International audience | Macrophytes are known to bioaccumulate metals, but a thorough understanding of tolerance strategies and molecular impact of metals in aquatic plants is still lacking. The present study aimed to compare Hg and Cd effects in a representative macrophyte, Elodea nuttallii using physiological endpoints and metabolite profiles in shoots and cytosol. Exposure 24 h to methyl-Hg (30 ng L), inorganic Hg (70 ng L) and Cd (280 μg L) did not affect photosynthesis, or antioxidant enzymes despite the significant accumulation of metals, confirming a sublethal stress level. In shoots, Cd resulted in a higher level of regulation of metabolites than MeHg, while MeHg resulted in the largest number of regulated metabolites and IHg treatment regulated no metabolites significantly. In cytosol, Cd regulated more metabolites than IHg and only arginine, histidine and mannose were reduced by MeHg exposure. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of data suggested that exposure to MeHg resulted in biochemical changes including aminoacyl-tRNA biosynthesis, glycine, serine and threonine metabolism, nitrogen metabolism, arginine and proline metabolism, cyanoamino acid metabolism, while the treatment of Cd stress caused significant variations in aminoacyl-tRNA biosynthesis and branched-chain amino acids pathways. Data supports an impact of MeHg on N homeostasis, while Cd resulted in an osmotic stress-like pattern and IHg had a low impact. Marked differences in the responses to MeHg and IHg exposure were evidenced, supporting different molecular toxicity pathways and main impact of MeHg on non-soluble compartment, while main impact of IHg was on soluble compartment. Metabolomics was used for the first time in this species and proved to be very useful to confirm and complement recent knowledge gained by transcriptomics and proteomics, highlighting the high interest of multi-omics approaches to identify early impact of environmental pollution.

Arsenic (As) is a highly toxic metalloid adversely affecting the environment, human health, and crop productivity. The present study assessed the synergistic effects of salinity and As on photosynthetic attributes, stomatal regulations, and metabolomics responses of the xero-halophyte Salvadora persica to decipher the As-salinity cross-tolerance mechanisms and to identify the potential metabolites/metabolic pathways involved in cross-tolerance of As with salinity. Salinity and As stress-induced significant stomatal closure in S. persica suggests an adaptive response to decrease water loss through transpiration. NaCl supplementation improved the net photosynthetic rate (by +39%), stomatal conductance (by +190%), water use efficiency (by +55%), photochemical quenching (by +37%), and electron transfer rate (54%) under As stress as compared to solitary As treatment. Our results imply that both stomatal and non-stomatal factors account for a reduction in photosynthesis under high salinity and As stress conditions. A total of 64 metabolites were identified in S. persica under salinity and/or As stress, and up-regulation of various metabolites support early As-salinity stress tolerance in S. persica by improving antioxidative defense and ROS detoxification. The primary metabolites such as polyphenols (caffeic acid, catechin, gallic acid, coumaric acid, rosmarinic acid, and cinnamic acid), amino acids (glutamic acid, cysteine, glycine, lysine, phenylalanine, and tyrosine), citrate cycle intermediates (malic acid, oxalic acid, and α-ketoglutaric acid), and most of the phytohormones accumulated at higher levels under combined treatment of As + NaCl compared to solitary treatment of As. Moreover, exogenous salinity increased glutamate, glycine, and cysteine, which may induce higher synthesis of GSH-PCs in S. persica. The metabolic pathways that were significantly affected in response to salinity and/or As include inositol phosphate metabolism, citrate cycle, glyoxylate and dicarboxylate metabolism, amino acid metabolism, and glutathione metabolism. Our findings indicate that inflections of various metabolites and metabolic pathways facilitate S. persica to withstand and grow optimally even under high salinity and As conditions. Moreover, the addition of salt enhanced the arsenic tolerance proficiency of this halophyte.

Carbendazim (CBZ), a broad-spectrum pesticide frequently detected in fruits and vegetables, could trigger potential toxic risks to mammals. To facilitate the assessment of health risks, this study aimed to characterize the cytochrome P450 (CYPs)-mediated metabolism profiles of CBZ by a combined experimental and computational study. Our results demonstrated that CYPs-mediated region-selective hydroxylation was a major metabolism pathway for CBZ in liver microsomes from various species including rat, mouse, minipig, dog, rabbit, guinea pig, monkey, cow and human, and the metabolite was biosynthesized and well-characterized as 6-OH-CBZ. CYP1A displayed a predominant role in the region-selective hydroxylation of CBZ that could attenuate its toxicity through converting it into a less toxic metabolite. Meanwhile, five other common pesticides including chlorpyrifos-methyl, prochloraz, chlorfenapyr, chlorpyrifos, and chlorothalonil could significantly inhibit the region-selective hydroxylation of CBZ, and consequently remarkably increased CBZ exposure in vivo. Furthermore, computational study clarified the important contribution of the key amino acid residues Ser122, and Asp313 in CYP1A1, as well as Asp320 in CYP1A2 to the hydroxylation of CBZ through hydrogen bonds. These results would provide some useful information for the metabolic profiles of CBZ by mammalian CYPs, and shed new insights into CYP1A-mediated metabolic detoxification of CBZ and its health risk assessment.

It is highly likely that the toxicity of water accommodated fractions (WAF) will influence marine microalgae, and consequently lead to potential risk for the marine ecological environment. However, it was often neglected whether WAF can influence the transformation of relative compounds in organisms. The metabolism of amino acids (AAs) can be used to track physiological changes in microalgae because amino acids are the basis of proteins and enzymes. In this study, using marine Chlorophyta Platymonas helgolandica as the test organism, the effects of different concentrations of WAF on AA compositions and stable carbon isotope ratios (δ¹³C) of individual AAs of Platymonas helgolandica were investigated. The results showed that the WAF of #180 fuel oil had an obvious suppressing effect on the growth and chlorophyll a content of microalgae. The growth inhibitory rate at 96 h was 80.66% at a WAF concentration of 0.50 mg L⁻¹ compared with the control. Furthermore, seven among the 16 AAs, including alanine, cysteine, proline, aspartic acid, lysine, histidine and tyrosine, had relatively high abundance. Under the glycolysis pathway, the cysteine abundance was higher than control, meaning that the biosynthesized pathway of alanine through cysteine as a precursor could be damaged. Phosphoenolpyruvate (PEP) was an important synthesis precursor of alanine (leucine) and aromatic AA family (Phenylalanine and tyrosine), and played an important role in δ¹³CAAₛ fractionation under the WAF stress. Under the TCA pathway, to protect cell metabolism activities under WAF stress, the δ¹³C value of threonine and proline abundance in microalgae with the increase in WAF stress. Therefore, δ¹³CAAₛ fractionation can be used as a novel method for toxicity evaluation of WAF on future.

The accumulation of cadmium (Cd) in rice grains is closely associated with the content of mineral nutrients and amino acid metabolism, but the causal link among them is unclear. Profiles of amino acids (AAs) and quantities of essential nutrients in grains from early and late rice cultivars grown at four sites with different Cd levels were analyzed in the present study. Hazard quotients (HQs) for consumers by intake of rice from late cultivars were much higher than that from early cultivars at sites with soil Cd content of 0.25, 0.61 and 0.84 mg kg⁻¹. Cadmium accumulation in grains resulted in a sharp reduction of total essential AAs and non-essential AAs in both early and late rice cultivars. High-Cd-accumulating (HCA) cultivars had significantly higher level of glutamate (Glu) than low-Cd-accumulating (LCA) cultivars when rice Cd content was less than 0.20 mg kg⁻¹. However, Glu level in grains dramatically declined with the accumulation of Cd, which subsequently leaded to the reduction of other AAs. Cadmium content was well predicted by five amino acids (i.e., Glu, Alanine, Phenylalanine, Glycine and Threonine) or four essential elements (Ca, Fe, Mn and Zn) when rice Cd was less than 0.80 mg kg⁻¹. Amino acids played more important roles than nutrients in Cd accumulation. When Cd content was in the range of 0.40–1.16 mg kg⁻¹, the Mn content in rice increased significantly with the increase of Cd content, while the Glu content dropped down synchronously. Remarkably, the ratio between Mn and Glu displayed the highest direct path coefficient on Cd accumulation than any single cation or amino acid. These results indicate that high capacity in synthesizing Glu and concentrating Mn is the determinant factor for Cd accumulation in rice grains, and abundant Glu in aleurone layer may alleviate Cd toxicity by forming Glu-Cd complex.

Technologies for cleaner production of rice in cadmium (Cd) contaminated field are being explored worldwide. In order to investigate the inhibition mechanism of phosphate on Cd transport in soil-plant system, controlled experiments were performed in this study. Experimental results showed that Cd levels in roots, flag leaves, rachises and grains of rice plants (Oryza sativa L.) were significantly reduced by supplement of 0.5–2.5 g kg⁻¹ calcium magnesium phosphate fertilizer (CMP). Path coefficient analysis revealed that phosphorous had significant negative direct effect on Cd, but positive indirect effect on essential and non-essential amino acids. Applying 2.5 g kg⁻¹ CMP made the Cd concentration decreased by 45.7% while free essential and non-essential amino acids increased by 28.0–28.6% in grains. Levels of the branched-chain amino acids in grains were much higher than other essential amino acids, and increased with the amount of CMP fertilization. After application of CMP, pH of soil solution and thickness of the iron plaque around roots increased significantly. Spectra from X-ray photoelectron spectrometer (XPS) showed that content of N, P and Fe increased apparently, C, O and Ca had no change, while S decreased by 74.2% in roots after application of 2.5 g kg⁻¹ CMP. Meanwhile, Cd concentration in protoplasts of root cells decreased by 39.5–80.1% with the increase of CMP. These results indicate that application of CMP can effectively inhibit Cd accumulation in root protoplasts by promoting iron plaque formation on the root surface, reduce Cd concentration and increase free amino acids in rice grains.

Different pH conditions have been demonstrated to affect the activities of dechlorinating populations participating in the successive dechlorination of trichloroethylene to ethylene. However, the mechanism of the effect of pH conditions on the assembly of dechlorinating populations and their relations to the structure, function, and dynamics of the microbiome are unclear. In this study, we evaluated the effects of pH on microbiomes assembled in anaerobic trichloroethylene-dechlorinating reactors under neutral (pH 7.2), acidic (pH 6.2), and alkaline (pH 8.2) conditions. The results revealed that among the reactors, the acidic reactor had the highest efficiency for dechlorination without accumulation of dechlorinated metabolites, even at high loading rates. The results of high-throughput sequencing of the 16S rRNA gene indicated that the microbiomes in the 3 reactors underwent varied dynamic succession. The acidic reactor harbored a higher degree of complex microbes, dechlorinator diversity, and abundance of the Victoria subgroup of Dehalococcoides (1.2 ± 0.1 × 10⁶ cell/mL), which were approximately 10–10²-fold higher than those at neutral and alkaline conditions. The pH settings altered species–species connectivity and complexity of microbial interaction networks, with more commensal interactions in the dechlorinators of the acidic reactor. As predicted, abundances of several functional gene categories were in strong linearity with pH values, and the microbiome possessed significantly more abundant functions in the acidic reactor (P < 0.001), such as potentially stimulating hydrogen production, cobalamin synthesis, cobalt transport, transport and metabolism of amino acids and secondary metabolites, cell motility, and transcription. All results of microbiomic analyses consistently revealed the observed superior dechlorination process and suggested an association of the reductive dechlorination process with the pH-dependent microbiome. The results of this study provide a new insight into the trichloroethylene dechlorination with regards to pH, and they will be useful for improving bioremediation and management of trichloroethylene-contaminated sites.

Triclosan is one of the most frequently detected emerging contaminants in aquatic environment. In this study, we investigated the biouptake, toxicity and biotransformation of triclosan in freshwater algae Cymbella sp. The influence of humic acid, as a representative of dissolved organic matter, was also explored. Results from this study showed that triclosan was toxic to Cymbella sp. with 72 h EC₅₀ of 324.9 μg L⁻¹. Humic acid significantly reduced the toxicity and accumulation of triclosan in Cymbella sp. SEM analysis showed that Cymbella sp. were enormously damaged under 1 mg L⁻¹ triclosan exposure and repaired after the addition of 20 mg L⁻¹ humic acid. Triclosan can be significantly taken up by Cymbella sp. The toxicity of triclosan is related to bioaccumulated triclosan as the algal cell numbers decreased when intracellular triclosan increased. A total of 11 metabolites were identified in diatom cells and degradation pathways are proposed. Hydroxylation, methylation, dechlorination, amino acids conjunction and glucuronidation contributed to the transformative reactions of triclosan in Cymbella sp., producing biologically active products (e.g., methyl triclosan) and conjugation products (e.g., glucuronide or oxaloacetic acid conjugated triclosan), which may be included in the detoxification mechanism of triclosan.

The ecological toxicity of hexabromocyclododecane (HBCD) on animals, including fish and mice, has been reported, but its effects in plants, particularly its toxic mechanism, have rarely been investigated. An untargeted metabolomics approach for comprehensive assessment was selected to study the alterations in the metabolic profiles in pak choi leaves induced by exposure to trace-level amounts of HBCD diastereomers over 30 days. A supervised orthogonal partial least-squares-discriminant analysis (OPLS-DA) was performed to investigate differences between the HBCD and control groups. The discriminating metabolites were identified using public databases. The results indicated that the toxicity of the HBCD diastereomers was ordered as γ-HBCD > α-HBCD > β-HBCD. 13 metabolites were identified as potential biomarkers to discriminate the presence of HBCD toxicity. The lipid, carbohydrate, nucleotide and amino acid metabolic pathways affected were found in accordance with animals and humans, and also HBCD could induce the interference of the secondary metabolite pathways. The system of the stress defences was activated, including signalling pathway, antioxidant defence system, shikimate and phenylpropanoid metabolism. The carbohydrate and amino acid metabolism were disturbed by HBCD intervention, and the lipid, amino acid and secondary metabolite metabolism were regulated for HBCD stress prevention. These results provide insights into the mechanism and degree of HBCD phytotoxicity.

Recently, concerns have been raised that residues of glyphosate-based herbicides may interfere with the homeostasis of the intestinal bacterial community and thereby affect the health of humans or animals. The biochemical pathway for aromatic amino acid synthesis (Shikimate pathway), which is specifically inhibited by glyphosate, is shared by plants and numerous bacterial species. Several in vitro studies have shown that various groups of intestinal bacteria may be differently affected by glyphosate. Here, we present results from an animal exposure trial combining deep 16S rRNA gene sequencing of the bacterial community with liquid chromatography mass spectrometry (LC-MS) based metabolic profiling of aromatic amino acids and their downstream metabolites. We found that glyphosate as well as the commercial formulation Glyfonova®450 PLUS administered at up to fifty times the established European Acceptable Daily Intake (ADI = 0.5 mg/kg body weight) had very limited effects on bacterial community composition in Sprague Dawley rats during a two-week exposure trial. The effect of glyphosate on prototrophic bacterial growth was highly dependent on the availability of aromatic amino acids, suggesting that the observed limited effect on bacterial composition was due to the presence of sufficient amounts of aromatic amino acids in the intestinal environment. A strong correlation was observed between intestinal concentrations of glyphosate and intestinal pH, which may partly be explained by an observed reduction in acetic acid produced by the gut bacteria. We conclude that sufficient intestinal levels of aromatic amino acids provided by the diet alleviates the need for bacterial synthesis of aromatic amino acids and thus prevents an antimicrobial effect of glyphosate in vivo. It is however possible that the situation is different in cases of human malnutrition or in production animals.