Toxicity testing of heavy metals with theRhizobium-legume symbiosis: High sensitivity to cadmium and arsenic compounds

Legume root nodules are the site of biological nitrogen fixation in theRhizobium-legame symbiosis. Nodules are structures unique to this symbiosis and they are morphologically as well as physiologically distinct from other plant organs. Organic substances affecting the macro- or microsymbionts vitality, such as PAHs (Wetzel et al., 1991), reduce nodulation even before visible damage to the plant can be detected. We present data that the formation of nodules (nodulation) may also serve for ecotoxicological evaluation of heavy metals in different binding states. Tests were performed in petri dishes with alfalfa (lucerne) seedlings inoculated withRhizobium meliloti. Cultivation took place in growth cabinets with carefully standardized and documented growth conditions. Data from stressed plants was recorded after 14 days of cultivation on contaminated substrate. A dose responsive decrease in nodulation was found after application of cadmium acetate, cadmium iodide, cadmium chloride, sodium salts of arsenate and arsenite, arsenic pentoxide, and lead nitrate, whereas lead acetate showed no effect up to a concentration of 3 μM. The dose response curves were used to calculate EC10, EC50 and EC90 values. EC50 values for cadmium compounds range from 1.5 to9.5 pM. Testing different arsenic compounds results in EC50 from 2.6 to 20.1 μM. EC50 of lead nitrate is 2.2 μM. The sensitivity, reproducibility and reliability of this test system is discussed compared to established biotests.