AzuR From the SmtB/ArsR Family of Transcriptional Repressors Regulates Metallothionein in Anabaena sp. Strain PCC 7120

Metallothioneins (MTs) are cysteine-rich, metal-sequestering cytosolic proteins that play a key role in maintaining metal homeostasis and detoxification. We had previously characterized NmtA, a MT from the heterocystous, nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 and demonstrated its role in providing protection against cadmium toxicity. In this study, we illustrate the regulation of Anabaena NmtA by AzuR (Alr0831) belonging to the SmtB/ArsR family of transcriptional repressors. There is currently no experimental evidence for any functional role of AzuR. It is observed that azuR is located within the znuABC operon but in the opposite orientation and remotely away from the nmtA locus. Sequence analysis of AzuR revealed a high degree of sequence identity with Synechococcus SmtB and a distinct α5 metal binding site similar to that of SmtB. In order to characterize AzuR, we overexpressed it in Escherichia coli and purified it by chitin affinity chromatography. Far-UV circular dichroism spectroscopy indicated that the recombinant AzuR protein possessed a properly folded structure. Glutaraldehyde cross-linking and size-exclusion chromatography revealed that AzuR exists as a dimer of ∼28 kDa in solution. Analysis of its putative promoter region [100 bp upstream of nmtA open reading frame (ORF)] identified the presence of a 12–2–12 imperfect inverted repeat as the cis-acting element important for repressor binding. Electrophoretic mobility shift assays (EMSAs) showed concentration-dependent binding of recombinant dimeric AzuR with the promoter indicating that NmtA is indeed a regulatory target of AzuR. Binding of AzuR to DNA was disrupted in the presence of metal ions like Zn2+, Cd2+, Cu2+, Co2+, Ni2+, Pb2+, and Mn2+. The metal-dependent dissociation of protein–DNA complexes suggested the negative regulation of metal-inducible nmtA expression by AzuR. Overexpression of azuR in its native strain Anabaena 7120 enhanced the susceptibility to cadmium stress significantly. Overall, we propose a negative regulation of Anabaena MT by an α5 SmtB/ArsR metalloregulator AzuR.

Changes in Metal Distribution, Vegetative Growth, Reactive Oxygen and Nutrient Absorption of Tagetes patula under Soil Cadmium Stress

Phytoremediation with hyperaccumulator plants has been recognized as a potential way for the clearing of cadmium (Cd)-contaminated soil. In this study, hyperaccumulator Tagetes patula was treated with seven concentrations of Cd, ranging from 0 to 300 mg kg−1. The Cd enrichment and nutrient contents in different organs during different growth phases were investigated. Under Cd concentrations ≤75 mg kg-1, the morphological growth of T. patula did not change significantly regardless of growth stage. However, when Cd concentration exceeded 150 mg kg−1, the morphological growth was remarkedly inhibited. The root/shoot ratio remained unchanged except for at 300 mg kg−1. In addition, Cd negatively influenced the flowering process at the concentration of 300 mg kg−1. Cd content increased significantly in Cd-treated plants. Nitrogen absorption was increased under Cd treatments, and phosphorus content was also increased under concentration ≤150 mg·kg−1. However, the potassium content in the flower was decreased under 300 mg kg−1. Furthermore, the contents of H2O2, O2− and malondialdehyde were increased during the seedling phase, especially when Cd concentration was ≥150 mg kg−1. In summary, T. patula showed a strong ability to tolerate Cd, and such ability might be explained by nutrient absorption and reactive oxygen clearness.

Cloning, Functional Characterization and Response to Cadmium Stress of the Thioredoxin-like Protein 1 Gene from Phascolosoma esculenta

Cadmium (Cd) is a heavy metal toxicant and is widely distributed in aquatic environments. It can cause excessive production of reactive oxygen species (ROS) in the organism, which in turn leads to a series of oxidative damages. Thioredoxin (Trx), a highly conserved disulfide reductase, plays an important role in maintaining the intracellular redox homeostasis in eukaryotes and prokaryotes. Phascolosoma esculenta is an edible marine worm, an invertebrate that is extensively found on the mudflats of coastal China. To explore the molecular response of Trx in mudflat organisms under Cd stress, we identified a new Trx isoform (Trx-like protein 1 gene) from P. esculenta for the first time, designated as PeTrxl. Molecular and structural characterization, as well as multiple sequence and phylogenetic tree analysis, demonstrated that PeTrxl belongs to the Trx superfamily. PeTrxl transcripts were found to be ubiquitous in all tissues, and the highest expression level occurred in the coelomic fluid. Exposure to three sublethal concentrations of Cd resulted in the upregulation and then downregulation of PeTrxl expression levels over time in coelomic fluid of P. esculenta. The significant elevation of PeTrxl expression after 12 and 24 h of Cd exposure at 6 and 96 mg/L, respectively, might reflect its important role in the resistance to Cd stress. Recombinant PeTrxl (rPeTrxl) showed prominent dose-dependent insulin-reducing and ABTS free radical-scavenging abilities. After exposure to 96 mg/L Cd for 24 h, the ROS level increased significantly in the coelomic fluid, suggesting that Cd induced oxidative stress in P. esculenta. Furthermore, the injection of rPeTrxl during Cd exposure significantly reduced the ROS in the coelomic fluid. Our data suggest that PeTrxl has significant antioxidant capacity and can protect P. esculenta from Cd-induced oxidative stress.

Foliar-Applied Selenium Nanoparticles Alleviate Cadmium Stress Through Changes in Physio-Biochemical Status and Essential Oil Profile of Coriander (Coriandrumsativum L.) Leaves

Since large areas of agricultural soils around the world are contaminated by Cd, a cost-effective and practical method is needed for the safe production of edible plants. The effective role of many nanomaterials to improve plant yield and alleviate environmental pollutions is addressed; however, selenium nanoparticles (Se-NPs) potential has not been well-known yet. The aim of this work was to investigate foliar application of Se-NPs on yield, water content, proline concentration, phenolic content, lipid peroxidation, and essential oil (EO) attributes of coriander (Coriandrum sativum L.) under Cd stress. This study consisted of Cd contamination (0, 4, and 8 mg L-1) and foliar application of Se-NPs (0, 20, 40, and 60 mg L-1) in a hydroponic system. The results showed enhanced Cd concentration of root and shoot of coriander under Cd stress; however, Se-NPs alleviate the uptake of Cd. There were observed the decreased shoot and root weight, chlorophyll (Chl) content and relative water content (RWC) by progressing Cd toxicity, but Se-NPs improved these attributes. Moderate Cd stress and 40 or 60 mg L-1 Se-NPs increased phenolic and flavonoid contents as well as EO yield. Proline concentration and malondialdehid (MDA) increased by enhancing Cd stress, but Se-NPs decreased MDA. The GC/MS analysis showed that the main EO constitutes were n-decanal (18.80-29.70%), 2E-dodecanal (14.23-19.87%), 2E-decanal (12.60-19.40%), and n-nonane (7.23-12.87%), representing different amounts under Cd pollution and Se-NPs. To sum up, 40-60 mg L-1 Se-NPs are effective in alleviating Cd stress.

Iminodisuccinic Acid Relieved Cadmium Stress in Rapeseed Leaf by Affecting Cadmium Distribution and Cadmium Chelation With Pectin

Rapeseed (Brassica napus. L) is a nutritious vegetable, while cadmium (Cd) pollution threatens the growth, productivity, and food security of rapeseed. By studying the effects of iminodisuccinic acid (IDS), an easily biodegradable and environmental friendly chelating agent, on Cd distribution at the organ and cellular level, we found IDS promoted dry matter accumulation of rapeseed and increased the contents of photosynthetic pigment in leaves. Inhibited root-shoot Cd transport resulted in higher activity of antioxidant enzymes and decreased hydrogen peroxide (H2O2) and malondialdehyde (MDA) accumulation in leaves, which indicated that IDS contributed to alleviating Cd-caused oxidative damage in leaf cells. Additionally, IDS increased Cd distribution in cell wall (CW) and relieved Cd toxicity in organelle of leaves, while IDS did not change the contents of different CW components. The improved Cd fixation in leaf CW was mainly attributed to enhanced demethylation of covalently bound pectin (CSP) and Cd chelation with CSP.