Wheat Leaf Antioxidative Status—Variety-Specific Mechanisms of Zinc Tolerance during Biofortification

In this study, we evaluated the leaf antioxidative responses of three wheat varieties (Srpanjka, Divana, and Simonida) treated with two different forms of zinc (Zn), Zn-sulfate and Zn-EDTA, in concentrations commonly used in agronomic biofortification. Zn concentration was significantly higher in the flag leaves of all three wheat varieties treated with Zn-EDTA compared to control and leaves treated with Zn-sulfate. Both forms of Zn increased malondialdehyde level and total phenolics content in varieties Srpanjka and Divana. Total glutathione content was not affected after the Zn treatment. Zn-sulfate increased the activities of glutathione reductase (GR) and guaiacol peroxidase (GPOD) in both Srpanjka and Divana, while glutathione S-transferase (GST) was only induced in var. Srpanjka. Chelate form of Zn increased the activities of GST and GPOD in both Simonida and Divana. Catalase activity was shown to be less sensitive to Zn treatment and was only induced in var. Srpanjka treated with Zn-EDTA where GPOD activity was not induced. Concentrations of Zn used for agronomic biofortification can induce oxidative stress in wheat leaves. The antioxidative status of wheat leaves could be a good indicator of Zn tolerance, whereas wheat genotype and chemical form of Zn are the most critical factors influencing Zn toxicity.

Changes in the Composition of the Soil Bacterial Community in Heavy Metal-Contaminated Farmland

The structural changes of microorganisms in soil are the focus of soil indicators research. The purpose of this study was to investigate the changes in the composition of the soil bacterial community in heavy metal-contaminated soil. A total of six soil samples (two sampling times) were collected from contaminated farmland at three different depths (surface, middle, and deep layer). The pH value was measured. The concentrations of heavy metals (Cr, Ni, Cu, Zn, Cd, and Pb) and the soil bacterial community were analyzed using ICP-OES and 16S rRNA gene sequencing. The results of the two samplings showed that the pH value in the deep layer decreased from 6.88 to 6.23, and the concentrations of Cu, Zn, Cd, and Pb, with a smaller ion radius, increased by 16–28%, and Shannon, Chao1 increased by ~13%. The bacteria community composition at the three depths changed, but Proteobacteria, Acidobacteria, and Actinobacteria were the dominant phyla. In the copper and zinc tolerance test, the isolated bacterium that was able to tolerate copper and zinc was Bacillus sp. We found that, the longer the heavy metal pollution was of concern, the higher the tolerance. These results can be used as references for the microbial remediation of heavy metal-contaminated farmland.

Isolation screening and molecular characterization of zinc solubilizing bacteria and their effect on the growth of wheat (Triticum aestivum)

Wheat (Triticum aestivum) is a major cereal crop grown worldwide. Most of the world population depends on wheat for their nutrient requirement. Zinc (Zn) is one of the most crucial elements required for the development of wheat plant. It is one of the micronutrients required in many biochemical cycles. It has been found that the concentration of Zn is below the required level in the soil and hence it remains deficient in the crops. To ameliorate the deficit, chemical fertilizers are added in the soil, where as biofertilizers are preferred over chemicals in sustainable agriculture. The paper describes the isolation, screening and molecular characterization of the zinc solubilizing bacteria (ZSB) to improve plant growth. A total of 100 soil samples were collected from the rhizospheric soil of wheat plants. ZSB were isolated by dilution plating on Bunt and Rovira media. The 50 isolates were selected and screened for their Zn solubilization. The zinc tolerance of all the isolates varied from 0.5% to 2% of insoluble Zn. Based on the Zn tolerance ability, 15 bacterial isolates were screened for Phosphate solubilization and further analyzed for the synthesis of IAA, NH3, siderophore production and chitinase activity. The three isolates were selected on the basis of the plant growth promoting characteristics for molecular characterization and were found to be homologous to Bacillus cereus, Pseudomonas aeruginosa and Bacillus tropicus. This study documented the establishment and survival of ZSB in the wheat rhizosphere and enhanced plant productivity, thus indicating the potential of isolates as commercial biofertilizers.

High-Zinc Supplementation of Weaned Piglets Affects Frequencies of Virulence and Bacteriocin Associated Genes Among Intestinal Escherichia coli Populations

To prevent economic losses due to post-weaning diarrhea (PWD) in industrial pig production, zinc (Zn) feed additives have been widely used, especially since awareness has risen that the regular application of antibiotics promotes buildup of antimicrobial resistance in both commensal and pathogenic bacteria. In a previous study on 179 Escherichia coli collected from piglets sacrificed at the end of a Zn feeding trial, including isolates obtained from animals of a high-zinc fed group (HZG) and a corresponding control group (CG), we found that the isolate collection exhibited three different levels of tolerance toward zinc, i.e., the minimal inhibitory concentration (MIC) detected was 128, followed by 256 and 512 μg/ml ZnCl2. We further provided evidence that enhanced zinc tolerance in porcine intestinal E. coli populations is clearly linked to excessive zinc feeding. Here we provide insights about the genomic make-up and phylogenetic background of these 179 E. coli genomes. Bayesian analysis of the population structure (BAPS) revealed a lack of association between the actual zinc tolerance level and a particular phylogenetic E. coli cluster or even branch for both, isolates belonging to the HZG and CG. In addition, detection rates for genes and operons associated with virulence (VAG) and bacteriocins (BAG) were lower in isolates originating from the HZG (41 vs. 65% and 22 vs. 35%, p < 0.001 and p = 0.002, resp.). Strikingly, E. coli harboring genes defining distinct pathotypes associated with intestinal disease, i.e., enterotoxigenic, enteropathogenic, and Shiga toxin-producing E. coli (ETEC, EPEC, and STEC) constituted 1% of the isolates belonging to the HZG but 14% of those from the CG. Notably, these pathotypes were positively associated with enhanced zinc tolerance (512 μg/ml ZnCl2 MIC, p < 0.001). Taken together, zinc excess seems to influence carriage rates of VAGs and BAGs in porcine intestinal E. coli populations, and high-zinc feeding is negatively correlated with enteral pathotype occurrences, which might explain earlier observations concerning the relative increase of Enterobacterales considering the overall intestinal microbiota of piglets during zinc feeding trials while PWD rates have decreased.

Indexes of Radicle are Sensitive and Effective for Assessing Copper and Zinc Tolerance in Germinating Seeds of Suaeda salsa

Euhalophytes, such as Suaeda salsa, are ideal candidates to remediate heavy metal-polluted saline soils. However, the metal tolerance ability of dimorphic seeds and subsequent seedlings is largely unknown. This study investigated the tolerance of S. salsa seeds to different concentrations of Cu2+ (0–300 mM) and Zn2+ (0–300 mM) during germination and seedling growth stages. Results showed that dimorphic seeds of S. salsa had high metal tolerance during germination, and even germinated under 300 mM Cu and Zn treatments. However, seedling growth was more sensitive to metal solutions and radicle growth was almost completely inhibited by Cu at 10 mM, and by Zn at 50 mM. Germinating seeds and seedlings of S. salsa had a higher metal toxicity threshold of Zn than that of Cu. In all indexes, indexes of radicle were the most sensitive and effective indicator of metal tolerance. Seeds of S. salsa germinated successfully and seedlings survived under high Zn and Cu stress. The results suggest that S. salsa could be sown directly in heavy metal-contaminated soils for phytoremediation.

Identification of the Genetic Requirements for Zinc Tolerance and Toxicity in Saccharomyces cerevisiae

Zinc is essential for almost all living organisms, since it serves as a crucial cofactor for transcription factors and enzymes. However, it is toxic to cell growth when present in excess. The present work aims to investigate the toxicity mechanisms induced by zinc stress in yeast cells. To this end, 108 yeast single-gene deletion mutants were identified sensitive to 6 mM ZnCl2 through a genome-wide screen. These genes were predominantly related to the biological processes of vacuolar acidification and transport, polyphosphate metabolic process, cytosolic transport, the process utilizing autophagic mechanism. A result from the measurement of intracellular zinc content showed that 64 mutants accumulated higher intracellular zinc under zinc stress than the wild-type cells. We further measured the intracellular ROS (reactive oxygen species) levels of 108 zinc-sensitive mutants treated with 3 mM ZnCl2. We showed that the intracellular ROS levels in 51 mutants were increased by high zinc stress, suggesting their possible involvement in regulating ROS homeostasis in response to high zinc. The results also revealed that excess zinc could generate oxidative damage and then activate the expression of several antioxidant defenses genes. Taken together, the data obtained indicated that excess zinc toxicity might be mainly due to the high intracellular zinc levels and ROS levels induced by zinc stress in yeast cells. Our current findings would provide a basis to understand the molecular mechanisms of zinc toxicity in yeast cells.