Evidences showing 2,4-D ethyl ester and pencycuron-induced DNA damage in cyanobacteria and detection by PCR

Abstract

The impact of 2,4-D ethyl ester and pencycuron in inducing DNA damage in three species of cyanobacteria-Anabaena fertilissima, Aulosira fertilissima, and Westiellopsis prolifica as evidenced by PCR-based assays: RAPD and 16S rRNA amplification was examined. Exposure of genomic DNA (in vitro) to pencycuron for 4 days did not produce severe damage in DNA fragments of all three cyanobacterial species whereas exposure to 2,4-D ethyl ester markedly inhibited the template activity of genomic DNA compared to untreated cultures of A. fertilissima. In A. fertilissima a single band of approximately 1000 bp was observed even after 16 days of exposure to 60 ppm pencycuron which suggests that certain segments of DNA are resistant to pencycuron DNA damaging effects. However, a significant effect was observed in the case of W. prolifica for 2,4-D ethyl ester and pencycuron where complete disappearance of fragments was not recorded even after 16 days of incubation and interestingly some new DNA bands were induced. Similar to the effects with RAPD profile, amplification of rRNA was significantly inhibited following exposure of genomic DNA to 2,4-D ethyl ester and pencycuron. Our findings clearly demonstrate that pesticide concentrations affected cyanobacterial DNA and lethality of these microbes might be due to irreversible DNA damage. Thus, it is postulated that PCR assays may be conveniently used for screening DNA damage produced by 2,4-D ethyl ester and pencycuron in all three cyanobacteria examined in this study.