15N metabolic test for the determination of phytotoxic effects of chemicals and contaminated environmental samples.

A stable isotope(15)N-nitrogen test (ESIMA = Ecotoxicological Stable Isotope Metabolic Assay) was developed to assess biological effects and the potential toxicological hazard of chemicals and contaminated environmental samples on plant metabolism. The assay measures the effect of toxicants on the incorporation of a(15)N labelled tracer into the total nitrogen fraction (both the nonprotein and protein fraction) of plants. Segments of Pisum arvense epicotyls are used as test substrates because of their high metabolic activity. The plant material is incubated under standardised conditions for two hours; subsequently(15)N incorporation is analysed by determining the(15)N abundance ((15)N atom-%) in the epicotyl segments. The effects of toxicants are evaluated by comparing the(15)N incorporation rates of control tissue and epicotyl segments exposed to individual chemicals or complex environmental samples. The specificity and sensitivity of effects as indicated by ESIMA were compared with effects as measured by two established ecotoxicological bioassays, the pollen tube growth test using pollen of Nicotiana sylvestris and the bacterial luminescence inhibition test using pollen of Photobacterium phosphoreum. The results of the study clearly indicate the suitability of ESIMA for assessing toxic impacts on plant nitrogen metabolism.