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Nickel (Ni) bioavailability in the soil can be alleviated by the integrated use of immobilizing agents. In the current study, it was observed that sole of combined use of biochar (BA), zeolite (ZT) and gravel sludge (GE) significantly influenced the Ni bioavailability in Ni stimulated soil compared to control. The bioavailable Ni contents were measured in DTPA, Ca(NO3)2 and extracts and soil water extract (SWE) with indicator plant red clover (Trifolium pratense L.). The impact of immobilizing agents and a bioavailable portion of Ni was assessed through physiology, growth and biochemical characters of red clover were recorded. Results showed that application of ZT significantly decreased the extractable Ni concentrations in all treatments, however, the combined use of BA and ZT significantly reduced the Ni and decreased the generation of (ROS) thus positively influence physiological, agronomic and biochemical attributes of red clover. Therefore, ZT and BA as sole or combination of both were selected the second experiment with the object to immobilize Ni in soil with both maize and sunflower as a test crop. Results revealed the treatment receiving ZT: BA with 1:3 significantly improved the growth, yield, chemical and biochemical attributes in both crops over control. Nickel-induced oxidative stress was significantly mitigated in this treatment compared to control. However, the highest decrease in uptake of Ni was observed where ZT: BA was 1:1 compared to control. Based on finds of the second study, the third study was designed with ZT: BA 1:1 selected from the previous study. Wheat (Triticum aestivum L.) grown and proline (PO) was exogenously applied according to the treatment plan. Proline acts as a stress alleviator and mitigates heavy metals stress. Proline was applied with ZT: BA 1:1 mixture to determine the efficiency of these amendments and PO to mitigate Ni stress and improvement in antioxidants activity in wheat. The finding suggested that the combined use of immobilizing agents with some stress alleviator could significantly reduce Ni bioavailability and increased the biomass, physiological and biochemical parameters. Ni induced oxidative stress was also ameliorated in wheat with ZT, BA, and PO, compared to control.
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