Freeze survival of the cyanobacteria Microcoleus chthonoplastes without cryoprotector

A Microcoleus chthonoplastes strain SC7B9002-1 isolated from microbial mats in tidal channels from San Carlos, Baja California Sur, Mexico was subjected to short- (15 days) and long-term (2 years) conservation assays in liquid nitrogen (-196 degrees C) using cryoprotective agents, such as 5% DMSO, 20% PVP-40, and 20% glycerol. Survival rate, chlorophyll a, protein, and nucleic acids content were observed in each case. Interesting growth and a significant increase in protein content was observed when no cryoprotectant was used during liquid nitrogen immersion. In the absence of a cryoprotectant, M. chthonoplastes lost their typical shape resembled spheroplasts, and recovery cultivation times after freezing were 5 and 25 days (short and long-term, respectively). Recovery from long-term preservation with 5% DMSO took 15 days. PVP and glycerol did not allow recovery of viable cells. The survival of M. chthonoplastes to freezing without cryoprotectant and the adaptive mechanisms that allow surviving under freezing conditions are discussed.