Juvenile sparids (Rhabdosargus holubi) consistently select structurally dense vegetated habitat in nursery seascapes.

Seagrass habitats provide structural complexity in coastal estuarine and marine environments, which offer fish optimal foraging grounds and refuge from predation. However, seagrasses are some of the most threatened ecosystems globally, with anthropogenic activities such as population growth and environmental degradation leading to the fragmentation, thinning, and loss of these habitats. Rhabdosargus holubi is one of only a few vegetation-associated marine fish species in South African estuaries. Although field studies have shown a strong association with seagrass over other aquatic vegetation for the juveniles of this species, habitat choice has never been empirically tested. Here, we used artificial vegetation units to test habitat choice (different structural complexities) for this species. We also tested whether habitat choice is influenced by a predatory threat, with fish preferentially selecting dense habitat in the presence of a predator and whether this effect may be more apparent in smaller individuals. We found that R. holubi significantly prefer greater structural complexity over less complex habitats, in both the absence and presence of a predator and for both small and large juveniles, showing that R. holubi actively choose more complex structures and are attracted to the structure per se irrespective of the threat of predation. This study highlights the importance of dense seagrass as nursery areas for this species and demonstrates how the loss of these habitats could impact the nursery function of estuaries.

Lipophilic Metabolites of Spartina maritima and Puccinellia maritima Involved in Their Tolerance to Salty Environments.

Spartina maritima and Puccinellia maritima are two fascinating but underexplored halophytic species, and herein, the chemical profile of their hexane extracts is described. Terpenoids and sterols were the most abundant chemical groups in both species. The second dominant class was alcohols and the third esters of fatty acids. The chemical lipophilic profile of both S. maritima and P. maritima is herein reported for the first time. Through the accomplished data, it is possible to conclude that these species are rich in essential compounds that can be relevant to endorse their use as nutraceuticals. Furthermore, through a principal component analysis, a clear differentiation between the taxa was achieved, which indicates that their response to salinity stress is different. That fact was confirmed by the pathway enrichment analysis, which showed that the induced changes in metabolic pathways vary in each species.

The chloroplast genome of the hexaploid Spartina maritima (Poaceae, Chloridoideae): Comparative analyses and molecular dating.

The history of many plant lineages is complicated by reticulate evolution with cases of hybridization often followed by genome duplication (allopolyploidy). In such a context, the inference of phylogenetic relationships and biogeographic scenarios based on molecular data is easier using haploid markers like chloroplast genome sequences. Hybridization and polyploidization occurred recurrently in the genus Spartina (Poaceae, Chloridoideae), as illustrated by the recent formation of the invasive allododecaploid S. anglica during the 19th century in Europe. Until now, only a few plastid markers were available to explore the history of this genus and their low variability limited the resolution of species relationships. We sequenced the complete chloroplast genome (plastome) of S. maritima, the native European parent of S. anglica, and compared it to the plastomes of other Poaceae. Our analysis revealed the presence of fast-evolving regions of potential taxonomic, phylogeographic and phylogenetic utility at various levels within the Poaceae family. Using secondary calibrations, we show that the tetraploid and hexaploid lineages of Spartina diverged 6-10 my ago, and that the two parents of the invasive allopolyploid S. anglica separated 2-4 my ago via long distance dispersal of the ancestor of S. maritima over the Atlantic Ocean. Finally, we discuss the meaning of divergence times between chloroplast genomes in the context of reticulate evolution.

Diversity and Ecological Characterization of Sporulating Higher Filamentous Marine Fungi Associated with Spartina maritima (Curtis) Fernald in Two Portuguese Salt Marshes.

Fungal communities associated with early stages of decomposition of Spartina maritima (Curtis) Fernald were assessed in two geographically distinct salt marshes in Portugal by direct observation of fungal sporulating structures. Twenty-three fungal taxa were identified from 390 plant samples, 11 of which were common to both study sites. Natantispora retorquens, Byssothecium obiones, Phaeosphaeria spartinicola, Phoma sp. 1 and Stagonospora sp. were the most frequent fungal taxa in the studied communities. The fungal species Anthostomella spissitecta, Camarosporium roumeguerii, Coniothyrium obiones, Decorospora gaudefroyi, Halosarpheia trullifera, Leptosphaeria marina and Stagonospora haliclysta were recorded for the first time on S. maritima plants; with the exception of C. roumeguerii and L. marina, all of these species were also new records for Portugal. The differences between species composition of the communities associated with S. maritima were attributed to differences in abiotic conditions of the salt marshes. Although the fungal taxa were distributed differently along the host plants, common species to both fungal communities were found on the same relative position, e.g. B. obiones, Lulworthia sp. and N. retorquens occurred on the basal plant portions, Buergenerula spartinae, Dictyosporium pelagicum and Phoma sp. 1 on the middle plant portions and P. spartinicola and Stagonospora sp. on the top plant portions. The distinct vertical distribution patterns reflected species-specific salinity requirements and flooding tolerance, but specially substrate preferences. The most frequent fungi in both communities also exhibited wider distribution ranges and produced a higher number of fruiting structures, suggesting a more active key role in the decay process of S. maritima.

Blue carbon and nutrient stocks in salt marsh and seagrass from an urban African estuary.

Halophytes in estuaries are effective sinks of carbon, nitrogen, and phosphorus. Blue carbon (BC) is carbon stored in coastal habitats such as mangroves, salt marshes and seagrass. The objectives of this study were to firstly assess the biomass and sediment C stocks in salt marsh (Spartina maritima and Salicornia tegetaria) and seagrass (Zostera capensis) habitats of the Swartkops Estuary, South Africa. Secondly, we applied the nutrient pollutant indicator (NPI) to assess the nutrient status of the estuary. Six replicate sediment cores of 1 m (summer) and 0.5 m (winter) per plant species were collected at six sites. Six replicates for biomass of each species were harvested at each site during summer and winter. Biomass and nutrient dynamics showed that there were distinct seasonal differences in the magnitude of C, N, and P stored in the plants. The sediment was the dominant C pool and differed spatially with creek sites storing more C. Out of the three species, S. maritima stored the most C (224 ± 19.1 Mg C ha-1 in sediment, 16.7 ± 1.2 Mg C ha-1 in biomass), followed by S. tegetaria (207 ± 3.5 Mg C ha-1 in sediment, 4.3 ± 0.4 Mg C ha-1 in biomass) and Z. capensis (180 ± 18.4 Mg C ha-1 in sediment, 2.1 ± 0.7 Mg C ha-1 in biomass). The N:biomass ratios were low in both seasons for Z. capensis, S. maritima and S. tegetaria (0.08, 0.02 and 0.04, respectively) indicating consistent eutrophic conditions in the estuary. Resultantly, the plants displayed a significantly lower below-ground standing biomass highlighting the potential variations of BC storage in eutrophic estuaries.

PGPR Reduce Root Respiration and Oxidative Stress Enhancing Spartina maritima Root Growth and Heavy Metal Rhizoaccumulation.

The present study aims to unravel ecophysiological mechanisms underlying plant-microbe interactions under natural abiotic stress conditions, specifically heavy metal pollution. Effect of plant growth promoting rhizobacteria (PGPR) bioaugmentation on Spartina maritima in vivo root respiration and oxidative stress was investigated. This autochthonous plant is a heavy metal hyperaccumulator cordgrass growing in one of the most polluted estuaries in the world. The association with native PGPR is being studied with a view to their biotechnological potential in environmental decontamination. As a novelty, the oxygen-isotope fractionation technique was used to study the in vivo activities of cytochrome oxidase (COX) and alternative oxidase (AOX) pathways. Inoculated plants showed decreased antioxidant enzymatic activities and in vivo root respiration rates. The reduction in respiratory carbon consumption and the stress alleviation may explain the increments observed in S. maritima root biomass and metal rhizoaccumulation after inoculation. For the first time, plant carbon balance and PGPR are interrelated to explain the effect of rhizobacteria under abiotic stress.

Comparative assessment of LECA and Spartina maritima to remove emerging organic contaminants from wastewater.

The present work aimed to evaluate the capacity of constructed wetlands (CWs) to remove three emerging organic contaminants with different physicochemical properties: caffeine (CAF), oxybenzone (MBPh), and triclosan (TCS). The simulated CWs were set up with a matrix of light expanded clay aggregates (LECA) and planted with Spartina maritima, a salt marsh plant. Controlled experiments were carried out in microcosms using deionized water and wastewater collected at a wastewater treatment plant (WWTP), with different contaminant mass ranges, for 3, 7, and 14 days. The effects of variables were tested isolatedly and together (LECA and/or S. maritima). The presence of LECA and/or S. maritima has shown higher removal (around 61-97%) of lipophilic compounds (MBPh and TCS) than the hydrophilic compound (CAF; around 19-85%). This was attributed to the fact that hydrophilic compounds are dissolved in the water column, whereas the lipophilic ones suffer sorption processes promoting their removal by plant roots and/or LECA. In the control (only wastewater), a decrease in the three contaminant levels was observed. Adsorption and bio/rhizoremediation are the strongest hypothesis to explain the decrease in contaminants in the tested conditions.

Phytochelatins and monothiols in salt marsh plants and their relation with metal tolerance.

Phytochelatins (PCs) and monothiols and their relation with trace element concentrations were studied in three plant species from two Portuguese salt marshes. Belowground tissues showed always higher element concentrations, while enhanced values of monothiols were found in aboveground biomass. Glutathione was usually the most abundant monothiol. The concentration of total PCs was higher in leaves or stems than in roots of Halimione portulacoides and Sarcocornia perennis, while in Spartina maritima the highest concentrations were reported in large roots. PC2 was synthesized by all tissues and species and was higher in large roots of S. maritima. PC4 and PC5 were in high levels in small roots of S. maritima. PC2 was positively correlated with As, Zn and Pb. Although being the first evidence of PCs and monothiols in these species under natural conditions, our results do not point to a simple relationship with elements concentrations, suggesting a complex mechanism involved.

Screening beneficial rhizobacteria from Spartina maritima for phytoremediation of metal polluted salt marshes: comparison of gram-positive and gram-negative strains.

The aim of our work was the isolation and characterization of bacteria from the rhizosphere of Spartina maritima in the metal contaminated Odiel estuary (Huelva, SW Spain). From 25 strains, 84 % were identified as gram-positive, particularly Staphylococcus and Bacillus. Gram-negative bacteria were represented by Pantoea and Salmonella. Salt and heavy metal tolerance, metal bioabsorption, plant growth promoting (PGP) properties, and biofilm formation were investigated in the bacterial collection. Despite the higher abundance of gram-positive bacteria, gram-negative isolates displayed higher tolerance toward metal(loid)s (As, Cu, Zn, and Pb) and greater metal biosorption, as deduced from ICP-OES and SEM-EDX analyses. Besides, they exhibited better PGP properties, which were retained in the presence of metals and the ability to form biofilms. Gram-negative strains Pantoea agglomerans RSO6 and RSO7, together with gram-positive Bacillus aryabhattai RSO25, were selected for a bacterial consortium aimed to inoculate S. maritima plants in metal polluted estuaries for phytoremediation purposes.

Nutrient levels modify saltmarsh responses to increased inundation in different soil types.

Saltmarshes have been depleted historically, and cumulative stressors threaten their future persistence. We examined experimentally how nutrient availability (high vs. low) affects the responses of Spartina maritima to increased inundation in two mineral soil types (low vs. medium organic). Increased inundation, one of the effects of accelerated sea level rise, had negative effects on most plant growth parameters, but the magnitude varied with soil and nutrient levels, and between plants from different locations. Average differences between inundation treatments were largest at high nutrient conditions in low organic matter soils. We conclude that saltmarsh vegetation would be more drastically affected by increased inundation in low than in medium organic matter soils, and especially in estuaries already under high nutrient availability. This knowledge enhances the prediction of changes at the foreshore of saltmarshes related to sea level rise, and the development of site-specific conservation strategies.

Scouting contaminated estuaries: heavy metal resistant and plant growth promoting rhizobacteria in the native metal rhizoaccumulator Spartina maritima.

Spartina maritima is a native endangered heavy metal rhizoaccumulator cordgrass naturally growing in southwest coasts of Spain, where is used as a biotool to rehabilitate degraded salt marshes. Fifteen bacterial strains were isolated from the rhizosphere of S. maritima growing in the estuary of the Tinto River, one of the most polluted areas in the world. A high proportion of bacteria were resistant towards several heavy metals. They also exhibited multiple plant growth promoting (PGP) properties, in the absence and the presence of Cu. Bacillus methylotrophicus SMT38, Bacillusaryabhattai SMT48, B. aryabhattai SMT50 and Bacilluslicheniformis SMT51 were selected as the best performing strains. In a gnobiotic assay, inoculation of Medicago sativa seeds with the selected isolates induced higher root elongation. The inoculation of S. maritima with these indigenous metal-resistant PGP rhizobacteria could be an efficient method to increase plant adaptation and growth in contaminated estuaries during restoration programs.

Endophytic Cultivable Bacteria of the Metal Bioaccumulator Spartina maritima Improve Plant Growth but Not Metal Uptake in Polluted Marshes Soils.

Endophytic bacterial population was isolated from Spartina maritima tissues, a heavy metal bioaccumulator cordgrass growing in the estuaries of Tinto, Odiel, and Piedras River (south west Spain), one of the most polluted areas in the world. Strains were identified and ability to tolerate salt and heavy metals along with plant growth promoting and enzymatic properties were analyzed. A high proportion of these bacteria were resistant toward one or several heavy metals and metalloids including As, Cu, and Zn, the most abundant in plant tissues and soil. These strains also exhibited multiple enzymatic properties as amylase, cellulase, chitinase, protease and lipase, as well as plant growth promoting properties, including nitrogen fixation, phosphates solubilization, and production of indole-3-acetic acid (IAA), siderophores and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. The best performing strains (Micrococcus yunnanensis SMJ12, Vibrio sagamiensis SMJ18, and Salinicola peritrichatus SMJ30) were selected and tested as a consortium by inoculating S. maritima wild plantlets in greenhouse conditions along with wild polluted soil. After 30 days, bacterial inoculation improved plant photosynthetic traits and favored intrinsic water use efficiency. However, far from stimulating plant metal uptake, endophytic inoculation lessened metal accumulation in above and belowground tissues. These results suggest that inoculation of S. maritima with indigenous metal-resistant endophytes could mean a useful approach in order to accelerate both adaption and growth of this indigenous cordgrass in polluted estuaries in restorative operations, but may not be suitable for rhizoaccumulation purposes.

Moving closer towards restoration of contaminated estuaries: Bioaugmentation with autochthonous rhizobacteria improves metal rhizoaccumulation in native Spartina maritima.

Spartina maritima is an ecosystem engineer that has shown to be useful for phytoremediation purposes. A glasshouse experiment using soil from a metal-contaminated estuary was designed to investigate the effect of a native bacterial consortium, isolated from S. maritima rizhosphere and selected owing to their plant growth promoting properties and multiresistance to heavy metals, on plant growth and metal accumulation. Plants of S. maritima were randomly assigned to three soil bioaugmentation treatments (without inoculation, one inoculation and repeated inoculations) for 30 days. Growth parameters and photosynthetic traits, together with total concentrations of several metals were determined in roots and/or leaves. Bacterial inoculation improved root growth, through a beneficial effect on photosynthetic rate (AN) due to its positive impact on functionality of PSII and chlorophyll concentration. Also, favoured intrinsic water use efficiency of S. maritima, through the increment in AN, stomatal conductance and in root-to-shoot ratio. Moreover, this consortium was able to stimulate plant metal uptake specifically in roots, with increases of up to 19% for As, 65% for Cu, 40% for Pb and 29% for Zn. Thus, bioaugmentation of S. maritima with the selected bacterial consortium can be claimed to enhance plant adaptation and metal rhizoaccumulation during marsh restoration programs.