Impact of phenanthrene exposure on activities of nitrate reductase, phosphoenolpyruvate carboxylase, vacuolar H+-pyrophosphatase and plasma membrane H+-ATPase in roots of soybean, wheat and carrot

• Phenanthrene exposure enhances nitrate reductase and vacuolar H+-pyrophosphatase.
• Phosphoenolpyruvate carboxylase is inhibited by phenanthrene.
• Plasma membrane H+-ATPase plays an essential role in phenanthrene uptake.
• The above four enzymes cannot explain the species difference in phenanthrene uptake.

Crops differ in uptake of polycyclic aromatic hydrocarbons (PAHs). However, the physiological mechanism underlying the differences in PAH uptake among crop species is unclear. As PAH transport by H+-coupled symporters may affect cytosolic pH, and intracellular pH is precisely regulated for metabolism, four enzymes linked with cytosolic pH regulation in root cells were determined at different concentrations of phenanthrene (PHE) and pHs. Activity of nitrate reductase (NRase) is stimulated by PHE and follows the order: wheat > carrot > soybean. The optimum pH of NRase is 7.5 in wheat and carrot, while 6.5 in soybean. The activity of phosphoenolpyruvate carboxylase (PEPCase) is generally decreased by PHE and always lower in soybean than in wheat and carrot. PEPCase peaks its maximum activity at pH 7.0 and wheat shows the highest value with high PHE (1.2 mg L−1) at pHs of 7.0 and 8.0 in comparison with carrot and soybean. Vacuolar H+-pyrophosphatase (V-H+-PPase) is promoted by PHE in wheat and carrot, while it does not change significantly in soybean (P > 0.05). V-H+-PPase reaches its maximum value at pH 7.5 and follows the order of wheat > carrot > soybean at alkaline medium with high PHE. Plasma membrane (PM) H+-ATPase is enhanced by K+ and follows the order: wheat > soybean > carrot. The orders of activity of these enzymes are not consistent with that of PHE uptake, indicating their various functions. However, in soybean, PM H+-ATPase activity under high PHE and the uptake of PHE are always higher than those in wheat and carrot, suggesting that PM H+-ATPase plays an essential role in PHE uptake. Therefore, the changes of PM H+-ATPase activity seem to be a direct consequence responding to PHE stress and those of NRase, PEPCase and V-H+-PPase might be the downstream accidents regulated by cytosoilc pH. Our study provides insight into the roles of these enzymes in non-photosynthetic tissues and supports their importance in the physiological responses to PHE stress among different crops.