Arsenite Mediates Selenite Resistance and Reduction in Enterobacter sp. Z1, Thereby Enhancing Bacterial Survival in Selenium Environments.

Arsenic (As) is widely present in the environment, and virtually all bacteria possess a conserved ars operon to resist As toxicity. High selenium (Se) concentrations tend to be cytotoxic. Se has an uneven regional distribution and is added to mitigate As contamination in Se-deficient areas. However, the bacterial response to exogenous Se remains poorly understood. Herein, we found that As(III) presence was crucial for Enterobacter sp. Z1 to develop resistance against Se(IV). Se(IV) reduction served as a detoxification mechanism in bacteria, and our results demonstrated an increase in the production of Se nanoparticles (SeNPs) in the presence of As(III). Tandem mass tag proteomics analysis revealed that the induction of As(III) activated the inositol phosphate, butanoyl-CoA/dodecanoyl-CoA, TCA cycle, and tyrosine metabolism pathways, thereby enhancing bacterial metabolism to resist Se(IV). Additionally, arsHRBC, sdr-mdr, purHD, and grxA were activated to participate in the reduction of Se(IV) into SeNPs. Our findings provide innovative perspectives for exploring As-induced Se biotransformation in prokaryotes.

Contributions of selenium-oxidizing bacteria to selenium biofortification and cadmium bioremediation in a native seleniferous Cd-polluted sandy loam soil.

Selenium (Se) is a trace element that is essential for human health. Daily dietary Se intake is governed by the food chain through soil-plant systems. However, the cadmium (Cd) content tends to be excessive in seleniferous soil, in which Se and Cd have complex interactions. Therefore, it is a great challenge to grow crops containing appreciable amounts of Se but low amounts of Cd. We compared the effects of five Se-transforming bacteria on Se and Cd uptake by Brassica rapa L. in a native seleniferous Cd-polluted soil. The results showed that three Se-oxidizing bacteria (LX-1, LX-100, and T3F4) increased the Se content of the aboveground part of the plant by 330.8%, 309.5%, and 724.3%, respectively, compared to the control (p < 0.05). The three bacteria also reduced the aboveground Cd content by 15.1%, 40.4%, and 16.4%, respectively (p < 0.05). In contrast, the Se(IV)-reducing bacterium ES2-45 and weakly Se-transforming bacterium LX-4 had no effect on plant Se uptake, although they did decrease the aboveground Cd content. In addition, the three Se-oxidizing bacteria increased the Se available in the soil by 38.4%, 20.4%, and 24.0%, respectively, compared to the control (p < 0.05). The study results confirm the feasibility of using Se-oxidizing bacteria to simultaneously enhance plant Se content and reduce plant Cd content in seleniferous Cd-polluted soil.

Selenium-oxidizing Agrobacterium sp. T3F4 decreases arsenic uptake by Brassica rapa L. under a native polluted soil.

Toxic arsenic (As) and trace element selenium (Se) are transformed by microorganisms but their complex interactions in soil-plant systems have not been fully understood. An As- and Se- oxidizing bacterium, Agrobacterium sp. T3F4, was applied to a native seleniferous As-polluted soil to investigate As/Se uptake by the vegetable Brassica rapa L. and As-Se interaction as mediated by strain T3F4. The Se content in the aboveground plants was significantly enhanced by 34.1%, but the As content was significantly decreased by 20.5% in the T3F4-inoculated pot culture compared to the control (P < 0.05). Similar result was shown in treatment with additional 5 mg/kg of Se(IV) in soil. In addition, the As contents in roots were significantly decreased by more than 35% under T3F4 or Se(IV) treatments (P<0.05). Analysis of As-Se-bacterium interaction in a soil simulation experiment showed that the bioavailability of Se significantly increased and As was immobilized with the addition of the T3F4 strain (P < 0.05). Furthermore, an As/Se co-exposure hydroponic experiment demonstrated that As uptake and accumulation in plants was reduced by increasing Se(IV) concentrations. The 50% growth inhibition concentration (IC50) values for As in plants were increased about one-fold and two-fold under co-exposure with 5 and 10 µmol/L Se(IV), respectively. In conclusion, strain T3F4 improves Se uptake but decreases As uptake by plants via oxidation of As and Se, resulting in decrease of soil As bioavailability and As/Se competitive absorption by plants. This provides a potential bioremediation strategy for Se biofortification and As immobilization in As-polluted soil.

Microbial oxidation of organic and elemental selenium to selenite.

Selenium (Se) is an essential trace element for life. Se reduction has attracted much attention in the microbial Se cycle, but there is less evidence for Se oxidation. In particular, it is unknown whether microorganisms oxidise organic Se(-II). In this study, four strains of bacteria, namely Dyella spp. LX-1 and LX-66, and Rhodanobacter spp. LX-99 and LX-100, isolated from seleniferous soil, were involved in the oxidation of selenomethionine (SeMet), selenocystine (SeCys2), selenourea and Se(0) to selenite (Se(IV)) in pure cultures. The oxidation rates of organic Se were more rapidly than those of Se(0) in liquid media. Then Se(0) and SeMet were used as examples, microbial oxidation was the predominant process for both additional Se(0) and SeMet in sterilised alkaline or acidic soils. The Se(IV) concentrations were significantly higher at pH 8.56 than at pH 5.25. In addition, water-soluble Se (SOLSe) and exchangeable and carbonate-bound Se (EXC-Se) fractions increased dramatically with these four Se-oxidising bacteria in unsterilised seleniferous soil. To our knowledge, this is the first study to find that various bacteria are involved in the oxidation of organic Se to Se oxyanions, bridging the gap of Se redox in the Se biogeochemical cycle.

Comparative efficacy of bio-selenium nanoparticles and sodium selenite on morpho-physiochemical attributes under normal and salt stress conditions, besides selenium detoxification pathways in Brassica napus L.

Selenium nanoparticles (SeNPs) have attracted considerable attention globally due to their significant potential for alleviating abiotic stresses in plants. Accordingly, further research has been conducted to develop nanoparticles using chemical ways. However, our knowledge about the potential benefit or phytotoxicity of bioSeNPs in rapeseed is still unclear. Herein, we investigated the effect of bioSeNPs on growth and physiochemical attributes, and selenium detoxification pathways compared to sodium selenite (Se (IV)) during the early seedling stage under normal and salt stress conditions. Our findings showed that the range between optimal and toxic levels of bioSeNPs was wider than Se (IV), which increased the plant's ability to reduce salinity-induced oxidative stress. BioSeNPs improved the phenotypic characteristics of rapeseed seedlings without the sign of toxicity, markedly elevated germination, growth, photosynthetic efficiency and osmolyte accumulation versus Se (IV) under normal and salt stress conditions. In addition to modulation of Na+ and K+ uptake, bioSeNPs minimized the ROS level and MDA content by activating the antioxidant enzymes engaged in ROS detoxification by regulating these enzyme-related genes expression patterns. Importantly, the main effect of bioSeNPs and Se (IV) on plant growth appeared to be correlated with the change in the expression levels of Se-related genes. Our qRT-PCR results revealed that the genes involved in Se detoxification in root tissue were upregulated upon Se (IV) treated seedlings compared to NPs, indicating that bioSeNPs have a slightly toxic effect under higher concentrations. Furthermore, bioSeNPs might improve lateral root production by increasing the expression level of LBD16. Taken together, transamination and selenation were more functional methods of Se detoxification and proposed different degradation pathways that synthesized malformed or deformed selenoproteins, which provided essential mechanisms to increase Se tolerance at higher concentrations in rapeseed seedlings. Current findings could add more knowledge regarding the mechanisms underlying bioSeNPs induced plant growth.

Microbial reduction and resistance to selenium: Mechanisms, applications and prospects.

Selenium is an essential trace element for humans, animals and microorganisms. Microbial transformations, in particular, selenium dissimilatory reduction and bioremediation applications have received increasing attention in recent years. This review focuses on multiple Se-reducing pathways under anaerobic and aerobic conditions, and the phylogenetic clustering of selenium reducing enzymes that are involved in these processes. It is emphasized that a selenium reductase may have more than one metabolic function, meanwhile, there are several Se(VI) and/or Se(IV) reduction pathways in a bacterial strain. It is noted that Se(IV)-reducing efficiency is inconsistent with Se(IV) resistance in bacteria. Moreover, we discussed the links of selenium transformations to biogeochemical cycling of other elements, roles of Se-reducing bacteria in soil, plant and digestion system, and the possibility of using functional genes involved in Se transformation as biomarker in different environments. In addition, we point out the gaps and perspectives both on Se transformation mechanisms and applications in terms of bioremediation, Se fortification or dietary supplementation.

Selenium-oxidizing Agrobacterium sp. T3F4 steadily colonizes in soil promoting selenium uptake by pak choi (Brassica campestris).

Selenium (Se) deficiency in soil is linked to its low content in edible crops, resulting in adverse impacts on the health of 15% of the global population. The crop mainly absorbs oxidized selenate and selenite from soil, then converts them into organic Se. However, the role of Se-oxidizing bacteria in soil Se oxidation, Se bioavailability and Se absorption into plants remains unclear. The strain Agrobacterium sp. T3F4, isolated from seleniferous soil, was able to oxidize elemental Se into selenite under pure culture conditions. The green fluorescent protein (gfp)-gene-marked strain (T3F4-GFP) and elemental Se or selenite (5 mg·kg-1) were added to pak choi (Brassica campestris ssp. chinensis) pot cultures. Observation of the fluorescence and viable counting indicated that GFP-expressing bacterial cells steadily colonized the soil in the pots and the leaves of the pak choi, reaching up to 6.6 × 106 and 2.0 × 105 CFU g-1 at 21 days post cultivation, respectively. Moreover, the total Se content (mostly organic Se) was significantly increased in the pak choi under T3F4 inoculated pot culture, with elemental Se(0) being oxidized into Se(IV), and soil Se(IV) being dissolved before being absorbed by the crop. After strain T3F4 was inoculated, no significant differences in microbial diversity were observed in the soils and roots, whereas the abundance of Rhizobium spp. was significantly increased. To our knowledge, this is the first time that Se-oxidizing Agrobacterium sp. T3F4 has been found to steadily colonize soil and plant tissues, and that its addition to soil increases the absorption of Se in plants. This study provides a potential strategy for Se biofortification.

The essentialness of glutathione reductase GorA for biosynthesis of Se(0)-nanoparticles and GSH for CdSe quantum dot formation in Pseudomonas stutzeri TS44.

Pseudomonas stutzeri TS44 was able to aerobically reduce Se(IV) into SeNPs and transform Se(IV)/Cd(II) mixture into CdSe-QDs. The SeNPs and CdSe-QDs were systematically characterized by surface feature analyses, and the molecular mechanisms of SeNPs and CdSe-QD formation in P. stutzeri TS44 were characterized in detail. In vivo, under 2.5 mmol/L Se(IV) exposure, GorA was essential for catalyzing of Se(IV) reduction rate decreased by 67% when the glutathione reductase gene gorA was disrupted, but it was not decreased in the glutathione synthesis rate-limiting gene gshA mutated strain compared to the wild type. The complemented strains restored the phenotypes. While under low amount of Se(IV) (0.5 mmol/L), GSH played an important role for Se(IV) reduction. In vitro, GorA catalyzed Se(IV) reduction with NADPH as the electron donor (Vmax of 3.947 ± 0.1061 µmol/min/mg protein under pH 7.0 and 28℃). In addition, CdSe-QDs were successfully synthesized by a one-step method in which Se(IV) and Cd(II) were added to bacterial culture simultaneously. GSH rather than GorA is necessary for CdSe-QD formation in vivo and in vitro. In conclusion, the results provide new findings showing that GorA functions as a selenite reductase under high amount Se(IV) and GSH is essential for bacterial CdSe-QD synthesis.

Paenibacillus flagellatus sp. nov., isolated from selenium mineral soil.

Strain DXL2T, a Gram-stain-negative, rod-shaped, endospore-forming, motile, aerobic bacterium, was isolated from selenium mineral soil. DXL2T had the highest 16S rRNA gene sequence similarities with those of Paenibacillus ginsengarviGsoil 139T (96.8 %), Paenibacillushemerocallicola DLE-12T (95.5 %) and Paenibacillus hodogayensisSGT (95.4 %). The genome size of DXL2T was 7.24 Mb, containing 6243 predicted protein-coding genes, with a DNA G+C content of 60.2 mol%. DXL2T contained meso-diaminopimelic acid in the cell-wall peptidoglycan. The major cellular fatty acids were anteiso-C15 : 0, iso-C16 : 0 and iso-C15 : 0. The major quinone was menaquinone 7. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two aminophospholipids, an unidentified aminolipid, phosphatidylmethylethanolamine, an unidentified glycolipid and an unidentified phospholipid. Compared with the other strains, DXL2T had a specific phospholipid and a specific aminolipid, it hydrolyzed Tween 40 and could not assimilate potassium gluconate. On the basis of the phenotypic, chemotaxonomic and phylogenetic results, strain DXL2T represents a novel species within the genus Paenibacillus, for which the name Paenibacillusflagellatus sp. nov. is proposed. The type strain is DXL2T (=KCTC 33976T=CCTCC AB 2018054T).

Proteins enriched in charged amino acids control the formation and stabilization of selenium nanoparticles in Comamonas testosteroni S44.

Elemental selenium nanoparticles (SeNPs) are useful in medicine, environmental remediation and in material science. Biosynthesized SeNPs (BioSeNPs) by bacteria are cheap, eco-friendly and have a lower cytotoxicity in comparison with chemically synthesized ones. Organic matters were found to cap on the surface of BioSeNPs, but the functions were still not entirely clear. The purified BioSeNPs were coated in a thick layer of organic substrates observed by transmission electron microscopy (TEM). Fourier Transform Infrared (FT-IR) and quantitative detection of the coating agents showed that one gram of purified BioSeNPs bound 1069 mg proteins, 23 mg carbohydrates and only very limited amounts of lipids. Proteomics of BioSeNPs showed more than 800 proteins bound to BioSeNPs. Proteins enriched in charged amino acids are the major factor thought to govern the formation process and stabilization of BioSeNPs in bacteria. In view of the results reported here, a schematic model for the molecular mechanism of BioSeNPs formation in bacteria is proposed. These findings are helpful for the artificial green synthesis of stable SeNPs under specific condition and guiding the surface modification of SeNPs for medicine application.

Reduction of selenite to Se(0) nanoparticles by filamentous bacterium Streptomyces sp. ES2-5 isolated from a selenium mining soil.

BACKGROUND: Selenium (Se) is an essential trace element in living systems. Microorganisms play a pivotal role in the selenium cycle both in life and in environment. Different bacterial strains are able to reduce Se(IV) (selenite) and (or) Se(VI) (selenate) to less toxic Se(0) with the formation of Se nanoparticles (SeNPs). The biogenic SeNPs have exhibited promising application prospects in medicine, biosensors and environmental remediation. These microorganisms might be explored as potential biofactories for synthesis of metal(loid) nanoparticles. RESULTS: A strictly aerobic, branched actinomycete strain, ES2-5, was isolated from a selenium mining soil in southwest China, identified as Streptomyces sp. based on 16S rRNA gene sequence, physiologic and morphologic characteristics. Both SEM and TEM-EDX analysis showed that Se(IV) was reduced to Se(0) with the formation of SeNPs as a linear chain in the cytoplasm. The sizes of the SeNPs were in the range of 50-500 nm. The cellular concentration of glutathione per biomass decreased along with Se(IV) reduction, and no SeNPs were observed in different sub-cellular fractions in presence of NADPH or NADH as an electron donor, indicating glutathione is most possibly involved in vivo Se(IV) reduction. Strain ES2-5 was resistant to some heavy metal(loid)s such as Se(IV), Cr(VI) and Zn(II) with minimal inhibitory concentration of 50, 80 and 1.5 mM, respectively. CONCLUSIONS: The reducing mechanism of Se(IV) to elemental SeNPs under aerobic condition was investigated in a filamentous strain of Streptomyces. Se(IV) reduction is mediated by glutathione and then SeNPs synthesis happens inside of the cells. The SeNPs are released via hypha lysis or fragmentation. It would be very useful in Se bioremediation if Streptomyces sp. ES2-5 is applied to the contaminated site because of its ability of spore reproduction, Se(IV) reduction, and adaptation in soil.

Paenibacillus selenii sp. nov., isolated from selenium mineral soil.

Strain W126(T), a Gram-reaction-positive, spore-forming, rod-shaped, facultatively anaerobic bacterium, motile by means of peritrichous flagella, was isolated from selenium mineral soil in Hubei province of China. 16S rRNA gene sequence analysis demonstrated that this isolate belonged to the genus Paenibacillus, with 97.9 % sequence similarity to Paenibacillus anaericanus MH21(T), while compared with the other species of the genus Paenibacillus, the 16S rRNA gene sequence similarities were less than 96.0%. DNA-DNA hybridization between strain W126(T) and Paenibacillus anaericanus DSM 15890(T) was 24%. The major isoprenoid menaquinone was menaquinone-7. Anteiso-C(15 : 0) was the major fatty acid. The DNA G+C content was 42.3 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, three unknown aminophospholipids and an unknown lipid. Strain W126(T) contained A1γ-meso-diaminopimelic acid in the cell-wall peptidoglycan. The phenotypic, chemotaxonomic and genotypic data indicate that strain W126(T) represents a novel species of the genus Paenibacillus, for which the name Paenibacillus selenii sp. nov. is proposed. The type strain is W126(T) ( = KCTC 33420(T) = CCTCC AB 2014003(T)).

Paenibacillus selenitireducens sp. nov., a selenite-reducing bacterium isolated from a selenium mineral soil.

A Gram-stain-positive, rod-shaped, facultatively anaerobic bacterium, designated strain ES3-24(T), was isolated from a selenium mineral soil. The isolate was endospore-forming, nitrate-reducing and motile by means of peritrichous flagella. The major menaquinone was menaquinone 7 (MK-7) and the predominant fatty acids (>5%) were anteiso-C15:0, iso-C16:0, C16:0 and anteiso-C17:0. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and two unknown aminophospholipids. Strain ES3-24(T) contained meso-diaminopimelic acid in the cell-wall peptidoglycan and the DNA G+C content was 49.6 mol%. According to phylogenetic analysis based on the 16S rRNA gene sequence, strain ES3-24(T) was most closely related to Paenibacillus terrigena A35(T), with 16S rRNA gene sequence identity of 98.3%, while the other members of the genus Paenibacillus had 16S rRNA gene sequence identities of less than 95.0%. DNA-DNA relatedness between strain ES3-24(T) and P. terrigena CCTCC AB206026(T) was 39.3 %. In addition, strain ES3-24(T) showed obvious differences from closely related species in major polar lipids, nitrate reduction and other physiological and biochemical characteristics. The data from our polyphasic taxonomic study reveal that strain ES3-24(T) represents a novel species of the genus Paenibacillus, for which the name Paenibacillus selenitireducens sp. nov. is proposed. The type strain is ES3-24(T) ( = KCTC 33157(T) = CCTCC AB2013097(T)).