Combined action of gamma radiation and exposure to copper ions on Lemna minor L.

PURPOSE: Under natural conditions, the reaction of living organisms to the action of acute gamma radiation depends on other stressors, including heavy metals. The aim of this work was to study changes in morphometric parameters, the content of photoassimilation pigments and the level of oxidative stress in irradiated duckweed at various copper concentrations in the culture medium. MATERIALS AND METHODS: As a model organism, we used Lemna minor L. Duckweed was exposed to acute γ-radiation at doses of 18, 42, 63 Gy. After irradiation, the plants were transferred into a medium containing 3, 5, 6.3 µmol/L Cu. On the 4th day of exposure, the levels of chlorophyll, carotenoids, malondialdehyde (MDA) were measured; after 7 days, the specific growth rate, the level of damage, the change in the frond area, copper concentration in plant tissues were determined. RESULTS: The action of γ-radiation (18, 42, 63 Gy) and copper ions (3, 5, 6.3 µmol/L) reduced the growth rate, increased the membrane lipid peroxidation, reduced the area of the fronds more significantly than under the separate action of the factors. The factors acted antagonistically on the specific growth rate. The content of copper in the tissues of irradiated plants (42, 63 Gy) increased. CONCLUSION: Irradiation of duckweed with acute doses of gamma radiation reduced the resistance of plants to excess copper in the environment.

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.

Active thermography for real time monitoring of UV-B plant interactions.

Although Ultraviolet-B (UV-B)-plant interactions have been extensively analysed in the past years, many physiological aspects of the complex plant response mechanisms still need to be elucidated. Depending on the energy dose, this part of the electromagnetic spectrum can induce detrimental or beneficial effects in plant and fruit. In the present work, active thermography is used to analyse in real time the response of plants under different doses of artificial UV-B. In particular, we investigated the temporal variations of the leaf surface temperature (LST) to UV-B exposure by Long Pulse and Lock-in thermography in Epipremnum aureum and in Arabidopsis plants overexpressing or knockout mutants of UVR8, the known UV-B photoreceptor. In both cases, UV-B irradiation triggers a cooling effect, namely a thermal response characterised by a LST lower respect to the initial value. Lock-in thermography demonstrated that the cooling effect is associated with an immediate mobilization and accumulation of water in the leaves. Also, we demonstrated that thermographic responses change according to the different capability of plants to tolerate high UV-B radiation. Our study highlights new physiological and physical aspects of the plants response to UV-B radiation and, more in general, it opens new opportunities for the use of the thermography as smart tool for real-time monitoring of plant environmental interactions.

Spectral quality of monochromatic LED affects photosynthetic acclimation to high-intensity sunlight of Chrysanthemum and Spathiphyllum.

Vertical farming using light-emitting diode offers potential for the early production phase (few weeks) of young ornamental plants. However, once transferred to the greenhouse, the photosynthetic acclimation of these young plants might depend on this initial light regime. To obtain insight about this acclimatization, Chrysanthemum (sun species) and Spathiphyllum (shade species) were preconditioned in growth chambers for 4 weeks under four light qualities: blue (B), red (R), red/blue (RB, 60% R) and white (W) at 100 µmol m-2 s-1 . Monochromatic light (R and B) limited leaf development of both species, which resulted in a lower leaf mass per area when compared to multispectral light (RB for Chrysanthemum, RB and W for Spathiphyllum). R-developed leaves had a lower photosynthetic efficiency in both species. After the light quality pretreatment, plants were transferred to the greenhouse with high-intensity natural light conditions. On the first day of transfer, R and B preconditioned leaves of both species had an inhibited photosynthesis. After 1 week in natural light condition, rapid light curve parameters of Chrysanthemum leaves that developed under B acclimated to sunlight had a similar level than RB-developed leaves unlike R-leaves. Spathiphyllum leaves showed a decrease in maximum electron transport rate and this was most pronounced for the R pretreatment. After 1 month, R-preconditioned Chrysanthemum had the lowest dry mass, while no effects on the dry weight of Spathiphyllum with respect to the pretreatments were observed. Light quality during preconditioning affected the leaf ability to acclimate to natural high light intensities in greenhouse environment.

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.

Gene coexpression analysis reveals dose-dependent and type-specific networks responding to ionizing radiation in the aquatic model plant Lemna minor using public data.

Ionizing radiations (IRs) are widespread damaging stresses to plant growth and development. However, the regulatory networks underlying the mechanisms of responses to IRs remains poorly understood. Here, a set of publicly available transcriptomic data (conducted by Van Hoeck et al. 2015a), in which Lemna minor plants were exposed to a series of doses of gamma, beta and uranium treatments was used to perform gene coexpression network analysis. Overall, the genes involved in DNA synthesis and chromatin structure, light signalling, photosynthesis, and carbohydrate metabolism were commonly responsive to gamma, beta and uranium treatments. Genes related to anthocyanin accumulation and trichome differentiation were specifically downregulated, andgenes related to nitrogen and phosphate nutrition, cell vesicle transport, mitochondrial electron transport and ATP synthesis were specifically upregulated in response to uranium treatment. While genes involved in DNA damage and repair, RNA processing and RNA binding were specifically downregulated and genes involved in calcium signalling, redox and degradation of carbohydrate metabolism were specifically upregulated responding to gamma radiation. These findings revealed both dose-dependent and typespecific networks responding to different IRs in L. minor, and can be served as a useful resource to better understand the mechanisms of responses to different IRs in other plants.

Influence of light intensity on chloroplast development and pigment accumulation in the wild-type and etiolated mutant plants of Anthurium andraeanum 'Sonate'.

Seedlings of wild-type and etiolate mutant plants of Anthurium andraeanum cultivar 'Sonate' were treated for 15 d with different light intensities (20, 100, and 400 µmol·m-2·s-1) to analyze leaf plastid development and pigment content. Significant changes appeared in treated seedlings, including in leaf color, plastid ultrastructure, chloroplast development gene AaGLK expression, chlorophyll and anthocyanin contents, and protoplast shape. Wild-type and etiolated plants exhibited different plastid structures under the same light condition. The results suggest that light intensity is a crucial environmental factor influencing plastid development and leaf color formation in the A. andraeanum cultivar 'Sonate'.

Leaf wound induced ultraweak photon emission is suppressed under anoxic stress: Observations of Spathiphyllum under aerobic and anaerobic conditions using novel in vivo methodology.

Plants have evolved a variety of means to energetically sense and respond to abiotic and biotic environmental stress. Two typical photochemical signaling responses involve the emission of volatile organic compounds and light. The emission of certain leaf wound volatiles and light are mutually dependent upon oxygen which is subsequently required for the wound-induced lipoxygenase reactions that trigger the formation of fatty acids and hydroperoxides; ultimately leading to photon emission by chlorophyll molecules. A low noise photomultiplier with sensitivity in the visible spectrum (300-720 nm) is used to continuously measure long duration ultraweak photon emission of dark-adapting whole Spathiphyllum leaves (in vivo). Leaves were mechanically wounded after two hours of dark adaptation in aerobic and anaerobic conditions. It was found that (1) nitrogen incubation did not affect the pre-wound basal photocounts; (2) wound induced leaf biophoton emission was significantly suppressed when under anoxic stress; and (3) the aerobic wound induced emission spectra observed was > 650 nm, implicating chlorophyll as the likely emitter. Limitations of the PMT photocathode's radiant sensitivity, however, prevented accurate analysis from 700-720 nm. Further examination of leaf wounding profile photon counts revealed that the pre-wounding basal state (aerobic and anoxic), the anoxic wounding state, and the post-wounding aerobic state statistics all approximate a Poisson distribution. It is additionally observed that aerobic wounding induces two distinct exponential decay events. These observations contribute to the body of plant wound-induced luminescence research and provide a novel methodology to measure this phenomenon in vivo.

Lemna minor plants chronically exposed to ionising radiation: RNA-seq analysis indicates a dose rate dependent shift from acclimation to survival strategies.

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 7days in a dose-rate dependent manner. In this study responses on molecular (gene expression) and physiological (photosynthetic) level are evaluated in L. minor plants exposed to ionising radiation. To enable this, we examined the gene expression profiles of irradiated L. minor plants by using an RNA-seq approach. The gene expression data reveal indications that L. minor 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 L. minor plants into survival strategies. Notably, the photosynthetic process seems to be unaffected in L. minor 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.

Silver nanoparticles induced reactive oxygen species via photosynthetic energy transport imbalance in an aquatic plant.

The rapid growth in silver nanoparticles (AgNPs) commercialization has increased environmental exposure, including aquatic ecosystem. It has been reported that the AgNPs have damaging effects on photosynthesis and induce oxidative stress, but the toxic mechanism of AgNPs is still a matter of debate. In the present study, on the model aquatic higher plant Spirodela polyrhiza, we found that AgNPs affect photosynthesis and significantly inhibit Photosystem II (PSII) maximum quantum yield (Fv/Fm) and effective quantum yield (ΦPSII). The changes of non-photochemical fluorescence quenching (NPQ), light-induced non-photochemical fluorescence quenching [Y(NPQ)] and non-light-induced non-photochemical fluorescence quenching [Y(NO)] showed that AgNPs inhibit the photo-protective capacity of PSII. AgNPs induce reactive oxygen species (ROS) that are mainly produced in the chloroplast. The activity of ribulose-1, 5-bisphosphate carboxylase-oxygenase (Rubisco) was also very sensitive to AgNPs. The internalized Ag, regardless of whether the exposure was Ag+ or AgNPs had the same capacity to generate ROS. Our results support the hypothesis that intra-cellular AgNP dissociate into high toxic Ag+. Rubisco inhibition leads to slowing down of CO2 assimilation. Consequently, the solar energy consumption decreases and then the excess excitation energy promotes ROS generation in chloroplast.

Recovery of Lemna minor after exposure to sulfadimethoxine irradiated and non-irradiated in a solar simulator.

Sulfonamides are the second most widely used group of veterinary antibiotics which are often detected in the environment. They are eliminated from freshwaters mainly through photochemical degradation. The toxicity of sulfadimethoxine (SDM) was evaluated with the use of Lemna minor before and after 1- and 4-h irradiation in a SunTest CPS+ solar simulator. Eight endpoints consisting of: number and total area of fronds, fresh weight, chlorophylls a and b, carotenoids, activity of catalase and guaiacol peroxidase, and protein content were determined. The total frond area and chlorophyll b content were the most sensitive endpoints with EC50 of 478 and 554 µg  L-1, respectively. The activity of guaiacol peroxidase and catalase increased at SDM concentrations higher than 125 and 500 µg  L-1, respectively. The SDM photodegradation rate for first order kinetics and the half-life were 0.259 h-1 and 2.67  h, respectively. The results show that the toxicity of irradiated solutions was caused by SDM only, and the photoproducts appeared to be either non-toxic or much less toxic to L. minor than the parent compound. To study the recovery potential of L. minor, after 7 days exposure in SDM solutions, the plants were transferred to fresh medium and incubated for the next 7 days. L. minor has the ability to regenerate, but a 7-day recovery phase is not sufficient for it to return to an optimal physiological state.

A mathematical model of non-photochemical quenching to study short-term light memory in plants.

Plants are permanently exposed to rapidly changing environments, therefore it is evident that they had to evolve mechanisms enabling them to dynamically adapt to such fluctuations. Here we study how plants can be trained to enhance their photoprotection and elaborate on the concept of the short-term illumination memory in Arabidopsis thaliana. By monitoring fluorescence emission dynamics we systematically observe the extent of non-photochemical quenching (NPQ) after previous light exposure to recognise and quantify the memory effect. We propose a simplified mathematical model of photosynthesis that includes the key components required for NPQ activation, which allows us to quantify the contribution to photoprotection by those components. Due to its reduced complexity, our model can be easily applied to study similar behavioural changes in other species, which we demonstrate by adapting it to the shadow-tolerant plant Epipremnum aureum. Our results indicate that a basic mechanism of short-term light memory is preserved. The slow component, accumulation of zeaxanthin, accounts for the amount of memory remaining after relaxation in darkness, while the fast one, antenna protonation, increases quenching efficiency. With our combined theoretical and experimental approach we provide a unifying framework describing common principles of key photoprotective mechanisms across species in general, mathematical terms.

The different response mechanisms of Wolffia globosa: Light-induced silver nanoparticle toxicity.

Silver nanoparticles (AgNPs) have emerged as a promising bactericide. Plants are a major point of entry of contaminants into trophic chains. Here, the physiological responses of Wolffia globosa to AgNPs have been probed using different light schemes, and these data may reveal new insights into the toxic mechanism of AgNPs. W. globosa was grown in culture medium and treated with different concentrations of AgNPs for 24h under pre- and post-illuminated conditions. However, fluorescence quenching, the accumulation of sugar and the reduction of Hill reaction activity were found in response to the AgNP-stresses. In the pre-illuminated condition, oxidative damage was obvious, as indicated by the higher malondialdehyde (MDA) content and an up-regulation of superoxide dismutase (SOD) activity. The maximum increases of MDA content and SOD activity were 1.14 and 2.52 times the respective controls when exposed to 10mg/L AgNPs. In contrast, in the post-illuminated condition, the alterations in photosynthetic pigment and soluble proteins content were more significant than the alterations in oxidative stress. The contents of chlorophyll a, carotenoids and soluble protein decreased to 77.7%, 66.2% and 72.9% of the controls after treatment with the highest concentration of AgNPs (10mg/L). Based on the different physiological responses, we speculated that in the pre-illuminated condition, oxidative stress was responsible for the decline in the oxygen evolution rate, while in the post-illuminated condition, the decrease in the Hill reaction activity could be attributed to the blocking of electron transfer and an insufficient proton supply. Our findings demonstrate that environmental factors regulate the physiological responses of plants to AgNPs through distinct mechanisms.

Influence of nutrient medium composition on uranium toxicity and choice of the most sensitive growth related endpoint in Lemna minor.

Uranium (U) toxicity is known to be highly dependent on U speciation and bioavailability. To assess the impact of uranium on plants, a growth inhibition test was set up in the freshwater macrophyte Lemna minor. First growth media with different compositions were tested in order to find a medium fit for testing U toxicity in L. minor. Following arguments were used for medium selection: the ability to sustain L. minor growth, a high solubility of U in the medium and a high percentage of the more toxic U-species namely UO2(2+). Based on these selection criteria a with a low phosphate concentration of 0.5 mg L(-1) and supplemented with 5 mM MES (2-(N-morpholino)ethanesulfonic acid) to ensure pH stability was chosen. This medium also showed highest U toxicity compared to the other tested media. Subsequently a full dose response curve for U was established by exposing L. minor plants to U concentrations ranging from 0.05 µM up to 150 µM for 7 days. Uranium was shown to adversely affect growth of L. minor in a dose dependent manner with EC10, EC30 and EC50 values ranging between 1.6 and 4.8 µM, 7.7-16.4 µM and 19.4-37.2 µM U, respectively, depending on the growth endpoint. Four different growth related endpoints were tested: frond area, frond number, fresh weight and dry weight. Although differences in relative growth rates and associated ECx-values calculated on different endpoints are small (maximal twofold difference), frond area is recommended to be used to measure U-induced growth effects as it is a sensitive growth endpoint and easy to measure in vivo allowing for measurements over time.

Characterizing dose response relationships: Chronic gamma radiation in Lemna minor induces oxidative stress and altered polyploidy level.

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.

ß-Radiation Stress Responses on Growth and Antioxidative Defense System in Plants: A Study with Strontium-90 in Lemna minor.

In the following study, dose dependent effects on growth and oxidative stress induced by ß-radiation were examined to gain better insights in the mode of action of ß-radiation induced stress in plant species. Radiostrontium (9°Sr) was used to test for ß-radiation induced responses in the freshwater macrophyte Lemna minor. The accumulation pattern of 90Sr 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 ß-radiation dose rates. Exposing L. minor plants for seven days to a 9°Sr activity concentration of 25 up to 25,000 kBq·L⁻¹ resulted in a dose rate between 0.084 ± 0.004 and 97 ± 8 mGy·h⁻¹. After seven days of exposure, root fresh weight showed a dose dependent decrease starting from a dose rate of 9.4 ± 0.5 mGy·h⁻¹. Based on these data, an EDR10 value of 1.5 ± 0.4 mGy·h⁻¹ was estimated for root fresh weight and 52 ± 17 mGy·h⁻¹ for frond fresh weight. Different antioxidative enzymes and metabolites were further examined to analyze if ß-radiation induces oxidative stress in L. minor.

Chloroplast thylakoid structure in evergreen leaves employing strong thermal energy dissipation.

In nature, photosynthetic organisms cope with highly variable light environments--intensities varying over orders of magnitudes as well as rapid fluctuations over seconds-to-minutes--by alternating between (a) highly effective absorption and photochemical conversion of light levels limiting to photosynthesis and (b) powerful photoprotective thermal dissipation of potentially damaging light levels exceeding those that can be utilized in photosynthesis. Adjustments of the photosynthetic apparatus to changes in light environment involve biophysical, biochemical, and structural adjustments. We used electron micrographs to assess overall thylakoid grana structure in evergreen species that exhibit much stronger maximal levels of thermal energy dissipation than the more commonly studied annual species. Our findings indicate an association between partial or complete unstacking of thylakoid grana structure and strong reversible thermal energy dissipation that, in contrast to what has been reported for annual species with much lower maximal levels of energy dissipation, is similar to what is seen under photoinhibitory conditions. For a tropical evergreen with tall grana stacks, a loosening, or vertical unstacking, of grana was seen in sun-grown plants exhibiting pronounced pH-dependent, rapidly reversible thermal energy dissipation as well as for sudden low-to-high-light transfer of shade-grown plants that responded with photoinhibition, characterized by strong dark-sustained, pH-independent thermal energy dissipation and photosystem II (PSII) inactivation. On the other hand, full-sun exposed subalpine confers with rather short grana stacks transitioned from autumn to winter via conversion of most thylakoids from granal to stromal lamellae concomitant with photoinhibitory photosynthetic inactivation and sustained thermal energy dissipation. We propose that these two types of changes (partial or complete unstacking of grana) in thylakoid arrangement are both associated with the strong non-photochemical quenching (NPQ) of chlorophyll fluorescence (a measure of photoprotective thermal energy dissipation) unique to evergreen species rather than with PSII inactivation per se.

The influence of light intensity and photoperiod on duckweed biomass and starch accumulation for bioethanol production.

Duckweed has been considered as a valuable feedstock for bioethanol production due to its high biomass and starch production. To investigate the effects of light conditions on duckweed biomass and starch production, Lemna aequinoctialis 6000 was cultivated at different photoperiods (12:12, 16:8 and 24:0h) and light intensities (20, 50, 80, 110, 200 and 400µmolm(-2)s(-1)). The results showed that the duckweed biomass and starch production was increased with increasing light intensity and photoperiod except at 200 and 400µmolm(-2)s(-1). Considering the light cost, 110µmolm(-2)s(-1) was optimum light condition for starch accumulation with the highest maximum growth rate, biomass and starch production of 8.90gm(-2)day(-1), 233.25gm(-2) and 98.70gm(-2), respectively. Moreover, the results suggested that high light induction was a promising method for duckweed starch accumulation. This study provides optimized light conditions for future industrial large-scale duckweed cultivation.

Pb-induced avoidance-like chloroplast movements in fronds of Lemna trisulca L.

Lead ions are particularly dangerous to the photosynthetic apparatus, but little is known about the effects of trace metals, including Pb, on regulation of chloroplast redistribution. In this study a new effect of lead on chloroplast distribution patterns and movements was demonstrated in mesophyll cells of a small-sized aquatic angiosperm Lemna trisulca L. (star duckweed). An analysis of confocal microscopy images of L. trisulca fronds treated with lead (15 µM Pb2+, 24 h) in darkness or in weak white light revealed an enhanced accumulation of chloroplasts in the profile position along the anticlinal cell walls, in comparison to untreated plants. The rearrangement of chloroplasts in their response to lead ions in darkness was similar to the avoidance response of chloroplasts in plants treated with strong white light. Transmission electron microscopy X-ray microanalysis showed that intracellular chloroplast arrangement was independent of the location of Pb deposits, suggesting that lead causes redistribution of chloroplasts, which looks like a light-induced avoidance response, but is not a real avoidance response to the metal. Furthermore, a similar redistribution of chloroplasts in L. trisulca cells in darkness was observed also under the influence of exogenously applied hydrogen peroxide (H2O2). In addition, we detected an enhanced accumulation of endogenous H2O2 after treatment of plants with lead. Interestingly, H2O2-specific scavenger catalase partly abolished the Pb-induced chloroplast response. These results suggest that H2O2 can be involved in the avoidance-like movement of chloroplasts induced by lead. Analysis of photometric measurements revealed also strong inhibition (but not complete) of blue-light-induced chloroplast movements by lead. This inhibition may result from disturbances in the actin cytoskeleton, as we observed fragmentation and disappearance of actin filaments around chloroplasts. Results of this study show that the mechanisms of the toxic effect of lead on chloroplasts can include disturbances in their movement and distribution pattern.

Microbial community and removal of nitrogen via the addition of a carrier in a pilot-scale duckweed-based wastewater treatment system.

Carriers were added to a pilot-scale duckweed-based (Lemna japonica 0223) wastewater treatment system to immobilize and enhance microorganisms. This system and another parallel duckweed system without carriers were operated for 1.5 years. The results indicated the addition of the carrier did not significantly affect the growth and composition of duckweed, the recovery of total nitrogen (TN), total phosphorus (TP) and CO2 or the removal of TP. However, it significantly improved the removal efficiency of TN and NH4(+)-N (by 19.97% and 15.02%, respectively). The use of 454 pyrosequencing revealed large differences of the microbial communities between the different components within a system and similarities within the same components between the two systems. The carrier biofilm had the highest bacterial diversity and relative abundance of nitrifying bacteria (3%) and denitrifying bacteria (24% of Rhodocyclaceae), which improved nitrogen removal of the system. An efficient N-removal duckweed system with enhanced microorganisms was established.