Effect of cryopreservation on the bacterial community structure of filamentous cyanobacteria, Trichormus variabilis (Nostocales, Cyanobacteria).

Cryopreservation is an efficient method used to preserve microorganisms for long periods of time, such as up to 30 years, without changes in genetic and physiological characteristics. As cyanobacteria and microalgae are usually maintained as both axenic and xenic cultures, knowledge of co-cultured bacteria and changes in their community structure is important for the successful maintenance of microbial culture collections. In this study, research on the changes in co-cultured bacterial community structure during cyanobacterial cryopreservation were investigated using three different experimental groups by next generation sequencing (NGS): 1) cultured Trichormus variabilis without cryopreservation (control group), 2) cultured T. variabilis after cryopreservation in 10% dimethyl sulfoxide (MeSO) for 14 days (cryo-cell group), and 3) cultured T. variabilis after cryopreservation in 10% MeSO for 14 days within alginate beads (cryo-bead group). The results showed that the abundance of Sphingomonas and Hydrogenophaga (belonging to phylum Proteobacteria) was significantly increased in the cryo-bead group (Sphingomonas, control: 0.25%, cryo-cell: 1.32%, cryo-bead: 41.70%; Hydrogenophaga, control: 5.47%, cryo-cell: 5.24%, cryo-bead: 12.32%). However, the abundance of the phylum Bacteroidetes was significantly decreased in the cryo-bead group compared to that in the other groups (control: 26.29%, cryo-cell: 38.84%, cryo-bead: 11.43%). Bacterial diversity was generally reduced after cryopreservation in the cryo-bead group, where the overgrowth of a few unique bacteria was observed in the co-cultured bacterial community. These results imply that changes in the co-cultured bacterial community during preservation should be considered as an important factor for the development of methods for cyanobacterial cryopreservation.