Ultraviolet B modulates gamma radiation-induced stress responses in Lemna minor at multiple levels of biological organisation.

Elevated levels of ionizing and non-ionizing radiation may co-occur and pose cumulative hazards to biota. However, the combined effects and underlying toxicity mechanisms of different types of radiation in aquatic plants remain poorly understood. The present study aims to demonstrate how different combined toxicity prediction approaches can collectively characterise how chronic (7 days) exposure to ultraviolet B (UVB) radiation (0.5 W m-2) modulates gamma (γ) radiation (14.9, 19.5, 43.6 mGy h-1) induced stress responses in the macrophyte Lemna minor. A suite of bioassays was applied to quantify stress responses at multiple levels of biological organisation. The combined effects (no-enhancement, additivity, synergism, antagonism) were determined by two-way analysis of variance (2 W-ANOVA) and a modified Independent Action (IA) model. The toxicological responses and the potential causality between stressors were further visualised by a network of toxicity pathways. The results showed that γ-radiation or UVB alone induced oxidative stress and programmed cell death (PCD) as well as impaired oxidative phosphorylation (OXPHOS) and photosystem II (PSII) activity in L. minor. γ-radiation also activated antioxidant responses, DNA damage repair and chlorophyll metabolism, and inhibited growth at higher dose rates (≥20 mGy h-1). When co-exposed, UVB predominantly caused non-interaction (no-enhancement or additive) effects on γ-radiation-induced antioxidant gene expression, energy quenching in PSII and growth for all dose rates, whereas antagonistic effects were observed for lipid peroxidation, OXPHOS, PCD, oxidative stress, chlorophyll metabolism and genes involved in DNA damage responses. Synergistic effects were observed for changes in photochemical quenching and non-photochemical quenching, and up-regulation of antioxidant enzyme genes (GST) at one or more dose rates, while synergistic reproductive inhibition occurred at all three γ-radiation dose rates. The present study provides mechanistic knowledge, quantitative understanding and novel analytical strategies to decipher combined effects across levels of biological organisation, which should facilitate future cumulative hazard assessments of multiple stressors.

Effects of artificial ultraviolet B radiation on the macrophyte Lemna minor: a conceptual study for toxicity pathway characterization.

MAIN CONCLUSION: UVB radiation caused irradiance-dependent and target-specific responses in non-UVB acclimated Lemna minor. Conceptual toxicity pathways were developed to propose causal relationships between UVB-mediated effects at multiple levels of biological organisation. Macrophytes inhabit waterways around the world and are used in hydroponics or aquaponics for different purposes such as feed and wastewater treatment and are thus exposed to elevated levels of UVB from natural and artificial sources. Although high UVB levels are harmful to macrophytes, mechanistic understanding of irradiance-dependent effects and associated modes of action in non-UVB acclimated plants still remains low. The present study was conducted to characterise the irradiance-dependent mechanisms of UVB leading to growth inhibition in Lemna minor as an aquatic macrophyte model. The L. minor were continuously exposed to UVB (0.008-4.2 W m-2) and constant UVA (4 W m-2) and photosynthetically active radiation, PAR (80 µmol m-2 s-1) for 7 days. A suite of bioassays was deployed to assess effects on oxidative stress, photosynthesis, DNA damage, and transcription of antioxidant biosynthesis, DNA repair, programmed cell death, pigment metabolism and respiration. The results showed that UVB triggered both irradiance-dependent and target-specific effects at multiple levels of biological organization, whereas exposure to UVA alone did not cause any effects. Inhibition of photosystem II and induction of carotenoids were observed at 0.23 W m-2, whereas growth inhibition, excessive reactive oxygen species, lipid peroxidation, cyclobutane pyrimidine dimer formation, mitochondrial membrane potential reduction and chlorophyll depletion were observed at 0.5-1 W m-2. Relationships between responses at different levels of biological organization were used to establish a putative network of toxicity pathways to improve our understanding of UVB effects in aquatic macrophytes under continuous UVB exposures. Additional studies under natural illuminations were proposed to assess whether these putative toxicity pathways may also be relevant for more ecologically relevant exposure scenarios.

Comparative sensitivity to gamma radiation at the organismal, cell and DNA level in young plants of Norway spruce, Scots pine and Arabidopsis thaliana.

MAIN CONCLUSION: Persistent DNA damage in gamma-exposed Norway spruce, Scots pine and Arabidopsis thaliana, but persistent adverse effects at the organismal and cellular level in the conifers only. Gamma radiation emitted from natural and anthropogenic sources may have strong negative impact on plants, especially at high dose rates. Although previous studies implied different sensitivity among species, information from comparative studies under standardized conditions is scarce. In this study, sensitivity to gamma radiation was compared in young seedlings of the conifers Scots pine and Norway spruce and the herbaceous Arabidopsis thaliana by exposure to 60Co gamma dose rates of 1-540 mGy h-1 for 144 h, as well as 360 h for A. thaliana. Consistent with slightly less prominent shoot apical meristem, in the conifers growth was significantly inhibited with increasing dose rate ≥ 40 mGy h-1. Post-irradiation, the conifers showed dose-rate-dependent inhibition of needle and root development consistent with increasingly disorganized apical meristems with increasing dose rate, visible damage and mortality after exposure to ≥ 40 mGy h-1. Regardless of gamma duration, A. thaliana showed no visible or histological damage or mortality, only delayed lateral root development after ≥ 100 mGy h-1 and slightly, but transiently delayed post-irradiation reproductive development after ≥ 400 mGy h-1. In all species dose-rate-dependent DNA damage occurred following ≥ 1-10 mGy h-1 and was still at a similar level at day 44 post-irradiation. In conclusion, the persistent DNA damage (possible genomic instability) following gamma exposure in all species may suggest that DNA repair is not necessarily mobilized more extensively in A. thaliana than in Norway spruce and Scots pine, and the far higher sensitivity at the organismal and cellular level in the conifers indicates lower tolerance to DNA damage than in A. thaliana.

No evidence of a protective or cumulative negative effect of UV-B on growth inhibition induced by gamma radiation in Scots pine (Pinus sylvestris) seedlings.

Exposure to ambient UV-B radiation may prime protective responses towards various stressors in plants, though information about interactive effects of UV-B and gamma radiation is scarce. Here, we aimed to test whether UV-B exposure could prime acclimatisation mechanisms contributing to tolerance to low-moderate gamma radiation levels in Scots pine seedlings, and concurrently whether simultaneous UV-B and gamma exposure may have an additive adverse effect on seedlings that had previously not encountered either of these stressors. Responses to simultaneous UV-B (0.35 W m-2) and gamma radiation (10.2-125 mGy h-1) for 6 days with or without UV-B pre-exposure (0.35 W m-2, 4 days) were studied across various levels of organisation, as compared to effects of either radiation type. In contrast to UV-B, and regardless of UV-B presence, gamma radiation at ≥42.9 mGy h-1 caused increased formation of reactive oxygen species and reduced shoot length, and reduced root length at 125 mGy h-1. In all experiments there was a gamma dose rate-dependent increase in DNA damage at ≥10.8 mGy h-1, generally with additional UV-B-induced damage. Gamma-induced growth inhibition and gamma- and UV-B-induced DNA damage were still visible 44 days post-irradiation, even at 20.7 mGy h-1, probably due to genomic instability, but this was reversed after 8 months. In conclusion, there was no evidence of a protective effect of UV-B on gamma-induced growth inhibition and DNA damage in Scots pine, and no additive adverse effect of gamma and UV-B radiation on growth in spite of the additional UV-B-induced DNA damage.

Modes of action and adverse effects of gamma radiation in an aquatic macrophyte Lemna minor.

High dose rates of ionizing radiation have been reported to cause adverse effects such as reduction in reproduction and growth, and damage to protein and lipids in primary producers. However, the relevant effects of ionizing radiation are still poorly understood in aquatic plants. This study was intended to characterize the biological effects and modes of action (MoAs) of ionizing radiation using gamma radiation as the prototypical stressor and duckweed Lemna minor as a model organism. Lemna minor was exposed to 1, 14, 24, 46, 70 mGy/h gamma radiation dose rates from a cobalt-60 source for 7 days following the testing principles of the OECD test guideline 221. A suite of bioassays was applied to assess the biological effects of gamma radiation at multiple levels of biological organization, including detection of reactive oxygen species (ROS), oxidative stress responses (total glutathione, tGSH; lipid peroxidation, LPO), DNA damage, mitochondrial dysfunctions (mitochondrial membrane potential, MMP), photosynthetic parameters (chlorophyll a, chl a; chlorophyll b, chl b; carotenoids; Photosystem II (PSII) performance; CO2 uptake), intercellular signaling (Ca2+ release) and growth. Gamma radiation increased DNA damage, tGSH level and Ca2+ content together with reduction in chlorophyll content, maximal PSII efficiency and CO2 uptake at dose rates between 1 and 14 mGy/h, whereas increases in cellular ROS and LPO, inhibition of MMP and growth were observed at higher dose rates (≥24 mGy/h). A network of toxicity pathways was proposed to portray the causal relationships between gamma radiation-induced physiological responses and adverse outcomes to support the development of Adverse Outcome Pathways (AOPs) for ionizing radiation-mediated effects in primary producers.

Daily light integral and day light quality: Potentials and pitfalls of nighttime UV treatments on cucumber powdery mildew.

Nighttime ultraviolet (UV) radiation, if applied properly, has a significant potential for management of powdery mildews in many crop species. In this study, the role of growth light duration, irradiance, a combination of both (daily light integral) and light spectral quality (blue or red) on the efficacy of UV treatments against powdery mildew caused by Podosphaera xanthii and the growth performance of cucumber plants was studied in growth chambers. Increasing daily light integral provided by high-pressure sodium lamps (HPS) decreased efficacy of nighttime UV treatments against P. xanthii, but it increased plant growth. Furthermore, the efficacy of nighttime UV decreased when day length was increased from 16 to 20h at a constant daily light integral. The efficacy of nighttime UV increased if red light was applied after UV treatment, showing the possibility of day length extension without reducing the effect of UV. Increasing the dose of blue light during daytime reduced the efficacy of nighttime UV in controlling the disease, whereas blue deficient growth light (<6% of blue) caused UV mediated curling of young leaves. Furthermore, application of blue light after nighttime UV reduced its disease control efficacy. This showed the importance of maintaining a minimum of blue light in the growth light before nighttime UV treatment. Findings from this study showed that optimization of nighttime UV for management of powdery mildew is dependent on the spectral composition of the photosynthetically active radiation.

Sensitivity of the green algae Chlamydomonas reinhardtii to gamma radiation: Photosynthetic performance and ROS formation.

The aquatic environment is continuously exposed to ionizing radiation from both natural and anthropogenic sources, making the characterization of ecological and health risks associated with radiation of large importance. Microalgae represent the main source of biomass production in the aquatic ecosystem, thus becoming a highly relevant biological model to assess the impacts of gamma radiation. However, little information is available on the effects of gamma radiation on microalgal species, making environmental radioprotection of this group of species challenging. In this context, the present study aimed to improve the understanding of the effects and toxic mechanisms of gamma radiation in the unicellular green algae Chlamydomonas reinhardtii focusing on the activity of the photosynthetic apparatus and ROS formation. Algal cells were exposed to gamma radiation (0.49-1677mGy/h) for 6h and chlorophyll fluorescence parameters obtained by PAM fluorometry, while two fluorescent probes carboxy-H2DFFDA and DHR 123 were used for the quantification of ROS. The alterations seen in functional parameters of C. reinhardtii PSII after 6h of exposure to gamma radiation showed modifications of PSII energy transfer associated with electron transport and energy dissipation pathways, especially at the higher dose rates used. Results also showed that gamma radiation induced ROS in a dose-dependent manner under both light and dark conditions. The observed decrease in photosynthetic efficiency seems to be connected to the formation of ROS and can potentially lead to oxidative stress and cellular damage in chloroplasts. To our knowledge, this is the first report on changes in several chlorophyll fluorescence parameters associated with photosynthetic performance and ROS formation in microalgae after exposure to gamma radiation.

The lichens Xanthoria elegans and Cetraria islandica maintain a high protection against UV-B radiation in Arctic habitats.

This study reports UV screening pigments in the upper cortices of two widespread lichens collected in three sun-exposed locations along a latitudinal gradient from the Arctic lowland to alpine sites of the Central European Alps. Populations from the Alps receive 3-5 times higher UV-B irradiance than their Arctic counterparts from Svalbard because of latitudinal and altitudinal gradients in UV-B irradiance. In Cetraria islandica, the screening capacity of melanin in the upper cortices was assessed by direct measurements of cortical transmittance (250-1,000 nm). A comparison of cortical transmittances in brown sun-exposed and pale shade-adapted forest C. islandica thalli showed that fungal melanins strongly absorb both UV-B and photosynthetically active radiation (PAR). For Xanthoria elegans cortical UV-B absorbing pigments, mainly the orange parietin, were extracted and quantified. Field experiments with extracted, parietin-deficient X. elegans thalli cultivated under various filters showed that UV-B was essential for the induction of parietin synthesis. The parietin resynthesis in these parietin-deficient samples increased with decreasing latitude of their location in which they had been sampled, which may imply that the synthesis of pigments is habitat specific. However, no latitudinal gradient in cortical screening capacity was detected for any of the two species investigated in the field. This implies that Arctic populations maintain a high level of screening pigments in spite of low ambient UV-B, and that the studied lichen species presumably may tolerate an increase in UV-B radiation due to the predicted thinning of the ozone layer over polar areas.