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Three ecotypes of Arabidopsis thaliana, ecotype Columbia (Wild type, Wt) and two mutants (pgr5 and ndf4), were used to evaluate the effects of diuron on photosynthetic activity of A. thaliana. It was found that diuron adversely affected the fresh weight and chlorophyll content of the plants. Chlorophyll fluorescence studies determined that the pgr5 mutant was more sensitive to diuron than Wt and the ndf4 mutant. Gene expression analysis revealed different roles for the two cyclic electron transfer (CET) pathways, NAD(P)H dehydrogenase (NDH) and proton gradient regulation (PGR5) pathways, in the plant after diuron treatment. For example, a gene in the NDH pathway, lhca5, was activated in the low dose (LD) group in the pgr5 mutant, but was down-regulated in the moderate dose (MD) group, along with two other NDH-related genes (ppl2 and ndhH). In the PGR5 pathway, the pgr5 gene was functional under conditions of increased stress (MD group), and was up-regulated to a greater extent in the ndf4 mutant than that in the Wt and pgr5 mutant. Our results suggest that the PGR5 pathway in plants is more important than the NDH pathway during resistance to environmental stress. Deficiencies in the PGR5 pathway could not be counteracted by the NDH pathway, but deficiencies in the NDH pathway could be overcome by stimulating PGR5., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The widely used solvent extractant, tributyl phosphate (TBP), primarily used as a solvent for the conventional processing of nuclear fuel, has come under scrutiny recently due to concerns surrounding potential environmental contamination and toxicity. In this study, we found that, in Phaeodactylum tricornutum, administration of TBP severely inhibited algal cell growth by reducing photosynthetic efficiency and inducing oxidative stress. We further explored the effect of TBP by examining the gene expression of the photosynthetic electron transport chain and its contribution to reactive oxygen species (ROS) burst. Our data revealed that TBP affected both fatty acid content and profile by regulating the transcription of genes related to glycolysis, fatty acid biosynthesis, and β-oxidation. These results demonstrated that TBP did in fact trigger the synthesis of ROS, disrupting the subcellular membrane structure of this aquatic organism. Our study brings new insight into the fundamental mechanism of toxicity exerted by TBP on the marine alga P. tricornutum.

Photosynthesis is a very important metabolic pathway for plant growth and crop yield. This report investigated the effect of the herbicide imazethapyr on photosynthesis in the Arabidopsis thaliana pnsB3 mutant (a defect in the NDH pathway) and pgr5 mutant (a defect in the PGR5 pathway) to determine which cyclic electron transport chain (CET) of the NDH and PGR5 pathways is more important for protecting the photosynthetic system under herbicide stress. The results showed that 20 μg/L imazethapyr markedly inhibited the growth of the three ecotypes of A. thaliana and produced more anthocyanins and reactive oxygen species (ROS), particularly in the pgr5 mutant. The chlorophyll fluorescence results showed that PSII was severely damaged in the pgr5 mutant. Additionally, the CET was significantly stimulated to protect the photosynthetic system from light damage in Wt and the pnsB3 mutant but not the pgr5 mutant. The real-time PCR analysis indicated that imazethapyr treatment considerably decreased the transcript levels of most photosynthesis-related genes in the three treated groups. Several genes in the PGR5 pathway were significantly induced in the pnsB3 mutant, but no genes in the NDH pathway were induced in the pgr5 mutant. The gene transcription analysis showed that the pgr5 mutant cannot compensate for the deficit in the PGR5 pathway by stimulating the NDH pathway, whereas the pnsB3 mutant can compensate for the deficit in the CET cycle by regulating the PGR5 pathway. The iTRAQ analyses also showed that the photosynthesis system, glycolysis, and TCA cycle suffered the most severe damage in the pgr5 mutant. All of these results showed that the PGR5 pathway is more critical for electron transfer around PSI than the NDH pathway to resist herbicide stress.

Atrazine (ATZ) is a commonly used herbicide that has recently come under scrutiny due to potential environmental toxicity and contamination. In this study, we found that the administration of ATZ indeed leads to reduction of photosynthesis and oxidative stress in Phaeodactylum tricornutum at the treated doses higher than 100 μg L(-1) after 48 h. We further explored the effect of ATZ on photosystem II (PSII) and gene expression of electron transport chain. Collectively, our results may suggest that ATZ entered the chloroplasts in alga depending on ATZ's liposolubility and directly attacked on the electron transport chain, especially PSII, contributing to reactive oxygen species (ROS) burst. The increasing ROS could act as signals to induce or disturb the expression of photosynthesis-related genes, resulting in the imbalance of antioxidation and pro-oxidation in the alga.

Effects of sublethal levels of the photosystem-interfering herbicides atrazine (Atr) and methyl viologen (MV) on photosynthetic electron transport were investigated in Arabidopsis thaliana mutants with defects in cyclic electron transfer (CET) activity. Analysis based on chlorophyll fluorescence parameters showed that pgr5 mutant (a defect in the PGR5 pathway) was more sensitive to both Atr and MV than wild type (Wt) and pnsB3 mutant (a defect in the NDH pathway). Real-time PCR (polymerase chain reaction) analysis of transcripts indicated that Wt plants showed marked increases in transcripts in the PRG5 and NDH pathways under treatment with either Atr or MV. In contrast, Atr increased the gene transcripts in CET, but MV decreased them in pnsB3 mutant plants. Atr did not increase the transcripts, while MV down-regulated them in pgr5 mutant. Immunoblot analysis partially supported the changes in the transcripts; that is, the protein levels of PGRL1 and PGR5 were increased in pnsB3 mutant, while no protein level was increased in pgr5 mutant after the herbicide treatment. The present results suggest that cyclic electron transport is very sensitive to photosystem-interference induced by chemicals and that the PGR5 pathway is very critical for regulation. Thus, pgr5 mutants may be useful plants for monitoring photosystem-interfering herbicides.

Imazethapyr (IM) is a chiral herbicide with reported enantioselective biological activities between its enantiomers. This report investigated the effect of enantioselectivity between R- and S-IM in Arabidopsis thaliana on chlorophyll synthesis and photosynthesis. The results suggest that R-IM inhibited the transcription of chlM to a greater extent than S-IM, which reduced chlorophyll synthesis. R-IM also showed a stronger inhibitory effect than S-IM on the transcription of photosynthesis-related genes, affecting linear electron transport and CO(2) fixation. IM stress enantioselectively induced transcriptional upregulation of the ndhH gene, a representative of the NDH complex. In contrast, the expression of pgr5 was downregulated, which demonstrated that IM stress enhanced adenosine 5'-triphosphate (ATP) synthesis by stimulating an NDH-dependent and not ferredoxin (FD)-independent route. This study suggested that R-IM has a greater toxic effect on photosynthesis than S-IM, affecting plant growth through chlorophyll synthesis.

Background: Hexachlorocyclohexane (HCH) has been used for plant protection and sanitation world-widely, and its isomers have been detected in water, soil, and air as well as in vegetation. As a sink for lipophilic pollutants, vegetation is very important for the degradation and fate of organic contamination; however, little was known about their phytotoxicity and mechanisms of toxic effect. In this study, the stereoselective phototoxicity of four isomers (α, β, γ, and δ) of HCHs mediated by independent as well as interconnecting systems of photosynthesis and enzymatic antioxidant defense system in Arabidopsis thaliana were assessed., Principal Findings: Our results revealed that all the HCHs not only stimulated the activities of catalase (CAT) and peroxidase (POD), but also inhibited the activity of superoxide dismutase (SOD). In photosynthesis system, the photosynthetic efficiency of PSI and PSII were all down regulated. Meanwhile, results from both systems showed that δ-HCH was the most toxic one, while α-HCH the least in Arabidopsis thaliana., Conclusions: For the first time, stereoselective effects of different isomers of HCH in plant were demonstrated. And the results suggest that it requires further research to fully elucidate the environmental toxicity and their mechanisms.

When the concentrations of ampicillin (Amp), atrazine (Atr) and cadmium chloride (Cd) reach excessive quantities, they become toxic to aquatic organisms. Due to the acceleration of the industrialization and the intensification of human activities, the incidence and concentrations of these types of pollutants in aquatic systems are increasing. The primary purpose of this study was to evaluate the short-term effects of Amp, Atr and Cd on the physiological indices and gene expression levels in Microcystis aeruginosa. These three pollutants significantly induced antioxidant activity but continuously accelerated the cellular oxidative damage in microalgae, which suggests an imbalance between the oxidant and the antioxidant systems. Amp, Atr and Cd also decreased the transcription of psaB, psbD1 and rbcL; the lowest transcription of these genes was only 38.1, 23.7 and 7% of the control, respectively. These three pollutants affected nitrogen (N) and phosphorous (P) uptake by inhibiting the transcription of N or P absorbing and transporting related genes, and they down regulated the transcription of microcystin-related genes, which caused a decrease of microcystin levels; and the lowest level of microcystin was only 42.4% of the control. Our results suggest that these pollutants may cause pleiotropic effects on algal growth and physiological and biochemical reactions, and they may even affect secondary metabolic processes.

Nonylphenol (NP) is regarded as a kind of persistent organic pollutant which exists ubiquitously in the environment. The objective of this study was to evaluate the effects of NP on Chlorella vulgaris physiological indices and gene transcription. The results showed that NP stress inhibited algal growth in short-term bioassay. NP also decreased chlorophyll content, including chl a, chl b, and total chlorophyll. NP caused oxidant hurt by overproducing reactive oxygen species (ROS), which might destroy the overall membrane system to cause malondialdehyde content increase. NP inhibited photosynthesis-related gene transcription in C. vulgaris after 24 to 48 h exposure. The lowest transcript levels of psaB, psbA, and rbcL in C. vulgaris decreased to only 18.5%, 7%, and 4% of the control, respectively. Taken together, our results demonstrate that NP is toxic to fresh algae growth by affecting the photosynthesis-related genes transcription and overproducing ROS to disrupt cell structure in a short period.

Chlorella vulgaris was tested to assess their toxicities in freshwater contaminated by the metal compounds of copper (Cu) and cadmium (Cd), both singly and combined. Exposure to 0.5 and 1.5 microM Cu or 1.0 and 2.0 microM Cd alone significantly decreased algal growth and chlorophyll content and increased reactive oxygen species (ROS) content. Two-way ANOVA analysis shows that the combination of these two metal compounds decreased cell growth, chlorophyll content and increased ROS content synergistically. The highest algal cell inhibition was 78.55%, the lowest levels of chl a, chl b and total-chl were 10.59%, 33.33% and 17.94% of the control, respectively. The highest increase in ROS was 9.15-fold greater than that of the control when exposed to Cu(1.5)+Cd(2.0). Real-time PCR shows that Cu and Cd reduced the transcript abundance of psbA and rbcL, but without a synergistic interaction, whereas Cu and Cd increased the transcript abundance of psaB synergistically. These results demonstrate that Cu and Cd independently inhibit PSII activity and CO(2) assimilation, but synergistically increase ROS content to disrupt chlorophyll synthesis and inhibit cell growth.

This study investigated the effects of paraquat, a widely used herbicide, on the aquatic unicellular alga Chlorella vulgaris through short-term toxicity tests at the physiological and gene transcriptional levels. Exposure to 0.5 microM paraquat increased the activities of the antioxidant enzymes superoxide dismutase, peroxidase, and catalase to levels 4.93, 3.19, and 3.09 times higher, respectively, than those of the control. Furthermore, exposure to 0.75 microM paraquat increased the activities of these antioxidant enzymes to even higher levels. The decrease in chlorophyll content and the increases in reactive oxygen species (ROS) and malondialdehyde content following exposure to paraquat suggest that the alga was severely damaged and cell growth was greatly inhibited. Real-time PCR showed that paraquat reduced the transcript abundance of psaB and rbcL to 7.09 and 29.83% of the control, respectively. Our results demonstrate that paraquat inhibited electron transport and CO2 assimilation, and also triggered the synthesis of ROS that disrupt cellular structure and inhibit cell growth.

This study investigated the effects of N-phenyl-2-naphthylamine, an effective allelochemical on aquatic unicellular algae Chlorella vulgaris at physiological gene transcription level. Exposure to 2.5 mg L(-1) of N-phenyl-2-naphthylamine increased the activities of the antioxidant enzymes, superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), which were 2.47, 3.24, and 4.27 times higher than that of the control, however, exposure to 4.0 mg L(-1) N-phenyl-2-naphthylamine decreased the activities of these antioxidant enzymes. An increase in malondialdehyde content and a decrease in chlorophyll content following exposure to N-phenyl-2-naphthylamine suggested that the alga was severely damaged and that cell growth was greatly inhibited. Electron microscopy showed that the plasma membrane was detached from the cell wall, the nucleus was condensed, and the structure of chloroplasts was disrupted, in response to N-phenyl-2-naphthylamine exposure. Real-time PCR showed that N-phenyl-2-naphthylamine reduced the transcript abundance of psaB and psbC to 3% and 1% of the control, respectively. These results demonstrated that N-phenyl-2-naphthylamine not only inhibited photosynthesis, but also triggered the synthesis of reactive oxygen species (ROS) to disrupt the subcellular structure of this aquatic organism.

Abstract Azoxystrobin (AZ) is an effective broad-spectrum fungicide. Due to its extensive application, AZ is detectable in aquatic ecosystems and thus influences aquatic organisms. In this study, the acute toxicity (96 h) of AZ at concentrations of 1.0 mg/L and 5.0 mg/L on the diatom Phaeodactylum tricornutum were examined. At the tested concentrations, AZ significantly inhibited P. tricornutum growth and destroyed its cellular structure. Furthermore, the mechanisms of AZ-induced toxicity on P. tricornutum changed as the exposure time extended. Forty-eight hours after exposure, AZ inhibited P. tricornutum growth primarily via inducing oxidative stress, which increased the activity of two main antioxidant enzymes, superoxide dismutase and peroxidase, and inhibited energy metabolism. However, after 96 h of treatment, the decline in the photosynthetic capacity of P. tricornutum demonstrated that the photosystem was the main AZ target. The pigment content and expression levels of genes related to photosynthetic electron transfer reactions were also significantly decreased. The present study describes AZ toxicity in P. tricornutum and is very valuable for assessing the environmental risk of AZ. Graphical abstract fx1 Highlights • A broad-spectrum fungicide, azoxystrobin, is toxic to non-target species. • Acute toxicity (96 h) of azoxystrobin to P. tricornutum was examined. • Azoxystrobin induced oxidative stress and inhibited energy metabolism. • Azoxystrobin lowered the photosynthetic capacity in P. tricornutum. [ABSTRACT FROM AUTHOR]

Linoleic acid (LA), is the product of secondary metabolism secreted from Microcystis aeruginosa , and it exhibits allelopathic activity against eukaryotic algae. However, information about on the mechanisms associated with the inhibition of algal activity by LA is limited. In this study, Chlorella pyrenoidosa was treated with LA (20–120 μg L −1 ) for 4 days, and its growth inhibition and physiological responses were examined for potential toxic mechanisms. The photosynthetic efficiency of C. pyrenoidosa was inhibited by LA treatments, and the F v /F m parameter decreased significantly compared to that of controls; however, the photosynthetic pigment content did not change significantly. Peroxidase activity was enhanced, relieving oxidative damage in algae after LA treatments. However, superoxide dismutase and catalase were suppressed, ultimately leading to the aggravation of lipid peroxidation. Transcriptome-based gene expression analysis revealed that the 120 μg L −1 LA treatment significantly inhibited the transcription of genes related to photosynthesis, carbon metabolism, and amino acid metabolism in C. pyrenoidosa , suggesting that these genes might be key LA targets in C. pyrenoidosa . Moreover, the expression of genes involved in vitamin, lipid, nitrogen cycling, terpenoid, and ascorbate metabolism was also affected, suggesting that LA inhibits algal cell growth through multiple pathways. The identification of LA-responsive genes in C. pyrenoidosa provides new insight into LA stress responses in eukaryotic algae. [ABSTRACT FROM AUTHOR]

Carbon nanotubes can be either toxic or beneficial to plant growth and can also modulate toxicity of organic contaminants through surface sorption. The complex interacting toxic effects of carbon nanotubes and organic contaminants in plants have received little attention in the literature to date. In this study, the toxicity of multiwall carbon nanotubes (MWCNT, 50 mg/L) and paraquat (MV, 0.82 mg/L), separately or in combination, were evaluated at the physiological and the proteomic level in Arabidopsis thaliana for 7–14 days. The results revealed that the exposure to MWCNT had no inhibitory effect on the growth of shoots and leaves. Rather, MWCNT stimulated the relative electron transport rate and the effective photochemical quantum yield of PSII value as compared to the control by around 12% and lateral root production up to nearly 4-fold as compared to the control. The protective effect of MWCNT on MV toxicity on the root surface area could be quantitatively explained by the extent of MV adsorption on MWCNT and was related to stimulation of photosynthesis, antioxidant protection and number and area of lateral roots which in turn helped nutrient assimilation. The influence of MWCNT and MV on photosynthesis and oxidative stress at the physiological level was consistent with the proteomics analysis, with various over-expressed photosynthesis-related proteins (by more than 2 folds) and various under-expressed oxidative stress related proteins (by about 2–3 folds). This study brings new insights into the interactive effects of two xenobiotics (MWCNT and MV) on the physiology of a model plant. [ABSTRACT FROM AUTHOR]

Diclofop-methyl (DM), a common post-emergence herbicide, is frequently used in agricultural production. Silver nanoparticles (AgNPs) are one of the most widely used nanoparticles, and as such, have been detected and monitored in several environmental systems. Here we investigated the interactive effects of DM and AgNPs on the physiological morphology, photosynthesis and antioxidant system of Arabidopsis thaliana . Our results demonstrated that a 1.0 mg/L DM treatment had no significant effect on the fresh weight of plant shoots and the content of total chlorophyll and anthocyanin. However, a 0.5 mg/L AgNPs treatment was found to significantly inhibit plant growth and chlorophyll synthesis, and was found to cause more severe oxidative damage in plants compared to the effects observed in a hydroponic suspension in which DM and AgNPs were jointly present. Meanwhile, the relative transcript levels of photosynthesis related genes ( psb A, rbc L, pgrl1 A and pgrl1 B) in the combined group were found to be slightly increased compared to transcript levels in the AgNPs group, in order to maintain ATP generation at relatively normal levels in order to repair light damage. One explanation for these observed antagonistic effects was that the existence of DM affects the stability of AgNPs and reduced Ag + release from AgNPs in the mixed solution. Thereupon, the Ag + -content was found to decrease in shoots and roots in the combined group by 15.2% and 9.4% respectively, compared to the AgNPs group. The coexistence of herbicides and nanomaterials in aquatic environments or soil systems will continue to exist due to their wide usages. Our current study highlights that the antagonistic effects between DM and AgNPs exerted a positive impact on A. thaliana growth. [ABSTRACT FROM AUTHOR]

Nano-aluminium oxide (nAl 2 O 3 ) is one of the most widely used nanomaterials. However, nAl 2 O 3 toxicity mechanisms and potential beneficial effects on terrestrial plant physiology remain poorly understood. Such knowledge is essential for the development of robust nAl 2 O 3 risk assessment. In this study, we studied the influence of a 10-d exposure to a total selected concentration of 98 μM nAl 2 O 3 or to the equivalent molar concentration of ionic Al (AlCl 3 ) (196 μM) on the model plant Arabidopsis thaliana on the physiology (e.g., growth and photosynthesis, membrane damage) and the transcriptome using a high throughput state-of-the-art technology, RNA-seq. We found no evidence of nAl 2 O 3 toxicity on photosynthesis, growth and lipid peroxidation. Rather the nAl 2 O 3 treatment stimulated root weight and length by 48% and 39%, respectively as well as photosynthesis opening up the door to the use of nAl 2 O 3 in biotechnology and nano agriculture. Transcriptomic analyses indicate that the beneficial effect of nAl 2 O 3 was related to an increase in the transcription of several genes involved in root growth as well as in root nutrient uptake (e.g., up-regulation of the root hair-specific gene family and root development genes, POLARIS protein). By contrast, the ionic Al treatment decreased shoot and root weight of Arabidopsis thaliana by 57.01% and 45.15%, respectively. This toxic effect was coupled to a range of response at the gene transcription level including increase transcription of antioxidant-related genes and transcription of genes involved in plant defense response to pathogens. This work provides an integrated understanding at the molecular and physiological level of the effects of nAl 2 O 3 and ionic Al in Arabidopsis . [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the protective effect of ammonium (NH) on Cr toxicity to the freshwater alga Chlorella vulgaris. We followed an array of cellular functions and biomolecules in C. vulgaris cells exposed to 50 or 100 μM Cr at three different initial NH concentrations (0.5, 3, and 10 mM). The results showed that Cr strongly inhibited cell yield of C. vulgaris, but 10 mM NH could decrease by more than two-fold Cr toxicity on cell yield compared to exposure to 0.5 mM NH. Cr toxicity on gene transcripts and cellular substructure was also much lower at high than at low NH. Our results suggest that this protecting effect of NH on intracellular Cr toxicity could be due to several factors, such as enhance uptake of phosphorus, increase in C and N assimilation efficiency, and increase transcription of photosynthesis-related genes. [ABSTRACT FROM AUTHOR]

Abstract: Algal blooms have been increasing in prevalence all over the world, destroying ecosystems and placing other organisms at risk. Chemical remediation is one of most important methods of controlling algal bloom formation. The effects of copper sulfate, hydrogen peroxide (H2O2) and N-phenyl-2-naphthylamine on photosynthesis-related and microcystin-related gene transcription and physiological changes of Microcystis aeruginosa were analyzed. The results suggest that transcription of psaB, psbD1 and rbcL was inhibited by the three algaecides, which blocked the electron transport chain, significantly enhanced reactive oxygen species (ROS) accumulation and overwhelmed the antioxidant system. The increase in ROS destroyed pigment synthesis and membrane integrity, which inhibited or killed the algal cells. Furthermore, H2O2 treatment down-regulated mcyD transcription, which indicated a decrease in the microcystin level in the cells. Our results demonstrate that H2O2 has the greatest potential as an algaecide because it not only inhibits algae growth but may reduce microcystin synthesis. [ABSTRACT FROM AUTHOR]

Copper nanoparticles (CuNPs) have been used in a broad range of applications. However, they are inevitably released into the marine environment, making it necessary to evaluate their potential effects on marine phytoplankton. In this study, the short-term (96 h) effects of CuNPs and CuSO 4 on Phacodactylum tricornutum growth, photosynthesis, reactive oxygen species production and transcription were assessed. It was found that high concentrations (40 μM) of CuNPs and CuSO 4 significantly inhibited the growth, photosynthesis and induced oxidative stress of P. tricornutum, while lower concentrations caused a hormetic response as indicated by a slight stimulation in algal growth. The high percentage of dissolved Cu (78–100%) in culture medium suggested that the dissolved Cu was the main driver of toxicity during CuNPs treatment. The algal cells upregulated electron transport chain-related genes to produce more energy and restore photosynthesis after 96 h of treatment with CuNPs and CuSO 4 . This study delineates the cellular mechanism behind the toxicity of CuNPs and CuSO 4 on marine diatoms. [ABSTRACT FROM AUTHOR]