artificial shading nets, in situ leaf growth was affected negatively from 94% shading onwards. This shading was observed to reduce the light intensity reaching the seagrass bed to a level below the reported range of light compensation points for Z. noltii. Cymodocea nodosa (Ucria) Ascherson on average had higher leaf area and relative growth rates than Z. noltii and much lower loss rates, resulting in a positive net increase in September. Standing stocks were also higher than for Z. noltii. A mixed seagrass bed containing the above two species and Halodule wrightii Ascherson had the highest observed total biomass: 335 g m^–2 ash-free dry weight.     

Dinamica dei sistemi a Cymodocea nodosa,Zostera marina e Zostera noltii nel Mediterraneo

Abstract

Data on geographical and depth distribution, sediment granulometry, salinity, biomass variability, below/aboveground biomass ratio, and reproductive strategies of seagrass communities in the Mediterranean Sea were analysed to describe their dynamics patterns.

Notwithstanding their different latitudinal distribution, they have a similar seasonal biomass variability, deriving both from extrinsic forcing (e.g. light and temperature) and intrinsic species-specific components (e.g. reproduction pattern), these latter being responsible for their different structure and seasonal dynamics.     

Characterization and seasonal variation of individual flavonoids in Zostera marina and Zostera noltii from Norwegian coastal waters

The flavonoid content in leaves of Zostera marina and the endangered Zostera noltii, including mono- and disulphated flavonoids, from different sample localities were characterized. Seasonal variation of both individual and total flavonoid, as well as rosmarinic acid concentration were revealed. Minor amounts of luteolin 7-(6″-malonyl)glucoside (6) and apigenin7-(6″-malonyl)glucoside (11) were identified in Z. noltii for the first time. The total flavonoid content was found to be higher in Z. noltii than in Z. marina at most of the examined localities, and the qualitative flavonoid content was somewhat different in the two species. The quantitative variation of flavonoids and rosmarinic acid was found to be relatively consistent from year to year in Z. marina during a period of three years. The two species appeared though to have a different flavonoid production in the various seasons at the West coast. While Z. marina had the highest content in young leaves in May or June, with a markedly decrease from June to September and the lowest measured content in February, Z. noltii had the lowest measured flavonoid content in May/June followed by an increase from June to September and the highest measured content during wintertime in February. The observed seasonal differences may be related to the fact that Z. noltii is considered a perennial, thermophilous species, and the increasing flavonoid production during the colder seasons from September to March/April in Norway may serve as a protective function.     

Seasonal variation in short-term survival of Zostera noltii transplants in a declining meadow in Portugal

In one decade (from 1986 to 1997), the area occupied by Zostera noltii in the Mondego estuary (western coast of Portugal) decreased from 150,000 to 200 m². Our purpose was to identify the most suitable season for transplantation of Z. noltii, as one of the methodologies included in a broader restoration plan of the local Z. noltii meadows. Each month a short-term transplantation experiment (20 days) was undertaken, where the net growth rate of transplants, the number of lost shoots and the number of surviving shoots was estimated. Transplant loss variation was markedly seasonal (ANOVA, F_1,12 = 13.11, p < 0.01). From spring to middle autumn (April–October), the amount of lost shoots was significantly higher (average = 63%) than during late autumn and winter (average = 26%). Multiple regression results suggest that the number of surviving shoots was negatively related with insolation and NH₄-N concentration. Additionally, the net growth rate of the transplants was negatively related with salinity.In conclusion, we recommend that at southern European latitudes, Z. noltii transplantation be undertaken during late autumn and winter. The success of transplantation at this time of the year seems to be related with the prevailing external conditions, namely, low temperatures and low photon flux densities. Furthermore, we hypothesise that the proportion of aboveground to belowground biomass in Z. noltii may also influence transplant survival. Transplantation success was higher whenever belowground biomass was higher than aboveground biomass. In the Mondego estuary, in autumn and winter, the belowground biomass of Z. noltii represents about 51% of the total plant biomass, while in spring and summer, this value decreases to 31% of the total biomass.     

The effects of the polychaete Nereis diversicolor on the distribution and transplanting success of Zostera noltii

Loss of saltmarsh vegetation in south-east England is a significant problem for conservation and coastal defence. The losses of vegetation began in the 1930s with the loss of intertidal Zostera marina and have continued more recently. Some preliminary trials at re-establishing Zostera in some estuaries of Essex have not been successful. This paper addresses the hypothesis that the infauna, particularly the polychaete Nereis diversicolor, may restrict natural colonisation by Zostera and reduce the success of transplanting trials. In field experiments, Z. noltii were transplanted into areas where Nereis were common, close to an established seagrass bed and into two other estuaries. The transplants protected from the effects of the polychaetes by netting had a higher survival rate, lower index of root damage and greater biomass at the end of the experiments than those that were unprotected. In laboratory experiments, Nereis reduced the survival of Z. noltii. They were observed grasping the leaves and pulling them into their burrows. These results indicate that herbivory and disturbance by N. diversicolor is responsible, at least in part, for the restriction of the distribution of Z. noltii and may have been important in limiting the success of previous transplanting experiments. A hypothesis is proposed which states that there are two stable states on the upper mud-flats. One state is dominated by plants, including Zostera spp., which prevent colonisation by burrowing infauna, and the other is dominated by infauna which prevent colonisation by plants. Managing these two states could be the key to re-establishing the early successional stages of saltmarsh development. Received: 15 June 1999 / Received in revised form: 6 July 1999 / Accepted: 8 July 1999     

Expressed sequence tags from heat-shocked seagrass Zostera noltii (Hornemann) from its southern distribution range

Predicted global climate change threatens the distributional ranges of species worldwide. We identified genes expressed in the intertidal seagrass Zostera noltii during recovery from a simulated low tide heat-shock exposure. Five Expressed Sequence Tag (EST) libraries were compared, corresponding to four recovery times following sub-lethal temperature stress, and a non-stressed control. We sequenced and analyzed 7009 sequence reads from 30 min, 2 h, 4 h and 24 h after the beginning of the heat-shock (AHS), and 1585 from the control library, for a total of 8594 sequence reads. Among 51 Tentative UniGenes (TUGs) exhibiting significantly different expression between libraries, 19 (37.3%) were identified as ‘molecular chaperones’ and were over-expressed following heat-shock, while 12 (23.5%) were ‘photosynthesis TUGs’ generally under-expressed in heat-shocked plants. A time course analysis of expression showed a rapid increase in expression of the molecular chaperone class, most of which were heat-shock proteins; which increased from 2 sequence reads in the control library to almost 230 in the 30 min AHS library, followed by a slow decrease during further recovery. In contrast, ‘photosynthesis TUGs’ were under-expressed 30 min AHS compared with the control library, and declined progressively with recovery time in the stress libraries, with a total of 29 sequence reads 24 h AHS, compared with 125 in the control. A total of 4734 TUGs were screened for EST-Single Sequence Repeats (EST-SSRs) and 86 microsatellites were identified.     

Diversity of bacteria and Archaea in sulphate-reducing enrichment cultures inoculated from serial dilution of Zostera noltii rhizosphere samples

We have analysed the diversity of culturable sulphate-reducing bacteria (SRB) in Zostera noltii colonized sediments from Bassin d'Arcachon (France). Four organic substrates have been tested as well as the combination of H2 and CO2 to select for lithotrophic SRB. All energy sources were supplied in parallel cultures that were amended with yeast extract plus NH4+ and prepared without a source of combined nitrogen, the latter to select for diazotrophic SRB. The 10 different enrichment media were inoculated from serial dilution of rhizosphere samples. The highest dilution cultures yielding positive growth (i.e. 10-7) were studied by molecular techniques (16S rDNA clone libraries, RISA and ARDRA). Lactate as a single organic substrate in combination with a source of combined nitrogen resulted in selection of members of the Desulfovibrionaceae. Surprisingly, when lactate was added without a source of combined nitrogen, Desulfobacteriaceae were selected. A strong influence of the presence or absence of combined nitrogen was also observed for the substrates sucrose and fructose. Whereas the liquid culture growing on sucrose and NH4+ systematically yielded 16S rDNA clones related to an environmental unidentified green sulphur bacterium (OPS185), on plates we were able to isolate a SRB related to Desulfovibrio dechloracetivorans, which likely represents a non-described species. Under diazotrophic conditions, sucrose selected for SRB clones related to the cluster formed by Desulfovibrio zosterae, Desulfovibrio salexigens and Desulfovibrio bastinii. The corresponding isolate obtained on plates showed only low sequence similarity with this closest neighbour (93.8%), and we suggest that it also represents a non-described species. Surprisingly, a 16S rDNA sequence corresponding to an archaeon, i.e. a non-extremophile Crenoarchaeota, was retrieved from several of the SRB enrichment cultures even after subsequent transfers.     

Characterization of microsatellite loci in the dwarf eelgrass Zostera noltii (Zosteraceae) and cross‐reactivity with Z. japonica

Zostera noltii is an important species of eelgrass occurring along European, north African, Mediterranean, Black Sea and Azov Sea coasts. Nine microsatellite loci were developed and no linkage disequilibrium was observed. Cross‐amplification was observed for all loci (polymorphic) in Z. japonica; only four loci amplified (monomorphic) in Z. marina.     

Seasonal dynamics of <Emphasis Type="Italic">Zostera noltii</Emphasis> Hornem. in two Mediterranean lagoons

The density, biomass and shoot morphology of two populations of Zostera noltii were monitored from January 1998 to July 1999 at two shallow Mediterranean lagoons of Biguglia and Urbino, which differ in hydro-morphological conditions and nutrient loading. Monitoring included the principal biological and foliar parameters (shoot density, aboveground and belowground biomass, length, width and number of leaves, LAI and coefficient A: percentage of leaves having lost their apex), the organic matter contents of the sediment and the environmental conditions (salinity, turbidity, temperature, nutrient concentrations and dissolved oxygen levels). The two populations of Z. noltii displayed seasonal changes in density (1600–19600 m²), aboveground biomass (11–153 g. DW. m⁻²), leaf length (33–255 mm), and leaf width (0.9–1.8 mm). Temperature and turbidity were significant environmental factors influencing the temporal changes observed in the Z. noltii meadows studied. Conversely, the belowground biomass, the number of leaves per shoot and the LAI did not undergo any seasonal changes. In the Biguglia lagoon, the functioning dynamics of the Z. noltii seagrass beds are determined by the catchment area and the inputs of nutrients derived from it, whereas in the Urbino lagoon the dynamics of the Z. noltiibeds depend on low levels of water turbidity.     

The Effect of Zostera noltii, Spartina maritima and Scirpus maritimus on Sediment Pore-water Profiles in a Temperate Intertidal Estuary

The objective of the present work was to study the effect of plants common in temperate latitudes (Zostera noltii, Spartina maritima and Scirpus maritimus) on sediment nutrient profiles, and to compare it to sand- and mud-flats without vegetation. The study focused on the organic matter contents, the concentration of dissolved inorganic nutrients (PO₄–P, NH₃–N, NO₃–N), an on the estimation of the total amount of these nutrients during day and night conditions and their potential net-fluxes. It was also hypothesised that in an estuarine system, different plants may have specific effects, and consequently different contributions to the system nutrient dynamics as a whole. Sediment profiles of loss on ignition (LOI) showed an increase of the organic matter contents from sand-flat, to Zostera, Spartina, mud-flat and Scirpus. Statistically, there were significant differences between sediment profiles of phosphate, ammonia and nitrate (Mann-Whitney test, p<0.05), during day and night periods. These results suggest that there is an intense mobility of nutrients in the sediment, showing a day-night variation of nutrient concentrations in the pore-water. In the plants’ rhizosphere, the day-night variation of nutrients seemed dependent on plant biomass and penetration of the roots. Additionally, coupling between plant and sediment seems to be a species-specific process. In spring, Scirpus salt marsh reaches the maximum density and biomass, and despite the higher organic matter contents in the plant covered sediment, Scirpus acts as a sink of nutrients. In contrast, the top 10 cm of the sediment in the Spartina salt marsh and in the Zostera beds may contribute to the efflux of nutrients during the night period, especially phosphate.     

Photosynthetic temperature acclimation in two coexisting seagrasses,Zostera marina L. and Ruppia maritima L.

The physiological responses to temperature were investigated in two coexisting seagrasses, Zostera marina L. and Ruppia maritima L. sensu lato from the lower Chesapeake Bay, Virginia. Seven plant collections were made from March to July, 1983 at ambient temperatures of 8–30°C. Both species maintained relatively constant fresh: dry weight ratios and chlorophyll a:b ratios over the five-month period. Total chlorophyll content remained constant in Z. marina while that of R. maritima doubled from March to July. P_max values for both species increased with increasing temperature and declined at temperatures above 19 and 23°C (Z. marina and R. maritima, respectively). P_max values were significantly higher for R. maritima compared to Z. marina at temperatures above 19°C. Both short-term (laboratory) and long-term (in situ) responses to temperature regimes affected estimates of the photosynthetic capacity of both species. Thus, temperature histories of experimental material should be carefully considered when interpreting temperature effects on photosynthesis. This study provides support of the hypothesis that seasonal community dynamics of Z. marina and R. maritima in Chesapeake Bay are regulated in part by different responses to light and temperature.     

Effects of CO 2 enrichment on photosynthesis,growth, and nitrogen metabolism of the seagrass Zostera noltii

Seagrass ecosystems are expected to benefit from the global increase in CO ₂ in the ocean because the photosynthetic rate of these plants may be C_i‐limited at the current CO ₂ level. As well, it is expected that lower external pH will facilitate the nitrate uptake of seagrasses if nitrate is cotransported with H⁺ across the membrane as in terrestrial plants. Here, we investigate the effects of CO ₂ enrichment on both carbon and nitrogen metabolism of the seagrass Zostera noltii in a mesocosm experiment where plants were exposed for 5 months to two experimental CO ₂ concentrations (360 and 700 ppm). Both the maximum photosynthetic rate (P_m) and photosynthetic efficiency (α) were higher (1.3‐ and 4.1‐fold, respectively) in plants exposed to CO ₂‐enriched conditions. On the other hand, no significant effects of CO ₂ enrichment on leaf growth rates were observed, probably due to nitrogen limitation as revealed by the low nitrogen content of leaves. The leaf ammonium uptake rate and glutamine synthetase activity were not significantly affected by increased CO ₂ concentrations. On the other hand, the leaf nitrate uptake rate of plants exposed to CO ₂‐enriched conditions was fourfold lower than the uptake of plants exposed to current CO ₂ level, suggesting that in the seagrass Z. noltii nitrate is not cotransported with H⁺ as in terrestrial plants. In contrast, the activity of nitrate reductase was threefold higher in plant leaves grown at high‐CO ₂ concentrations. Our results suggest that the global effects of CO ₂ on seagrass production may be spatially heterogeneous and depend on the specific nitrogen availability of each system. Under a CO ₂ increase scenario, the natural levels of nutrients will probably become limiting for Z. noltii. This potential limitation becomes more relevant because the expected positive effect of CO ₂ increase on nitrate uptake rate was not confirmed.     

Sulphate reduction and nitrogen fixation rates associated with roots, rhizomes and sediments from Zostera noltii and Spartina maritima meadows

Sulphate reduction rates (SRR) and nitrogen fixation rates (NFR) associated with isolated roots, rhizomes and sediment from the rhizosphere of the marine macrophytes Zostera noltii and Spartina maritima , and the presence and distribution of Bacteria on the roots and rhizomes, were investigated. Between 1‰ and 3‰ of the surface area of the roots and rhizomes of both macrophytes were colonized by Bacteria. Bacteria on the surfaces of S. maritima roots and rhizomes were evenly distributed, while the distribution of Bacteria on Z. noltii roots and rhizomes was patchy. Root- and rhizome-associated SRR and NFR were always higher than rates in the bulk sediment. In particular, nitrogen fixation associated with the roots and rhizomes was 41–650-fold higher than in the bulk sediment. Despite the fact that sulphate reduction was elevated on roots and rhizomes compared with bulk sediment, the contribution of plant-associated sulphate reduction to overall sulphate reduction was small (≤ 11%). In contrast, nitrogen fixation associated with the roots and rhizomes accounted for 31% and 91% of the nitrogen fixed in the rhizosphere of Z. noltii and S. maritima respectively. In addition, plant-associated nitrogen fixation could supply 37–1613% of the nitrogen needed by the sulphate-reducing community. Sucrose stimulated nitrogen fixation and sulphate reduction significantly in the root and rhizome compartments of both macrophytes, but not in the bulk sediment.     

An evaluation of small-scale genetic diversity and the mating system in Zostera noltii on an intertidal sandflat in the Wadden Sea

Background and Aims

The dwarf eelgrass, Zostera noltii, is a predominant inhabitant of soft-bottom intertidal regions along the coasts of northern Europe. It is a monoecious, protogynous angiosperm in which the potential for self-fertilization and inbreeding are high, especially if clone sizes exceed pollen dispersal distances. The aim of the present study was to determine the relationship between mating system and clonal structure, examine the relative roles of geitonogamous selfing and biparental inbreeding, measure pollen availability (multiple paternities) and estimate pollen dispersal.

Methods

A 100-m² plot was established in a large, intertidal Z. noltii meadow on the island of Sylt in the German Wadden Sea. A total of 256 adult shoots was sampled: one from the centre of 100 fixed 1-m² quadrats (large scale resolution) and an additional 156 from within eight randomly selected 1-m² sub-quadrats (small-scale resolution). DNA was extracted from seeds and leaf tissue of all samples and genotyped with nine microsatellite loci.

Key Results

Mating system analysis revealed high multilocus and single locus outcrossing rates. Average pollen dispersal distance was nearly the same as the average genet (clone) size. Multiple paternity was common and 20–30 % of mature seeds originated from matings within the plot. Among inbred seeds, most resulted from geitonogamy rather than biparental inbreeding.

Conclusions

Moderate disturbances intrinsic to the intertidal habitat appear to facilitate seed recruitment by gap formation. Pollen dispersal distances are sufficient to maintain outcrossing and high clonal diversity.