Effects of Ni²⁺ and Cu²⁺ on K⁺ and H⁺ currents in lily pollen protoplasts

Heavy metals affect plant development and reproduction if they are present in excessive amounts, a situation that is becoming increasingly common. Pollen is a convenient object for pollution assessment as it is in most cases a 2- or 3-cellular organism exposed to the environment. At the same time, pollen is a key stage in the life cycle of seed plants; pollen viability and efficiency of germination are crucial for reproductive success and crop yield. In the present study we reveal for the first time, to our knowledge, targets for heavy metals (Cu2+ and Ni2+) in the pollen grain plasma membrane using the patch-clamp technique. Ni2+ dramatically decreases K+ current in pollen grain protoplasts, whereas Cu2+ does not alter the current density. Instead, Cu2+ strongly enhances H+ current driven by H+-ATPase, whereas Ni2+ fails to affect this current. The short-term treatment with Cu2+ also leads to reactive oxygen species (ROS) accumulation in pollen grain protoplasts but intracellular pH and membrane potential remain unchanged. Ni2+ had no significant effect on ROS content or membrane potential. Thus, plasmalemma K+ channels in pollen grains are sensitive to Ni2+ and H+-ATPase is sensitive to Cu2+, possibly, in a ROS-mediated way. Both metals leave pollen viable since membrane potential is maintained at the control level.