Resistance of Surface Phosphohydrolases of Barley Root Cells to Nickel Salts Is One of Factors of Plant Adaptation to Excessive Nickel in the Environment

Abstract

We studied the effect of nickel ions on the activity of ecto-phosphohydrolases (acid phosphatase and Ca-stimulated nucleotidase) from root surface of etiolated barley seedlings as well as from root microsomal fraction. The presence of nickel nitrate (25 microM) proved to stop root growth and insignificantly (on average by 20%) decreased specific hydrolytic activity of both enzymes determined on root surface as well as in the root microsomal fraction. At the same time, direct addition of nickel to the incubation mixture when measuring the substrate hydrolysis demonstrated high resistance of the microsomal fraction enzymes to the salts. A significant decrease in Ca-stimulated nucleotidase activity was observed only for nickel nitrate concentrations above 100 microM, reaching 50-60% for 3 mM Ni(NO3)2. The presence of an activator ion as well as extended duration of the microsomal fraction pretreatment with nickel nitrate (2.5 h) did not increase its effect on the enzyme activity. The pattern of nickel effect on acid phosphatase activity depended on the presence of magnesium ions in the mixture but did not change after extended duration of the microsomal fraction pretreatment (3 h). Inhibition of acid phosphatase activity in the presence of magnesium was observed only for nickel nitrate concentrations above 500 microM being no more than 20% for 3 mM Ni(NO3)2. Hence, the hydrolytic enzymes of the apoplast of plant root cells have different tolerance to to nickel salts. We propose that an insignificant decrease in specific activity of surface hydrolases of plant roots grown on a medium containing nickel salts in concentrations inhibiting growth processes (25 microM) is not related to direct effect of Ni on the apoplastic enzymes. The significance of hydrolytic enzyme resistance in plant adaptation to high nickel content in the soil is discussed.