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Plants withstand pathogen attacks by recruiting beneficial bacteria to the rhizosphere and passing their legacy on to the next generation. However, the underlying mechanisms involved in this process remain unclear. In our study, we combined microbiomic and transcriptomic analyses to reveal how the rhizosphere microbiome assembled through multiple generations and defense-related genes expressed in Arabidopsis thaliana under pathogen attack stress. Our results showed that continuous exposure to the pathogen Pseudomonas syringae pv tomato DC3000 led to improved growth and increased disease resistance in a third generation of rps2 mutant Arabidopsis thaliana . It could be attributed to the enrichment of specific rhizosphere bacteria, such as Bacillus and Bacteroides . Pathways associated with plant immunity and growth in A . thaliana , such as MAPK signaling pathways, phytohormone signal transduction, ABC transporter proteins, and flavonoid biosynthesis, were activated under the influence of rhizosphere bacterial communities. Our findings provide a scientific basis for explaining the relationship between beneficial microbes and defense-related gene expression. Understanding microbial communities and the mechanisms involved in plant responses to disease can contribute to better plant management and reduction of pesticide use.

The rhizosphere is one of the key determinants of plant health and productivity. Mixtures of pesticides are commonly used in intensified agriculture. However, the combined mechanisms underlying their impacts on soil microbiota remain unknown. The present study revealed that the rhizosphere microbiota was more sensitive to azoxystrobin and oxytetracycline, two commonly used pesticides, than was the microbiota present in bulk soil. Moreover, the rhizosphere microbiota enhanced network complexity and stability and increased carbohydrate metabolism and xenobiotic biodegradation as well as the expression of metabolic genes involved in defence against pesticide stress. Co-exposure to azoxystrobin and oxytetracycline had antagonistic effects on Arabidopsis thaliana growth and soil microbial variation by recruiting organic-degrading bacteria and regulating ABC transporters to reduce pesticide uptake. Our study explored the composition and function of soil microorganisms through amplicon sequencing and metagenomic approaches, providing comprehensive insights into the synergistic effect of plants and rhizosphere microbiota on pesticides and contributing to our understanding of the ecological risks associated with pesticide use., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Knowledge of the dissemination and emergence of antibiotic resistance genes (ARGs) in the plant rhizosphere is essential for evaluating the risk of the modern ARGs in soil planetary health. However, little is known about the selection mechanism in the plant rhizosphere. Here, we firstly analyzed the dynamic changes in the rhizosphere antibiotic resistome during the process of three passage enrichment of the rhizosphere microbiome in Arabidopsis thaliana (Col-0) and found evidence that plants directionally enriched levels of beneficial functional bacteria with many ARGs. Using the metagenome, we next evaluated the enrichment potential of the resistome in four common crops (barley, indica rice, japonica rice, and wheat) and found that the wheat rhizosphere harbored more abundant ARGs. Therefore, we finally cultivated the rhizosphere microbiome of wheat for three generations and found that approximately 60 % of ARGs were associated with beneficial bacteria enriched in the wheat rhizosphere, which might enter the soil food web and threaten human health, despite also performing beneficial functions in the plant rhizosphere. Our study provides new insights into the dissemination of ARGs in the plant rhizosphere, and the obtained data may be useful for sustainable and ecologically safe agricultural development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

It has been widely reported that probiotic consortia in the rhizosphere can enhance the plant resistance to pathogens. However, the general composition and functional profiles of bacterial community in soils which suppress multiple diseases for various plants remain largely unknown. Here, we combined metadata analysis with machine learning to identify the general patterns of bacterial-community composition in disease-suppressive soils. Disease-suppressive soils significantly enriched Firmicutes and Actinobacteria but showed a decrease in Proteobacteria and Bacteroidetes. Our machine-learning models accurately identified the disease-conducive and -suppressive soils with 54 biomarker genera, 28 of which were potentially beneficial. We further carried out a successive passaging experiment with the susceptible rps2 mutant of Arabidopsis thaliana invaded by Pseudomonas syringae pv. tomato DC3000 (avrRpt2) for functional verification of potential beneficial bacteria. The disease-suppressive ability of Kribbella, Nocardioides and Bacillus was confirmed, and they positively activated the pathogen-associated molecular patterns-triggered immunity pathway. Results also showed that chemical control by pesticides in agricultural production decreased the disease-suppressive ability of soil. This study provides a method for accurately predicting the occurrence of multiple diseases in soil and identified potential beneficial bacteria to guide a wide range of multiple-strain biological control strategies in agricultural management., (© 2022 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Dichlorprop (2-(2,4-dichlorophenoxy) propionic acid, DCPP), a commonly used herbicide for weed control, can be residually detected in soil. It is still unclear whether chiral DCPP exerts an enantioselective adverse effect on plant metabolism and the microbial community of the phyllosphere. In this study, we selected Arabidopsis thaliana as a model plant to explore the effects of R- and S-DCPP enantiomers on plant physiological activities, metabolism, and associated changes in the phyllosphere microbial community. Results indicated that the fresh weight of plants decreased by 37.6% after R-DCPP treatment, whereas it increased by 7.6% after S-DCPP treatment. The R-DCPP enantiomer also caused stronger disturbance to leaf morphology, mesophyll cell structure, and leaf metabolites compared with S-DCPP. GC-MS analysis of DCPP-treated Arabidopsis leaves pointed out a differential profile mostly in carbohydrates, organic acids, and fatty acids, between S-DCPP and R-DCPP treatments. The diversity of phyllospheric microorganisms decreased and the stability of microbial community in the phyllosphere increased after R-DCPP treatment, whereas the opposite result was detected after S-DCPP exposure. The correlation analysis revealed that chiral herbicides may affect microbial communities in the phyllosphere by influencing leaf metabolism, while sugars and terpenoids were considered the main factors in reshaping the microbial community structure in the phyllosphere. Our study provides a new perspective for evaluating the effect of residual DCPP enantiomers on plant physiology and corresponding phyllosphere microorganism changes via the regulation of leaf metabolism, and clarifies the ecological risk of DCPP enantiomer application in agriculture., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Over 40% of herbicides used today are chiral. Dichlorprop (2, 4-DCPP) is a widely used typical broad-spectrum chiral aryloxyphenoxy propionic acid (AOPP) herbicide. However, the molecular mechanism of the enantioselectivity of DCPP enantiomers (S-DCPP and R-DCPP) and their effects on non-target organisms are remain unclear. In the present study, the model plant Arabidopsis thaliana was treated by DCPP enantiomers to directly reveal the effects of DCPP enantiomers on plant growth, as well as metabolic profile. Results showed that the enantioselectivity embodied in that R-DCPP treatment led to the decrease of shoot weight, the significantly variation on morphology of shoot and root, oxidative damage, et al., while the plant morphology also changes to a certain extent associated oxidative damage after treated by S-DCPP. By using metabolomic analysis, it was found that R-DCPP had significant effects on A. thaliana leaf metabolism, including lactose metabolism, starch and sucrose metabolism, TCA cycle, fatty acid biosynthesis pathway and pentose phosphate pathway, and accumulated a lot of antioxidants in plant leaves, while the amino acids and some terpenoids increased in S-DCPP group. Our study provides a new direction to explore the relationship between chiral herbicides on leaf metabolism, and the effect of this relationship on the plant growth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Chemical exposure can indirectly affect leaf microbiota communities, but the mechanism driving this phenomenon remains largely unknown. Results revealed that the co-exposure of glyphosate and multi-carbon nanotubes (CNTs) caused a synergistic inhibitory effect on the growth and metabolism of Arabidopsis thaliana shoots. However, only a slight inhibitory effect was induced by nanotubes or glyphosate alone at the tested concentrations. Several intermediate metabolites of nitrogen metabolism and fatty acid synthesis pathways were upregulated under the combined treatment, which increased the amount of energy required to alleviate the disruption caused by the combined treatment. Additionally, compared with the two individual treatments, the glyphosate/nanotube combination treatment induced greater fluctuations in the phyllosphere bacterial community members with low abundance (relative abundance (RA) <1%) at both the family and genus levels, and among these bacteria some plant growth promotion and nutrient supplement related bacteria were markable increased. Strikingly, strong correlations between phyllosphere bacterial diversity and metabolites suggested a potential role of leaf metabolism, particularly nitrogen and carbohydrate metabolism, in restricting the range of leaf microbial taxa. These correlations between phyllosphere bacterial diversity and leaf metabolism will improve our understanding of plant-microbe interactions and the extent of their drivers of variation and the underlying causes of variability in bacterial community composition., (Copyright © 2021. Published by Elsevier B.V.)

The interaction between plants and microorganisms directly affects plant health and sustainable agricultural development. Leaves represent a wide-area habitat populated by a variety of microorganisms, whose impact on host environmental adaptability could influence plant growth and function. The driving factors for phyllosphere microbiota assemblage are the focus of current research. Here, we investigated the effect of growth stage (i.e., bolting, flowering, and maturation) and genotype of Arabidopsis thaliana (wild-type and the two photosynthetic mutants ndf4 and pgr5) on the composition of phyllosphere microbiota. Our results show that species abundance varied significantly between the three genotypes at different growth stages, whereas species richness and evenness varied only for ndf4. The leaf surface shared a core microbiota dominated by Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes in all tested growth stages and genotypes. Phyllosphere specificity varied more with respect to growth stage than to genotype. In summary, both the growth stage and genotype of A. thaliana are crucial in shaping phyllosphere bacterial composition, with the former being a stronger driver. Our findings provide a novel for investigating whether the host properties influence the phyllosphere community and favor healthy development of plants., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Agricultural chemicals have the potential to become pollutants that adversely affect plant growth. Interactions between these compounds are likely, but potential synergies are under-researched. Multiwall carbon nanotubes are increasingly finding novel uses in agriculture, as delivery mechanisms and as slow-release fertilizers. There is potential for nanotubes to interact with other agricultural chemicals in unpredictable ways. To investigate this possibility, we examined interactions with glyphosate, a widely used herbicide that is also attracting increasing concern over its potential for non-target effects. Here we examined potential synergistic effects on hydroponically grown Arabidopsis thaliana. Single treatments did not affect plant growth significantly, or did only mildly. However, combined treatment significantly affected both plant root and shoot growth. High-level content of malondialdehyde and up-regulated of metabolic antioxidant molecules in plant indicated that combined group caused the strong oxidative damage, while the decreased of antioxidant enzyme activities indicated an imbalance between reactive oxygen species (ROS)and the antioxidant defense system due to the continuously generated ROS. Besides, several intermediate metabolites of unsaturated fatty acids synthesis pathways were up-regulated in combined treatment, which clarified that combined group changed membrane components. The increase of intermediate metabolites in combined group also reflected more energy consumption in the repairment of the disrupt of combined treatment. The synergistic effect observed was attributed to the accumulation of glyphosate resulting from permeability and transportability of the carbon nanotubes. Overall, the risk of nanotube-herbicide interaction suggests a caution use of nanotubes in agricultural applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Imazethapyr (IM) is a widely used acetolactate synthase-inhibiting chiral herbicide. It has long-term residuals that may be absorbed by the human body through the edible parts of plants, such as vegetable leaves or fruits. Here, we selected a model plant, Arabidopsis thaliana, to determine the effects of R-IM and S-IM on its leaf structure, photosynthetic efficiency, and metabolites, as well as the structures of microorganisms in the phyllosphere, after 7 days of exposure. Our results indicated enantiomeric differences in plant growth between R-IM and S-IM; 133 µg/kg R-IM showed heavier inhibition of photosynthetic efficiency and greater changes to subcellular structure than S-IM. R-IM and S-IM also had different effects on metabolism and leaf microorganisms. S-IM mainly increased lipid compounds and decreased amino acids, while R-IM increased sugar accumulation. The relative abundance of Moraxellaceae human pathogenic bacteria was increased by R-IM treatment, indicating that R-IM treatment may increase leaf surface pathogenic bacteria. Our research provides a new perspective for evaluating the harmfulness of pesticide residues in soil, phyllosphere microbiome changes via the regulation of plant metabolism, and induced pathogenic bacterial accumulation risks., Competing Interests:    , (Copyright © 2020. Published by Elsevier B.V.)

Imazethapyr (IM) is a chiral herbicide with two enantiomers (R-IM and S-IM). Here, we determined the enantioselective effects of IM on Arabidopsis thaliana biomass and chlorophyll content, root exudates and rhizosphere microbes after 7 days of exposure. The results suggested that 133 μg/kg R-IM enantiomer in soil slightly inhibited plant biomass but S-IM did not exert significant inhibitory effects. The rhizosphere microorganism composition was also found to have enantiomeric differences between R- and S-IM. The relative abundance of beneficial rhizosphere microbes such as Bacillus and Ramlibacter increased much more with R-IM treatment than with S-IM treatment, indicating that the rhizosphere recruited some beneficial microbes to resist the herbicide stress. The IM enantiomers exerted a significant influence on root exudates with enantioselectivity. R-IM resulted in higher levels of most amino acids, organic acids, sugars and other metabolites after 7 days of exposure; few metabolites were increased by only the S-IM treatment. The correlation analyses between compounds (sugars, amino acids and organic acid) and microbes at the genus level revealed that the number of microbes was more positively correlated with organic acids than other compounds, indicating that organic acids can attract more microbes than amino acids and sugars. Some organic acids, such as 3-hydroxybutyric acid, may be a carbon source for the beneficial microbe Ramlibacter. This study increases the understanding of the differences in IM enantiomer toxicity with respect to plant physiological activity and soil microorganisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Many studies have considered silver nanoparticles (AgNPs) cytotoxicity to mammalian and human cell lines and plant growth. However, only few studies considered toxic effects of AgNPs on plant offspring, especially on flowering. Arabidopsis thaliana was treated with 12.5 mg/kg AgNPs employing parental-(P-AgNPs) and offspring-generation (O-AgNPs) exposure to study the effects of AgNPs on flowering and floral development. Exposure to P-AgNPs was found to significantly decrease petal and pollen viability and subsequently reduced pod production. The inhibition of A. thaliana vegetative growth caused by P-AgNPs exposure was transferred to offspring and even became more severe in the O-AgNPs group. Further, the transcription of genes related to flowering and floral organ development in P-AgNPs and O-Con plants was downregulated by approximately 10-40% compared to the transcription in P-Con plants and showed a stronger decrease in the O-AgNPs group to 30-50% of that in the P-AgNPs group. This resulted in a delay in flowering of 4, 3 and 8 days in P-AgNPs, O-Con and O-AgNPs plants, respectively. Our research shows that the negative effects on floral development can be transferred to the offspring in A. thaliana, which may have significant implications with regard to the risks posed by NPs to food safety and security., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Imazethapyr (IM) is an acetolactate synthase (ALS)-inhibiting herbicide that has been widely used in recent years. However, IM spraying can lead to the accumulation of herbicide residues in leaves. Here, we determined the effects of IM spraying on the plant growth and leaf surface microbial communities of Arabidopsis thaliana after 7 and 14 days of exposure. The results suggested that IM spraying inhibited plant growth. Fresh weight decreased to 48% and 26% of the control value after 7 and 14 days, respectively, of 0.035 kg/ha IM exposure. In addition, anthocyanin content increased 9.2-fold and 37.2-fold relative to the control content after 7 and 14 days of treatment, respectively. Furthermore, IM spraying destroyed the cell structures of the leaves, as evidenced by increases in the number of starch granules and the stomatal closure rate. Reductions in photosynthetic efficiency and antioxidant enzyme activity were observed after IM spraying, especially after 14 days of exposure. The diversity and evenness of the leaf microbiota were not affected by IM treatment, but the composition of community structure at the genus level was altered by IM spraying. Imazethapyr application increased the abundance of Pseudomonas, a genus that includes species pathogenic to plants and humans, indicating that IM potentially increased the abundance of pathogenic bacteria on leaves. Our findings increase our understanding of the relationships between herbicide application and the microbial community structures on plant leaves, and they provide a new perspective for studying the ecological safety of herbicide usage., (Copyright © 2019. Published by Elsevier B.V.)

Background: Plant phenology has crucial biological, physical, and chemical effects on the biosphere. Phenological drivers have largely been studied, but the role of plant microbiota, particularly rhizosphere microbiota, has not been considered., Results: We discovered that rhizosphere microbial communities could modulate the timing of flowering of Arabidopsis thaliana. Rhizosphere microorganisms that increased and prolonged N bioavailability by nitrification delayed flowering by converting tryptophan to the phytohormone indole acetic acid (IAA), thus downregulating genes that trigger flowering, and stimulating further plant growth. The addition of IAA to hydroponic cultures confirmed this metabolic network., Conclusions: We document a novel metabolic network in which soil microbiota influenced plant flowering time, thus shedding light on the key role of soil microbiota on plant functioning. This opens up multiple opportunities for application, from helping to mitigate some of the effects of climate change and environmental stress on plants (e.g. abnormal temperature variation, drought, salinity) to manipulating plant characteristics using microbial inocula to increase crop potential.

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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

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 (psbA, rbcL, pgrl1A and pgrl1B) 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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The widespread application of copper oxide nanoparticles (nCuO) results in ecological risk when nanoparticles enter the environment. This study clarifies the mechanism of nCuO toxicity in Arabidopsis thaliana seedlings via comparison with copper (Cu) ion bioeffects. Under the same culture conditions, Cu 2+ ion exposure exerted a stronger inhibitory effect on plant fresh weight and growth and caused stronger oxidative disruption (measured by malondialdehyde, MDA) than nCuO exposure. The Cu 2+ ions also showed a stronger induction effect than did nCuO on the activity of antioxidant enzymes and the transcription of antioxidant-related genes. Dissolved Cu 2+ ions contributed a minority of the toxicity of nCuO, implying that nCuO itself showed relative strong phytotoxicity. The work presented here will help increase our understanding of the toxicity of metal nanoparticles in plants.

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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

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.)

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.

Both 2-[(2,4-dichlorophenoxy)acetoxy](methy)lmethyl-5,5-dimethyl-1,3,2-dioxaphosphinan-2-one (termed as IIa) and 2-[(4-chloro-2-methyl-phenoxy)-acetoxy](methyl)methyl-5,5-dimethyl-1,3,2-dioxaphosphinan-2-one (termed as IIr) are novel herbicide candidates that positively affect herbicidal activity via the introduction of a phosphorus-containing heterocyclic ring. This report investigated the mechanism of IIa and IIr on weed control in the model plant Arabidopsis thaliana at physiological, ultrastructural and molecular levels. IIa and IIr significantly inhibited the growth of A. thaliana and altered its root structure by inhibiting energy metabolism and lipid or protein biosynthesis. These compounds also significantly affected the absorption of nitrogen and phosphorus by down-regulating the transcripts of nitrate transporter-related genes, ammonium transporter-related genes and phosphorus transporter-related genes., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Understanding how herbicides affect plant reproduction and growth is critical to develop herbicide toxicity model and refine herbicide risk assessment. Although our knowledge of herbicides toxicity mechanisms at the physiological and molecular level in plant vegetative phase has increased substantially in the last decades, few studies have addressed the herbicide toxicity problematic on plant reproduction. Here, we determined the long-term (4-8 weeks) effect of a chiral herbicide, imazethapyr (IM), which has been increasingly used in plant crops, on floral organ development and reproduction in the model plant Arabidopsis thaliana. More specifically, we followed the effect of two IM enantiomers (R- and S-IM) on floral organ structure, seed production, pollen viability and the transcription of key genes involved in anther and pollen development. The results showed that IM strongly inhibited the transcripts of genes regulating A. thaliana tapetum development (DYT1: DYSFUNCTIONAL TAPETUM 1), tapetal differentiation and function (TDF1: TAPETAL DEVELOPMENT AND FUNCTION1), and pollen wall formation and developments (AMS: ABORTED MICROSPORES, MYB103: MYB DOMAIN PROTEIN 103, MS1: MALE STERILITY 1, MS2: MALE STERILITY 2). Since DYT1 positively regulates 33 genes involved in cell-wall modification (such as, TDF1, AMS, MYB103, MS1, MS2) that can catalyze the breakdown of polysaccharides to facilitate anther dehiscence, the consistent decrease in the transcription of these genes after IM exposure should hamper anther opening as observed under scanning electron microscopy. The toxicity of IM on anther opening further lead to a decrease in pollen production and pollen viability. Furthermore, long-term IM exposure increased the number of apurinic/apyrimidinic sites (AP sites) in the DNA of A. thaliana and also altered the DNA of A. thaliana offspring grown in IM-free soils. Toxicity of IM on floral organs development and reproduction was generally higher in the presence of the R-IM enantiomer than of the S-IM enantiomer. This study unraveled several IM toxicity targets and mechanisms at the molecular and structural level linked to the toxicity of IM trace concentrations on A. thaliana reproduction.

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.

Plant growth and development are strongly affected by environmental pollutants, such as herbicides. Widely used herbicides can remain in soil or aquatic systems for long periods of time. Herbicide pollutants have been reported to heavily affect global plant growth and pose a significant challenge to agriculture. However, it is unclear whether herbicides affect plant flowering. Here, we demonstrated that imazethapyr (IM), a chiral herbicide, can enantioselectively promote flowering in Arabidopsis thaliana. We clarified the possible mechanism by which IM promotes flowering and found that the photoperiod pathway may play an important role in propagating the IM stress signal. IM enantiomers decreased the amplitude of core oscillators (CIRCADIAN CLOCK ASSOCIATED 1 and LATE ELONGATED HYPOCOTYL) and utilized the up-regulation of the GIGANTEA-(CONSTANS)-FLOWERING LOCUS T pathway to induce floral gene, APETALA1 over-expression enantioselectively; this treatment ultimately caused early flowering. Our findings provide new insight into the method by which plants control reproductive timing in response to herbicide stress. Flowering time is an important trait in crops and affects the life cycles of pollinator species. The persistence of herbicides in the biosphere will alter plant life cycles and diversity., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Silver nanoparticles (AgNPs) are one of the most widely used nanomaterials, but the mechanism of AgNP toxicity in terrestrial plants is still unclear. We compared the toxic effects of AgNPs and Ag+ on Arabidopsis thaliana at the physiological, ultrastructural and molecular levels. AgNPs did not affect seed germination; however, they showed stronger inhibitory effect on root elongation than Ag+. The results of transmission electron microscopy and metal content analysis showed that AgNPs could be accumulated in leaves. These absorbed AgNPs disrupted the thylakoid membrane structure and decreased chlorophyll content, which can inhibit plant growth. By comparison, a small amount of Ag+ was absorbed by seedlings, and it did not pronouncedly affect chloroplast structure and other metal ion absorption as AgNPs did. Compared with Ag+, AgNPs could alter the transcription of antioxidant and aquaporin genes, indicating that AgNPs changed the balance between the oxidant and antioxidant systems, and also affected the homeostasis of water and other small molecules within the plant body. All the data from physiological, ultrastructural and molecular levels suggest that AgNPs were more toxic than Ag+.

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.

Diclofop-methyl (DM) is a widely used chiral herbicide, which rapidly hydrolyzes to its major metabolite diclofop acid (DC) after application. With a carbon chiral center, DC not only is an important ingredient of herbicidal activity, but also has a long half-life in soil. Studies so far have only considered the activity of racemic DM in target organisms, and the enantioselective toxicity in nontarget plants of DM and DC has yet to be explored. In this study, the enantioselective phytotoxicity of DC mediated by oxidative stress and the key enzyme ACCase in the fatty acid synthesis system on the model plant Arabidopsis thaliana was investigated. Significant differences between the two enantiomers were observed in phytotoxicity including growth inhibition, oxidative damage and alteration of key genes expression of ACCase, with R-DC showing greater toxicity to Arabidopsis thaliana than S-DC. The results of molecular docking showed that there was a stronger affinity between R-DC and the target enzyme carboxyltransferase domain of ACCase, likely leading to the enantioselective phytotoxicity of DC. This study suggested that chirality of both parent compounds and metabolites should be considered to improve our understanding of the environmental fate and risks of chiral pesticides.

Background: The enantiomers of a chiral compound possess different biological activities, and one of the enantiomers usually shows a higher level of toxicity. Therefore, the exploration of the causative mechanism of enantioselective toxicity is regarded as one of primary goals of biological chemistry. Imazethapyr (IM) is an acetolactate synthase (ALS)-inhibiting chiral herbicide that has been widely used in recent years with racemate. We investigated the enantioselectivity between R- and S-IM to form reactive oxygen species (ROS) and to regulate antioxidant gene transcription and enzyme activity., Results: Dramatic differences between the enantiomers were observed: the enantiomer of R-IM powerfully induced ROS formation, yet drastically reduced antioxidant gene transcription and enzyme activity, which led to an oxidative stress. The mechanism by which IM affects carbohydrate metabolism in chloroplasts has long remained a mystery. Here we report evidence that enantioselectivity also exists in starch metabolism. The enantiomer of R-IM resulted in the accumulation of glucose, maltose and sucrose in the cytoplasm or the chloroplast and disturbed carbohydrates utilization., Conclusion: The study suggests that R-IM more strongly retarded plant growth than S-IM not only by acting on ALS, but also by causing an imbalance in the antioxidant system and the disturbance of carbohydrate metabolism with enantioselective manner.