The study of biodiversity of phenol-degrading bacteria by rep-PCR

Recent progress in molecular microbial ecology hasrevealed that traditional culturing methods fail torepresent the scope of microbial diversity in nature,since only a small proportion of viable microorganismsin a sample are recovered by culturing techniques. Todevelop methods to investigate the full extent of microbialdiversity, we used metagenomic studies for phenoldegrading bacteria in a petroleum contaminated soil.Metagenomics is an application of modern genomicstechniques to the study of communities of microbialorganisms directly in their natural environments,bypassing the need for isolation and lab cultivation ofindividual species .Therefore, in this metagenomic study,and to demonstratethe diversity of isolated bacteria,genomic fingerprinting was investigated byrepetitiveextragenic palindromic sequence PCR (rep-PCR).However, bacteria were isolated from contaminated soilby plating after enrichment in batch cultures.Then, totalbacterial DNA was extracted by set-buffer method andrep-PCR performed with primers REP1R-I and REP2-I.Different patterns were obtained by separation of PCRproducts by agarose gel electrophoresis.However,the rep-PCR analysis was repeated several timesto determine the reproducibilityof the method. Thebacterial cells isolated were classified into 18 distinctgroups by a repetitive extragenic palindromic sequence PCR analysis. It is established that molecular approacheshaveexpanded our knowledge of the diversity anddistributionof microbial populations in the environment.On the other hand, the REP PCR could becomea powerfultool for the molecular genetic analysis of bacteriaand forbacterial taxonomy, since it allows the fingerprintingofindividual genera, species, and strains and couldhelpdetermine phylogenetic relationships. The same isexpected in metagenomics

کلیدواژه ها:

metagenomics ، biodiversity ، rep-PCR ، biodegradation