Alterations in expression of stress-responsive genes in Arabidopsis thaliana (L.) Heynh. roots induced by toxic Ni²⁺ levels and joint application of Ni²⁺ and L-histidine

Nickel (Ni) is an essential trace element for higher plants, being a part of urease, glyoxylases and hydrogenases. However, in high concentrations, this metal exerts a pronounced toxic effect. Levels of Ni²⁺ that are toxic to plants (>10^–5 M) are typical for soils formed from volcanic rocks, near mining and metallurgical plants, landfills, waste disposal sites. Elevated nickel levels are also recorded in the soils of Belarus. In the presented work, we examined the effect of a range of toxic Ni²⁺ concentrations (0.3–2 mM NiCl₂) on the expression of a number of important stress response genes in model plants Arabidopsis thaliana (L.) Heynh., as well as the effect of a natural plant protective agent, L-histidine (His), which is intensively synthesized by plants under nickel stress and is able to bind Ni²⁺, on this process. In the experiments conducted using the Real-Time Polymerase Chain Reaction (RT-PCR) method, it was found that the introduction of Ni²⁺ into the culture medium caused a dose-dependent increase in the relative expression of genes encoding glutathione reductase (GR1), NADPH oxidase (RBOHC), Ca²⁺-dependent protein kinase (CPK6), catalase (CAT2) and outward-rectifying K⁺ channel (GORK1). The maximum increase was observed upon treatment with 2 mM Ni²⁺, relative transcript levels were 5.9, 5.0, 3.0, 2.8 and 2.2 times higher than in control for RBOHC, GR1, CPK6, CAT2 and GORK1 respectively. In the case of the genes encoding poly(ADP-ribose)-polymerase (PARP1), cyclin B2 (CYCB2), and Cu/Zn-superoxide dismutase (CSD2), the transcript levels increased at low Ni²⁺ concentrations and then decreased at higher Ni²⁺ concentrations in the medium. The introduction of His, in conjunction with Ni²⁺, prevented the Ni²⁺-induced change in gene expression. Thus, it was demonstrated that A. thaliana plants respond to excess Ni²⁺ by inducing the expression of enzymatic antioxidants, proteins involved in redox- and Ca²⁺-mediated cellular signaling. This response is accompanied by alterations in the systems of control cell division and DNA replication, which can be regulated by exogenous His.

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