Role of SulP, a nuclear-encoded chloroplast sulfate permease, in sulfate transport and H2 evolution in Chlamydomonas reinhardtii.

Antisense technology was applied to the green alga Chlamydomonas reinhardtiito probe the function of a novel nuclear gene encoding a chloroplast-envelope localized sulfate permease (SulP; GenBank Accession Numbers AF467891 and AF481828). Analysis showed that antiSulP transformants are impaired in sulfate uptake, a consequence of repression in the SulP gene expression. Antisense antiSulP transformants exhibited a sulfur-deprivation phenotype, strong induction of arylsulfatase activity, and global induction of sulfate assimilation gene expression. In sealed cultures, opposite to the wild-type control, antiSulP strains photo-evolved H2, underlining the notion of sulfate uptake limitation by the chloroplast, a slow-down in the rate of oxygen evolution, establishment of anaerobiosis due to internal respiration and spontaneous expression of the [Fe]-hydrogenase in these strains. It is concluded that antiSulP strains are promising as tools to limit the supply of sulfates to the chloroplast, leading to a down-regulation of H2O-oxidation and O2-evolution activity, to the constitutive expression of the [Fe]-hydrogenase and continuous H2-photoproduction in Chlamydomonas reinhardtii.Thus, antisulPstrains might permit a study of the biochemistry of H2 metabolism in this green alga under constitutive anaerobic oxygenic photosynthesis conditions.