Micropropagation of Myriophyllum alterniflorum (Haloragaceae) for stream rehabilitation: first in vitro culture and reintroduction assays of a heavy-metal hyperaccumulator immersed macrophyte.

Nowadays, submersed aquatic macrophytes play a key role in stream ecology and they are often used as biomonitors of freshwater quality. So, these plants appear as natural candidates to stream rehabilitation experiments. Among them, the stream macrophyte Myriophyllum alterniflorum is used recently as biomonitor and is potentially useful for the restoration of heavy-metal contaminated localities. The best way to obtain a mass production of watermilfoil plants is micropropagation. We developed in vitro culture of M. alterniflorum and the effects of five media on the plant development were assessed. Five morphological and four physiological endpoints were examined leading to the recommendation of the Murashige and Skoog medium for ecotoxicological studies on chlorophyllous parts, and of the Gaudet medium for root cytotoxicity and phytoremediation studies. Micropropagated clones were acclimatized in a synthetic medium and in situ reintroduction was performed efficiently. This is the first report of micropropagated plants transplantation in streams. The successful establishment of watermilfoil beds even in polluted areas strongly suggested that ecological restoration using micropropagated watermilfoil is a promising biotechnology for phytoremediation and rehabilitation of degraded areas. Moreover, high bioconcentration factors evidenced that watermilfoil hyperaccumulates Cd and Cu, and could be potentially used in phytoremediation studies.