Cryopreservation of anoxygenic phototrophic Fe(II)-oxidizing bacteria

Abstract

Preservation and storage of microbial stock cultures is desirable since the risk of contamination or loss of living cultures is immanent while over long periods mutations accumulate. Generally, it is rather difficult to preserve photosynthetic bacteria due to their sensitive photosynthetic membranes. Phototrophic Fe(II)-oxidizing bacteria face an additional challenge; since they are exposed to light and Fe(II) during growth, they have to cope with radicals from Fenton reactions of Fe(II)-species, light and water. Therefore, phototrophic Fe(II)-oxidizing strains are thought to be especially susceptible to genetic modifications. Here, we provide a simple and fast protocol using glycerol as cryo-protectant to cryopreserve three strains of anoxygenic phototrophic Fe(II)-oxidizing bacteria from different taxa: alpha-proteobacteria, gamma-proteobacteria and chloroflexi. All three strains investigated could be revived after 17 months at -72 degrees C. This suggests that a long-term storage of phototrophic Fe(II)-oxidizing strains is possible.