Stable transformation of the unicellular lichenic green alga Coccomyxa solorinae-saccatae via electroporation

Abstract

In this study, we have demonstrated the applicability of electroporation for the stable nuclear transformation of Coccomyxa solorinae-saccatae. An antibiogram revealed that Hygromycin B and G418 are the most effective selective agents among eight different antibiotics tested. We have shown that a plasmid vector containing the hptII gene, coding for hygromycin B phosphotransferase, with expression driven by the strong cauliflower mosaic virus CaMV35S promoter ensures sufficient protection of transformed algal cells against high concentrations of Hygromycin B. The ability to drive transgene expression in the alga C. solorinae-saccatae offers unique opportunities to study the physiology of lichenic algae, as it is one of the symbiotic strains of the Coccomyxa simplex group. Furthermore, our findings demonstrate that electroporation is a convenient and effective technique for the transformation of algae in the Coccomyxa genus.