Toxic effects of Ni(2+) on the chloroplasts of the two duckweed species Spirodela polyrhiza, clone SJ and Lemna minor, clone St were investigated according to the ISO 20079 protocol. Ni(2+) induced a transition from chloroplasts to chloro-amyloplasts and amylo-chloroplasts, but not to gerontoplasts, as shown by electron microscopy. The contents of the chlorophylls a and b decreased strongly, whereas that of carotenoids remained approximately constant. Most striking was, however, the accumulation of transitory starch. Bell-shaped dose-response curves showed that Spirodela and Lemna amassed maximum starch contents of approximately 10% and 7%, respectively, on a fresh weight basis. Because Ni(2+) in the concentrations applied does not stimulate photosynthesis, the Ni(2+)-induced starch accumulation indicates that the export of carbohydrates out of the plastids decreased, most probably due to the lower demand of the rest of the cells as a result of the Ni(2+)-dependent inhibition of growth. The half-maximal concentrations for inhibition of the fresh weight increase over the 7-day test period were 3.7 microM and 6.6 microM for Spirodela and Lemna, respectively: Spirodela was thus somewhat more sensitive to the heavy metal. Both species accumulated approximately 3g of Ni(2+) per kg of dry weight after application of 100 microM NiCl(2). Because of their high sensitivity to phytotoxic effects, however, Spirodela and Lemna do not appear to be particularly suitable for phytoremediation of Ni(2+)-contaminated waste water. The high sensitivity to Ni(2+) makes them instead a suitable system for ecotoxicological testing in accordance with the ISO 20079 protocol.