Genetic Polymorphism and Transplantation Success in the Mediterranean Seagrass Posidonia oceanica

We studied the role of genetic variability of donor beds in establishing transplantation criteria for the Mediterranean seagrass Posidonia oceanica. Horizontal rhizomes, collected from three geographically distinct populations, were transplanted into a common bed at a highly human‐impacted locality. The transplantation site was located near one of the donor populations. After three years, the shoots collected in the population adjacent to the transplanting site showed the lowest growth performance. Genetic variability, assessed through the analysis of hypervariable microsatellite regions, and growth performance followed a similar trend. The shoots growing and branching at the highest rate were those collected from populations with the highest heterozygosity values, despite greater geographic distance. No genetic differences were found between the transplanted shoots and shoots from donor meadows, as expected due to the low rate of sexual reproduction in P. oceanica and the short time that had passed since the transplants. The problem of affecting the local gene pool by the introduction of foreign genotypes could arise, but introduction of new alleles could balance the degradation of genetic variability caused by human impact. In general our study suggests that the genetic variability of source material is an important aspect to consider in the development of seagrass restoration strategies.     

Anthesis effects on Posidonia oceanica (L.) Delile phenology in the Bay of Calvi (Corsica,Mediterranean Sea)

In Posidonia oceanica (L.) Delile, anthesis induces a decrease in the number of juvenile leaves resulting in a significant reduction in the number of leaves on the flowering shoots. All the leaves of the flowering shoots are narrower than the leaves of nonflowering shoots. A modification of the leaf growth also appears in flowering shoots: the oldest leaves are longer and the leaves induced during or after anthesis are shorter. At 10 m depth, in the Bay of Calvi, anthesis lasts roughly 3 months and the flowering is induced 7 months before anthesis.     

Vicariance patterns in the Mediterranean Sea: east–west cleavage and low dispersal in the endemic seagrass Posidonia oceanica

Aim The seagrass, Posidonia oceanica is a clonal angiosperm endemic to the Mediterranean Sea. Previous studies have suggested that clonal growth is far greater than sexual recruitment and thus leads to low clonal diversity within meadows. However, recently developed microsatellite markers indicate that there are many different genotypes, and therefore many distinct clones present. The low resolution of markers used in the past limited our ability to estimate clonality and assess the individual level. New high‐resolution dinucleotide microsatellites now allow genetically distinct individuals to be identified, enabling more reliable estimation of population genetic parameters across the Mediterranean Basin. We investigated the biogeography and dispersal of P. oceanica at various spatial scales in order to assess the influence of different evolutionary factors shaping the distribution of genetic diversity in this species. Location The Mediterranean. Methods We used seven hypervariable microsatellite markers, in addition to the five previously existing markers, to describe the spatial distribution of genetic variability in 34 meadows spread throughout the Mediterranean, on the basis of an average of 35.6 (± 6.3) ramets sampled. Results At the scale of the Mediterranean Sea as a whole, a strong east–west cleavage was detected (amova) . These results are in line with those obtained using previous markers. The new results showed the presence of a putative secondary contact zone at the Siculo‐Tunisian Strait, which exhibited high allelic richness and shared alleles absent from the eastern and western basins. F statistics (pairwise θ ranges between 0.09 and 0.71) revealed high genetic structure between meadows, both at a small scale (about 2 to 200 km) and at a medium scale within the eastern and western basins, independent of geographical distance. At the intrameadow scale, significant spatial autocorrelation in six out of 15 locations revealed that dispersal can be restricted to the scale of a few metres. Main conclusions A stochastic pattern of effective migration due to low population size, turnover and seed survival is the most likely explanation for this pattern of highly restricted gene flow, despite the importance of an a priori seed dispersal potential. The east–west cleavage probably represents the outline of vicariance caused by the last Pleistocene ice age and maintained to this day by low gene flow. These results emphasize the diversity of evolutionary processes shaping the genetic structure at different spatial scales.     

Genetic structure in the Mediterranean seagrass Posidonia oceanica: disentangling past vicariance events from contemporary patterns of gene flow

The Mediterranean Sea is a two‐basin system, with the boundary zone restricted to the Strait of Sicily and the narrow Strait of Messina. Two main population groups are recognized in the Mediterranean endemic seagrass Posidonia oceanica, corresponding to the Western and the Eastern basins. To address the nature of the East–West cleavage in P. oceanica, the main aims of this study were: (i) to define the genetic structure within the potential contact zone (i.e. the Strait of Sicily) and clarify the extent of gene flow between the two population groups, and (ii) to investigate the role of present water circulation patterns vs. past evolutionary events on the observed genetic pattern. To achieve these goals, we utilized SSR markers and we simulated, with respect to current regime, the possible present‐day dispersal pattern of Posidonia floating fruits using 28‐day numerical Lagrangian trajectories. The results obtained confirm the presence of the two main population groups, without any indices of reproductive isolation, with the break zone located at the level of the Southern tip of Calabria. The populations in the Strait of Sicily showed higher affinity with Western than with Eastern populations. This pattern of genetic structure probably reflects historical avenues of recolonization from relict glacial areas and past vicariance events, but seems to persist as a result of the low connectivity among populations via marine currents, as suggested by our dispersal simulation analysis.     

Lulwoana sp., a dark septate endophyte in roots of Posidonia oceanica (L.) Delile seagrass

Posidonia oceanica is the most common, widespread and important monocotyledon seagrass in the Mediterranean Basin, and hosts a large biodiversity of species, including microorganisms with key roles in the marine environment. In this study, we ascertain the presence of a fungal endophyte in the roots of P. oceanica growing on different substrata (rock, sand and matte) in two Sicilian marine meadows. Staining techniques on root fragments and sections, in combination with microscope observations, were used to visualise the fungal presence and determine the percentage of fungal colonisation (FC) in this tissue. In root fragments, statistical analysis of the FC showed a higher mean in roots anchored on rock than on matte and sand. In root sections, an inter‐ and intracellular septate mycelium, producing intracellular microsclerotia, was detected from the rhizodermis to the vascular cylinder. Using isolation techniques, we obtained, from both sampling sites, sterile, slow‐growing fungal colonies, dark in colour, with septate mycelium, belonging to the dark septate endophytes (DSEs). DNA sequencing of the internal transcribed spacer (ITS) region identified these colonies as Lulwoana sp. To our knowledge, this is the first report of Lulwoana sp. as DSE in roots of P. oceanica. Moreover, the highest fungal colonisation, detected in P. oceanica roots growing on rock, suggests that the presence of the DSE may help the host in several ways, particularly in capturing mineral nutrients through lytic activity.     

Effects of Dredging Activities on Population Dynamics of Posidonia oceanica (L.) Delile in the Mediterranean Sea: The Case Study of Capo Feto (SW Sicily, Italy)

Between 1981 and 1993 a methane pipeline was deployed between Sicily (Italy) and Tunisia. This involved the construction of a pipeline trench, which damaged the Posidonia oceanica (L.) Delile meadow at Capo Feto (SW Sicily, Italy) and disturbed the surrounding meadow. Seagrass growth and population dynamics were examined at different depth ranges and at increasing distances from the construction site outer limit (5, 15, 30, 50 m). Results showed significant differences between the shallow (10±3.3 m) and the deep (20±4.6 m) meadow as well as differences among distances. The age structure of P. oceanica varied along the distance gradient and with depth. The mortality rate decreased with distance from the trench at all depth ranges, showing that the plants close to the excavation suffered a higher level of disturbance. Turnover and annual gross shoot recruitment rate (R_gross) were higher in the shallow portion of the meadow than in the deep range. Forecast of future meadow development (R_net) close to the trench indicates that, if present conditions are maintained, shoot density will be reduced by 50% over the next 6 to 17 yrs.     

Detrital stocks and dynamics of the seagrass Posidonia oceanica (L.) Delile in the Spanish Mediterranean

Previous studies have shown that most leaf production (>90%) of the seagrass Posidonia oceanica is shed after senescence and that a substantial percentage (up to 80%) may thereafter be exported off the seagrass meadows by waves and currents. It has also been reported that P. oceanica meadows can accumulate large stocks of belowground detritus due to slow decomposition rates. However, the generality of these results across broad spatial scales is poorly known. In this report, we examine the fate of leaf production and the magnitude and dynamics of belowground detritus in 16 P. oceanica meadows distributed along the Spanish Mediterranean. Herbivores removed a small percentage of leaf production in all the meadows (≤13%), with most leaf production (>85%) being shed after senescence. Most shed leaves (>90%) were exported off the meadows by physical agents, such as waves and currents. The amount of belowground detritus stored within 10–15 cm from the sediment surface varied from ca. 70 to 7500 g DW m⁻² among the meadows examined, and they accumulated at rates ranging from ca. 65 to 650 g DW m⁻² per year. These values are large when compared to other communities of aquatic and terrestrial macrophytes. Our results show that P. oceanica meadows in the Spanish Mediterranean support high values of secondary production in other systems by exporting large amounts of leaf detritus as well as acting as substantial carbon sinks by accumulating large reservoirs of belowground detritus. Therefore, the increasing anthropogenic threats on P. oceanica could entail an important loss of secondary production and carbon storage in Mediterranean coastal ecosystems.     

Genetic variability of Posidonia oceanica (L.) Delile in relation to local factors and biogeographic patterns

To evaluate genetic differences of Posidonia oceanica (L.) Delile both at smaller (within a meadow) and larger scale (Mediterranean basin), plants of P. oceanica were analyzed by PCR technique and compared using random amplified polymorphic DNA (RAPD) markers. Results were associated to known differences in phenology. At the small-scale level, P. oceanica shoots collected in the bay of Monterosso al Mare (Liguria, NW Mediterranean Sea) showed genetic differences among sampling stations, with a decrease in genetic diversity along an anthropogenic disturbance gradient. At basin level, genetic differences were detected among 11 P. oceanica shoots coming from different regions of the Mediterranean, and transplanted to the Port-Cros National Park (France) between 1989 and 1991: Izmir, Turkey; Athens, Greece; Taranto, Italy; Ischia Island, Italy; Lavezzi, France; Port-Cros, France; Banyuls, France; Palma de Majorca, Balearic Islands, Spain; Marsa Bay, Algiers. By cluster analysis two major Mediterranean groups were distinguished, the Eastern Mediterranean Group (EMG) and the Western Mediterranean Group (WMG). This suggests that eastern and western populations of P. oceanica have diverged during the colonization of the Mediterranean (after near extinction of the Mediterranean biota in the Messinian period, approximately 5.6 million years ago), and have experienced little gene flow between them. Cluster analysis also indicated that previously described phenological differences among P. oceanica populations in different sectors of the Mediterranean are not mere phenotypic responses to different climatic and hydrological conditions but may well have a genetic basis.     

Consequences of Mediterranean warming events in seagrass (Posidonia oceanica) flowering records

Posidonia oceanica, a seagrass endemic to the Mediterranean forms extended and extremely persistent meadows. It is a clonal plant with an apparently irregular pattern of flowering events. An extensive bibliographic review allowed the reconstruction of past flowering events of this species around the Mediterranean, with a high degree of confidence for the last 30 years. The data series on annual flowering prevalence (FP, flowering records per total records) and flowering intensity (FI, fraction of flowering shoots) produced have been compared with four series on Sea Surface annual Temperature maxima (SST_max) obtained for the NW Mediterranean (averaged from the local data series of l'Estartit and Villefranche: 1957–2005) and for the Eastern, Western basin and the whole Mediterranean sea (extracted from NCEP Reynolds interpolated SST maps: 1982–2005). Significant warming trends are detected in the Mediterranean SST_max series, at a rate of (mean+SE) 0.04±0.01°C yr⁻¹ (R²=0.24, P<0.01, N=24 years), in the Eastern basin series (0.06±0.01°C yr⁻¹, R²=0.43, P<0.001, N=24 years) and in the long SST_max series of the NW Mediterranean (0.02±0.01 C yr⁻¹, R²=0.12, P<0.02, N=49 years). The magnitudes of the SST_max anomalies around the absolute warming trend do not increase with time in any SST_max series. Peaks of FP and FI in the Mediterranean seem to occur each 9–11 years, and coincide with peaks of annual SST_max. Annual FP and FI increase with the residuals of annual SST_max warming trend in all Mediterranean basins (FP_MED: R²=0.27, P<0.01, N=23; FP_NW: R²=0.34, P<0.01, N=31; FP_E: R²=0.20; P<0.10, N=23). An outstanding event of P. oceanica flowering across the Mediterranean has been registered in Autumn 2003; 1 month after the highest annual SST_max recorded in the series. The hypothesis of flowering induction by thermal stress as the possible cause of this relationship is discussed, as well as the potential use of P. oceanica flowering record as early indicator of biological change induced by global sea warming in Mediterranean marine ecosystems.     

Experimental Evaluation of the Restoration Capacity of a Fish‐Farm Impacted Area with Posidonia oceanica (L.) Delile Seedlings

Marine aquaculture is an activity that has induced severe local losses of seagrass meadows along the coastal areas. The purpose of this study was to evaluate the capacity of an area degraded by fish‐farm activities to support Posidonia oceanica seedlings. In the study site, a bay in the southeast coast of Spain where part of a meadow disappeared by fish‐farm activities, seedlings inside mesh‐pots were planted in three areas. Two plots were established in each area, one in P. oceanica dead matte and another inside a P. oceanica meadow. To evaluate if sediment conditions were adequate for the life of the seedlings, half of them were planted in direct contact with the sediment and the other half were planted above the surface of the sediment in each plot. Monitoring during 1 year showed that there were large differences in seedling survival between the dead matte and the P. oceanica meadow. While seedlings planted in dead matte had a high survivorship after 1 year (75%), seedlings planted in P. oceanica progressively died (survivorship of 20% after 1 year). The average leaf length of the seedlings surviving in the two substrata was not different, but the leaf area per seedling was lower in the seedlings growing inside the P. oceanica meadow during most part of the year. Seedling survivorship and vegetative development were not affected by the level of planting and suggest that the sediment conditions are adequate for the life of P. oceanica seedlings.     

Genetic differentiation of white sea bream within the Lion's Gulf and the Ligurian Sea (Mediterranean Sea)

An allozymic study was conducted on 190 individuals of white sea bream, Diplodus sargus sampled on five locations throughout the Lion's Gulf and the Ligurian Sea (Mediterranean Sea), in order to identify genetic structure. Electrophoretic surveys carried out on muscle and liver tissues identified 25 loci of which 12 were polymorphic ( P 0.95). G -test analysis shows significant differences on allelic frequencies between the five stations at six loci. Examination of the spatial structure was performed using Nei's distances and F statistics, and indicated significant genetic differences between three groups. A group which clustered Blanes (Spain), Marseille (France) and Livorno (Italy) where the absence of genetic difference can be explained by migration of larvae and adults along a coastal 'continuum'. The Elba sample (Italy) shows genetic divergence from other samples and this difference may result from isolation due to limited migration of larvae and adults. Banyuls (France), differs from all other stations. Several hypotheses are proposed to explain genetic patterns including local current systems, larval dispersal, geographic isolation and historical effects, and variation in the size classes of sampled individuals between sites.     

Detection of environmental impacts by bottom trawling onPosidonia oceanica (L.) Delile meadows: sensitivity of fish and macroinvertebrate communities

Along the Mediterranean coast,Posidonia oceanica (L.) Delile meadows have a great ecological and economical importance. However, there is a general regression of these meadows due to human activities such as illegal bottom trawling, may be affecting to overall ecosystem health. We examined changes in the community structure of mobile fauna associated withP. oceanica meadows at different spatial scales and taxonomic levels. The aim of this paper was to identify the most efficient taxonomic level to use in environmental impact studies of bottom trawling. At the macroscale level (10 to 100 m), there were significant differences between sites in the densities of some fish species and also the total fish assemblage structure, at both family and species taxonomical levels. At the microscale (0.1 to 1 m), some species of amphipods and isopods showed significant differences in their population densities. In the overall analysis of community structure, the coarse taxonomical levels, such as phyla and class, did not show significant differences, however amphipods and isopods showed significant differences at family and species levels. From these results, both study scales are required to detect changes onPosidonia meadows' fauna. Monitoring of some fish species such asDiplodus annularis (Linnaeus, 1758) and the overall fish assemblage as well as the structure of the amphipod and isopod communities appears to be the most efficient tool in the assessment of environmental impacts by bottom trawling onP. oceanica meadows.     

A meta‐analysis reveals a positive correlation between genetic diversity metrics and environmental status in the long‐lived seagrass Posidonia oceanica

The seagrass Posidonia oceanica is a key engineering species structuring coastal marine systems throughout much of the Mediterranean basin. Its decline is of concern, leading to the search for short‐ and long‐term indicators of seagrass health. Using ArcGIS maps from a recent, high‐resolution (1–4 km) modelling study of 18 disturbance factors affecting coastal marine systems across the Mediterranean (Micheli et al. 2013, http://globalmarine.nceas.ucsb.edu/mediterranean/ ), we tested for correlations with genetic diversity metrics (allelic diversity, genotypic/clonal diversity and heterozygosity) in a meta‐analysis of 56 meadows. Contrary to initial predictions, weak but significantly positive correlations were found for commercial shipping, organic pollution (pesticides) and cumulative impact. This counterintuitive finding suggests greater resistance and resilience of individuals with higher genetic and genotypic diversity under disturbance (at least for a time) and/or increased sexual reproduction under an intermediate disturbance model. We interpret the absence of low and medium levels of genetic variation at impacted locations as probable local extinctions of individuals that already exceeded their resistance capacity. Alternatively, high diversity at high‐impact sites is likely a temporal artefact, reflecting the mismatch with pre‐environmental impact conditions, especially because flowering and sexual recruitment are seldom observed. While genetic diversity metrics are a valuable tool for restoration and mitigation, caution must be exercised in the interpretation of correlative patterns as found in this study, because the exceptional longevity of individuals creates a temporal mismatch that may falsely suggest good meadow health status, while gradual deterioration of allelic diversity might go unnoticed.     

Investigation of the sorption mechanisms of metal-complexed dye onto Posidonia oceanica (L.) fibres through kinetic modelling analysis

This research deals with the exploitation of highly available and renewable marine biomass, Posidonia oceanica (L.) fibres as low cost biosorbent for the removal of metal-complexed textile dyestuff from aqueous medium, and the investigation of the probably involved physiochemical mechanisms. Experiments were carried out in batch reactor. Firstly, the adsorption process was studied as a function of initial solution pH and contact time under different initial dye concentration. The results showed that the highest dye adsorption capacity was found at pH 2 under a constant temperature of 30 °C, and the equilibrium state was reached within 48 h of exposure time. Secondly, several adsorption kinetic models were applied to fit the experimental data, namely Lagergren irreversible first-order, Reversible first-order, Pseudo-second-order, Elovich, Ritchie and intraparticle diffusion models. The proposed explanations were deduced from the theoretical assumptions behind the most appropriate model(s), which could satisfactorily describe the present biosorption phenomenon. The interpretation of the related results have shown that, with R² of about 99%, the pseudo-second order model is the most suitable dynamic theory describing the biosorption of metal complex dye onto P. oceanica fibres, predicting therefore a chemisorption process.     

Planktonic larval stages of Brachyura in the Gulf of Tigullio (Ligurian Sea,Italy)

Summary

Horizontal planktonic catches made in the Gulf of Tigullio (Genoa, Ligurian Sea) have been analyzed with the aim of identifying the zoeae of Brachyura, whose knowledge is poor in all the northern Mediterranean Sea. Forty-four taxa (42 species plus Liocarcinus spp. and Brachynotus spp.) have been identified, 12 of which were present with all the larval stages. The seasonal distribution of the taxa has been analyzed, and it was possible to recognize species present throughout the year as well as spring/summer, summer/autumn, and winter/summer species. Problems regarding zoeae of dubious identification are discussed.     

A catastrophic mass-mortality episode of gorgonians and other organisms in the Ligurian Sea (North-western Mediterranean), summer 1999

In the late summer of 1999, an extensive mortality of gorgonians and other epi-benthic organisms was observed in the Ligurian Sea (Mediterranean Sea) from the Tuscan Archipelago to Marseille. Quantitative data from Tino Island and Portofino Promontory indicated that the proportion of affected gorgonians ranged from 60% to 100% in populations having a density of 9–27.8 colonies m⁻², suggesting that millions of sea fans died along the coast of Liguria. This mass mortality episode coincided with a sudden increase of sea water temperature down to more than 50 m depth. Laboratory analyses showed that the colonies stressed by high temperature also underwent extensive attack by microrganisms (protozoans and fungi), which are interpreted as opportunistic pathogens.     

Unexpected response of the seagrass Posidonia oceanica to a warm-water episode in the north western Mediterranean Sea

The response of Posidonia oceanica (Linnaeus) Delile to the warm-water episode of summer 1999 was studied by means of the technique of lepidochronology. Study sites include three sites affected by the mass mortality event of benthic invertebrates and one not affected. The results showed a significant decline in some parameters (number of leaves and/or rhizome growth) for the three sites affected by the mass mortality event for the year following the warm-water episode (1999-2000). A similar decline was not observed for the unaffected site. The fact that high temperatures could have a negative impact on deep Posidonia oceanica near its cold limit of distribution is an unexpected result.     

Detecting environmental impacts on the Mediterranean seagrass Posidonia oceanica (L.) Delile: the use of reconstructive methods in combination with 'beyond BACI' designs

Interannual changes in leaf production and rhizome elongation rates of the seagrass Posidonia oceanica have been evaluated by means of reconstructive methods at the Travello meadow (Ligurian Sea, NW Mediterranean, Italy) to provide evidence of responses to the putative impact of a beach replenishment made with terrigenous materials in 1993. Two additional meadows (Genoa-Quinto and Noli) were sampled as controls. An asymmetrical sampling design ('beyond BACI': Before/After, Control/Impact) was thus used to detect the impact on the basis of a single sampling as dating methods obviated the lack of pre-impact data. At all three meadows investigated, leaf production and rhizome elongation rates were reconstructed for 12 previous years (from 1988 to 1999). A marked decrease in the leaf production rate (around 20%) was assessed only at Travello immediately after the putative disturbance. Control meadows, instead, did not display any significant variations in the pattern of change of this variable from before to after the putative impact. With regard to rhizome growth rates, no significant changes in space and time attributable to the putative impact have been detected. The present data also suggest that the impact studied may be considered as a pulse disturbance, since leaf production appeared to recover over a comparatively short-time scale (around 2-3 years) if compared to the low turn-over and high longevity of P. oceanica. The high potential of asymmetrical sampling designs in combination with dating methods is discussed in the light of the results presented here.     

Spatial distribution of the copepod Centropages typicus in Ligurian Sea (NW Mediterranean). Role of surface currents estimated by Topex-Poseidon altimetry

A particle-tracking model was used to simulate the dispersion and development of the planktonic copepod Centropages typicus during spring in Ligurian Sea. We show that mesoscale current structure, with a coastal jet and eddies, plays a key role in the transport and dispersion of C. typicus during its life cycle. Although, in the north, offshore Nice, cohorts can be advected southwestward out of Ligurian basin, more to the south others are retained in the central eddy and may give the start to the spring bloom of this species. However, input of individuals from the south through the Corsican Channel and along the west coast of Corsica may also be important in spring. This study shows that the ambit of C. typicus population is larger than the Ligurian Sea.