The potential contamination of the food chain is the most important aspect of arsenic (As) pollution, since it is highly toxic to all organisms. Thus, the search for As hyperaccumulators suitable to remove As from contaminated soils appears to be a vital task. Horseradish (Armoracia rusticana), a crop plant with a high potential to accumulate heavy metals, can also serve to study the physiological processes that accompany arsenic stress. The significant adverse effect caused by arsenic exposure is an oxidative stress. Plants have developed a highly organized system to quench free radicals, which includes the action of both enzymatic and non-enzymatic quenching. Saccharides are proposed to possess outstanding antioxidant activity in vitro, and thus, they are likely to effectively quench free radicals also in plant tissues. In this study, root cultures (hairy root type) of horseradish were grown in vitro on media with different concentrations of arsenic (5-60 µg l-1). Arsenic slowed down the growth, nevertheless up to three-fold biomass increase was achieved at the highest dose. Moreover, root tissues were able to remove as much as 75% of arsenic from the cultivation medium within 7 days. We also evaluated diverse oxidative-stress-related features: contents of reactive oxygen species, the activities of key antioxidant enzymes, and the contents of important antioxidant molecules, such as glutathione, proline, phenolic compounds and non-structural carbohydrates. At all arsenic treatments, we observed a significant proline content increase and enhanced antioxidant enzymes (peroxidase, catalase and glutathione-S-transpherase) activities peaking, however, at different doses. Soluble carbohydrates contents also significantly increased after 7-day treatment a then decreased nearly to the original levels. This study points to efficient antioxidant system of horseradish hairy roots enabling good growth and substantial As accumulation even under high As exposure. Providing that horseradish shares these important features with this model system, we could propose that horseradish is a promising candidate to exploit in arsenic phytoremediation.

Department of Experimental Plant Biology Faculty of Science Charles University Viničná 5 Prague 2 128 44 Czech Republic

Department of Genetics and Microbiology Faculty of Science Charles University Viničná 5 Prague 2 128 44 Czech Republic

Faculty of Agrobiology Food and Natural Resources Czech University of Life Science Kamýcká 961 129 Prague 6 165 00 Czech Republic

Laboratory of Plant Biotechnologies Institute of Experimental Botany AS CR Rozvojová 313 Prague 6 165 02 Czech Republic

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