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Fungi are central to every terrestrial and many aquatic ecosystems, but the mechanisms underlying fungal tolerance to mercury, a global pollutant, remain unknown. Here, we show that the plant symbiotic fungus Metarhizium robertsii degrades methylmercury and reduces divalent mercury, decreasing mercury accumulation in plants and greatly increasing their growth in contaminated soils. M. robertsii does this by demethylating methylmercury via a methylmercury demethylase (MMD) and using a mercury ion reductase (MIR) to reduce divalent mercury to volatile elemental mercury. M. robertsii can also remove methylmercury and divalent mercury from fresh and sea water even in the absence of added nutrients. Overexpression of MMD and MIR significantly improved the ability of M. robertsii to bioremediate soil and water contaminated with methylmercury and divalent mercury. MIR homologs, and thereby divalent mercury tolerance, are widespread in fungi. In contrast, MMD homologs were patchily distributed among the few plant associates and soil fungi that were also able to demethylate methylmercury. Phylogenetic analysis suggests that fungi could have acquired methylmercury demethylase genes from bacteria via two independent horizontal gene transfer events. Heterologous expression of MMD in fungi that lack MMD homologs enabled them to demethylate methylmercury. Our work reveals the mechanisms underlying mercury tolerance in fungi, and may provide a cheap and environmentally friendly means of cleaning up mercury pollution.
Assuntos
Mercúrio , Metarhizium , Compostos de Metilmercúrio , Biodegradação Ambiental , Água , Mercúrio/toxicidade , Filogenia , Ecossistema , Metarhizium/genética , SoloRESUMO
The effects of vegetable planting on soil loss and nutrient loss, runoff, soil erosion, and nitrogen (ammonium nitrogen and nitrate nitrogen) losses under individual rainfalls of fruit- and leaf-vegetable fields between April to October in 2021 were observed using in-situ observation testing. The results showed that: â the runoff, erosion, and nitrogen loss of the fruit-vegetable pattern (eggplant-chili) were 1.27-2.00 times those under the leaf-vegetable pattern (leaf lettuce-sweet potato leaves), especially under the second season vegetable period. Those losses under the second season vegetable accounted for 50.86%-68.83% of the total losses under different vegetable patterns, which were approximately 1.03-2.04 times those under the first season vegetable. The runoff, erosion, and nutrient loss of vegetable fields under different treatments were both concentrated in June and July, and the nitrogen loss was mainly in the form of nitrate nitrogen with surface runoff. â¡ The runoff, erosion, and nutrient losses under individual rainfalls of vegetable fields under different treatments fluctuated among the vegetable growing season, and the losses were mainly concentrated in several typical rainfall events. On the whole, the loss and concentration of nitrate and ammonium nitrogen in runoff and erosion sediment of vegetables in the first season were lower than those in the second season. The runoff, erosion, and loss of ammonium nitrogen and nitrate nitrogen of fruit-vegetable were higher than those of leaf-vegetable. ⢠Both rainfall amount and maximum 30 min rainfall intensity had significantly positive effects on runoff, soil loss, and nitrogen loss. Runoff, erosion, and nutrient losses under different vegetable patterns were mainly generated by moderate rain, heavy rain, and heavy rainstorms, which accounted for 29.58%-46.68%, 24.54%-36.79%, and 24.01%-39.13% of the total losses, respectively. The results also showed that soil erosion and nutrient losses generated by different rainfall grades were obviously different for the fruit- and leaf-vegetable treatments. The results indicated that the vegetable pattern had significant impacts on soil loss and nutrient loss, and the leaf-vegetable pattern could reduce soil erosion and nutrient loss compared with the fruit-vegetable pattern. Furthermore, for different vegetable patterns and vegetable growing seasons, the effects of rainfall on soil loss and nutrient loss were quite different. The results of this study were helpful in clarifying the soil erosion and nutrient loss characteristics of vegetable fields in South China.
Assuntos
Erosão do Solo , Verduras , Nitrogênio/análise , Nitratos , Fósforo/análise , Movimentos da Água , Solo , Chuva , ChinaRESUMO
Human lysozyme (hLYZ), known for its bacteriolytic activity, is widely applied in the food and pharmaceutical industries as an antimicrobial agent. However, its extensive application was limited by its low large-scale production efficiency. In this study, a combinational method of integrating codon optimization, multiple gene copies, and ER molecular chaperone co-expression was developed to improve the heterologous production of hLYZ in Pichia pastoris GS115. Our results showed that increasing the copy number of the optimized hLYZ gene in P. pastoris could enhance its secretory production level up to 1.57-fold. The recombinant opt-hLYZ-6C strain that contains six copies of opt-hLYZ gene exhibited the highest mRNA transcription levels, giving the highest production of 0.22 ± 0.02 mg/mL of hLYZ in the medium supernatant with a bacteriolytic activity of 14,680 ± 300 U/mL against Micrococcus lysodeikticus in the shaking flask experiment. Moreover, co-overexpression of ER retention molecular chaperones, such as Pdi1 or Ero1, in the recombinant opt-hLYZ-6C strain both presented positive effects on the secretory production of hLYZ. Our further characterization indicated that tandem co-expression of Ero1 and Pdi1 together presented an added-up effect. The secretory production of hLYZ in the medium supernatant reached 0.34 ± 0.02 mg/mL of the recombinant opt-hLYZ-6C-EP strain in the shaking flask experiment, with a bacteriolytic activity of 21,200 ± 400 U/mL. Compared to the recombinant opt-hLYZ-1C strain, these final improvements were calculated as 2.43-fold and 2.30-fold on secretory protein levels and antibacterial activity, respectively. Finally, the recombinant opt-hLYZ-6C-EP strain was applied for high-density cultivation in 5 L of fermenter, in which the secretory yield of hLYZ reached 2.34 ± 0.02 mg/mL in the medium supernatant, with a bacteriolytic activity of 1.76 ± 0.02 × 105 U/mL against M. lysodeikticus. All these numbers presented the highest heterologous production levels of hLYZ in microbial systems.
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INTRODUCTION: Teeth with a palatal radicular groove are challenging to diagnose, treat, and save. We classified this condition using cone-beam computed tomographic (CBCT) cross-sectional configurations combined with radiographic and morphologic observations during an intentional replantation procedure to identify and analyze the characteristics of palatal radicular grooves comprehensively as a reference for diagnosis, treatment planning, and prognosis evaluation of this condition. METHODS: Eight cases with palatal radicular grooves present in permanent maxillary incisors were investigated by radiography and CBCT imaging before intentional replantation. RESULTS: The palatal radicular grooves were classified into 3 types based on groove depth and cross-sectional shape on CBCT images: type I, with a shallow groove depth, corresponding to a normal, simple, and single root canal; type II, with a medium groove depth, corresponding to a C-shaped canal system; and type III, with a deep groove depth, almost bisecting the root of the tooth, simultaneously present with 2 independent root canals and an apex with normal shape, corresponding to a labial groove connecting with a palatal groove. The extracted teeth and their radiographic images showed corresponding characteristics. CONCLUSIONS: The CBCT cross-sectional images allowed the best visualization of the depth of the grooves and highlighted the difficulties entailed in treating such teeth. Intentional replantation is an effective therapeutic decision for this type of deformity and can provide better prognosis estimation, especially in teeth with type II and III grooves.