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Uranium, a heavy metal and primordial radionuclide, is present in surface waters and soils both naturally and due to industrial activities. Uranium is known to be toxic to plants and its uptake and toxicity can be influenced by multiple factors such as pH and the presence of different ions. However, the precise role of the different ions in uranium uptake is not yet known. Here we investigated whether calcium influences uranium uptake and toxicity in the terrestrial plant Arabidopsis thaliana. To this end, A. thaliana plants were exposed to different calcium and uranium concentrations and furthermore, calcium channels were blocked using the calcium channel blocker lanthanum chloride (LaCl

This work was supported by a PhD grant for Jackline Kariuki from the Belgian Nuclear Research Centre (SCK . CEN).

Ectomycorrhizal (EM) fungi form symbioses with dominant tree families in boreal, temperate and tropical ecosystems and are important drivers of ecosystem function. EM fungal hyphae extend over a large area making them susceptible to enhanced radiation levels from naturally occurring or anthropogenically originating radioisotopes in the rhizosphere. In this study, the in-vitro effects of ionizing radiation on the growth and biomass of EM fungi Suillus luteus, S. bovinus and Rhizopogon luteolus were investigated. EM fungal cultures were exposed to gamma radiation from a 137Cs source for 137 h in darkness at 21 °C at dose rates of 404, 108.5 and 54.9 mGy h−1 resulting in total absorbed doses of 55.21, 14.82 and 7.50 Gy respectively. Cultures grown in the dark at 21 °C but not exposed to the 137Cs source served as the control. Our results show that EM fungi vary in their sensitivity to ionizing radiation. EM fungi used in this study produced melanin and reactive oxygen species scavenging enzymes such as catalase and superoxide dismutase as a response to ionizing radiation.

This study aimed to compare the potential of Lemna minor, Spirodela sp., Eichhornia crassipes and Pistia stratiotes to remove 60Co from a realistic aquatic environment. Although all four plant species performed similarly well after 3 days of exposure to 50 kBq L−1 60Co, Lemna minor and Spirodela sp. came forward as having higher 60Co removal potential. This conclusion is, in first instance, based on the high 60Co removal percentage obtained after a short contact time (e.g. more than 95% could be removed after 6 h by Spirodela sp.). Additionally, Lemna minor and Spirodela sp. accumulated a high amount of 60Co per gram of biomass. For example, Lemna minor accumulated over three times more 60Co per gram of biomass compared to Pistia stratiotes and Eichhornia crassipes. Both plants also performed well in the pH range 5–9. We used Lemna minor to test the influence of the initial 60Co concentration (10, 50, 100 and 200 kBq L−1 60Co) on its phytoremediation capacity but no differences could be observed in removal percentage. In addition, it was shown that by optimising the initial amount of biomass, radioactive waste production can be minimised whilst maintaining high 60Co removal rates. Our study shows that these aquatic plants can be used for phytoremediation of 60Co from contaminated water and can be considered as a “green” addition or alternative for conventional remediation techniques.

Pollution of surface waters is a worldwide problem for people and wildlife. Remediation and phytoremediation approaches can offer a solution to deal with specific scenarios. Lemna minor, commonly known as duckweed, can absorb and accumulate pollutants in its biomass. To evaluate if L. minor could be applied for phytoremediation purposes, it is necessary to further investigate its remediation capability and to identify which parameters affect the remediation process. Such a model must include both plant growth and pollutant exchange. A remediation model based on a robust experimental study can help to evaluate L. minor as a proper remediation strategy and to predict the outcome of a L. minor based remediation system. To set up this model, this paper focusses on a detailed experimental study and a comprehensive mathematical modelling approach to represent L. minor growth as a function of biomass, temperature, light irradiation and variable nutrient concentrations. The influence of environmental conditions on L. minor growth was studied, by composing 7 days growth curves. Plants were grown under predefined environmental conditions (25°C, 14h photoperiod, 220 μmol m−2 s−1 light intensity and a modified Hoagland solution with 23.94 mg N L−1 and 3.10 mg P L−1 (N:P ratio of 7.73)) as standard for all experiments. The influence of different temperatures (6, 10, 15, 20, 25, 30 and 35°C), light intensities (63, 118, 170, 220 and 262 μmol m−2 s−1), photoperiods (12h and 14h) and N:P ratios (1.18, 3.36, 7.73 and 29.57) were tested in the model. As a result, a growth model was optimised using separate datasets for temperature, light intensity, photoperiod and nutrients and validated by further integrated testing. The growth model is a stable platform for application in phytoremediation of radionuclides in contaminated water, to be extended in future studies with information of pollutant uptake, pollutant-nutrient interactions and transfer to the biomass.

The mutants Atnoa1 and Atnia1nia2noa1-2 having a defective chloroplast developmental process, showed enhanced chlorophyll levels when they were grown on Murashige and Skoog (MS) medium and on exposure with uranium (U) on Hoagland medium. Thus we hypothesized that these mutants probably produced NO in MS medium and on exposure with U. Wild-type Col-0, Atnoa1, Atnia1nia2noa1-2 plants were cultured on modified Hoagland and 1/10 MS media and NO generation in the roots of these mutants was monitored using NO selective fluorescent dyes, DAF-2DA and Fl2E. Both Atnoa1 and Atnia1nia2noa1-2 triple mutants produced NO as observed by increases in DAF-2T and Fl2E fluorescence when these mutants were grown on MS medium but not on Hoagland medium. In presence of NO scavenger, methylene blue (MB, 200 μM), DAF-2T and Fl2E fluorescence was completely abolished. On the other hand treatment of the plants with 25 μM U triggered NO generation. U-treated Atnoa1 and Atnia1nia2noa1-2 plants upregulated genes (POR B, POR D, CHL D) involved in the chlorophyll biosynthesis. From these results it was concluded that Atnoa1 and Atnia1nia2noa1-2 are conditional NO producers and it appears that NO generation in plants substantially depends on growth medium and NIA1, NIA2 or NOA1 does not appear to be really involved in NO generation in MS medium or after U exposure.

This study aimed to compare the potential of Lemna minor, Spirodela sp., Eichhornia crassipes and Pistia stratiotes to remove

Uranium (U) is a nonessential and toxic heavy metal and radioactive element. Nitric oxide (NO) and hydrogen peroxide (H 2 O 2 ) are intimately involved in the growth, development and metabolic function of plant cells. In addition to possible toxic activities, a role for these signalling molecules in the regulation of plant responses to toxic metals has been proposed. Here, we investigated the synthesis of NO and H 2 O 2 in Arabidopsis thaliana seedlings exposed to 25 μM U. We demonstrated that U induces NO and H 2 O 2 production in the roots and leaves. Addition of the NO releasing compound, sodium nitroprusside (SNP) as well as the NO synthase inhibitor (L-NAME) to the plants, resulted in a decrease of the U-induced NO production and both alleviated U toxicity as indicated by plant growth (leaf area and biomass). Treatment with L-NAME further resulted in the disappearance of the U-induced H 2 O 2 in the shoots of the plants. It appears that generation of NO and H 2 O 2 is intimately linked in the toxicity response of U-exposed A. thaliana plants.

The biological effects and interactions of different radiation types in plants are still far from understood. Among different radiation types, external gamma radiation treatments have been mostly studied to assess the biological impact of radiation toxicity in organisms. Upon exposure of plants to gamma radiation, ionisation events can cause, either directly or indirectly, severe biological damage to DNA and other biomolecules. However, the biological responses and oxidative stress related mechanisms under chronic radiation conditions are poorly understood in plant systems. In the following study, it was questioned if the Lemna minor growth inhibition test is a suitable approach to also assess the radiotoxicity of this freshwater plant. Therefore, L. minor plants were continuously exposed for seven days to 12 different dose rate levels covering almost six orders of magnitude starting from 80 μGy h(-1) up to 1.5 Gy h(-1). Subsequently, growth, antioxidative defence system and genomic responses of L. minor plants were evaluated. Although L. minor plants could survive the exposure treatment at environmental relevant exposure conditions, higher dose rate levels induced dose dependent growth inhibitions starting from approximately 27 mGy h(-1). A ten-percentage growth inhibition of frond area Effective Dose Rate (EDR10) was estimated at 95 ± 7 mGy h(-1), followed by 153 ± 13 mGy h(-1) and 169 ± 12 mGy h(-1) on fresh weight and frond number, respectively. Up to a dose rate of approximately 5 mGy h(-1), antioxidative enzymes and metabolites remained unaffected in plants. A significant change in catalase enzyme activity was found at 27 mGy h(-1) which was accompanied with significant increases of other antioxidative enzyme activities and shifts in ascorbate and glutathione content at higher dose rate levels, indicating an increase in oxidative stress in plants. Recent plant research hypothesized that environmental genotoxic stress conditions can induce endoreduplication events. Here an increase in ploidy level was observed at the highest tested dose rate. In conclusion, the results revealed that in plants several mechanisms and pathways interplay to cope with radiation induced stress.

Long-lived radionuclides such as (90)Sr and (137)Cs can be naturally or accidentally deposited in the upper soil layers where they emit β/γ radiation. Previous studies have shown that arbuscular mycorrhizal fungi (AMF) can accumulate and transfer radionuclides from soil to plant, but there have been no studies on the direct impact of ionizing radiation on AMF. In this study, root organ cultures of the AMF Rhizophagus irregularis MUCL 41833 were exposed to 15.37, 30.35, and 113.03 Gy gamma radiation from a (137)Cs source. Exposed spores were subsequently inoculated to Plantago lanceolata seedlings in pots, and root colonization and P uptake evaluated. P. lanceolata seedlings inoculated with non-irradiated AMF spores or with spores irradiated with up to 30.35 Gy gamma radiation had similar levels of root colonization. Spores irradiated with 113.03 Gy gamma radiation failed to colonize P. lanceolata roots. P content of plants inoculated with non-irradiated spores or of plants inoculated with spores irradiated with up to 30.35 Gy gamma radiation was higher than in non-mycorrhizal plants or plants inoculated with spores irradiated with 113.03 Gy gamma radiation. These results demonstrate that spores of R. irregularis MUCL 41833 are tolerant to chronic ionizing radiation at high doses.

In the following study, dose dependent effects on growth and oxidative stress induced by -radiation were examined to gain better insights in the mode of action of -radiation induced stress in plant species. Radiostrontium (Sr-90) was used to test for -radiation induced responses in the freshwater macrophyte Lemna minor. The accumulation pattern of Sr-90 was examined for L. minor root and fronds separately over a seven-day time period and was subsequently used in a dynamic dosimetric model to calculate -radiation dose rates. Exposing L. minor plants for seven days to a Sr-90 activity concentration of 25 up to 25,000 kBqL(-1) resulted in a dose rate between 0.084 +/- 0.004 and 97 +/- 8 mGyh(-1). After seven days of exposure, root fresh weight showed a dose dependent decrease starting from a dose rate of 9.4 +/- 0.5 mGyh(-1). Based on these data, an EDR10 value of 1.5 +/- 0.4 mGyh(-1) was estimated for root fresh weight and 52 +/- 17 mGyh(-1) for frond fresh weight. Different antioxidative enzymes and metabolites were further examined to analyze if -radiation induces oxidative stress in L. minor. The authors thank the Research foundation-Flanders (FWO) (G.A040.11N) and the European Commission Contract Fission-2010-3.5.1-269672 Strategy for Allied Radioecology (www.star-radioecology.org) for financial support of this work. Belgian nuclear research institute (SCK.CEN) is further thanked for funding the PhD of A. Van Hoeck.

As photosynthesis is already known to be affected under various metal stresses, the aim of this study is to investigate uranium effects on photosynthetic parameters. Therefore, 18-day-old Arabidopsis thaliana seedlings were exposed to 50 μM uranium during 1–96 h. Uranium uptake, effects on growth parameters of roots and leaves and further responses on photosynthesis, pigment concentrations and lipid peroxidation in leaves were investigated. Uranium was highly taken up by the roots (50,352 ± 3383 μg g−1 DW at 96 h) causing complete growth arrest of the plants. Although uranium concentrations in the leaves remained low (15.0 ± 4.0 μg g−1 DW at 96 h), a remarkable photosynthetic response mechanism was observed. By chlorophyll fluorescence measurements it was observed that the potential photosynthetic efficiency (Fv/Fm) remained maximal while the effective efficiency of photosystem II (φPSII), which is a measure for the proportion of light absorbed by PSII used in photochemistry, even increased due to a decrease in non-photochemical quenching (NPQ), which indicates the conversion of excess energy into heat, but no alterations in non-regulated energy dissipation (NO). When measuring rapid light curves (RLC), giving the increase of the electron transport rate as function of irradiance, no differences were observed for the maximal electron transport rate (ETRmax) but an increase in α, representing the photosynthetic rate in the light-limited region of the RLC, was observed under uranium stress. We concluded that plant leaves start increasing their photosynthetic efficiency and decreasing their non-photochemical quenching under uranium stress.

Ecotoxicological research provides knowledge on ionising radiation-induced responses in different plant species. However, the sparse data currently available are mainly extracted from acute exposure treatments. To provide a better understanding of environmental exposure scenarios, the response to stress in plants must be followed in more natural relevant chronic conditions. We previously showed morphological and biochemical responses in Lemna minor plants continuously exposed for 7 days in a dose-rate dependent manner. In this study responses on molecular (gene expression) and physiological (photosynthetic) level are evaluated in Leminor plants exposed to ionising radiation. To enable this, we examined the gene expression profiles of irradiated Leminor plants by using an RNA-seq approach. The gene expression data reveal indications that Leminor plants exposed at lower dose rates, can tolerate the exposure by triggering acclimation responses. In contrast, at the highest dose rate tested, a high number of genes related to antioxidative defense systems, DNA repair and cell cycle were differentially expressed suggesting that only high dose rates of ionising radiation drive Leminor plants into survival strategies. Notably, the photosynthetic process seems to be unaffected in Leminor plants among the tested dose rates. This study, supported by our earlier work, clearly indicates that plants shift from acclimation responses towards survival responses at increasing dose rates of ionising radiation. (C) 2017 Elsevier B.V. All rights reserved.

When terrestrial environments get contaminated with long-lived gamma emitting radionuclides, plants that grow in these contaminated areas are exposed to gamma radiation during consecutive generations. Therefore it is important to evaluate the gamma induced stress response in plants in and between generations. The objective of this research is to reveal differences at the level of the antioxidative stress response between generations with a different radiation history. An experiment was conducted in which 7-days old Arabidopsis thaliana plants were exposed for 14 days to four different gamma dose rates: 22 mGy/h, 38 mGy/h, 86 mGy/h and 457 mGy/h. Two different plant groups were used: plants that were not exposed to gamma radiation before (P0) and plants that received the aforementioned gamma treatment during their previous generation (S1). Growth, the concentration of the antioxidants ascorbate and glutathione, a number of antioxidative enzyme activities and their gene transcript levels were analysed. A dose-rate dependent induction was seen for catalase (CAT) and guaiacol peroxidase (GPX) in the roots and for syringaldazine peroxidase (SPX) in the shoots. Differences between the two generations were observed for CAT and GPX in the roots, where a significantly higher activity of these reactive oxygen species (ROS) detoxifying enzymes was observed in the S1 generation. For SPX in the shoots, a dose dependent upregulation was observed in the PO generation. However, high SPX activities were present for all doses in the S1 generation. These differences in enzyme activity between generations for SPX and GPX and the involvement of these enzymes in cell wall biosynthesis, suggest an important role for cell wall strengthening in the response to gamma irradiation. (C) 2016 Elsevier Ltd. All rights reserved. This project was supported by the European project COMET (7th PCRD EURATOM Contract Number: Fission-2012-3.4.1-604794) (www.comet-radioecology.org) and the Belgian nuclear research centre (SCK CEN).

Common duckweed (Lemna minor L.) is ideally suited to test the impact of metals on freshwater vascular plants. Literature on cadmium (Cd) and uranium (U) oxidative responses in L. minor are sparse or, for U, non-existent. It was hypothesised that both metals impose concentration-dependent oxidative stress and growth retardation on L. minor. Using a standardised 7-day growth inhibition test, the adverse impact of these metals on L. minor growth was confirmed, with EC50 values for Cd and U of 24.1 2.8 and 29.5 1.9 lM, respectively, and EC10 values of 1.5 0.2 and 6.5 0.9 lM, respectively. The metal-induced oxidative stress response was compared through assessing the activity of different antioxidative enzymes [catalase, glutathione reductase, superoxide dismutase (SOD), ascorbate peroxidase (APOD), guaiacol peroxidase (GPOD) and syringaldizyne peroxidase (SPOD)]. Significant changes in almost all antioxidative enzymes indicated their importance in counteracting the U- and Cd-imposed oxidative burden. However, some striking differences were also observed. For activity of APODs and SODs, a biphasic but opposite response at low Cd compared to U concentrations was found. In addition, Cd (0.520 lM) strongly enhanced plant GPOD activity, whereas U inhibited it. Finally, in contrast to Cd, U up to 10 lM increased the level of chlorophyll a and b and carotenoids. In conclusion, although U and Cd induce similar growth arrest in L. minor, the U-induced oxidative stress responses, studied here for the first time, differ greatly from those of Cd.

As the environment is inevitably exposed to ionizing radiation from natural and anthropogenic sources, it is important to evaluate gamma radiation induced stress responses in plants. The objective of this research is therefore to investigate radiation effects in Arabidopsis thaliana on individual and subcellular level by exposing 2-weeks-old seedlings for 7 days to total doses of 3.9 Gy, 6.7 Gy, 14.8 Gy and 58.8 Gy and evaluating growth, photosynthesis, chlorophyll a, chlorophyll b and carotenoid concentrations and antioxidative enzyme capacities. While the capacity of photosystem II (PSII measured as Fv/Fm) remained intact, plants started optimizing their photosynthetic process at the lower radiation doses by increasing the PSII efficiency (φPSII) and the maximal electron transport rate (ETRmax) and by decreasing the non-photochemical quenching (NPQ). At the highest radiation dose, photosynthetic parameters resembled those of control conditions. On subcellular level, roots showed increased superoxide dismutase (SOD) and ascorbate peroxidase (APX) capacities under gamma irradiation but catalase (CAT), syringaldazine peroxidase (SPX) and guaiacol peroxidase (GPX) activities, on the other hand, decreased. In the leaves no alterations were observed in SOD, CAT and SPX capacities, but GPX was highly affected. Based on these results it seems that roots are more sensitive for oxidative stress under gamma radiation exposure than leaves.

The edible cyanobacterium Arthrospira is resistant to ionising radiation. The cellular mechanisms underlying this radiation resistance are, however, still largely unknown. Therefore, additional molecular analysis was performed to investigate how these cells can escape from, protect against, or repair the radiation damage. Arthrospira cells were shortly exposed to different doses of 60Co gamma rays and the dynamic response was investigated by monitoring its gene expression and cell physiology at different time points after irradiation. The results revealed a fast switch from an active growth state to a kind of 'survival modus' during which the cells put photosynthesis, carbon and nitrogen assimilation on hold and activate pathways for cellular protection, detoxification, and repair. The higher the radiation dose, the more pronounced this global emergency response is expressed. Genes repressed during early response, suggested a reduction of photosystem II and I activity and reduced tricarboxylic acid (TCA) and Calvin-Benson-Bassham (CBB) cycles, combined with an activation of the pentose phosphate pathway (PPP). For reactive oxygen species detoxification and restoration of the redox balance in Arthrospira cells, the results suggested a powerful contribution of the antioxidant molecule glutathione. The repair mechanisms of Arthrospira cells that were immediately switched on, involve mainly proteases for damaged protein removal, single strand DNA repair and restriction modification systems, while recA was not induced. Additionally, the exposed cells showed significant increased expression of arh genes, coding for a novel group of protein of unknown function, also seen in our previous irradiation studies. This observation confirms our hypothesis that arh genes are key elements in radiation resistance of Arthrospira, requiring further investigation. This study provides new insights into phasic response and the cellular pathways involved in the radiation resistance of microbial cells, in particularly for photosynthetic organisms as the cyanobacterium Arthrospira.