Leaf metabolic influence of glyphosate and nanotubes on the Arabidopsis thaliana phyllosphere.

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.

Acute toxicity of the fungicide azoxystrobin on the diatom Phaeodactylum tricornutum.

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.

Enantioselective Oxidative Stress Induced by S- and Rac-metolachlor in Wheat (Triticum aestivum L.) Seedlings.

The unfounded use of chiral pesticides has caused widespread concern. In this study, the enantioselective effects of S- and racemic (Rac)-metolachlor on the oxidative stress of wheat seedlings was determined based on physiological and gene transcription differences. Growth inhibition increased with increasing concentrations of tested metolachlor, and S-metolachlor had a stronger inhibitory effect than did Rac-metolachlor. Root growth was also significantly inhibited, but no enantioselective effects from the tested concentrations of the metolachlor enantiomers were observed. At a concentration of 5 mg L-1, the maximal fresh weight inhibition reached 63.7% and 53.8% for S-metolachlor and Rac-metolachlor, respectively. In response to the S-metolachlor treatment, the maximum level of superoxide anions and malondialdehyde (MDA) increased to 1.73 and 2.55 times that in response to the control treatment, both of which were greater than those in response to the Rac-metolachlor treatment. The activity of superoxide dismutase (SOD) also increased in response to the S-metolachlor treatment, but the activity of peroxidase (POD) decreased. Real-time polymerase chain reaction (PCR) revealed that, compared with the Rac-metolachlor treatment, the S-metolachlor treatment attenuated the expression of several antioxidant genes. Together, these results demonstrate that S-metolachlor has a greater effect than does Rac-metolachlor on wheat seedlings.

Effects of chiral herbicide dichlorprop on Arabidopsis thaliana metabolic profile and its implications for microbial communities in the phyllosphere.

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.

Enantioselective metabolomic modulations in Arabidopsis thaliana leaf induced by the herbicide dichlorprop.

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.

Synergistic effects of glyphosate and multiwall carbon nanotubes on Arabidopsis thaliana physiology and metabolism.

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.

Offspring toxicity of silver nanoparticles to Arabidopsis thaliana flowering and floral development.

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.

Comparative molecular and metabolic responses of wheat seedlings (Triticum aestivum L.) to the imazethapyr enantiomers S-IM and R-IM.

The enantioselective effects of imazethapyr (IM) enantiomers on wheat seedlings in a hydroponic medium were studied. R-IM at 0.05mg/L exerted a stronger inhibitory effect on shoot weight and root weight than 0.05mg/L S-IM, suggesting that R-IM more severely inhibited growth. Oxidative damage, based on the anthocyanin content, malondialdehyde (MDA) content, antioxidant enzyme activities and transcript levels of antioxidant enzyme genes, were studied together with the cellular ultrastructure of wheat leaves. The anthocyanin and MDA contents in the R-IM treatment group were significantly increased compared with those in the control group, but no significant changes were observed in the S-IM treatment group. The antioxidant enzyme activities of CAT and SOD were inhibited by 0.32- and 0.73-fold, respectively, in the 14day R-IM treatment group compared to those in the control. However, the transcript levels of antioxidant enzyme genes, including CuZnSOD, POD and CAT, were downregulated in the 14day R-IM exposure group, but those of DHAR were not. The number and size of starch granules increased and chloroplast swelling was observed in wheat leaf cells after R-IM exposure, which showed that photosynthetic functions were potentially disturbed. These results directly or indirectly imply that R-IM exposure causes more oxidative stress and exerts a stronger negative effect on wheat than S-IM. A metabolomics approach revealed that the tricarboxylic acid cycle was heavily suppressed by R-IM treatment. Some amino acids (proline, threonine, lysine, valine) were increased by only the R-IM treatment, indicating the activation of antioxidant pathways. The decrease in a series of fatty acids implied that the cell membrane composition changed in response to R-IM. These results provide a deeper understanding of the enantioselective effects of IM enantiomers on the molecular and metabolic responses in wheat seedlings.

The fungicide azoxystrobin promotes freshwater cyanobacterial dominance through altering competition.

BACKGROUND: Sharp increases in food production worldwide are attributable to agricultural intensification aided by heavy use of agrochemicals. This massive use of pesticides and fertilizers in combination with global climate change has led to collateral damage in freshwater systems, notably an increase in the frequency of harmful cyanobacterial blooms (HCBs). The precise mechanisms and magnitude of effects that pesticides exert on HCBs formation and proliferation have received little research attention and are poorly constrained. RESULTS: We found that azoxystrobin (AZ), a common strobilurin fungicide, can favor cyanobacterial growth through growth inhibition of eukaryotic competitors (Chlorophyta) and possibly by inhibiting cyanobacterial parasites (fungi) as well as pathogenic bacteria and viruses. Meta-transcriptomic analyses identified AZ-responsive genes and biochemical pathways in eukaryotic plankton and bacteria, potentially explaining the microbial effects of AZ. CONCLUSIONS: Our study provides novel mechanistic insights into the intertwined effects of a fungicide and eutrophication on microbial planktonic communities and cyanobacterial blooms in a eutrophic freshwater ecosystem. This knowledge may prove useful in mitigating cyanobacteria blooms resulting from agricultural intensification.

Impact of copper nanoparticles and ionic copper exposure on wheat (Triticum aestivum L.) root morphology and antioxidant response.

Copper nanoparticles (nCu) are widely used in industry and in daily life, due to their unique physical, chemical, and biological properties. Few studies have focused on nCu phytotoxicity, especially with regard to toxicity mechanisms in crop plants. The present study examined the effect of 15.6 µM nCu exposure on the root morphology, physiology, and gene transcription levels of wheat (Triticum aestivum L.), a major crop cultivated worldwide. The results obtained were compared with the effects of exposing wheat to an equivalent molar concentration of ionic Cu (Cu2+ released from CuSO4) and to control plants. The relative growth rate of roots decreased to approximately 60% and the formation of lateral roots was stimulated under nCu exposure, possibly due to the enhancement of nitrogen uptake and accumulation of auxin in lateral roots. The expression of four of the genes involved in the positive regulation of cell proliferation and negative regulation of programmed cell death decreased to 50% in the Cu2+ treatment compared to that of the control, while only one gene was down-regulated to about half of the control in nCu treatment. This explained the decreased root cell proliferation and higher extent of induced cell death in Cu2+- than in nCu-exposed plants. The increased methane dicarboxylic aldehyde accumulation (2.17-fold increase compared with the control) and decreased antioxidant enzyme activities (more than 50% decrease compared with the control) observed in the Cu2+ treatment in relation to the nCu treatment indicated higher oxidative stress in Cu2+- than in nCu-exposed plants. Antioxidant (e.g., proline) synthesis was pronouncedly induced by nCu to scavenge excess reactive oxygen species, alleviating phytotoxicity to wheat exposed to this form of Cu. Overall, oxidative stress and root growth inhibition were the main causes of nCu toxicity.