基于454和Illumina高通量测序平台对长江口邻近海域沉积物微生物群落的比较分析
Comparative analysis of microbial community in the sediments from the adjacent area of Yangtze Estuary by 454 pyrosequencing and illumina sequencing
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摘要: 作为第二代测序技术的主要平台, 454和Illumina测序方法在微生物核酸分析方面有着广泛应用。本研究以16S rRNA基因为标靶, 分别采用454和Illumina测序技术对长江口邻近海域表层沉积物中的微生物DNA进行分析, 通过对序列长度、微生物群落多样性及组成进行比较, 分析两种方法在沉积物微生物群落结构研究方面的适用性。结果表明, Illumina测序获得的序列数和OTU数要多于454, 其中获得的OTUs中以非优势菌群为主。Illumina与454相比能够检出更多微生物菌群, 且检测到的稀有菌群占有较大比例。综上可知, Illumina测序技术在分析微生物群落多样性及物种组成方面要优于454测序技术, 454的长序列优势并未能体现出来, 且454测序技术可能低估了微生物组成及其多样性。Abstract: As the main methods of the second generation high-throughput sequencing technologies, 454 pyrosequencing and Illumina sequencing have been widely utilized in the microbial nucleic acid analysis.In this study, sequence length, microbial compositions, abundance and diversity in the sediments from the adjacent area of Yangtze Estuary were analyzed using 16S rRNA gene-based 454 pyrosequencing and Illumina sequencing technology to compare the advantages and disadvantages of two techniques.The results showed that more sequences and OTUs were obtained by Illumina sequencing, which included many rare species.Illumina sequencing technology had an advantage of microbiological community analysis over 454 pyrosequencing technology, especially in the detection of rare species.In summary, Illumina sequencing technology was better than 454 pyrosequencing technology in the analysis of microbial diversity and community composition.454 pyrosequencing didn't reflect its advantage in long read length, and it probably underestimated the microbial composition and diversity.