Interception of radionuclides by planophile crops: A simple semi-empirical modelling approach in case of nuclear accident fallout.

Shortly after an atmospheric release, the interception of radionuclides by crop canopies represents the main uptake pathway leading to food chain contamination. The food chain models currently used in European emergency decision support systems require a large number of input parameters, which inevitably leads to high model complexity. In this study, we have established a new relationship for wet deposited radionuclides to simplify the current modelling approaches. This relationship is based on the hypothesis that the stage of plant development is the key factor governing the interception of radionuclides by crops having horizontally oriented leaves (planophile crops). The interception fraction (f) and the leaf area index normalized (fLAI) and mass normalized (fB) interception fractions were assessed for spinach (Spinacia oleracea) and radish (Raphanus sativus) at different stages of plant development and for different contamination treatments and plant densities. A database of 191 f values for Cs-137 and Th-229 was built and complemented with existing literature covering various radionuclides and crops with similar canopy structure. The overall f increased with the plant growth, while the reverse was observed for fB. The fLAI significantly decreased by doubling the contaminated rainfall deposited. Fitting a multiple linear regression to predict the f value as a function of the standing biomass (B), and the radionuclide form (anion and cation) led to a better estimation of the interception (R2 = 81%) than the ECOSYS-87 model (R2 = 35%). Hence, the simplified modelling approach here proposed seems to be a suitable risk assessment tool as fewer parameters will minimize the model complexity and facilitate the decision-making procedures in case of emergencies, when countermeasures need to be identified and implemented promptly.

Radiosensitivity and transgenerational effects in non-human species.

The ALLIANCE working group on effects of ionising radiation on wildlife brings together European researchers to work on the topics of radiosensitivity and transgenerational effects in non-human biota. Differences in radiation sensitivity across species and phyla are poorly understood, but have important implications for understanding the overall effects of radiation and for radiation protection; for example, sensitive species may require special attention in monitoring and radiation protection, and differences in sensitivity between species also lead to overall effects at higher levels (community, ecosystem), since interactions between species can be altered. Hence, understanding the mechanisms of interspecies radiation sensitivity differences may help to clarify mechanisms underpinning intraspecies variation. Differences in sensitivity may only be revealed when organisms are exposed to ionising radiation over several generations. This issue of potential long-term or hereditary effects for both humans and wildlife exposed to low doses of ionising radiation is a major concern. Animal and plant studies suggest that gamma irradiation can lead to observable effects in the F1 generation that are not attributable to inheritance of a rare stable DNA mutation. Several studies have provided evidence of an increase in genomic instability detected in germ or somatic cells of F1 organisms from exposed F0 organisms. This can lead to induced radiosensitivity, and can result in phenotypic effects or lead to reproductive effects and teratogenesis. In particular, studies have been conducted to understand the possible role of epigenetic modifications, such as DNA methylation, histone modifications, or expression of non-coding RNAs in radiosensitivity, as well as in adaptation effects. As such, research using biological models in which the relative contribution of genetic and epigenetic processes can be elucidated is highly valuable.

Chronic radiation exposure as an ecological factor: Hypermethylation and genetic differentiation in irradiated Scots pine populations.

Genetic and epigenetic changes were investigated in chronically irradiated Scots pine (Pinus sylvestris L.) populations from territories that were heavily contaminated by radionuclides as result of the Chernobyl Nuclear Power Plant accident. In comparison to the reference site, the genetic diversity revealed by electrophoretic mobility of AFLPs was found to be significantly higher at the radioactively contaminated areas. In addition, the genome of pine trees was significantly hypermethylated at 4 of the 7 affected sites.

The functional role of the photosynthetic apparatus in the recovery of Brassica napus plants from pre-emergent metazachlor exposure.

Metazachlor is a chloroacetamide herbicide, frequently used in Brassica napus cultivations around the world. Its primary target is the inhibition of very long chain fatty acid biosynthesis. This study included a morphological and physiological screening of hydroponically grown B. napus, exposed to a concentration range of 0, 0.25, 0.50, 0.75 and 1.0kg metazachlor per hectare. The results indicate that within a month after application, growth and development of B. napus are severely affected by low metazachlor doses. At intermediate metazachlor concentrations, loss of phosphorous and potassium from the plant tissues suggests destabilisation of cellular membranes, which may be a direct consequence of metazachlor application. This membrane instability could be indirectly linked with alterations of electron transport and a reduction of carbon assimilation. At increased metazachlor doses of 0.75kga.i.ha(-1), pigment concentrations are strongly reduced. However, chlorophyll fluorescence parameters seem to remain unaffected at metazachlor doses up to 0.75kga.i.ha(-1). At a metazachlor concentration of 1.0kga.i.ha(-1), negative effects are observed on all tested parameters, resulting in limited survival. The results indicate photosynthesis is assured at intermediate metazachlor concentrations for the cost of growth and development. It is clear that photosynthesis plays a key role in the survival strategy of young plants to overcome initially induced chemical stress.

Uranium and cadmium provoke different oxidative stress responses in Lemna minor L.

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 µm, respectively, and EC10 values of 1.5 ± 0.2 and 6.5 ± 0.9 µm, 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 (0.5-20 µm) strongly enhanced plant GPOD activity, whereas U inhibited it. Finally, in contrast to Cd, U up to 10 µm 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.

Joint research towards a better radiation protection-highlights of the Fifth MELODI Workshop.

MELODI is the European platform dedicated to low-dose radiation risk research. From 7 October through 10 October 2013 the Fifth MELODI Workshop took place in Brussels, Belgium. The workshop offered the opportunity to 221 unique participants originating from 22 countries worldwide to update their knowledge and discuss radiation research issues through 118 oral and 44 poster presentations. In addition, the MELODI 2013 workshop was reaching out to the broader radiation protection community, rather than only the low-dose community, with contributions from the fields of radioecology, emergency and recovery preparedness, and dosimetry. In this review, we summarise the major scientific conclusions of the workshop, which are important to keep the MELODI strategic research agenda up-to-date and which will serve to establish a joint radiation protection research roadmap for the future.

An invitation to contribute to a strategic research agenda in radioecology.

With intentions of integrating a portion of their respective research efforts into a trans-national programme that will enhance radioecology, eight European organisations recently formed the European Radioecology ALLIANCE (www.er-alliance.org). The ALLIANCE is an Association open to other organisations throughout the world with similar interests in promoting radioecology. The ALLIANCE members recognised that their shared radioecological research could be enhanced by efficiently pooling resources among its partner organizations and prioritising group efforts along common themes of mutual interest. A major step in this prioritisation process was to develop a Strategic Research Agenda (SRA). An EC-funded Network of Excellence in Radioecology, called STAR (Strategy for Allied Radioecology), was formed, in part, to develop the SRA. This document is the first published draft of the SRA. The SRA outlines a suggested prioritisation of research topics in radioecology, with the goal of improving research efficiency and more rapidly advancing the science. It responds to the question: "What topics, if critically addressed over the next 20 years, would significantly advance radioecology?" The three Scientific Challenges presented within the SRA, with their 15 associated research lines, are a strategic vision of what radioecology can achieve in the future. Meeting these challenges will require a directed effort and collaboration with many organisations the world over. Addressing these challenges is important to the advancement of radioecology and in providing scientific knowledge to decision makers. Although the development of the draft SRA has largely been a European effort, the hope is that it will initiate an open dialogue within the international radioecology community and its stakeholders. This is an abbreviated document with the intention of introducing the SRA and inviting contributions from interested stakeholders. Critique and input for improving the SRA are welcomed via a link on the STAR website (www.star-radioecology.org).

Dehydroascorbate and glucose are taken up into Arabidopsis thaliana cell cultures by two distinct mechanisms.

The possible involvement of glucose (Glc) carriers in the uptake of vitamin C in plant cells is still a matter of debate. For the first time, it was shown here that plant cells exclusively take up the oxidised dehydroascorbate (DHA) form. DHA uptake is not affected by 6-bromo-6-deoxy-ascorbate, an ascorbate (ASC) analogue, specifically demonstrating ASC uptake in animal cells. There is no competition between Glc and DHA uptake. Moreover, DHA and Glc carriers respond in the opposite manner to different inhibitors (cytochalasin B, phloretin and genistein). In conclusion, the plant plasma membrane DHA carrier is distinct from the plant Glc transporters.

Accumulation of tocopherols and tocotrienols during seed development of grape (Vitis vinifera L. cv. Albert Lavallée).

Tocopherols and tocotrienols are present in mature seeds. Yet, little is known about the physiological role and the metabolism of these compounds during seed development. Here we present data on tocopherol and tocotrienol accumulation during seed development in Vitis vinifera L. cv. Albert Lavallée (Royal). This species was chosen for its ability to synthesize both tocopherols and tocotrienols. It is shown here for the first time that during seed development there are significant differences in localization and accumulation kinetics of tocopherols and tocotrienols. Tocopherols are found homogeneously dispersed throughout all tissues of the seed, in concentrations ranging from 20 to 100 microg tocopherol per g dry weight. Tocopherol levels decrease gradually during seed development. In contrast, tocotrienols are only found in the endosperm of the seeds, accumulating in a sigmoid fashion during the maturation period of seed development. Tocotrienol levels were found to be (54+/-7.4) microg/g dry seed in 90-day-old seeds of V. vinifera L. Furthermore, tocotrienol biosynthesis is demonstrated in these seeds during tocotrienol accumulation and in an endosperm fraction isolated at 75 days after flowering.

Chlorophyll a fluorescence imaging of ozone-stressed Brassica napus L. plants differing in glucosinolate concentrations.

Brassicaceae are characterised by glucosinolates (GS), which appear to be involved not only in biotic but also in abiotic stress responses of plants. We investigated the effect of O (3) stress on leaf GS concentrations in two lines of BRASSICA NAPUS L., differing in GS content. Ozone fumigation decreased GS concentrations in leaves of B. NAPUS of one line. In control conditions, chlorophyll content, rates of saturating photosynthesis, and quantum yield of photosystem 2 differed between the two BRASSICA lines, but differences were smaller in O (3)-stress conditions, suggesting that the relationship between leaf GS concentration and sensitivity to abiotic stress merits further research. In agreement with other ecophysiological measurements, chlorophyll fluorescence imaging clearly distinguished both lines and in some cases also treatments. A method for analysis of fluorescence images accounting for the two-dimensional leaf heterogeneity is presented.

Copper-mediated oxidative burst in Nicotiana tabacum L. cv. Bright Yellow 2 cell suspension cultures.

In cell suspension cultures of Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) a rapid and concentration-dependent accumulation of H(2)O(2) is induced by excess concentrations of copper (up to 100 microM). This specific and early response towards copper stress was shown to be extracellular. Addition of 300 U of catalase per ml decreased the level of H(2)O(2). Superoxide dismutase (5 U/ml) induced an increase in H(2)O(2) production by 22.2%. This indicates that at least part of the H(2)O(2) is produced by dismutation of superoxide. Pretreatment of the cell cultures with the NAD(P)H oxidase inhibitors diphenylene iodonium (2 and 10 microM) and quinacrine (1 and 5 mM) prevented the generation of H(2)O(2) under copper stress for 90%. The influence of the pH on the H(2)O(2) production revealed the possible involvement of cell-wall-dependent peroxidases in the generation of reactive oxygen species after copper stress.

Transport and action of ascorbate at the plant plasma membrane.

The plasmalemma is both a bridge and a barrier between the cytoplasm and the outside world. It is a dynamic interface that perceives and transmits information concerning changes in the environment to the nucleus to modify gene expression. In plants, ascorbate is an essential part of this dialogue. The concentration and ratio of reduced to oxidized ascorbate in the apoplast, for example, possibly modulates cell division and growth. The leaf apoplast contains millimolar amounts of ascorbate that protect the plasmalemma against oxidative damage. The apoplastic ascorbate-dehydroascorbate redox couple is linked to the cytoplasmic ascorbate-dehydroascorbate redox couple by specific transporters for either or both metabolites. Although evidence about the mechanisms driving ascorbate or dehydroascorbate transport remains inconclusive, these carrier proteins potentially regulate the level and redox status of ascorbate in the apoplast. The redox coupling between compartments facilitated by these transport systems allows coordinated control of key physiological responses to environmental cues.

Carrier mediated uptake of dehydroascorbate into higher plant plasma membrane vesicles shows trans-stimulation.

The activity of the ascorbate (Asc) carrier of purified Phaseolus plasma membranes is demonstrated to be highly stimulated when membrane vesicles are preloaded with Asc. Asc transport is inhibited by DTT but is not affected by glutathione or ferricyanide, indicating that dehydroascorbate (DHA) is the preferred species for uptake. Asc transport in the loaded vesicles showed saturable kinetics with an apparent affinity constant of 24 microM and maximal uptake rate of 94 pmol/mg/min. Addition of DHA stimulated the efflux of Asc molecules from the loaded vesicles. Together these results suggest the presence of an Asc/DHA exchange mechanism in higher plant plasma membranes.

The Ascorbate Carrier of Higher Plant Plasma Membranes Preferentially Translocates the Fully Oxidized (Dehydroascorbate) Molecule.

Recently, the uptake of 14C-labeled ascorbate (ASC) into highly purified bean (Phaseolus vulgaris L.) plasma membrane vesicles was demonstrated in our laboratory. However, the question of the redox status of the transported molecule (ASC or dehydroascorbate [DHA]) remained unanswered. In this paper we present evidence that DHA is transported through the plasma membrane. High-performance liquid chromatography analysis of the redox status of ASC demonstrated that freshly purified plasma membranes exhibit a high ASC oxidation activity. Although it is not yet clear whether this activity is enzymatic, it complicates the interpretation of ASC-transport experiments in vitro and in vivo. In an attempt to correlate the ASC redox status to transport of the molecule, the ability of different compounds to reduce DHA was analyzed and their effect on ASC-transport activity tested. Administering of various reductants resulted in different levels of inhibition of ASC uptake (dithiothreitol > dithioerythritol > [beta]-mercaptoethanol > [beta]-mercaptopropanol). Glutathione, cysteine, dithionite, and thiourea did not significantly affect ASC transport. Statistical analysis indicated a strong correlation of the Spearman rank correlation coefficient (Rs) of 0.919 (P = 0.0005, n = 9) between the level of ASC oxidation and the amount of transported molecules into the vesicles. The administering of ASC oxidants such as ferricyanide and ASC oxidase resulted in a stimulated ASC uptake into the plasma membrane vesicles. Together, our results demonstrate that a vitamin C carrier in purified bean plasma membranes translocates DHA from the apoplast to the cytosol.

The Role of Ascorbate Free Radical as an Electron Acceptor to Cytochrome b-Mediated Trans-Plasma Membrane Electron Transport in Higher Plants.

The action of ascorbate free radical as an electron acceptor to cytochrome b-mediated trans-plasma membrane electron transport is demonstrated. Addition of ascorbate free radical to ascorbate-loaded plasma membrane vesicles caused a rapid oxidation of the cytochrome, followed by a slower re-reduction. The fully reduced dehydroascorbate was ineffective.

Transmembrane electron transport in ascorbate-loaded plasma membrane vesicles from higher plants involves a b-type cytochrome.

The possible involvement of a high-potential b-type cytochrome in plasma membrane electron transport was tested using ascorbate-loaded membrane vesicles. Absorption spectra demonstrated that the cytochrome was about 89% reduced in these preparations. Use of ascorbate oxidase and washing of the vesicles further indicated that reduction was mediated by intra-vesicular ascorbate. Addition of low concentrations of ferricyanide caused a rapid cytochrome oxidation followed by a slower re-reduction. The kinetics of this response indicate that the electron acceptor was fully reduced before re-reduction of the cytochrome occurred. These observations suggest that the b-type cytochrome mediates transmembrane electron transfer.