Different responses of Sinorhizobium sp. upon Pb and Zn exposure: Mineralization versus complexation.

Lead (Pb) and zinc (Zn) have been discharged into environment and may negatively impact ecological security. Rhizobia has gained attention due to their involvement in the restoration of metal polluted soils. However, little is known about the responses of rhizobia under Pb and Zn stress, especially the roles played by extracellular polymeric substances (EPS) in the resistance of these two metals. Here, Sinorhizobium sp. C10 was isolated from soil around a mining area and was exposed to a series of Pb/Zn treatments. The cell morphology and surface mineral crystals, EPS content and fluorescent substances were determined. In addition, the extracellular polysaccharides and proteins were characterized by attenuated total reflection infrared spectroscopy (ATR-IR) and X-ray photoelectron spectroscopy (XPS). The results showed that Zn stress induced the synthesis of EPS by C10 cells. Functional groups of polysaccharides (CO) and proteins (C-O/C-N) were involved in complexation with Zn. In contrast, C10 resisted Pb stress by forming lead phosphate (Pb3(PO4)2) on the cell surface. Galactose (Gal) and tyrosine played key roles in resistance to the Zn toxicity, whereas glucosamine (N-Glc) was converted to glucose in large amounts during extracellular Pb precipitation. Together, this study demonstrated that C10 possessed different strategies to detoxify the two metals, and could provide basis for bioremediation of Pb and Zn polluted sites.

Copper and zinc interact significantly in their joint toxicity to Chlamydomonas reinhardtii: Insights from physiological and transcriptomic investigations.

Copper (Cu) and zinc (Zn) often discharge simultaneously from industrial and agricultural sectors and cause stress to aquatic biota. Although microalgae have been extensively investigated for their responses to Cu or Zn exposure, how they cope with the mixtures of two metals, especially at transcriptomic level, remains largely unknown. In this study, Chlamydomonas reinhardtii was exposed to environmentally relevant concentrations of two metals. It was found that Zn promoted the entry of Cu into the algal cells. With the increase of combined toxicity, extracellular polymeric substances (EPS) and cell wall functional groups immobilized significant amounts of Cu and Zn. Furthermore, C. reinhardtii adjusted resistance strategies internally, including starch consumption and synthesis of chlorophyll and lipids. Upon high level of Cu and Zn coexistence, synergistic effects were observed in lipid peroxidation and catalase (CAT) activity. Under 1.05 mg/L Cu + 0.87 mg/L Zn, 256 differentially expressed genes (DEGs) were mainly involved in oxidative phosphorylation, ribosome, nitrogen metabolism; while 4294 DEGs induced by 4.21 mg/L Cu + 3.48 mg/L Zn were mainly related to photosynthesis, citric acid cycle, etc. Together, this study revealed a more comprehensive understanding of mechanisms of Cu/Zn detoxification in C. reinhardtii, emphasizing critical roles of photosynthetic carbon sequestration and energy metabolism in the metal resistance.

A comparative study of the accumulation and detoxification of copper and zinc in Chlamydomonas reinhardtii: The role of extracellular polymeric substances.

Extracellular polymeric substances (EPS) form an interface between microalgae and the surrounding water environment. Copper (Cu) and zinc (Zn) are essential micronutrients but may negatively affect microbial growth when their concentrations reach toxic thresholds. However, how EPS affect the accumulation and resistance of Cu and Zn in microalgae remains largely unknown. Here, we investigated EPS production upon Cu/Zn exposure and compared the tolerance strategies to the two metals by Chlamydomonas reinhardtii with and without EPS. Microalgal EPS synthesis was induced by Cu/Zn treatments, and the functional groups of polysaccharides and proteins were involved in complexation with metal ions. The extraction of EPS aggravated the toxicity and reduced the removal of metals from solution, but the effect was more pronounced for Cu than for Zn. Copper bound on the cell surface accounted for 54.6 ± 2.0 % of the Cu accumulated by C. reinhardtii, whose EPS components strongly correlated with Cu adsorption. In contrast, 74.3 ± 3.0 % of accumulated Zn was absorbed in cells, and glutathione synthesis was significantly induced. Redundancy and linear correlation analyses showed that the polysaccharide, protein and DNA contents in EPS were significantly correlated with Cu accumulation, absorption and adsorption but not with Zn. Data fitted to a Michaelis-Menten model further showed that the EPS-intact cells had higher binding capacity for Cu2+ but not for Zn2+. These differential impacts of EPS on Cu/Zn sorption and detoxification contribute to a more comprehensive understanding of the roles of microalgal EPS in the biogeochemical cycle of metals.

Physiological and proteomic responses of Chlamydomonas reinhardtii to arsenate and lead mixtures.

Arsenic (As) and lead (Pb) are frequently emitted from various sources into environment, but microbial responses to their combined toxicity have not been systematically investigated. In this study, Chlamydomonas reinhardtii was exposed to two levels of arsenate (As (V), 50, 500 µg/L), Pb (II) (500, 5000 µg/L) and their mixture (50 µg/L As (V) + 500 µg/L Pb (II); 500 µg/L As (V) + 5000 µg/L Pb (II)). The growth of C. reinhardtii was inhibited more remarkably by As (V) than by Pb (II). The As stress was alleviated by Pb in the 50 µg/L As (V) + 500 µg/L Pb (II) treatment, but was enhanced upon the 500 µg/L As (V) + 5000 µg/L Pb (II) exposure, with more pronounced changes in a number of physiological parameters of the algal cells. Proteomic results showed that 71 differently expressed proteins (DEPs) in the treatment of 50 µg/L As (V) + 500 µg/L Pb (II), and 167 DEPs were identified in that of 500 µg/L As (V) + 5000 µg/L Pb (II). These proteins were involved in energy metabolism, photosynthetic carbon fixation, reactive oxygen scavenging and defense, and amino acid synthesis. Taken together, these physiological and proteomic data demonstrated that C. reinhardtii could resist the As (V) and Pb (II) combined treatments through extracellular complexation and intracellular pathways.