RESUMEN
An arsenate reductase (Car1) from the Bacteroidetes species Rufibacter tibetensis 1351T 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 Ni2+. 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.
Asunto(s)
Arseniato Reductasas , Bacteroidetes , Dominio Catalítico , Oxidación-Reducción , Arseniato Reductasas/metabolismo , Arseniato Reductasas/genética , Arseniato Reductasas/química , Bacteroidetes/enzimología , Bacteroidetes/genética , Arseniatos/metabolismo , Cinética , Monoéster Fosfórico Hidrolasas/metabolismo , Monoéster Fosfórico Hidrolasas/genética , Monoéster Fosfórico Hidrolasas/química , Catálisis , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/química , Arsenitos/metabolismoRESUMEN
Pathogen detection from biological and environmental samples is important for global disease control. Despite advances in pathogen detection using deep learning, current algorithms have limitations in processing long genomic sequences. Through the deep cross-fusion of cross, residual and deep neural networks, we developed DCiPatho for accurate pathogen detection based on the integrated frequency features of 3-to-7 k-mers. Compared with the existing state-of-the-art algorithms, DCiPatho can be used to accurately identify distinct pathogenic bacteria infecting humans, animals and plants. We evaluated DCiPatho on both learned and unlearned pathogen species using both genomics and metagenomics datasets. DCiPatho is an effective tool for the genomic-scale identification of pathogens by integrating the frequency of k-mers into deep cross-fusion networks. The source code is publicly available at https://github.com/LorMeBioAI/DCiPatho.
Asunto(s)
Algoritmos , Programas Informáticos , Humanos , Redes Neurales de la Computación , Genoma , GenómicaRESUMEN
Soil contamination with arsenic (As) can cause phytotoxicity and reduce crop yield. The mechanisms of As toxicity and tolerance are not fully understood. In this study, we used a forward genetics approach to isolate a rice mutant, ahs1, that exhibits hypersensitivity to both arsenate and arsenite. Through genomic resequencing and complementation tests, we identified OsLPD1 as the causal gene, which encodes a putative lipoamide dehydrogenase. OsLPD1 was expressed in the outer cell layer of roots, root meristem cells, and in the mesophyll and vascular tissues of leaves. Subcellular localization and immunoblot analysis demonstrated that OsLPD1 is localized in the stroma of plastids. In vitro assays showed that OsLPD1 exhibited lipoamide dehydrogenase (LPD) activity, which was strongly inhibited by arsenite, but not by arsenate. The ahs1 and OsLPD1 knockout mutants exhibited significantly reduced NADH/NAD+ and GSH/GSSG ratios, along with increased levels of reactive oxygen species and greater oxidative stress in the roots compared with wild-type (WT) plants under As treatment. Additionally, loss-of-function of OsLPD1 also resulted in decreased fatty acid concentrations in rice grain. Taken together, our finding reveals that OsLPD1 plays an important role for maintaining redox homeostasis, conferring tolerance to arsenic stress, and regulating fatty acid biosynthesis in rice.
Asunto(s)
Arsénico , Dihidrolipoamida Deshidrogenasa , Ácidos Grasos , Homeostasis , Oryza , Proteínas de Plantas , Estrés Fisiológico , Adaptación Fisiológica/efectos de los fármacos , Adaptación Fisiológica/genética , Arsénico/toxicidad , Arsenitos/toxicidad , Dihidrolipoamida Deshidrogenasa/metabolismo , Dihidrolipoamida Deshidrogenasa/genética , Ácidos Grasos/biosíntesis , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Mutación/genética , Oryza/genética , Oryza/efectos de los fármacos , Oryza/metabolismo , Oxidación-Reducción/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Plastidios/metabolismo , Plastidios/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Estrés Fisiológico/efectos de los fármacos , Estrés Fisiológico/genéticaRESUMEN
The concentration, chemical speciation, and spatial distribution of essential and toxic mineral elements in cereal seeds have important implications for human health. To identify genes responsible for element uptake, translocation, and storage, high-throughput phenotyping methods are needed to visualize element distribution and concentration in seeds. Here, we used X-ray fluorescence microscopy (µ-XRF) as a method for rapid and high-throughput phenotyping of seed libraries and developed an ImageJ-based pipeline to analyze the spatial distribution of elements. Using this method, we nondestructively scanned 4,190 ethyl methanesulfonate (EMS)-mutagenized M1 rice (Oryza sativa) seeds and 533 diverse rice accessions in a genome-wide association study (GWAS) panel to simultaneously measure concentrations and spatial distribution of elements in the embryo, endosperm, and aleurone layer. A total of 692 putative mutants and 65 loci associated with the spatial distribution of elements in rice seed were identified. This powerful method provides a basis for investigating the genetics and molecular mechanisms controlling the accumulation and spatial variations of mineral elements in plant seeds.
Asunto(s)
Estudio de Asociación del Genoma Completo , Oryza , Humanos , Rayos X , Semillas/genética , Minerales , Microscopía Fluorescente , Oryza/genéticaRESUMEN
Copper (Cu) is an essential micronutrient for all living organisms but is also highly toxic in excess. Cellular homoeostasis of Cu is maintained by various transporters and metallochaperones. Here, we investigated the biological function of OsCOPT7, a member of the copper transporters (COPT) family, in Cu homoeostasis in rice. OsCOPT7 was mainly expressed in the roots and the expression was upregulated by Cu deficiency. OsCOPT7 was localized at the tonoplast and the endoplasmic reticulum. Knockout of OsCOPT7 increased Cu accumulation in the roots but decreased Cu concentrations in the shoots and grain. The knockout mutants contained higher concentrations of Cu in the roots cell sap but markedly lower concentrations of Cu in the xylem sap than wild-type plants. Seed setting and grain yield were reduced significantly in the knockout mutants grown in a low Cu soil. Knockout mutants were more tolerant to Cu toxicity. Yeast two-hybrid and bimolecular fluorescence complementation assays showed that OsCOPT7 interacts physically with the rice Cu chaperone antioxidant protein 1 (OsATX1). Taken together, our results indicate that OsCOPT7 is a specific Cu transporter functioning to export Cu from the vacuoles and the ER and plays an important role in controlling the root-to-shoot Cu translocation in rice.
Asunto(s)
Cobre , Retículo Endoplásmico , Regulación de la Expresión Génica de las Plantas , Oryza , Proteínas de Plantas , Transporte Biológico , Proteínas de Transporte de Catión/metabolismo , Proteínas de Transporte de Catión/genética , Cobre/metabolismo , Grano Comestible/metabolismo , Grano Comestible/genética , Retículo Endoplásmico/metabolismo , Técnicas de Inactivación de Genes , Oryza/metabolismo , Oryza/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Raíces de Plantas/metabolismo , Brotes de la Planta/metabolismo , Semillas/metabolismo , Semillas/genética , Vacuolas/metabolismoRESUMEN
Cadmium (Cd) is a toxic metal that poses serious threats to human health. Rice is a major source of dietary Cd but how rice plants transport Cd to the grain is not fully understood. Here, we characterize the function of the ZIP (ZRT, IRT-like protein) family protein, OsZIP2, in the root-to-shoot translocation of Cd and intervascular transfer of Cd in nodes. OsZIP2 is localized at the plasma membrane and exhibited Cd2+ transport activity when heterologously expressed in yeast. OsZIP2 is strongly expressed in xylem parenchyma cells in roots and in enlarged vascular bundles in nodes. Knockout of OsZIP2 significantly enhanced root-to-shoot translocation of Cd and alleviated the inhibition of root elongation by excess Cd stress; whereas overexpression of OsZIP2 decreased Cd translocation to shoots and resulted in Cd sensitivity. Knockout of OsZIP2 increased Cd allocation to the flag leaf but decreased Cd allocation to the panicle and grain. We further reveal that the variation of OsZIP2 expression level contributes to grain Cd concentration among rice germplasms. Our results demonstrate that OsZIP2 functions in root-to-shoot translocation of Cd in roots and intervascular transfer of Cd in nodes, which can be used for breeding low Cd rice varieties.
RESUMEN
Cadmium (Cd) is highly toxic to plants, but the targets and modes of toxicity remain unclear. We isolated a Cd-hypersensitive mutant of Arabidopsis thaliana, Cd-induced short root 2 (cdsr2), in the background of the phytochelatin synthase-defective mutant cad1-3. Both cdsr2 and cdsr2 cad1-3 displayed shorter roots and were more sensitive to Cd than their respective wild type. Using genomic resequencing and complementation, IAR4 was identified as the causal gene, which encodes a putative mitochondrial pyruvate dehydrogenase E1α subunit. cdsr2 showed decreased pyruvate dehydrogenase activity and NADH content, but markedly increased concentrations of pyruvate and alanine in roots. Both Cd stress and IAR4 mutation decreased auxin level in the root tips, and the effect was additive. A higher growth temperature rescued the phenotypes in cdsr2. Exogenous alanine inhibited root growth and decreased auxin level in the wild type. Cadmium stress suppressed the expression of genes involved in auxin biosynthesis, hydrolysis of auxin-conjugates and auxin polar transport. Our results suggest that auxin homeostasis is a key target of Cd toxicity, which is aggravated by IAR4 mutation due to decreased pyruvate dehydrogenase activity. Decreased auxin level in cdsr2 is likely caused by increased auxin-alanine conjugation and decreased energy status in roots.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Cadmio/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Homeostasis , Mutación , Ácidos Indolacéticos/metabolismo , Alanina , Piruvatos/metabolismo , Piruvatos/farmacología , Oxidorreductasas/metabolismo , Raíces de Plantas/metabolismoRESUMEN
Rice is a dominant source of inorganic arsenic (As) exposure for populations consuming rice as a staple food. Decreasing As accumulation in rice grain is important for improving food safety. Arsenite [As(III)], the main form of As in paddy soil porewater, is taken up inadvertently by OsLsi1 and OsLsi2, the two key transporters for silicon (Si) uptake in rice roots. Here, we investigated whether editing OsLsi1 or OsLsi2 can decrease As accumulation in rice grain without compromising grain yield. We used the CRISPR-Cas9 technology to edit the promoter region of OsLsi1 and the C-terminal coding sequence of OsLsi1 and OsLsi2, and we generated a total of 27 mutants. Uptake and accumulation of Si and As were evaluated in both short-term hydroponic experiments and in a paddy field. Deletion of 1.2-2 kb of the OsLsi1 promoter suppressed OsLsi1 expression in roots and Si uptake markedly and did not affect As(III) uptake or grain As concentration. Some of the OsLsi1 and OsLsi2 coding sequence mutants showed large decreases in the uptake of Si and As(III) as well as large decreases in Si accumulation in rice husks. However, only OsLsi2 mutants showed significant decreases (by up to 63%) in the grain total As concentration. Editing OsLsi2 mainly affected the accumulation of inorganic As in rice grain with little effect on the accumulation of dimethylarsenate (DMA). Grain yields of the OsLsi2 mutants were comparable to those of the wild type. Editing OsLsi2 provides a promising way to reduce As accumulation in rice grain without compromising the grain yield.
Asunto(s)
Arsénico , Oryza , Contaminantes del Suelo , Silicio/metabolismo , Oryza/genética , Proteínas de Transporte de Membrana , Transporte Biológico , SueloRESUMEN
Flooding of paddy fields during the rice growing season enhances arsenic (As) mobilization and greenhouse gas (e.g., methane) emissions. In this study, an adsorbent for dissolved organic matter (DOM), namely, activated carbon (AC), was applied to an arsenic-contaminated paddy soil. The capacity for simultaneously alleviating soil carbon emissions and As accumulation in rice grains was explored. Soil microcosm incubations and 2-year pot experimental results indicated that AC amendment significantly decreased porewater DOM, Fe(III) reduction/Fe2+ release, and As release. More importantly, soil carbon dioxide and methane emissions were mitigated in anoxic microcosm incubations. Porewater DOM of pot experiments mainly consisted of humic-like fluorophores with a molecular structure of lignins and tannins, which could mediate microbial reduction of Fe(III) (oxyhydr)oxides. Soil microcosm incubation experiments cospiking with a carbon source and AC further consolidated that DOM electron shuttling and microbial carbon source functions were crucial for soil Fe(III) reduction, thus driving paddy soil As release and carbon emission. Additionally, the application of AC alleviated rice grain dimethylarsenate accumulation over 2 years. Our results highlight the importance of microbial extracellular electron transfer in driving paddy soil anaerobic respiration and decreasing porewater DOM in simultaneously remediating As contamination and mitigating methane emission in paddy fields.
Asunto(s)
Arsénico , Carbono , Oryza , Suelo , Arsénico/metabolismo , Suelo/química , Contaminantes del Suelo , Carbón Orgánico/química , MetanoRESUMEN
Polyploidy is a prominent feature for genome evolution in many animals and all flowering plants. Plant polyploids often show enhanced fitness in diverse and extreme environments, but the molecular basis for this remains elusive. Soil salinity presents challenges for many plants including agricultural crops. Here we report that salt tolerance is enhanced in tetraploid rice through lower sodium uptake and correlates with epigenetic regulation of jasmonic acid (JA)-related genes. Polyploidy induces DNA hypomethylation and potentiates genomic loci coexistent with many stress-responsive genes, which are generally associated with proximal transposable elements (TEs). Under salt stress, the stress-responsive genes including those in the JA pathway are more rapidly induced and expressed at higher levels in tetraploid than in diploid rice, which is concurrent with increased jasmonoyl isoleucine (JA-Ile) content and JA signaling to confer stress tolerance. After stress, elevated expression of stress-responsive genes in tetraploid rice can induce hypermethylation and suppression of the TEs adjacent to stress-responsive genes. These induced responses are reproducible in a recurring round of salt stress and shared between two japonica tetraploid rice lines. The data collectively suggest a feedback relationship between polyploidy-induced hypomethylation in rapid and strong stress response and stress-induced hypermethylation to repress proximal TEs and/or TE-associated stress-responsive genes. This feedback regulation may provide a molecular basis for selection to enhance adaptation of polyploid plants and crops during evolution and domestication.
Asunto(s)
Metilación de ADN , Epigénesis Genética , Regulación de la Expresión Génica de las Plantas , Oryza/genética , Tolerancia a la Sal/genética , Ciclopentanos/metabolismo , Elementos Transponibles de ADN , Isoleucina/análogos & derivados , Isoleucina/metabolismo , Oxilipinas/metabolismo , TetraploidíaRESUMEN
Methylarsenite (MAs(III)), a product of arsenic biomethylation or bioreduction of methylarsenate (MAs(V)), has been proposed as a primitive antibiotic. However, the antibacterial property and the bactericidal mechanism of MAs(III) remain largely unclear. In this study, we found that MAs(III) is highly toxic to 14 strains of bacteria, especially against 9 strains of Gram-positive bacteria with half maximal inhibitory concentration (IC50) in the sub micromolar range for Staphyloccocus aureus, Microbacterium sp., Pseudarthrobacter siccitolerans and several Bacillus species. In a co-culture of B. subtilis 168 and MAs(III)-producer Enterobacter sp. CZ-1, the later reduced non-toxic MAs(V) to highly toxic MAs(III) to kill the former and gain a competitive advantage. MAs(III) induced autolysis of B. subtilis 168. Deletion of the autolysins LytC, LytD, LytE, and LytF suppressed MAs(III)-induced autolysis in B. subtilis 168. Transcriptomic analysis showed that MAs(III) downregulated the expression of the major genes involved in the biosynthesis of the cell wall peptidoglycan. Overexpression of an UDP-N-acetylglucosamine enolpyruvyl transferase gene murAA alleviated MAs(III)-induced autolysis in B. subtilis 168. MAs(III) disrupted the membrane potential of B. subtilis 168 and caused severe membrane damage. The results suggest that MAs(III) is a broad-spectrum antibiotic preferentially against Gram-positive bacteria by disrupting the cell wall biosynthesis pathway and cell membrane potential.
Asunto(s)
Antibacterianos , Arsénico , Antibacterianos/farmacología , Antibacterianos/metabolismo , Potenciales de la Membrana , Bacterias/metabolismo , Arsénico/metabolismo , Pared Celular/metabolismo , Bacillus subtilis/genéticaRESUMEN
Microbially mediated arsenic redox transformations are key for arsenic speciation and mobility in rice paddies. Whereas anaerobic anoxygenic photosynthesis coupled to arsenite (As(III)) oxidation has been widely examined in arsenic-replete ecosystems, it remains unknown whether this light-dependent process exists in paddy soils. Here, we isolated a phototrophic purple bacteria, Rhodobacter strain CZR27, from an arsenic-contaminated paddy soil and demonstrated its capacity to oxidize As(III) to arsenate (As(V)) using malate as a carbon source photosynthetically. Genome sequencing revealed an As(III)-oxidizing gene cluster (aioXSRBA) encoding an As(III) oxidase. Functional analyses showed that As(III) oxidation under anoxic phototrophic conditions correlated with transcription of the large subunit of the As(III) oxidase aioA gene. Furthermore, the non-As(III) oxidizer Rhodobacter capsulatus SB1003 heterologously expressing aioBA from strain CZR27 was able to oxidize As(III), indicating that aioBA was responsible for the observed As(III) oxidation in strain CZR27. Our study provides evidence for the presence of anaerobic photosynthesis-coupled As(III) oxidation in paddy soils, highlighting the importance of light-dependent, microbe-mediated arsenic redox changes in paddy arsenic biogeochemistry.
Asunto(s)
Arsénico , Arsenitos , Rhodobacter/genética , Ecosistema , Oxidación-Reducción , Oxidorreductasas , Bacterias , SueloRESUMEN
Rice is a major dietary source of inorganic arsenic (iAs), a highly toxic arsenical that accumulates in rice and poses health risks to rice-based populations. However, the availability of detection methods for iAs in rice grains is limited. In this study, we developed a novel approach utilizing a natural bacterial biosensor, Escherichia coli AW3110 (pBB-ArarsR-mCherry), in conjunction with amylase hydrolysis for efficient extraction, enabling high-throughput and quantitative detection of iAs in rice grains. The biosensor exhibits high specificity for arsenic and distinguishes between arsenite [As(III)] and arsenate [As(V)] by modulating the concentration of PO43- in the detection system. We determined the iAs concentrations in 19 rice grain samples with varying total As concentrations and compared our method with the standard technique of microwave digestion coupled with HPLC-ICP-MS. Both methods exhibited comparable results, without no significant bias in the concentrations of As(III) and As(V). The whole-cell biosensor demonstrated excellent reproducibility and a high signal-to-noise ratio, achieving a limit of detection of 16 µg kg-1 [As(III)] and 29 µg kg-1 [As(V)]. These values are considerably lower than the maximum allowable level (100 µg kg-1) for infant rice supplements established by the European Union. Our straightforward sensing strategy presents a promising tool for detecting iAs in other food samples.
Asunto(s)
Arsénico , Arsenicales , Oryza , Humanos , Lactante , Arsénico/análisis , Contaminación de Alimentos/análisis , Reproducibilidad de los Resultados , Arsenicales/análisisRESUMEN
Whole-genome duplication generates a tetraploid from a diploid. Newly created tetraploids (neo-tetraploids) of Arabidopsis (Arabidopsis thaliana) have elevated leaf potassium (K), compared to their diploid progenitor. Micro-grafting has previously established that this elevated leaf K is driven by processes within the root. Here, mutational analysis revealed that the K+-uptake transporters K+ TRANSPORTER 1 (AKT1) and HIGH AFFINITY K+ TRANSPORTER 5 (HAK5) are not necessary for the difference in leaf K caused by whole-genome duplication. However, the endodermis and salt overly sensitive and abscisic acid-related signaling were necessary for the elevated leaf K in neo-tetraploids. Contrasting the root transcriptomes of neo-tetraploid and diploid wild-type and mutants that suppress the neo-tetraploid elevated leaf K phenotype allowed us to identify a core set of 92 differentially expressed genes associated with the difference in leaf K between neo-tetraploids and their diploid progenitor. This core set of genes connected whole-genome duplication with the difference in leaf K between neo-tetraploids and their diploid progenitors. The set of genes is enriched in functions such as cell wall and Casparian strip development and ion transport in the endodermis, root hairs, and procambium. This gene set provides tools to test the intriguing idea of recreating the physiological effects of whole-genome duplication within a diploid genome.
Asunto(s)
Arabidopsis , Tetraploidía , Potasio , Redes Reguladoras de Genes , Arabidopsis/genética , Ploidias , Hojas de la Planta/genéticaRESUMEN
Soil contamination with trace metals and metalloids can cause toxicity to plants and threaten food safety and human health. Plants have evolved sophisticated mechanisms to cope with excess trace metals and metalloids in soils, including chelation and vacuolar sequestration. Sulfur-containing compounds, such as glutathione and phytochelatins, play a crucial role in their detoxification, and sulfur uptake and assimilation are regulated in response to the stress of toxic trace metals and metalloids. This review focuses on the multi-level connections between sulfur homeostasis in plants and responses to such stresses, especially those imposed by arsenic and cadmium. We consider recent progress in understanding the regulation of biosynthesis of glutathione and phytochelatins and of the sensing mechanism of sulfur homeostasis for tolerance of trace metals and metalloids in plants. We also discuss the roles of glutathione and phytochelatins in controlling the accumulation and distribution of arsenic and cadmium in plants, and possible strategies for manipulating sulfur metabolism to limit their accumulation in food crops.
Asunto(s)
Arsénico , Metaloides , Humanos , Cadmio/metabolismo , Arsénico/metabolismo , Metaloides/metabolismo , Fitoquelatinas/metabolismo , Glutatión/metabolismo , Productos Agrícolas/metabolismo , Azufre/metabolismoRESUMEN
Rice is an important source of calories and mineral nutrients for more than half of the world's population. The accumulation of essential and toxic mineral elements in rice grain affects its nutritional quality and safety. However, the patterns and processes by which different elements progressively accumulate during grain filling remain largely unknown. In the present study, we investigated temporal changes in dry matter, elemental concentrations, and the transcriptome in the grain of field-grown rice. We also investigated the effects of seed setting rate and the position of the grain within the rice panicle on element accumulation. Three different patterns of accumulation were observed: (i) elements including K, Mn, B, and Ca showed an early accumulation pattern; (ii) dry matter and elements including N, P, S, Mg, Cu, Zn, Mo, As, and Cd showed a mid accumulation pattern; and (iii) elements such as Fe showed a gradual increase pattern. These different accumulation patterns can be explained by the differences in the biogeochemical behavior of the various elements in the soil, as well as differences in plant nutrient redistribution, gene expression, and the sink-source relationship. These results improve our knowledge of the dynamics of elemental accumulation in rice grain and are helpful for identification of functional genes mediating the translocation of elements to grain.
Asunto(s)
Oryza , Oryza/genética , Oryza/metabolismo , Transcriptoma , Minerales/metabolismo , Grano Comestible/genética , Grano Comestible/metabolismo , Nutrientes , SueloRESUMEN
It is often expected that Zn decreases Cd accumulation in plants due to competition for the same transporters. Here, we found that increasing Zn supply markedly increased the root-to-shoot translocation of Cd in rice. RNA sequencing showed that high Zn up-regulated expression of genes involved in glutathione biosynthesis and metabolism and the Zn/Cd transporter gene OsHMA2, but down-regulated expression of genes related to Zn uptake. Knockout of the iron or Zn transporter genes OsIRT1, OsIRT2, or OsZIP9 did not affect the Zn promotional effect on Cd translocation. Knockout of the manganese/Cd transporter gene OsNRAMP5 greatly reduced Cd uptake but did not affect the Zn promotional effect. Variation in the tonoplast transporter gene OsHMA3 affected Cd translocation but did not change the Zn promotional effect. Knockout of the Zn/Cd transporter gene OsHMA2 not only decreased Cd and Zn translocation, but also abolished the Zn promotional effect. Increased expression of OsHMA2 under high Zn conditions supports the hypothesis that this transporter participates in the promotional effect of Zn on Cd translocation. The results also show that OsIRT1, OsIRT2, and OsZIP9 made only small contributions to Cd uptake under low Zn conditions but not under high Zn conditions, whereas the dominant role of OsNRAMP5 in Cd uptake diminished under low Zn conditions.
Asunto(s)
Cadmio , Oryza , Cadmio/metabolismo , Zinc/metabolismo , Oryza/genética , Oryza/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Transporte Biológico , Translocación Genética , Raíces de Plantas/genética , Raíces de Plantas/metabolismoRESUMEN
Dimethylarsenate [DMAs(V)] is a common methylated As species in soils and plants and can cause the physiological disorder straighthead disease in rice. Because DMAs(V) is relatively noncytotoxic, we hypothesize that phytotoxicity of DMAs(V) may arise from trivalent dimethylarsenite [DMAs(III)]. DMAs(III) has been detected in human urine samples but not in environmental samples, likely due to its instability under oxic conditions. We first established methods for preservation and detections of DMAs(III) in soil and plant samples. We showed that DMAs(III) was a major As species in soil solution from an anoxic paddy soil. Enrichment cultures for fermentative, sulfate-reducing, and denitrifying bacteria from the paddy soil could reduce DMAs(V) to DMAs(III). Twenty-two strains of anaerobic bacteria isolated from the soil showed some ability to reduce DMAs(V). Rice plants grown in hydroponic culture with DMAs(V) also showed the ability to reduce DMAs(V) to DMAs(III). Rice plants and grains grown in a flooded paddy soil contained both DMAs(V) and DMAs(III); their concentrations were higher in the spikelets with straighthead disease than those without. DMAs(III) was much more toxic to the protoplasts isolated from rice plants than DMAs(V). Taken together, the ability to reduce DMAs(V) to highly toxic DMAs(III) is common to soil anaerobes and rice plants.
Asunto(s)
Arsénico , Oryza , Contaminantes del Suelo , Humanos , Ácido Cacodílico , SueloRESUMEN
Microbial oxidation of environmental antimonite (Sb(III)) to antimonate (Sb(V)) is an antimony (Sb) detoxification mechanism. Ensifer adhaerens ST2, a bacterial isolate from a Sb-contaminated paddy soil, oxidizes Sb(III) to Sb(V) under oxic conditions by an unknown mechanism. Genomic analysis of ST2 reveals a gene of unknown function in an arsenic resistance (ars) operon that we term arsO. The transcription level of arsO was significantly upregulated by the addition of Sb(III). ArsO is predicted to be a flavoprotein monooxygenase but shows low sequence similarity to other flavoprotein monooxygenases. Expression of arsO in the arsenic-hypersensitive Escherichia coli strain AW3110Δars conferred increased resistance to Sb(III) but not arsenite (As(III)) or methylarsenite (MAs(III)). Purified ArsO catalyzes Sb(III) oxidation to Sb(V) with NADPH or NADH as the electron donor but does not oxidize As(III) or MAs(III). The purified enzyme contains flavin adenine dinucleotide (FAD) at a ratio of 0.62 mol of FAD/mol protein, and enzymatic activity was increased by addition of FAD. Bioinformatic analyses show that arsO genes are widely distributed in metagenomes from different environments and are particularly abundant in environments affected by human activities. This study demonstrates that ArsO is an environmental Sb(III) oxidase that plays a significant role in the detoxification of Sb(III).
Asunto(s)
Antimonio , Arsénico , Humanos , Antimonio/química , Antimonio/metabolismo , Flavina-Adenina Dinucleótido/metabolismo , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Oxidorreductasas/metabolismo , Oxidación-Reducción , Escherichia coli/metabolismoRESUMEN
Plants take up a wide range of trace metals/metalloids (hereinafter referred to as trace metals) from the soil, some of which are essential but become toxic at high concentrations (e.g., Cu, Zn, Ni, Co), while others are non-essential and toxic even at relatively low concentrations (e.g., As, Cd, Cr, Pb, and Hg). Soil contamination of trace metals is an increasing problem worldwide due to intensifying human activities. Trace metal contamination can cause toxicity and growth inhibition in plants, as well as accumulation in the edible parts to levels that threatens food safety and human health. Understanding the mechanisms of trace metal toxicity and how plants respond to trace metal stress is important for improving plant growth and food safety in contaminated soils. The accumulation of excess trace metals in plants can cause oxidative stress, genotoxicity, programmed cell death, and disturbance in multiple physiological processes. Plants have evolved various strategies to detoxify trace metals through cell-wall binding, complexation, vacuolar sequestration, efflux, and translocation. Multiple signal transduction pathways and regulatory responses are involved in plants challenged with trace metal stresses. In this review, we discuss the recent progress in understanding the molecular mechanisms involved in trace metal toxicity, detoxification, and regulation, as well as strategies to enhance plant resistance to trace metal stresses and reduce toxic metal accumulation in food crops.