Seasonal changes and diversity of bacteria in Bohai Bay by RFLP analysis of PCR-amplified 16S rDNA gene fragments

Information about seasonal bacterial composition and diversity is of great value for exploitation of marine biological resources and improvement of ecological environment. Here PCR-amplified restriction fragment length polymorphism (PCR–RFLP) of 16S rRNA genes was used to evaluate seasonal bacterial diversity and community composition in Bohai Bay. A total of 24 bacterial communities were sampled from seawater and sediment of three representative sites in a whole seasonal cycle: spring (April), summer (July), autumn (October), and winter (January). Bacterial Genomic DNA was extracted and PCR-amplified to obtain 16S rDNA fragments which were cloned to construct 24 16s rDNA libraries. Clones of each library were selected randomly for PCR–RFLP analysis of rDNA fragments, and eventually 101 genotypes were identified by RFLP fingerprintings. These 101 genotypes were sequenced and their respective phylotype was identified through the Blast tool of NCBI (similarity 96–100%) and phylogenetic analyses. Among our phylotypes, 80.2% belonged to the genera α-Proteobacteria, β-proteobacteria,γ-Proteobacteria, δ-Proteobacteria, e-proteobacteria, Flavobacteria, Cytophaga-Flavobacteria-Bacteroides, Verrucomicrobia, Firmicutes and Actinobacteria. Sequence analyses revealed that 47.5% (48) of clone sequences were similar to those of uncultured marine bacteria in the environment. In addition, bacterial diversity and composition clearly displayed seasonal variety. More genera were discovered in summer than any other seasons, and some special species appeared only in specific season.