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首页> 外文期刊>World Journal of Microbiology & Biotechnology >Seasonal changes and diversity of bacteria in Bohai Bay by RFLP analysis of PCR-amplified 16S rDNA gene fragments
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Seasonal changes and diversity of bacteria in Bohai Bay by RFLP analysis of PCR-amplified 16S rDNA gene fragments

机译：PCR扩增16S rDNA基因片段的RFLP分析渤海湾细菌的季节性变化和多样性

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摘要

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 alpha -Proteobacteria, beta -proteobacteria, gamma -Proteobacteria, delta -Proteobacteria, epsilon -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.
机译：有关季节性细菌组成和多样性的信息对于开发海洋生物资源和改善生态环境具有重要价值。本文利用PCR扩增的16S rRNA基因的限制性片段长度多态性（PCR-RFLP）来评估渤海湾的季节性细菌多样性和群落组成。在整个季节周期中，从三个代表性地点的海水和沉积物中采样了总共24个细菌群落：春季（4月），夏季（7月），秋季（10月）和冬季（1月）。提取细菌基因组DNA并PCR扩增以获得16S rDNA片段，将其克隆以构建24个16s rDNA文库。随机选择每个文库的克隆进行rDNA片段的PCR-RFLP分析，最终通过RFLP指纹图谱鉴定出101个基因型。对这101个基因型进行了测序，并通过NCBI的Blast工具（相似性96-100％）和系统发育分析鉴定了它们各自的系统型。在我们的系统型中，80.2％属于α-变形细菌，β-变形细菌，γ-变形细菌，δ-变形细菌，ε-变形细菌，黄细菌，食丝藻-黄细菌-拟杆菌，疣状微生物，菌毛和放线菌。序列分析表明，有47.5％（48）的克隆序列与环境中未培养的海洋细菌相似。此外，细菌的多样性和组成清楚地显示出季节性变化。夏季发现的属比其他任何季节都多，一些特殊物种仅在特定季节出现。

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