Resistance to UV-B induced DNA damage in extreme-tolerant cryptoendolithic Antarctic fungi: detection by PCR assays

Abstract

Cryptoendolithic Antarctic black fungi are adapted to the harshest terrestrial conditions as in the ice-free area of the McMurdo Dry Valleys. Recently, surviving space simulated conditions proves their bewildering extremotolerance. In order to investigate the potential DNA damage and their response after UV-B exposition, two strains of Antarctic cryptoendolithic black fungi, Cryomyces antarcticus CCFEE 534 and Cryomyces minteri CCFEE 5187, were irradiated at different UV-B doses. Since conventional methods cannot be applied to these organisms, the effect on the genome was assessed by RAPD and rDNA amplification PCR based assays; the results were compared with the responses of Saccharomyces pastorianus DBVPG 6283 treated with the same conditions. Results showed that template activity was drastically inhibited in S. pastorianus after irradiation. Dramatic changes in the RAPD profiles showed after 30 min of exposure while the rDNA amplification of SSU, LSU, and ITS portions failed after 30, 60, and 90 min of exposure respectively. No alteration was detected in the templates of the Antarctic strains where both RAPD profiles and rDNA PCR amplifications were unaffected even after 240 min of exposure. The electroferograms of the rDNA portions of Cryomyces strains were perfectly readable and conserved whilst the analyses revealed a marked alteration in S. pastorianus confirming the high resistance of the Antarctic strains to UV-B exposure.