TiO₂ nanoparticles and salinity stress in relation to artemisinin production and ADS and DBR2 expression in Artemisia absinthium L.

Nanopartículas de TiO₂ e estresse de salinidade em relação à produção de artemisinina e expressão de ADS e DBR2 em Artemisia absinthium L.

Abstract

Artemisia absinthium L. is an important herb that is widely cultivated in different parts of the world for its medicinal properties. The present study evaluated the effects of four concentrations of nanoparticles treatment (0, 10, 20 and 30 mg L^-1) and NaCl salinity stress (0, 50, 100 and 150 mM NaCl) and their interactions with respect to the expression of two key genes, i.e. DBR2 and ADS, in the biosynthesis pathway of artemisinin in A. absinthium. Total RNA was extracted and a relative gene expression analysis was carried out using Real-Time PCR. The amount of artemisinin was also determined by HPLC. All the experiments were performed as factorial in a completely randomized design in three replications. The results revealed that salinity stress and nanoparticles treatment and their interaction affected the expressions of these genes significantly. The highest levels of ADS gene expression were observed in the 30 mg L^-1 nanoparticles–treated plants in the presence of 150 mM salinity stress and the lowest levels in the 10 mg L^-1 nanoparticles–treated plants under 50 mM salinity stress. The maximum DBR2 gene expression was recorded in the 10 mg L^-1 nanoparticles–treated plants in the absence of salinity stress and the minimum expression in the 100 mM salinity-stressed plants in the absence of nanoparticles treatment. Moreover, the smallest amounts of artemisinin were observed in the 150 mM salinity-stressed plants in the absence of nanoparticles and the highest amounts in the 30 mg L^-1 nanoparticles–treated plants. The maximum amounts of artemisinin and ADS gene expression were reported from the plants in the same nanoparticles treatment and salinity stress conditions. In this regard, the amount of artemisinin was decreased by half in the plants containing the highest DBR2 gene expression. Meanwhile, no significant correlation was observed between these gene expressions and the artemisinin amount in the other nanoparticles–treated plants under different levels of salinity stress. The biosynthetic pathway of secondary metabolites appears to be very complex and dose not directly dependent on these gene expressions.

Keywords:
artemisinin; gene expression; salinity stress; titanium dioxide nanoparticles; wormwood