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使用桑格测序法和Illumina MiSeq测序法对孙德尔本斯红树林生态区浮游细菌群落结构的洞察:一项比较分析。

Insights into bacterioplankton community structure from Sundarbans mangrove ecoregion using Sanger and Illumina MiSeq sequencing approaches: A comparative analysis.

作者信息

Ghosh Anwesha, Bhadury Punyasloke

机构信息

Integrative Taxonomy and Microbial Ecology Research Group, Department of Biological Sciences, Indian Institute of Science Education and Research Kolkata, Mohanpur, 741246 Nadia, West Bengal, India.

出版信息

Genom Data. 2016 Nov 24;11:39-42. doi: 10.1016/j.gdata.2016.11.017. eCollection 2017 Mar.

Abstract

Next generation sequencing using platforms such as Illumina MiSeq provides a deeper insight into the structure and function of bacterioplankton communities in coastal ecosystems compared to traditional molecular techniques such as clone library approach which incorporates Sanger sequencing. In this study, structure of bacterioplankton communities was investigated from two stations of Sundarbans mangrove ecoregion using both Sanger and Illumina MiSeq sequencing approaches. The Illumina MiSeq data is available under the BioProject ID PRJNA35180 and Sanger sequencing data under accession numbers KX014101-KX014140 (Stn1) and KX014372-KX014410 (Stn3). Proteobacteria-, Firmicutes- and Bacteroidetes-like sequences retrieved from both approaches appeared to be abundant in the studied ecosystem. The Illumina MiSeq data (2.1 GB) provided a deeper insight into the structure of bacterioplankton communities and revealed the presence of bacterial phyla such as Actinobacteria, Cyanobacteria, Tenericutes, Verrucomicrobia which were not recovered based on Sanger sequencing. A comparative analysis of bacterioplankton communities from both stations highlighted the presence of genera that appear in both stations and genera that occur exclusively in either station. However, both the Sanger sequencing and Illumina MiSeq data were coherent at broader taxonomic levels. , , and like sequences were the abundant bacterial genera found in the studied ecosystem. Both the sequencing methods showed broad coherence although as expected the Illumina MiSeq data helped identify rarer bacterioplankton groups and also showed the presence of unassigned OTUs indicating possible presence of novel bacterioplankton from the studied mangrove ecosystem.

摘要

与传统分子技术(如采用桑格测序的克隆文库方法)相比,使用Illumina MiSeq等平台进行的下一代测序能够更深入地洞察沿海生态系统中浮游细菌群落的结构和功能。在本研究中,采用桑格测序和Illumina MiSeq测序方法,对孙德尔本斯红树林生态区两个站点的浮游细菌群落结构进行了调查。Illumina MiSeq数据可在生物项目编号PRJNA35180下获取,桑格测序数据的登录号为KX014101 - KX014140(站点1)和KX014372 - KX014410(站点3)。从两种方法中检索到的变形菌门、厚壁菌门和拟杆菌门样序列在研究的生态系统中似乎很丰富。Illumina MiSeq数据(2.1GB)更深入地洞察了浮游细菌群落的结构,并揭示了放线菌门、蓝细菌门、柔膜菌门、疣微菌门等细菌门类的存在,而基于桑格测序未检测到这些门类。对两个站点浮游细菌群落的比较分析突出了在两个站点都出现的属以及仅在其中一个站点出现的属。然而,在更广泛的分类水平上,桑格测序和Illumina MiSeq数据是一致的。 、 、 和 样序列是研究生态系统中丰富的细菌属。两种测序方法显示出广泛的一致性,尽管正如预期的那样,Illumina MiSeq数据有助于识别更罕见的浮游细菌群体,还显示出未分类的操作分类单元的存在,这表明研究的红树林生态系统中可能存在新的浮游细菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/44a2/5133466/5a08a8ac4aee/gr1a.jpg

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