Maintaining the cultivation of vegetables with low Pb accumulation while remediating the soil of an allotment garden (Nantes, France) by phytoextraction.

Lead (Pb) is commonly found in urban soils and can transfer to vegetables. This entails a health risk for consumers of garden crops. The increasing demand of gardening on urban soil linked to the population increase and concentration in urban areas induces an increase in the risk, as people could be forced to cultivate contaminated soils. The aim of this study was to evaluate the performance of a cropping system that allows simultaneously (i) growing eatable vegetables that accumulate few Pb and (ii) cleaning up the soil with other plants by phytoextraction. The tests were carried out in an allotment garden (Nantes, France) where soils are moderately enriched by Pb from geogenic origin (178 mg.kg-1 of dry soil on average). Four vegetables known to accumulate slightly Pb (Solanum lycopersicum, Brassica oleracea cv. "Capitata," Solanum tuberosum, and Phaseolus vulgaris) were grown. The in situ ability of Brassica juncea L. to progressively absorb the phytoavailable Pb of the soil was assessed during four seasons. Analyses of the edible parts of the four vegetables confirmed that they can all be safely cultivated. The accumulation of Pb in B. juncea shoots was too low (ca. 1 mg.kg-1 of dry matter at best) for phytoextraction purposes. Our results confirm that it is possible to grow very low Pb-accumulating vegetables on soils moderately contaminated with Pb, although it was not possible to reduce phytoavailable Pb rapidly enough with B. juncea. This study identifies possible avenues of research to improve this cropping system by using appropriate vegetables that will allow food production to continue on moderately contaminated soil while cleaning it up.

Haslea silbo, A Novel Cosmopolitan Species of Blue Diatoms.

Specimens of a new species of blue diatoms from the genus Haslea Simonsen were discovered in geographically distant sampling sites, first in the Canary Archipelago, then North Carolina, Gulf of Naples, the Croatian South Adriatic Sea, and Turkish coast of the Eastern Mediterranean Sea. An exhaustive characterization of these specimens, using a combined morphological and genomic approach led to the conclusion that they belong to a single new to science cosmopolitan species, Haslea silbo sp. nov. A preliminary characterization of its blue pigment shows similarities to marennine produced by Haslea ostrearia, as evidenced by UV-visible spectrophotometry and Raman spectrometry. Life cycle stages including auxosporulation were also observed, providing data on the cardinal points of this species. For the two most geographically distant populations (North Carolina and East Mediterranean), complete mitochondrial and plastid genomes were sequenced. The mitogenomes of both strains share a rare atp6 pseudogene, but the number, nature, and positions of the group II introns inside its cox1 gene differ between the two populations. There are also two pairs of genes fused in single ORFs. The plastid genomes are characterized by large regions of recombination with plasmid DNA, which are in both cases located between the ycf35 and psbA genes, but whose content differs between the strains. The two sequenced strains hosts three plasmids coding for putative serine recombinase protein whose sequences are compared, and four out of six of these plasmids were highly conserved.

Bioaugmentation coupled with phytoextraction for the treatment of Cd and Sr, and reuse opportunities for phosphogypsum rare earth elements.

The environmental and health impacts caused by phosphogypsum (PG) make it necessary to carefully manage these wastes. Bioaugmentation of a PG-compost mix with Bacillus cereus was associated with Trifolium pratense or Helianthus annuus for the phytoextraction of metal trace elements (MTE). In hydroponics, MTE concentrations in sunflower shoots are higher than in clover; however, as opposed to clover, it regulates their accumulation. The MTE accumulation levels by plants cultivated in pots with the PG-compost mix are much lower than in hydroponics due to lower concentration in available MTE. The bacteria-plant coupling has served to raise MTE concentrations, especially for rare earth elements (REE), i.e., Ce, La, Nd, Y, in the AP of sunflower, by factors of 4.4, 38.3, 3.4 and 21, respectively, compared to non-bioaugmented control. The translocation factor was also increased for all MTE and is ranged between 1.1 for Sr and 6.8 for Y. Moreover, the presence of bacteria raises plant biomass by a factor of 3.7 for shoots and 2.9 for the roots as regards clover. Results showed that in addition to phytoextraction of REE elements, all providing the promise of some kind of economic opportunity, the dispersion of PG stockpiles dust and erosion should be reduced.

Isolation and screening of indigenous bacteria from phosphogypsum-contaminated soils for their potential in promoting plant growth and trace elements mobilization.

Bacteria isolated from soils in the vicinity of phosphogypsum (PG) stockpiles were studied for their potential use in bioaugmentation-assisted phytoextraction. Quick, miniaturized biochemical tests were performed in the presence of metal trace elements (MTE), including rare earth elements (Cd, Sr, Ce, La, Nd and Y), corresponding to their bioavailable concentrations in PG. The intention herein was to assess the capacity of bacteria to: i) grow in PG; ii) produce indole acetic acid and ACC deaminase to promote plant growth and reduce stress; and iii) produce siderophores, including pyoverdine, to mobilize MTE. Results showed that even at maximum PG concentration (10 g/L and pH 3.40), 7 out of 32 isolates were able to grow. The biochemical tests showed differences in the presence or absence of MTE. The presence of MTE seems to promote the production of IAA by a factor of 3.25. On the contrary, it inhibits ACC deaminase and siderophore production, including pyoverdine. According to a scoring method applied, the two most efficient isolates exhibiting maximum metabolite production were identified as Bacillus sp.

Chelate-assisted phytoextraction of lead using Fagopyrum esculentum: laboratory vs. field experiments.

The development of more sustainable remediation techniques has been receiving greater attention, as an alternative to soil excavation plan in urban gardens. An in situ phytoextraction experiment with buckwheat (Fagopyrum esculentum) was performed with a 5 mmol kg-1 citric acid (CA) application. Joint experiments under laboratory conditions were conducted using various cultivars of F. esculentum in two soils with a Pb contamination of either geogenic or anthropogenic origin and various chelate concentrations. Results show that a minimum dose of 50 mmol kg-1 of CA is required to lower soil pH and raise the concentration of mobile Pb-CaCl2 for both soils. Consequently, Pb shoot uptake is increased from 6.3 to 8.9 times depending on soil type. Phytoextraction efficiency is found to be 1.3 to 2.0 times higher in the anthropogenic contaminated soil than in the soil with geogenic Pb. A scale effect has also been identified since Pb root accumulation under laboratory conditions was 2.4 times higher than in the field experiment. Despite an increase in the Pb extraction rate with CA, buckwheat appears to lack the efficiency needed to remove Pb in moderately contaminated soils. The calculated remediation period would last 166 years to remove the mobile Pb fraction.

Fate and transport of metal trace elements from phosphogypsum piles in Tunisia and their impact on soil bacteria and wild plants.

The phosphate industry in Tunisia generates large amounts of phosphogypsum (PG) with more than 107 t per year. Environmental impact of this solid waste was studied. Cd, Ce, La, Nd, Sr and Y were analyzed from soils near PG stockpiles (Sfax and M'dhilla) and sediments from marine discharge (Gabes). Their impacts on the bacterial community structure and wild plants were investigated. Metal trace elements (MTE) concentrations (in mg Kg-1 DM) were much higher in contaminated soil than in the control (at 12 km from PG stockpiles). Highest concentrations were recorded in top soil and decreased with depth. A low bacterial diversity was shown (impacted by plants more than by MTE). The MTE concentrations in aerial parts (AP) and roots varied according to the plant species and were higher in contaminated sites. Sr, La and Cd in the AP ranged 33.10-657.56, 2.22-11.05 and 0.21-14.20 mg Kg-1 DM respectively. Plants exhibiting the maximal metal concentrations in AP (in mg Kg-1 DM) were the following: Zygophylum album for Sr (657.56) >Zygophylum album for Cd (14.20) >Zygophylum album (11.05) for La >Conyza canadensis (1.11) for Ce >Conyza canadensis (0.75) for Nd >Arthrocemum inducum (0.72) for Y. Kochia indica showed the highest bioconcentration factor (1.60) for Cd, while Zygophylum album exhibited the highest translocation factor (6.12) for La. Zygophylum album would be the most suitable candidate for MTE phytoextraction. CAPSULE: Phosphogypsum contaminates soils near stockpiles with metal trace elements including rare earth element and selects wild plants able to be used for phytostabilization and phytomining.

2D Image Quantification of Microbial Iron Chelators (Siderophores) Using Diffusive Equilibrium in Thin Films Method.

Siderophores are natural metal chelating agents that strongly control the biogeochemical metal cycles such as Fe in the environment. This article describes a new methodology to detect and quantify at the micromolar concentration the spatial distribution at millimeter scale of siderophores within the root's system. The "universal" CAS assay originally designed for bacterial siderophores detection and later designed for fungus was adapted here for diffusive equilibrium in thin film gel techniques (DET). The method was calibrated against the marketed desferrioxamine mesylate (DFOM) siderophore and applied with experiments performed with sunflower ( Helianthus annuus) and wheat ( Triticum aestivum) cultivated on free iron agar medium plates. We present here the first results with 2D images of the siderophores distribution in the vicinity of the root system of plants. With this technique we detected (i) the production of siderophores on bacteria inoculated ( Pseudomonas fluorescens) environments and (ii) hotspots of natural iron binding ligands production up to 50 µM in the wheat rhizosphere. The lower detection limit in our experiment was 2.5 µmol/L. This new technique offers a unique opportunity to investigate the siderophore production in two dimensions in a wide range of applications from laboratory experiments to natural systems very likely using an in situ and nondestructive tool.

Effect of Pseudomonas fluorescens and pyoverdine on the phytoextraction of cesium by red clover in soil pots and hydroponics.

With the aim of improving the phytoextraction rate of cesium (Cs), the effect of Pseudomonas fluorescens ATCC 17400 and its siderophore pyoverdine (PVD) on the uptake of Cs by red clover was studied in soil pots. This work also provides a mechanistic understanding of the Cs-bacteria (or PVD)-illite-plant interactions by using a simplified experimental design, i.e., hydroponics with either Cs in solution or Cs-spiked illite in suspension. For soil spiked with 11.2 mmol kg-1 (1480 mg kg-1) of Cs, 0.43% of total Cs was taken up by red clover in 12 days (119 µmol g-1 (16 mg g-1) of Cs dry matter in roots and 40 µmol g-1 (5 mg g-1) in shoots). In hydroponics with Cs in solution (0.1 mmol L-1 or 13 mg L-1), 75% of Cs was taken up vs. only 0.86% with Cs-spiked illite suspension. P. fluorescens and PVD did not increase Cs concentrations in aboveground parts and roots of red clover and even decreased them. The damaging effect of PVD on red clover growth was demonstrated with the biomass yielding 66% of the control in soil pots (and 100% mortality after 12 days of exposition) and only 56% in hydroponics (78% with illite in suspension). Nonetheless, PVD and, to a lesser extent, P. fluorescens increased the translocation factor up to a factor of 2.8. This study clearly showed a direct damaging effect of PVD and to a lower extent the retention of Cs by biofilm covering both the roots and illite, both resulting in the lower phytoextraction efficiency.

Growth form defines physiological photoprotective capacity in intertidal benthic diatoms.

In intertidal marine sediments, characterized by rapidly fluctuating and often extreme light conditions, primary production is frequently dominated by diatoms. We performed a comparative analysis of photophysiological traits in 15 marine benthic diatom species belonging to the four major morphological growth forms (epipelon (EPL), motile epipsammon (EPM-M) and non-motile epipsammon (EPM-NM) and tychoplankton (TYCHO)) found in these sediments. Our analyses revealed a clear relationship between growth form and photoprotective capacity, and identified fast regulatory physiological photoprotective traits (that is, non-photochemical quenching (NPQ) and the xanthophyll cycle (XC)) as key traits defining the functional light response of these diatoms. EPM-NM and motile EPL showed the highest and lowest NPQ, respectively, with EPM-M showing intermediate values. Like EPL, TYCHO had low NPQ, irrespective of whether they were grown in benthic or planktonic conditions, reflecting an adaptation to a low light environment. Our results thus provide the first experimental evidence for the existence of a trade-off between behavioural (motility) and physiological photoprotective mechanisms (NPQ and the XC) in the four major intertidal benthic diatoms growth forms using unialgal cultures. Remarkably, although motility is restricted to the raphid pennate diatom clade, raphid pennate species, which have adopted a non-motile epipsammic or a tychoplanktonic life style, display the physiological photoprotective response typical of these growth forms. This observation underscores the importance of growth form and not phylogenetic relatedness as the prime determinant shaping the physiological photoprotective capacity of benthic diatoms.

Biochemical and behavioural responses of the marine polychaete Hediste diversicolor to cadmium sulfide quantum dots (CdS QDs): waterborne and dietary exposure.

Cadmium sulfide (CdS) quantum dots are widely used in medical imaging. The aim of this study was to examine toxicity effects of CdS engineered nanoparticles (CdS NPs) compared to soluble Cd, on marine ragworms (Hediste diversicolor) exposed for 14 d to these contaminants (10 µg Cd L(-1)) in seawater or via their food (contaminated worm tissue). In our experimental media, Dynamic Light Scattering studies showed that the majority of CdS remained in the nanoscale (1-10 nm) with the exception of few aggregates (100-300 nm). Labile Cd fractions released from CdS NPs were estimated by diffusive gradient in thin films, showing that about 50% of CdS NPs remained in nanoparticulate form. Ragworms accumulated Cd in both soluble Cd and CdS NPs in waterborne exposures only. Greater significant changes of biochemical responses were observed in worms exposed to CdS NPs in seawater compared to contaminated food. Catalase and glutathione-S-transferase activities were the most sensitive biochemical biomarkers responding to both Cd treatments for waterborne exposure. Inductions of CAT were higher in diet-exposed worms to Cd as NPs vs soluble form suggesting a specific "nano" effect. Caspase activities increased in worms exposed to soluble Cd and Cd NPs for the two routes of exposure compared to controls. Defences, may be insufficient to prevent reactive oxygen species generation and the associated apoptosis. Behaviour of invertebrates inside sediment showed impairments of body movements in worms exposed to CdS NPs. This study points out oxidative processes as the main consequences of exposure to Cd based NPs in worms.

Inheritance of mitochondrial DNA in the Pennate diatom Haslea ostrearia (Naviculaceae) during auxosporulation suggests a uniparental transmission.

We present the first study examining mtDNA transmission in diatoms, using sexual progeny of the pennate species Haslea ostrearia (Naviculaceae). A fragment of the cytochrome oxidase subunit I gene (cox1) with 7 nucleic substitutions between parental clones was used as a parental tracer in 16 F1 clones obtained from two pairs of mating crosses. Each cross involved a parental clone isolated from France (Bay of Bourgneuf) and Sweden (Kattegat Bay). We determined that all progeny possessed only one cox1 parental haplotype. These results suggest that the mitochondrial DNA transmission in H. ostrearia is uniparental. Implications and new topics of investigation are discussed.

Biochemical and behavioural responses of the endobenthic bivalve Scrobicularia plana to silver nanoparticles in seawater and microalgal food.

Because of their bactericidal effects, Ag nanoparticles (Ag NPs) have promising industrial development but could lead to potential ecological risks. The aim of this study was to examine the uptake and effect of silver (soluble or as lactate Ag NPs of 40 nm) at low concentrations (10 µg L(-1)) in the endobenthic bivalve Scrobicularia plana exposed, for 14 days, directly (water) or via the diet (microalgae). The stability of Ag NPs in seawater was examined using dynamic light scattering. Release of soluble Ag from Ag NPs in the experimental media was quantified by using diffusive gradient in thin film. Bioaccumulation of Ag in bivalves was measured by electrothermal atomic absorption spectrometry. Behavioural and biochemical biomarkers were determined in bivalves. Aggregation of Ag NPs and the release of soluble Ag from Ag NPs were observed in the experimental media. For both forms of Ag, bioaccumulation was much more important for waterborne than for dietary exposure. The response of oxidative stress biomarkers (catalase, glutathion S-transferase, superoxide dismutase) was more important after dietary than waterborne exposure to Ag (soluble and NPs). These defences were relatively efficient since they led to a lack of response of damage biomarkers. Burrowing was not affected for bivalves exposed directly or through the diet to both Ag forms but feeding behaviour was impaired after 10 days of dietary exposure. Since no differences of responses to Ag either soluble or nanoparticulate were observed, it seems that labile Ag released from Ag NPs was mainly responsible for toxicity.

Biological activities of purified marennine, the blue pigment responsible for the greening of oysters.

Marennine, the blue pigment produced by the diatom Haslea ostrearia , exists in two different forms, the intra- and extracellular forms. We investigated the antibacterial, antiviral, and antiproliferative properties of both of these forms. Both forms of marennine inhibited the development of marine bacteria, in particular the pathogenic organism Vibrio aesturianus , at concentrations as low as 1 µg/mL, but they did not display any effect on a wide range of pathogenic bacteria that are relevant for food safety. Both forms of the pigment produced by H. ostrearia also exhibited antiviral activity against the HSV1 herpes virus, with intra- and extracellular marennine having EC(50) values of 24.0 and 27.0 µg/mL, respectively. These values are 2 orders of magnitude higher than the value for the reference drug, Zovirax. Moreover, both forms of marennine were effective in slowing or inhibiting the proliferation of cancer cells. This study confirms the potential of marennine as a biologically active organic molecule, which could have a protective effect on bivalves, which filter seawater and fix the pigment on their gills. Moreover, marennine could be used in food engineering and chemistry as a natural blue pigment. However, despite that it is eaten and possibly assimilated by green oyster consumers, it also deserves in depth evaluation before being considered for use as a nutraceutical.

Light is a key factor in triggering sexual reproduction in the pennate diatom Haslea ostrearia.

Sexual reproduction is an obligatory phase in the life cycle of most diatoms, as cell size decreases with successive vegetative divisions and the maximal cell size is only restored by a specialized cell, the auxospore, which follows zygote formation as a result of sexual reproduction. While in pennate diatoms the induction of sexual reproduction depends primarily on cell-cell interactions, the importance of different external factors for the induction of sexual reproduction is less well known. Here, we investigated the effects of light on sexualization in the marine benthic pennate diatom Haslea ostrearia (Gaillon) R. Simonsen. Compatible clones were crossed and exposed to different combinations of light levels, qualities, and photoperiods. Light was found to be a key factor for sexualization, and to a certain extent, to control auxosporulation in H. ostrearia. The light conditions most favorable for sexual reproduction were low irradiances (<50 micromolphotons m(-2) s(-1)) and short photoperiods (6-10 h), conditions that prevail during winter, and to a lesser extent, the higher irradiances and longer photoperiods that correspond to the spring and fall, when blooms of this organism form in the natural environment. Auxospore formation was very rare in continuous light, and maximum in presence of red radiation, while it was never observed in darkness or in radiation other than red.