Genetic structure of two Thalassia testudinum populations from the south Texas Gulf coast

The south Texas Gulf coast is a unique ecosystem that contains a number of different bay systems. We used random amplification of polymorphic DNA (RAPD) markers to assess genetic diversity, differentiation and genetic distance between populations from two different bays that differed significantly in terms of flowering rate and disturbance. We found that while each bay contained a number of unique RAPD profiles, the average genetic diversity in each population was low. Genetic distance between the two populations was also low (F_st = 0.084) and the majority (92%) of the genetic variation was attributed to differences between individuals within populations. The population from the Laguna Madre location, however, was polymorphic for a larger number of markers, had a higher average genetic diversity and a larger number of unique RAPD profiles. The higher level of flowering at this location most likely accounts for the higher diversity.     

Experimental analysis of the response and recovery of Zostera marina (L.) and Halodule wrightii (Ascher.) to repeated light-limitation stress

Seagrasses are considered important indicators of decline in water quality resulting in increased light attenuation that negatively influences their growth and survival. Chronic light-limitation interspersed with unpredictable acute attenuation events have had poorly understood effects on seagrass recovery dynamics. Zostera marina (eelgrass) and Halodule wrightii (shoalgrass) were subject to a matrix of light-deprivation events followed by recovery periods to mimic repeated acute shading events. Plant survival, morphology, biomass, chlorophyll content, and Fv/Fm were assessed over time to determine recovery. At the end of the experiment, all plants were harvested and species-specific treatment effects were determined. Significant differences due to treatments were noted in all parameters measured. In general, responses were similar for both life-stages and between species, suggesting similar physiological tolerance to repeated acute light-attenuation events. Only plants in treatments where light-deprivation was followed by a recovery interval of at least the same duration showed signs of long-term survival. Chlorophyll fluorescence (Fv/Fm) was an important metric for assessing recovery, but it failed to detect the onset of mortality in many plants. Other metrics of plant condition need to be assessed and coupled with chlorophyll fluorescence data to assess seagrass “health”. This is of particular importance in field studies, where the history of the plants is largely unknown.     

Effect of nutrient availability, grazer assemblage and seagrass source population on the interaction between Thalassia testudinum (turtle grass) and its algal epiphytes

Seagrass leaves are often densely covered by epiphytic algae which can suppress seagrass productivity and has been implicated in declines of seagrass meadows worldwide. The net effect of epiphytes on seagrass growth and morphology depends on the independent and interactive effects of a variety of factors, including nutrient availability and the intensity of grazing on epiphytes. Here I report the results of a mesocosm experiment designed to test the effects of nutrient addition and within-functional group variation (grazer species composition and the source population of seagrass) on the strength of the interactions among grazers, epiphytes, and turtle grass (Thalassia testudinum). Turtle grass ramets from two sites in the northern Gulf of Mexico were cleared of epiphytes and transplanted into common-garden mesocosms. Replicate ramets were grown in a split-split plot design with two levels of dissolved nutrients and four different grazer species combinations (Tozeuma carolinense alone, Pagurus maclaughlinae alone, both species together, and no grazers present). As expected, grazers had a significant negative effect on epiphyte biomass/leaf area and a significant positive effect on turtle grass growth in the mesocosms. The two species were more similar in their direct effects on epiphyte biomass than in their indirect effects on turtle grass growth; this may reflect differences in epiphyte community composition under different grazer treatments. The effect of nutrient addition on turtle grass growth depended critically on the intensity of grazing: in the presence of grazers, turtle grass tended to produce a greater biomass of new leaf tissue in the tanks with nutrients added than in the control tanks. However, when grazers were absent, the direction of the effect was reversed, and plants with nutrients added grew less than the control plants. The two source populations of turtle grass differed significantly in epiphyte biomass/leaf area accrued in the mesocosms as well as in the strength of the effect of grazers on turtle grass growth. This suggests that population differentiation in seagrass interactions with epiphytes, as well as spatial and temporal variation in resources and grazer community composition, can greatly effect the role of epiphytes in limiting seagrass productivity.     

State of Thalassia testudinum Banks ex König meadows in the Veracruz Reef System,Veracruz, México

The vegetative development of the seagrass Thalassia testudinum was assessed in three reefs (Hornos, isla Sacrificios, isla de Enmedio) of the Veracruz Reef System (Veracruz, México) which are located at increasing distances from the mainland coast. Leaf nutrient content of T. testudinum suggests that the availability of nitrogen and phosphorus increases from Hornos reef to isla Sacrificios and isla de Enmedio, that is, as the distance to the coast increases. The total biomass of this species tended to increase similarly as a result of the large increase of the biomass of rhizomes and roots but not of leaves. In contrast to previous knowledge of the response of seagrasses along gradients of nutrient availability, the rhizome and roots to shoot biomass ratio increased by a factor of two as nutrient availability increased. The density of T. testudinum shoots and their mass, the LAI and leaf productivity, and the average number of leaves produced by a T. testudinum shoot in 1 year were lowest in the reef closest to shore (Hornos). Our results show that the vegetative development of T. testudinum in Hornos reef is restrained when compared with that in isla Sacrificios and isla de Enmedio and suggest that the differences of vegetative development of this species in the Veracruz Reef System might be driven by factors other than nutrient availability.     

Thalassia testudinum seedling responses to changes in salinity and nitrogen levels

The dominant seagrass in Florida Bay, Thalassia testudinum Banks ex König, is a stenohaline species with optimum growth around marine salinity (30-40 PSU). Previous studies have examined the responses of mature short shoots of T. testudinum to environmental stresses. Our goal was to assess responses of seedlings to changes in water chemistry in Florida Bay that might occur as part of the Comprehensive Everglades Restoration Plan (CERP). Specifically, we examined seedling survival, growth, photosynthesis, respiration and osmolality in response to hypo- and hyper-salinity conditions, as well as possible synergistic effects of depleted and elevated ammonium concentrations. The study was conducted in mesocosms on T. testudinum seedlings collected during August 2003 near Florida Bay. Hyper- and hypo-saline conditions were detrimental to the fitness of T. testudinum seedlings. Plants at 0 and 70 PSU exhibited 100% mortality and a significant decrease in survival was observed in the 10, 50 and 60 PSU treatments. Increased levels of ammonium further decreased growth in the lower salinity treatments. Seedlings in 30 and 40 PSU had the greatest growth. Quantum yield and relative electron transport rate, measured using PAM fluorometry, showed a decrease in photosynthetic performance on either side of the 30-40 PSU optimum. Tissue osmolality decreased significantly with decreased salinity but tissue remained consistently hyperosmotic to the media across all salinity treatments. Maintaining negative water potential and allocating more energy to osmoregulation may decrease the productivity of this species in salinity-stress conditions. Our results suggest that the salinity-tolerance limits of this seagrass at the seedling stage are not as broad as those reported for mature plants. Increased fresh water inflow, especially if co-occurring with an increase in water-column ammonium, could negatively affect successful recruitment of T. testudinum seedlings in northern regions of Florida Bay.     

Assessment of turtle grass (Thalassia testudinum ex Banks Konig) community structure in a UNESCO Biosphere Reserve

Tropical turtle grass beds (Thalassia testudinum) were investigated in an UNESCO Biosphere Reserve in St. Johns Island, United States Virgin Islands. Four coastal bays were studied to assess floral characteristics, benthic invertebrate density and secondary production. These bays varied with respect to oceanic exposure and anthropogenic stresses. Results indicate that Coral Bay Harbor, the most anthropogenetically impacted site, had the highest T. testudinum biomass, but the lowest floral diversity. Its faunal community was dominated by small polychaetes with significantly lower secondary production. The most protected site (Hurricane Hole) maintained the highest floral and faunal species richness, faunal density, faunal diversity, and secondary production. The other two bays, Great Lameshur and Little Lameshur, demonstrated intermediate plant biomass and species richness, faunal density and secondary production. Each of these bays, however, had high oceanic exposure due to their orientation which also demonstrated a sediment size shift to larger particles compared to the other sites. One unique finding was a significant increase in the shoot:root ratio in Little Lameshur where green turtle (Chelonia mydas) grazing was frequently observed, suggesting a potential top-down structuring force in this bay.     

Nutrient content of seagrasses and epiphytes in the northern Gulf of Mexico: Evidence of phosphorus and nitrogen limitation

We examined C:N:P ratios of seagrass leaves and epiphytic algae from the eastern shoreline of Grand Bay (Alabama, USA) and the entire shoreline of Big Lagoon (Florida, USA) during the summer of 2001 and March 2003, and used contour plotting of N:P ratios in both locations to examine spatial trends in our data. Results indicated phosphorus limitation for seagrass and epiphytes in each bay. In addition, C:N, C:P, and N:P ratios in both locations showed differences between summer and wintertime values for seagrasses; however, the only epiphytic elemental ratios to differ were C:P and N:P ratios in Grand Bay. Within Grand Bay, phosphorus limitation was stronger in epiphytes than seagrasses, with the largest amount of variation in N:P ratios occurring adjacent to the only developed land on the shoreline. In Big Lagoon, two distinct areas were present in N:P contour plots: the eastern end of the bay that was influenced by water from the Gulf of Mexico and Santa Rosa Sound, and the western end of the bay that was most influenced by Perdido Bay and a developed area along the northern shoreline. Detection of phosphorus limitation within Big Lagoon was not surprising, as both input sources to Big Lagoon are known to be low in phosphorus. However, phosphorus limitation in Grand Bay was unexpected, as both “feeder systems” (Mobile Bay and the Mississippi Sound) have high ambient phosphorus levels. As a result, C:N:P ratios from seagrasses and epiphytes may not accurately reflect ambient nutrient levels in Grand Bay due to decreased availability of some forms of phosphorus or increased competition for the uptake of phosphorus. Overall, our C:N:P analysis suggested that not only was P limitation greater than N limitation in Grand Bay and Big Lagoon, but patterns of nutrient limitation varied both temporally and geographically for inter- and intra-bay comparisons.     

Species-specific effects of elevated CO2 on resource allocation in Plantago maritima and Armeria maritima

We investigated the effects of increased atmospheric CO₂ on the biomass, photosynthesis, protein and phenolic concentrations and content of Plantago maritima and Armeria maritima. This enabled us to test the protein competition model (PCM) for predicting C allocation to phenolics. Three contrasting responses to elevated CO₂ (600 μmol CO₂ mol⁻¹) between the two study species were observed. (1) In P. maritima, plant biomass increased and the maximum carboxylation rate of Rubisco (V_c,max) was decreased. However, in A. maritima, shoot biomass decreased and the V_c,max of Rubisco was unchanged. (2) The total phenolic content increased in P. maritima but decreased in A. maritima. (3) Protein concentrations and content decreased in P. maritima and root protein concentrations and content increased in A. maritima. We conclude that C and N allocation to phenolics and proteins is species- and organ-specific and the PCM predictions were correct when phenolics and proteins were expressed on a per plant content basis.     

Synergistic effects of high temperature and sulfide on tropical seagrass

To examine the synergism of high temperature and sulfide on two dominant tropical seagrass species, a large-scale mesocosm experiment was conducted in which sulfide accumulation rates (SAR) were increased by adding labile carbon (glucose) to intact seagrass sediment cores across a range of temperatures. During the initial 10 d of the 38 d experiment, porewater SAR in cores increased 2- to 3-fold from 44 and 136 μmol L^− 1 d^− 1 at 28-29 °C to 80 and 308 μmol L^− 1 d^− 1 at 34-35 °C in Halodule wrightii and Thalassia testudinum cores, respectively. Labile C additions to the sediment resulted in SAR of 443 and 601 μmol L^− 1 d^− 1 at 28-29 °C and 758 to 1,557 μmol L^− 1 d^− 1 at 34-35 °C in H. wrightii and T. testudinum cores, respectively. Both T. testudinum and H. wrightii were highly thermal tolerant, demonstrating their tropical affinities and potential to adapt to high temperatures. While plants survived the 38 d temperature treatments, there was a clear thermal threshold above 33 °C where T. testudinum growth declined and leaf quantum efficiencies (Fv/Fm) fell below 0.7. At this threshold temperature, H. wrightii maintained shoot densities and leaf quantum efficiencies. Although H. wrightii showed a greater tolerance to high temperature, T. testudinum had a greater capacity to sustain biomass and short shoots under thermal stress with labile C enrichment, regardless of the fact that sulfide levels in the T. testudinum cores were 2 times higher than in the H. wrightii cores. Tropical seagrass tolerance to elevated temperatures, predicted in the future with global warming, should be considered in the context of the sediment-plant complex which incorporates the synergism of plant physiological responses and shifts in sulfur biogeochemistry leading to increased plant exposure to sulfides, a known toxin.     

Northern range extension of the seagrasses Halophila johnsonii and Halophila decipiens along the east coast of Florida,USA

The northern geographic limit for Halophila johnsonii and Halophila decipiens has been reported as Sebastian Inlet, within the Indian River Lagoon, Florida. Surveys conducted in August 2007 determined the new northern limit to be 21.5 km north of the previously known limit. This new northern limit is a 10% range extension for H. johnsonii, a federally threatened species. We conclude that these range extensions are recent, based on (1) the small size of patches; (2) unusually good water clarity conditions due to a recent drought; (3) recent mild/warmer winters; and (4) a recent mechanism for transporting propagules, the numerous hurricanes of 2004. Although this recent range extension is considered ephemeral, similar range extensions may have occurred in the past and may occur again in the future under favorable conditions given the high capacity of these two species for dispersal to favorable sites. The northern limits of these species should not be viewed as static locations; rather, they must be considered dynamic features.     

Chloroplast sequences reveal a diversity gradient in the Mediterranean Ruppia cirrhosa species complex

Two subcosmopolitan species Ruppia maritima and Ruppia cirrhosa are recognized throughout Europe, whereas Ruppia drepanensis is endemic to SW Europe. We aimed at characterizing the geographic structure of the chloroplast DNA haplotype diversity of 56 Ruppia populations across the European part of the Mediterranean. On the basis of five cpDNA markers (total length of 2300 bp) 16 haplotypes were obtained for 1546 individuals. Three populations of R. maritima showed a single haplotype and differed in five insertions/deletions and 16 substitutions from a highly variable R. cirrhosa species complex, which included R. drepanensis. The haplotype diversity of this species complex was much higher in the western Mediterranean basin than in the eastern basin. Analysis of molecular variance (AMOVA) showed significant differentiation of R. cirrhosa between the two basins and also within the western Mediterranean thereby suggesting the latter as an important centre of Ruppia diversity. An isolation-by-distance (IBD) pattern was stronger between the West-East basin populations than within basins. A PCO analysis of the western basin populations indicated a diversity gradient with those of Sardinia as polymorph intermediates. The low diversity within the eastern basin suggests that the observed gradient could be hypothesized as a historical dispersal of only a limited number of haplotypes from west to east.     

Growth,Na, K,osmolyte accumulation and lipid membrane peroxidation of two provenances of Cakile maritima during water deficit stress and subsequent recovery

The effects of water deficit stress on growth, Na⁺, K⁺ and osmolyte accumulation in the halophyte species Cakile maritima were investigated. Two Tunisian provenances, Tabarka and Chaffar, belonging to different bioclimatic stages, humid and arid, respectively, were compared. After germination, thirty-day-old seedlings were cultivated for 4 weeks under optimal or limiting water supply, at 100% and 25% of field capacity (FC), respectively. A subset of stressed plants was thereafter rehydrated. The final harvest was carried out after 60 days of treatment. Upon water deficit stress, Chaffar provenance showed significantly lower reduction in biomass production, net CO₂ assimilation and stomatal conductance as well as of leaf water content. Leaf malondialdehyde (MDA) content was significantly increased in the two provenances but this effect was more pronounced in Tabarka plants than in Chaffar ones. Several criteria seem to be associated with the relative tolerance of Chaffar to water deficit: a slow growth rate, a greater ability to control photosynthetic gas exchange, a high ability to preferentially allocate photoassimilates to its roots, and a greater capacity for osmotic adjustment ensured by K⁺ and some compatible solutes such as proline and glycine betaine, but not soluble sugars. The superiority of Chaffar provenance also appeared at the level of its ability to recover after a severe water deficit stress (irrigation at 25% FC only during one month). The data suggest that compatible osmolytes (proline and glycine betaine) accumulated upon water deficit stress play important roles in this halophyte, being involved not only in osmotic adjustment but probably serving also in preservation of the structural and functional integrity at the cellular level during water deficit.     

Nutrient impacts on epifaunal density and species composition in a subtropical seagrass bed

The capacity of epifauna to control algal proliferation following nutrient input depends on responses of both grazers and upper trophic level consumers to enrichment. We examined the responses of Thalassia testudinum (turtle grass) epifaunal assemblages to nutrient enrichment at two sites in Florida Bay with varying levels of phosphorus limitation. We compared epifaunal density, biomass, and species diversity in 2 m² plots that had either ambient nutrient concentrations or had been enriched with nitrogen and phosphorus for 6 months. At the severely P-limited site, total epifaunal density and biomass were two times higher in enriched than in unenriched plots. Caridean shrimp, grazing isopods, and gammarid amphipods accounted for much of the increase in density; brachyuran crabs, primary predatory fish, and detritivorous sea cucumbers accounted for most of the increase in biomass. At the less P-limited site, total epifaunal density and biomass were not affected by nutrient addition, although there were more caridean shrimp and higher brachyuran crab and pink shrimp biomass in enriched plots. At both sites, some variation in epifaunal density and biomass was explained by features of the macrophyte canopy, such as T. testudinum and Halodule wrightii percent cover, suggesting that enrichment may change the refuge value of the macrophyte canopy for epifauna. Additional variation in epifaunal density and biomass was explained by epiphyte pigment concentrations, suggesting that enrichment may change the microalgal food resources that support grazing epifauna. Increased epifaunal density in enriched plots suggests that grazers may be able to control epiphytic algal proliferation following moderate nutrient input to Florida Bay. Electronic supplementary material Electronic supplementary material is available for this article at and accessible for authorised users.     

Note on organic dormancy of estuarine Ruppia maritima L. seeds

Information on seed dormancy is one of the primary requirements for successful seedling propagation of submerged aquatics and seagrass. Studies on Ruppia maritima seed germination have been done, but the presence, requirements, and the types of dormancy have not been well understood. A laboratory study was conducted to understand presence and types of organic dormancy of the seeds of estuarine R. maritima collected from Lake Pontchartrain, Louisiana, USA. Our study results indicate that the brackish estuarine R. maritima population produces seeds that do not have any noticeable initial morphological, physical, and physiological dormancy. Although dry stratification reduced seed viability and final germination rates, drying seems to induce an earlier germination in R. maritima. Desiccation also appears to induce an environmental dormancy that can be disrupted by exposure to water. Further study on environmental dormancy is needed to provide information to develop methods for long-term seed storage that can be employed in greenhouse seedling propagation. Handling editor: P. Viaroli     

Septal pore complex morphology in the Agaricomycotina (Basidiomycota) with emphasis on the Cantharellales and Hymenochaetales

The ultrastructure of septa and septum-associated septal pore caps are important taxonomic markers in the Agaricomycotina (Basidiomycota, Fungi). The septal pore caps covering the typical basidiomycetous dolipore septum are divided into three main phenotypically recognized morphotypes: vesicular-tubular (including the vesicular, sacculate, tubular, ampulliform, and globular morphotypes), imperforate, and perforate. Until recently, the septal pore cap-type reflected the higher-order relationships within the Agaricomycotina. However, the new classification of Fungi resulted in many changes including revision of existing and addition of new orders. Therefore, the septal pore cap ultrastructure of more than 325 species as reported in literature was related to this new classification. In addition, the septal pore cap ultrastructures of Rickenella fibula and Cantharellus formosus were examined by transmission electron microscopy. Both fungi have dolipore septa associated with perforate septal pore caps. These results combined with data from the literature show that the septal pore cap-type within orders of the Agaricomycotina is generally monomorphic, except for the Cantharellales and Hymenochaetales.It appears from the fungal phylogeny combined with the septal pore cap ultrastructure that the vesicular-tubular and the imperforate type both may have arisen from endoplasmic reticulum. Thereafter, the imperforate type eventually gave rise to the perforate septal pore cap-type.     

A simple assay for determining activities of phosphopentomutase from a hyperthermophilic bacterium Thermotoga maritima

Phosphopentomutase (PPM) catalyzes the interconversion of α-d-(deoxy)-ribose 1-phosphate and α-d-(deoxy)-ribose 5-phosphate. We developed a coupled or uncoupled enzymatic assay with an enzyme nucleoside phosphorylase for determining PPM activities on d-ribose 5-phosphate at a broad temperature range from 30 to 90 °C. This assay not only is simple and highly sensitive but also does not require any costly special instrument. Via this technology, an open reading frame TM0167 from a thermophilic bacterium Thermotoga maritima putatively encoding PPM was cloned. The recombinant PPM was overexpressed in Escherichia coli Rosetta. This enzyme has the highest activity at 90 °C. MnCl₂ (0.1 mM) and 50 μM α-d-glucose 1,6-bisphosphate are cofactors. The kinetic parameters of K_m and k_cat are 1.2 mM and 185 s⁻¹ at 90 °C_, respectively. The enzyme has a half-life time of up to 156 min at 90 °C. This enzyme is the most active and thermostable PPM reported to date.     

The morphology of the larval stages of three species (Arrenurus mediorotundatus Thor, 1898; Arrenurus conicus Piers 1894; and Arrenurus cylindratus Piers, 1896) of the Megaluracarus subgenus (Acari: Hydrachnidia: Arrenuridae)

The morphology of the larval stages of three species (Arrenurus mediorotundatus, Arrenurus conicus and Arrenurus cylindratus) belonging to the Megaluracarus subgenus is described. Particular attention is paid to the common characters of the three species in which they differ from the other species belonging to the other subgenera. These are: the shape of the excretory pore plate, the shape of the dorsal plate, the length of coxal plate II medial margin and the C1-CpI, C4-CpIII distances. Other characters differ between the three species also. These are: the shape of the dorsal plate, the shape of coxal plate III, the shape of excretory pore plate and the presence or absence of secondary setae of the IFe3 setae.