Rossellomorea arthrocnemi sp. nov., a novel plant growth-promoting bacterium used in heavy metal polluted soils as a phytoremediation tool.

Strain EAR8T is a root endophyte isolated from Arthrocnemum macrostachyum plants collected from the Odiel marshes, Huelva (Spain). It presented in vitro plant growth-promoting properties and improved the plant growth and heavy metal accumulation in polluted soils playing an important role in phytoremediation strategies. Phenotypically, strain EAR8T cells were Gram-positive, aerobic and non-motile rods with terminal oval endospores and non-swollen sporangia which form beige, opaque, butyrous, raised and irregular colonies with undulate margins. The strain was able to grow between 15-45 °C, at pH 6.0-9.0 and tolerated 0-25 % NaCl (w/v) showing optimal growth conditions on trypticase soy agar plates supplemented with 2.5 % NaCl (w/v) at pH 7.0 and 37 °C for 24 h. Chemotaxonomic analyses showed that the isolate has meso-diaminopimelic acid as the peptidoglycan in the cell wall and MK-7 as the major respiratory quinone. The predominant fatty acids were anteiso-C15 : 0 and iso-C15 : 0 and the polar lipid profile was composed of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Phylogenetic analyses based on the whole proteomes of closest sequenced relatives confirmed that strain EAR8T is affiliated to the genus Rossellomorea and forms a clade with Rossellomorea vietnamensis 15-1T with maximum support. Genome analyses showed that EAR8T has indole-3-acetic acid and siderophore biosynthesis and transporters genes and genes related to resistance against heavy metals. Phenotypic and phylogenomic comparative studies suggested that strain EAR8T is a new representative of the genus Rossellomorea and the name Rossellomorea arthrocnemi sp. nov. is proposed. Type strain is EAR8T (=CECT 9072T=DSM 103900T).

Pseudoalteromonas rhizosphaerae sp. nov., a novel plant growth-promoting bacterium with potential use in phytoremediation.

Strain RA15T was isolated from the rhizosphere of the halophyte plant Arthrocnemum macrostachyum growing in the Odiel marshes (Huelva, Spain). RA15T cells were Gram stain-negative, non-spore-forming, aerobic rods and formed cream-coloured, opaque, mucoid, viscous, convex, irregular colonies with an undulate margin. Optimal growth conditions were observed on tryptic soy agar (TSA) plates supplemented with 2.5 % NaCl (w/v) at pH 7.0 and 28 °C, although it was able to grow at 4-32 °C and at pH values of 5.0-9.0. The NaCl tolerance range was from 0 to 15 %. The major respiratory quinone was Q8 but Q9 was also present. The most abundant fatty acids were summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C17 : 1 ω8c and C16 : 0. The polar lipids profile comprised phosphatidylglycerol and phosphatidylethanolamine as the most abundant representatives. Phylogenetic analyses confirmed the well-supported affiliation of strain RA15T within the genus Pseudoalteromonas, close to the type strains of Pseudoalteromonas neustonica, Pseudoalteromonas prydzensis and Pseudoalteromonas mariniglutinosa. Results of comparative phylogenetic and phenotypic studies between strain RA15T and its closest related species suggest that RA15T could be a new representative of the genus Pseudoalteromonas, for which the name Pseudoalteromonas rhizosphaerae sp. nov. is proposed. The type strain is RA15T (=CECT 9079T=LMG 29860T). The whole genome has 5.3 Mb and the G+C content is 40.4 mol%.

Kushneria phyllosphaerae sp. nov. and Kushneria endophytica sp. nov., plant growth promoting endophytes isolated from the halophyte plant Arthrocnemum macrostachyum.

Two endophytic bacteria (EAod3T and EAod7T) were isolated from the aerial part of plants of Arthrocnemum macrostachyum growing in the Odiel marshes (Huelva, Spain). Phylogenetic analysis based on 16S rRNA gene sequences indicated their affiliation to the genus Kushneria. 16S rRNA gene sequences of strains EAod3T and EAod7T showed the highest similarity to Kushneria marisflavi DSM 15357T (99.0 and 97.6 %, respectively). Digital DNA-DNA hybridization studies between the draft genomes of strain EAod3T and K. marisflavi DSM 15357T corresponded to 28.5 % confirming the novel lineage of strain EAod3T in the genus Kushneria. Cells of both strains were Gram-staining-negative, aerobic and motile rods able to grow at 4-37 °C, at pH 5.0-8.0 and tolerate 0.5-25 % NaCl (w/v). They presented ubiquinone Q9 and C16 : 0, C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c as the major fatty acids. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Based on the phenotypic and phylogenetic results, strains EAod3T (=CECT 9073T=LMG 29856T) and EAod7T (=CECT 9075T=LMG 29858T) are proposed as new representatives of the genus Kushneria, and the proposed names are Kushneria phyllosphaerae sp. nov. and Kushneria endophytica sp. nov., respectively. The whole genome sequence of strain EAod3T has a total length of 3.8 Mbp and a G+C content of 59.3 mol%.

Population Dynamic of the Annual Halophyte Salicornia ramosissima in Salt Pans: Towards a Sustainable Exploitation of Its Wild Populations.

Halophyte species growing under stressful conditions, such as the annual species of the Salicornia genus, have been recognized as a source of metabolites of pharmacological and nutraceutical interest. Therefore, planning the extraction of individual plants from wild populations in a sustainable way is especially important in the case of annual species. We studied the environmental matrix and population dynamic of four Salicornia ramosissima populations growing at two elevations in salt pans under a Mediterranean climate. In elevated areas, S. ramosissima populations presented maximum plant densities of between 628-6288 plants m-2 that remained almost constant until fruiting. In contrast, populations in depressed zones presented five-times greater soil-seed-bank densities and maximum plant densities than populations in elevated zones. In this context, populations in depressed zones lost c. 60% of their maximum plant densities from the end of spring and through summer. In whatever way the environmental matrix seemed to control the population dynamic of S. ramosissima in depressed zones, the effects of a stressful environment would interact with plant densities. In this sense, we recorded the density-dependent mortality for the densest population (max. 51,558 plants m-2). Our results are useful for planning a sustainable harvesting of natural populations of S. ramosissima.

Metal effects on germination and seedling development in closely-related halophyte species inhabiting different elevations along the intertidal gradient.

Seed germination and seedling establishment are very sensitive plant stages to metal pollution. Many halophyte species colonizing salt marshes are able to germinate and establish in highly contaminated habitats and low marsh halophyte species seem to show higher tolerance to metals than high marsh species. We analyzed the effects of copper, zinc and nickel in concentrations up to 2000 µM on seed germination and seedling growth in two closely related species of Sarcocornia, S. perennis, a low marsh species, and S. fruticosa, a high marsh species. Germination of both halophytes was not affected by any metal concentration, and their seedling growth, mainly radicle length, was reduced by increasing metal concentrations. Seedlings of S. perennis showed higher tolerance to the three metals than those of S. fruticosa. Our results are useful for designing ecotoxicological bioassays and planning phytoremediation projects in salt marshes.

Intraspecific variation in the response of the estuarine European isopod Cyathura carinata (Krøyer, 1847) to ocean acidification.

In the present study the model isopod, Cyathura carinata were exposed to four pHNIST treatments (control: 7.9; 7.5, 7, 6.5) in order to determine the tolerance and pH threshold value this estuarine species withstand under future acidification scenarios. Seawater acidification significantly affected the lifespan of C. carinata, where population density was remarkably reduced at the lowest pH treatment. The longevity, survivorship and swimming activity (related to the acquisition of energy) of these isopods decreased with decreasing pH. Furthermore, to determine the possible metabolic plasticity of this species, the swimming activity, the Na+/K + -ATPase activity (relevant for osmoregulation process), and the RNA:DNA ratio (an indicator of fitness) were measure from two populations of C. carinata, one inhabiting a stable environment (pHNIST 7.5-8.0) and one inhabiting a fluctuating pCO2 regimes (pH 3.3-8.5) subjected to three pH treatments (7.9, 7.0 and 6.5). The population from high fluctuating pCO2 conditions showed capacity to withstand to pH 6.5, as well as higher longevity and metabolic plasticity, when compared with the population from the habitat with slight pCO2 variation. These results indicate that Cyathura population from stable environments could be vulnerable to ocean acidification because it could trigger detrimental effects on its survival energy budget, and growth. However, ocean acidification has limited effect on the energy budget and survival of C. carinata population from highly variable habitats, suggesting that they are able to cope with the elevated energy demand. The difference showed between populations is likely an indication of genetic differentiation in tolerance to ocean acidification, possibly attributable to local adaptations, which could provide the raw material necessary for adaptation to future conditions. In addition, our results suggest that when assessing marine crustacean responses to changing environments on a global scale, variability in population and metabolic responses need to be considered.

Differential Effects of Increasing Salinity on Germination and Seedling Growth of Native and Exotic Invasive Cordgrasses.

Soil salinity is a key environmental factor influencing germination and seedling establishment in salt marshes. Global warming and sea level rise are changing estuarine salinity, and may modify the colonization ability of halophytes. We evaluated the effects of increasing salinity on germination and seedling growth of native Spartina maritima and invasive S. densiflora from wetlands of the Odiel-Tinto Estuary. Responses were assessed following salinity exposure from fresh water to hypersaline conditions and germination recovery of non-germinated seeds when transferred to fresh water. The germination of both species was inhibited and delayed at high salinities, while pre-exposure to salinity accelerated the speed of germination in recovery assays compared to non-pre-exposed seeds. S. densiflora was more tolerant of salinity at germination than S. maritima. S. densiflora was able to germinate at hypersalinity and its germination percentage decreased at higher salinities compared to S. maritima. In contrast, S. maritima showed higher salinity tolerance in relation to seedling growth. Contrasting results were observed with differences in the tidal elevation of populations. Our results suggest S. maritima is a specialist species with respect to salinity, while S. densiflora is a generalist capable of germination of growth under suboptimal conditions. Invasive S. densiflora has greater capacity than native S. maritima to establish from seed with continued climate change and sea level rise.