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Arsenic (As) poisoning in aquifers is a serious problem worldwide, especially in the middle-Gangetic Plain (MGP) of India. Microbially-mediated As speciation in such aquifers is governed by the arsenate-reductase enzyme, ArsC, encoded by the arsC gene of As-metabolizing bacteria. In this study, ArsC1 (119 aa) and ArsC2 (141 aa) of a highly resistant strain to arsenic, Citrobacter youngae IITK SM2 (CyIITKSM2), isolated from a mixed-oxic MGP groundwater were biochemically characterized. Coupled-arsenate-reductase assay and IC-ICP-MS analysis confirmed that ArsC2 showed higher As(V) reduction than ArsC1 in the dissolved phase, which was consistent with the prominent structural changes in ArsC2 as identified through circular dichroism spectroscopy. Furthermore, the two ArsCs were able to mobilize arsenic from solid-bound arsenate [As(V)-loaded goethite, AsG] predominantly as As(III). However, the total arsenic released in the presence of ArsC2 was ∼38 % and ∼88 % higher, respectively, as compared to the ArsC1-containing and ArsC-free conditions. A process-based model that considered ArsC-mediated As(V) reduction to As(III) in the dissolved phase, and surface complexation of As(V) and As(III) on goethite, suggested that the extent of arsenate binding with ArsC was not affected by whether As(V) was dissolved or was sorbed. However, the catalytic reduction rate was at least an order of magnitude lower in sorbed As(V) than in dissolved As(V). Mutants of ArsC2 exhibited variable but reduced efficiencies compared to the wild-type ArsC2. This reduction may be attributed to the C-terminal loop observed in the AlphaFold predicted structure of ArsC2, which was absent in ArsC1. This comprehensive biochemical and biophysical analysis of the arsenate reductases in Citrobacter youngae could enhance our understanding of the role these microbes play in arsenic mobilization within MGP aquifers., Competing Interests: Declaration of competing interest The authors declare that no competing financial interests exist, and this work has been carried out in compliance with ethical standards., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Selenium-enhanced arsenic uptake by As-hyperaccumulators Pteris vittata and Pteris cretica is known, but how it impacts antimony (Sb) uptake and associated mechanisms are unclear. Here, we investigated the effects of 2.5 μM selenate (Se 2.5 ) on Sb uptake by two plants after growing for 10 days under hydroponics containing 10 or 50 μM antimonate (SbV) (Sb 10 or Sb 50 ). Both plants were efficient in taking up SbV, which was reduced to SbIII (17-40 %) and mainly accumulated in the roots (86-97 %). The addition of Se increased the Sb contents by 78-97 and 29-33 % to 242-1358 and 132-697 mg kg -1 in P. vittata and P. cretica roots. Compared with the Sb 10 and Sb 50 treatments, addition of Se increased the SbV reduction, with more increase in P. vittata than P. cretica roots (181-273 % vs. 17-29 %). Enhanced GSH-GSSG cycle mediated by glutathione peroxidase (GPX) and glutathione reductase (GR) may play an important role in SbV reduction in the roots. Compared with the Sb treatments, addition of Se increased the GPX and GR activity by 71-97 and 2-50 % in P. vittata roots, and 59-153 and 22-63 % in P. cretica roots. Besides, Se upregulated the expression of arsenate reductases PvHAC1 and PvACR2 in P. vittata roots by 1.7-3.4 folds but not in P. cretica. Se-enhanced SbV reduction in P. vittata explains why it was more effective in Sb accumulation than P. cretica. Taken together, Se is effective in increasing the Sb uptake in both plants probably by promoting SbV reduction via GSH-GSSG cycle and/or PvHAC1/PvACR2, suggesting that Se may be used to enhance phytostabilization of Sb-contaminated soils., 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 Elsevier B.V. All rights reserved.)

Background: Arsenic (As) metabolism by soil microorganisms has an impact on As geochemical cycling in paddy soils, which in turn affects As uptake in rice. However, little is known about the key microorganisms involved in this process in Japanese paddy soil., Results: Total RNA was extracted from Japanese paddy soils with different levels of dissolved As under flooded conditions, and the transcription of As metabolic genes (arrA, ttrA and arsM) was analyzed via a metatranscriptomic approach. The results showed that ttrA was the predominant respiratory arsenate reductase gene transcribed in these soils rather than arrA, suggesting that ttrA contributes to the reductive dissolution of As. The predominant taxa expressing ttrA differed among soils but were mostly associated with genera known for their iron- and/or sulfate-reduction activity. In addition, a wide variety of microorganisms expressed and upregulated arsM approximately 5.0- to 13.2-fold at 9 d compared with 3 d of incubation under flooded conditions in flasks., Conclusions: Our results support the involvement of microbial activity in the geochemical cycling of As in Japanese paddy soils and suggest that ttrA may be one of the key genes involved in the formation of arsenite, an inorganic species taken up by rice., (© 2024. The Author(s).)

Citrobacter sp. XT1-2-2, a functional microorganism with potential utilization, has the ability to immobilize soil cadmium. In this study, the regulatory gene cysH, as a rate-limiting enzyme in the sulfur metabolic pathway, was selected for functional analysis affecting cadmium immobilization in soil. To verify the effect of APS reductase on CdS formation, the ΔAPS and ΔAPS-com strains were constructed by conjugation transfer. Through TEM analysis, it was found that the adsorption of Cd 2+ was affected by the absence of APS reductase in XT1-2-2 strain. The difference analysis of biofilm formation indicated that APS reductase was necessary for cell aggregation and biofilm formation. The p-XRD, XPS and FT-IR analysis revealed that APS reductase played an important role in the cadmium immobilization process of XT1-2-2 strain and promoting the formation of CdS. According to the pot experiments, the cadmium concentration of roots, culms, leaves and grains inoculated with ΔAPS strain was significantly higher than that of wild-type and ΔAPS-com strains, and the cadmium removal ability of ΔAPS strain was significantly lower than that of wild-type strain. The study provided insights into the exploration of new bacterial assisted technique for the remediation and safe production of rice in cadmium-contaminated paddy soils., 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 Inc. All rights reserved.)

An arsenate reductase (Car1) from the Bacteroidetes species Rufibacter tibetensis 1351 T was isolated from the Tibetan Plateau. The strain exhibits resistance to arsenite [As(III)] and arsenate [As(V)] and reduces As(V) to As(III). Here we shed light on the mechanism of enzymatic reduction by Car1. AlphaFold2 structure prediction, active site energy minimization, and steady-state kinetics of wild-type and mutant enzymes give insight into the catalytic mechanism. Car1 is structurally related to calcineurin-like metallophosphoesterases (MPPs). It functions as a binuclear metal hydrolase with limited phosphatase activity, particularly relying on the divalent metal Ni 2+ . As an As(V) reductase, it displays metal promiscuity and is coupled to the thioredoxin redox cycle, requiring the participation of two cysteine residues, Cys74 and Cys76. These findings suggest that Car1 evolved from a common ancestor of extant phosphatases by incorporating a redox function into an existing MPP catalytic site. Its proposed mechanism of arsenate reduction involves Cys74 initiating a nucleophilic attack on arsenate, leading to the formation of a covalent intermediate. Next, a nucleophilic attack of Cys76 leads to the release of As(III) and the formation of a surface-exposed Cys74-Cys76 disulfide, ready for reduction by thioredoxin., (© 2024 John Wiley & Sons Ltd.)

As sessile organisms, plants encounter dynamic and challenging environments daily, including abiotic/biotic stresses. The regulation of carbon and nitrogen allocations for the synthesis of plant proteins, carbohydrates, and lipids is fundamental for plant growth and adaption to its surroundings. Light, one of the essential environmental signals, exerts a substantial impact on plant metabolism and resource partitioning (i.e., starch). However, it is not fully understood how light signaling affects carbohydrate production and allocation in plant growth and development. An orphan gene unique to Arabidopsis thaliana, named QUA-QUINE STARCH (QQS) is involved in the metabolic processes for partitioning of carbon and nitrogen among proteins and carbohydrates, thus influencing leaf, seed composition, and plant defense in Arabidopsis. In this study, we show that PHYTOCHROME-INTERACTING bHLH TRANSCRIPTION FACTORS (PIFs), including PIF4, are required to suppress QQS during the period at dawn, thus preventing overconsumption of starch reserves. QQS expression is significantly de-repressed in pif4 and pifQ, while repressed by overexpression of PIF4, suggesting that PIF4 and its close homologs (PIF1, PIF3, and PIF5) act as negative regulators of QQS expression. In addition, we show that the evening complex, including ELF3 is required for active expression of QQS, thus playing a positive role in starch catabolism during night-time. Furthermore, QQS is epigenetically suppressed by DNA methylation machinery, whereas histone H3 K4 methyltransferases (e.g., ATX1, ATX2, and ATXR7) and H3 acetyltransferases (e.g., HAC1 and HAC5) are involved in the expression of QQS. This study demonstrates that PIF light signaling factors help plants utilize optimal amounts of starch during the night and prevent overconsumption of starch before its biosynthesis during the upcoming day., (© 2023 Society for Experimental Biology and John Wiley & Sons Ltd.)

Arsenic (As) has become natural health hazard for millions of people across the world due to its distribution in the food chain. Naturally, it is present in different oxidative states of inorganic [As(V) and As(III)] and organic (DMA, MMA and TMA) forms. Among different mitigation approaches, microbe mediated mitigation of As toxicity is an effective and eco-friendly approach. The present study involves the characterization of bacterial strains containing arsenite methyltransferase (Pseudomonas oleovorans, B4.10); arsenate reductase (Sphingobacterium puteale, B4.22) and arsenite oxidase (Citrobacter sp., B5.12) activity with plant growth promoting (PGP) traits. Efficient reduction of grain As content by 61 % was observed due to inoculation of methyltransferase containing B4.10 as compared to B4.22 (47 %) and B5.12 (49 %). Reduced bioaccumulation of As in root (0.339) and shoot (0.166) in presence of B4.10 was found to be inversely related with translocation factor for Mn (3.28), Fe (0.073), and Se (1.82). Bioaccumulation of these micro elements was found to be associated with the modulated expression of different mineral transporters (OsIRT2, OsFRO2, OsTOM1, OsSultr4;1, and OsZIP2) in rice shoot. Improved dehydrogenase (407 %), and β-glucosidase (97 %) activity in presence of P. oleovorans (B4.10) as compared to arsenate reductase (198 and 50 %), and arsenite oxidase (134 and 69 %) containing bacteria was also observed. Our finding confers the potential of methyltransferase positive P. oleovorans (B4.10) for As stress amelioration. Reduced grain As uptake was found to be mediated by improved plant growth and nutrient uptake associated with enhanced soil microbial activity., Competing Interests: Declaration of competing interest The authors have no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Selenate enhances arsenic (As) accumulation in As-hyperaccumulator Pteris vittata , but the associated molecular mechanisms are unclear. Here, we investigated the mechanisms of selenate-induced arsenic accumulation by exposing P. vittata to 50 μM arsenate (AsV 50 ) and 1.25 (Se 1.25 ) or 5 μM (Se 5 ) selenate in hydroponics. After 2 weeks, plant biomass, plant As and Se contents, As speciation in plant and growth media, and important genes related to As detoxification in P. vittata were determined. These genes included P transporters PvPht1;3 and PvPht1;4 (AsV uptake), arsenate reductases PvHAC1 and PvHAC2 (AsV reduction), and arsenite (AsIII) antiporters PvACR3 and PvACR3;2 (AsIII translocation) in the roots, and AsIII antiporters PvACR3;1 and PvACR3;3 (AsIII sequestration) in the fronds. The results show that Se 1.25 was more effective than Se 5 in increasing As accumulation in both P. vittata roots and fronds, which increased by 27 and 153% to 353 and 506 mg kg -1 . The As speciation analyses show that selenate increased the AsIII levels in P. vittata , with 124-282% more AsIII being translocated into the fronds. The qPCR analyses indicate that Se 1.25 upregulated the gene expression of PvHAC1 by 1.2-fold, and PvACR3 and PvACR3;2 by 1.0- to 2.5-fold in the roots, and PvACR3;1 and PvACR3;3 by 0.6- to 1.1-fold in the fronds under AsV 50 treatment. Though arsenate enhanced gene expression of P transporters PvPht1;3 and PvPht1;4 , selenate had little effect. Our results indicate that selenate effectively increased As accumulation in P. vittata , mostly by increasing reduction of AsV to AsIII in the roots, AsIII translocation from the roots to fronds, and AsIII sequestration into the vacuoles in the fronds. The results suggest that selenate may be used to enhance phytoremediation of As-contaminated soils using P. vittata .

Histone acetyltransferases (HATs) are involved in the epigenetic positive control of gene expression in eukaryotes. CREB-binding proteins (CBP)/p300, a subfamily of highly conserved HATs, have been shown to function as acetylases on both histones and non-histone proteins. In the model plant Arabidopsis thaliana among the five CBP/p300 HATs, HAC1, HAC5 and HAC12 have been shown to be involved in the ethylene signaling pathway. In addition, HAC1 and HAC5 interact and cooperate with the Mediator complex, as in humans. Therefore, it is potentially difficult to discriminate the effect on plant development of the enzymatic activity with respect to their Mediator-related function. Taking advantage of the homology of the human HAC catalytic domain with that of the Arabidopsis, we set-up a phenotypic assay based on the hypocotyl length of Arabidopsis dark-grown seedlings to evaluate the effects of a compound previously described as human p300/CBP inhibitor, and to screen previously described cinnamoyl derivatives as well as newly synthesized analogues. We selected the most effective compounds, and we demonstrated their efficacy at phenotypic and molecular level. The in vitro inhibition of the enzymatic activity proved the specificity of the inhibitor on the catalytic domain of HAC1, thus substantiating this strategy as a useful tool in plant epigenetic studies.

Plants perceive volatiles emitted from herbivore-damaged neighboring plants to urgently adapt or prime their defense responses to prepare for forthcoming herbivores. Mechanistically, these volatiles can induce epigenetic regulation based on histone modifications that alter the transcriptional status of defense genes, but little is known about the underlying mechanisms. To understand the roles of such epigenetic regulation of plant volatile signaling, we explored the response of Arabidopsis (Arabidopsis thaliana) plants to the volatile β-ocimene. Defense traits of Arabidopsis plants toward larvae of Spodoptera litura were induced in response to β-ocimene, through enriched histone acetylation and elevated transcriptional levels of defense gene regulators, including ethylene response factor genes (ERF8 and ERF104) in leaves. The enhanced defense ability of the plants was maintained for 5 d but not over 10 d after exposure to β-ocimene, and this coincided with elevated expression of those ERFs in their leaves. An array of histone acetyltransferases, including HAC1, HAC5, and HAM1, were responsible for the induction and maintenance of the anti-herbivore property. HDA6, a histone deacetylase, played a role in the reverse histone remodeling. Collectively, our findings illuminate the role of epigenetic regulation in plant volatile signaling., (© American Society of Plant Biologists 2022. All rights reserved. For permissions, please email: journals.permissions@oup.com.)