Isolation of putative type 2 metallothionein encoding sequences and spatial expression pattern in the seagrass Posidonia oceanica

Metallothioneins (MTs) are a group of proteins with low molecular mass and high cysteine content that bind to heavy metals and are thought to play a role in their metabolism and detoxification. Genes encoding MT-like proteins have been isolated in a number of plants. In this work we isolated nine MT-like sequences from copper- or cadmium-exposed plants of the seagrass Posidonia oceanica, a marine Angiosperm playing a major role in maintaining infralittoral ecosystems in the Mediterranean sea. These sequences, together with two other MTgenes previously isolated from this species, show high similarities with genes encoding type 2 MTs. Neighbour-joining analysis, at both deduced protein and 30-UTR sequence level, indicates that at least two subgroups occur within Posidonia type 2 MTs, showing, however, a strong sequence uniformity. Southern analysis of two type 2 MT-encoding sequences (Pomt2b and Pomt2f) belonging to the two different subgroups showed distinct hybridisation patterns. For both type 2 MTs, we have determined, by in situ technique, the expression domain in Posidonia plants. The members of these two MT subgroups show differences in their histological expression, with Pomt2b associated with proliferative tissues whereas Pomt2f is associated with lignified or suberized cell wall.