Screening of PAH-degrading bacteria in a mangrove swamp using PCR-RFLP

There are abundant PAH-degrading bacteria in mangrove sediments, and it is very important to screen the high efficiency degraders in order to perform bioremediation of PAH polluted environments. In order to obtain the more highly efficient PAH-degrading bacteria from a mangrove swamp, we first obtained 62 strains of PAH-degrading bacteria using traditional culture methods and based on their morphological characteristics. We then used the modern molecular biological technology of PCR-RFLP, in which the 16S rDNA of these strains were digested by different enzymes. Based on differences in the PCR-RFLP profiles, we obtained five strains of phenanthrene-degrading bacteria, five strains of pyrene-degrading bacteria, four strains of fluoranthene-degrading bacteria, five strains of benzo[a]pyrene-degrading bacteria and two strains of mixed PAH-degrading bacteria (including phenanthrene, pyrene, fluoranthene and benzo[a]pyrene). Finally, a total of 14 different PAH-degrading bacteria were obtained. The 16S rDNA sequences of these strains were aligned with the BLAST program on the NCBI website and it was found that they belonged to the alpha-proteobacteria and gamma-proteobacteria, including four strains, where the similarities were no more than 97% and which were suspected therefore to be new species. This study indicated that PCR-RFLP was a very important method to screen degrading-bacteria, and also a significant molecular biological tool for the rapid classification and accurate identification of many different strains. On the other hand, it also showed that rich bacterial resources existed in mangrove areas, and that exploring and developing the functional microorganism from these mangrove areas would have wide use in the study of bioremediation of contaminated environments in the future. (C) 2010 Elsevier Ltd. All rights reserved.