Assessment of nickel bioavailability through chemical extractants and red clover (Trifolium pratense L.) in an amended soil: Related changes in various parameters of red clover

Application of immobilizing agents may efficiently reduce the bioavailability of nickel (Ni) in the soil. Here we report the effect of biochar (BC), gravel sludge (GS) and zeolite (ZE) as a sole treatment and their combinations on the bioavailability of Ni after their application into a Ni-polluted soil. The bioavailability of Ni after the application of immobilizing agents was assessed through an indicator plant (red clover) and chemical indicators of bioavailability like soil water extract (SWE), DTPA and Ca(NO3)2 extracts. Additionally, the effects of Ni bioavailability and immobilizing agents on the growth, physiological and biochemical attributes of red clover were also observed. Application of ZE significantly reduced Ni concentrations in all chemical extracts compared to rest of the treatments. Similarly, the combined application of BC and ZE (BC+ ZE) significantly reduced Ni concentrations, reactive oxygen species (ROS) whereas, significant enhancement in the growth, physiological and biochemical attributes along with an improvement in antioxidant defence machinery of red clover plant, compared to rest of the treatments, were observed. Furthermore, BC+ ZE treatment significantly reduced bioconcentration factor (BCF) and bioaccumulation factor (BAF) of Ni in red clover, compared to rest of the treatments. The Ni concentrations in red clover leaves individually reflected a good correlation with Ni concentrations in the extracts (SWE at R2=0.79, DTPA extract at R2=0.84 and Ca(NO3)2 extracts at R2=0.86). Our results indicate that combined application of ZE and BC can significantly reduce the Ni bioavailability in the soil while in parallel improve the antioxidant defence mechanism in plants.