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Polycyclic aromatic hydrocarbons (PAH) have become a threat for the conservation of wetlands worldwide. The halophyte Spartina densiflora has shown to be potentially useful for soil phenanthrene phytoremediation, but no studies on bacteria-assisted hydrocarbon phytoremediation have been carried out with this halophyte. In this work, three phenanthrene-degrading endophytic bacteria were isolated from S. densiflora tissues and used for plant inoculation. Bacterial bioaugmentation treatments slightly improved S. densiflora growth, photosynthetic and fluorescence parameters. But endophyte-inoculated S. densiflora showed lower soil phenanthrene dissipation rates than non-inoculated S. densiflora (30% below) or even bulk soil (23% less). Our work demonstrates that endophytic inoculation on S. densiflora under greenhouse conditions with the selected PAH-degrading strains did not significantly increase inherent phenanthrene soil dissipation capacity of the halophyte. It would therefore be advisable to provide effective follow-up of bacterial colonization, survival and metabolic activity during phenanthrene soil phytoremediation., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Pollution of coasts by toxic metals and metalloids is a worldwide problem for which phytoremediation using halophytes and associated microbiomes is becoming relevant. Metal(loid) excess is a constraint for plant establishment and development, and plant growth promoting rhizobacteria (PGPR) mitigate plant stress under these conditions. However, mechanisms underlying this effect remain elusive. The effect of toxic metal(loid)s on activity and gene expression of ROS-scavenging enzymes in roots of the halophyte Spartina densiflora grown on real polluted sediments in a greenhouse experiment was investigated. Sediments of the metal-polluted joint estuary of Tinto and Odiel rivers and control, unpollutred samples from the Piedras estuary were collected and submitted to ICP-OES. Seeds of S. densiflora were collected from the polluted Odiel marshes and grown in polluted and unpolluted sediments. Rhizophere biofilm-forming bacteria were selected based on metal tolerance and inoculated to S. densiflora and grown for 4 months. Fresh or frozen harvested plants were used for enzyme assays and gene expression studies, respectively. Metal excess induced SOD (five-fold increase), whereas CAT and ascorbate peroxidase displayed minor induction (twofold). A twofold increase of TBARs indicated membrane damage. Our results showed that metal-resistant PGPR (P. agglomerans RSO6 and RSO7 and B. aryabhattai RSO25) contributed to alleviate metal stress, as deduced from lower levels of all antioxidant enzymes to levels below those of non-exposed plants. The oxidative stress index (OSI) decreased between 50 and 75% upon inoculation. The results also evidenced the important role of PAL, involved in secondary metabolism and/or lignin synthesis, as a pathway for metal stress management in this halophyte upon inoculation with appropriate PGPR, since the different inoculation treatments enhanced PAL expression between 3.75- and five-fold. Our data confirm, at the molecular level, the role of PGPR in alleviating metal stress in S. densiflora and evidence the difficulty of working with halophytes for which little genetic information is available., (© 2018 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

Metal contamination of estuaries is a severe environmental problem, for which phytoremediation is gaining momentum. In particular, the associations between halophytes-autochthonous rhizobacteria have proven useful for metal phytostabilization in salt marshes. In this work, three bacterial strains (gram-negative and gram-positive) were used for Spartina densiflora inoculation. All three bacteria, particularly Pantoea strains, promoted plant growth and mitigated metal stress on polluted sediments, as revealed from functionality of the photosynthetic apparatus (PSII) and maintenance of nutrient balance. Pantoea strains did not significantly affect metal accumulation in plant roots, whereas the Bacillus strain enhanced it. Metal loading to shoots depended on particular elements, although in all cases it fell below the threshold for animal consumption. Our results confirm the possibility of modulating plant growth and metal accumulation upon selective inoculation, and the suitability of halophyte-rhizobacteria interactions as biotechnological tools for metal phytostabilization in salt marshes, preventing metal transfer to the food chain., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

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.

The design of effective phytoremediation programs is severely hindered by poor seed germination on metal polluted soils. The possibility that inoculation with plant growth promoting rhizobacteria (PGPR) could help overcoming this problem is hypothesized. Our aim was investigating the role of PGPR in Spartina densiflora seed germination on sediments with different physicochemical characteristics and metal pollution degrees. Gram negative Pantoea agglomerans RSO6 and RSO7, and gram positive Bacillus aryabhattai RSO25, together with the consortium of the three strains, were used for independent inoculation experiments. The presence of metals (As, Cu, Pb and Zn) in sediments reduced seed germination by 80%. Inoculation with Bacillus aryabhattai RSO25 or Pantoea agglomerans RSO6 and RSO7 enhanced up to 2.5 fold the germination rate of S. densiflora in polluted sediments regarding non-inoculated controls. Moreover, the germination process was accelerated and the germination period was extended. The consortium did not achieve further improvements in seed germination., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2015 Elsevier B.V. All rights reserved.)

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., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In the salt marshes of the joint estuary of Tinto and Odiel rivers (SW Spain), one of the most polluted areas by heavy metals in the world, Spartina densiflora grows on sediments with high concentrations of heavy metals. Furthermore, this species has shown to be useful for phytoremediation. The total bacterial population of the rhizosphere of S. densiflora grown in two estuaries with different levels of metal contamination was analyzed by PCR denaturing gradient gel electrophoresis. Results suggested that soil contamination influences bacterial population in a greater extent than the presence of the plant. Twenty-two different cultivable bacterial strains were isolated from the rhizosphere of S. densiflora grown in the Tinto river estuary. Seventy percent of the strains showed one or more plant growth-promoting (PGP) properties, including phosphate solubilization and siderophores or indolacetic acid production, besides a high resistance towards Cu. A bacterial consortium with PGP properties and very high multiresistance to heavy metals, composed by Aeromonas aquariorum SDT13, Pseudomonas composti SDT3, and Bacillus sp. SDT14, was selected for further experiments. This consortium was able to two-fold increase seed germination and to protect seeds against fungal contamination, suggesting that it could facilitate the establishment of the plant in polluted estuaries.