Molecular, biochemical and morphological data suggest an affiliation of <i>Spongiochrysis hawaiiensis</i> with the Trentepohliales (Ulvophyceae, Chlorophyta)

The aeroterrestrial, unicellular green alga Spongiochrysis hawaiiensis had been included in the ulvophycean order Cladophorales based on small subunit (SSU) rDNA sequence data, and represents so far the only fully terrestrial member of this order. Other characteristics of S.?hawaiiensis that are atypical for Cladophorales include the presence of large amounts of carotenoids and a budding-like mode of cell division. As the position of this terrestrial, unicellular alga in an order of aquatic, multicellular green algae is unusual, we re-evaluated the phylogenetic relationships of this enigmatic organism based on supplementary SSU rDNA sequences as well as novel large ribosomal subunit (LSU) rDNA and internal transcribed spacer (ITS rDNA) sequences. Additionally, we examined several morphological characters of S.?hawaiiensis, as well as low molecular weight carbohydrate (LMWC) patterns of S.?hawaiiensis and members of the Cladophorales and Trentepohliales as potential chemotaxonomic markers. We found S.?hawaiiensis to be uninucleate. The analysis of the LMWC content detected the presence of the polyol erythritol in S.?hawaiiensis and in the Trentepohliales, while this compound was missing in the Cladophorales. The phylogenetic analyses of the novel sequences placed S.?hawaiiensis in the terrestrial Trentepohliales. This placement is supported by the aeroterrestrial habitat, the presence of large amounts of carotenoids, the uninucleate cells, and the presence of the polyol erythritol as a protective compound against water loss