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利用一组新引物对主要衣壳蛋白基因进行高通量测序揭示了不同水生环境中肌尾噬菌体的高度多样性

High Diversity of Myocyanophage in Various Aquatic Environments Revealed by High-Throughput Sequencing of Major Capsid Protein Gene With a New Set of Primers.

作者信息

Hou Weiguo, Wang Shang, Briggs Brandon R, Li Gaoyuan, Xie Wei, Dong Hailiang

机构信息

State Key Laboratory of Biogeology and Environmental Geology, China University of Geosciences, Beijing, China.

CAS Key Laboratory of Environmental Biotechnology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences (CAS), Beijing, China.

出版信息

Front Microbiol. 2018 May 3;9:887. doi: 10.3389/fmicb.2018.00887. eCollection 2018.

Abstract

Myocyanophages, a group of viruses infecting cyanobacteria, are abundant and play important roles in elemental cycling. Here we investigated the particle-associated viral communities retained on 0.2 μm filters and in sediment samples (representing ancient cyanophage communities) from four ocean and three lake locations, using high-throughput sequencing and a newly designed primer pair targeting a gene fragment (∼145-bp in length) encoding the cyanophage gp23 major capsid protein (MCP). Diverse viral communities were detected in all samples. The fragments of 142-, 145-, and 148-bp in length were most abundant in the amplicons, and most sequences (>92%) belonged to cyanophages. Additionally, different sequencing depths resulted in different diversity estimates of the viral community. Operational taxonomic units obtained from deep sequencing of the MCP gene covered the majority of those obtained from shallow sequencing, suggesting that deep sequencing exhibited a more complete picture of cyanophage community than shallow sequencing. Our results also revealed a wide geographic distribution of marine myocyanophages, i.e., higher dissimilarities of the myocyanophage communities corresponded with the larger distances between the sampling sites. Collectively, this study suggests that the newly designed primer pair can be effectively used to study the community and diversity of myocyanophage from different environments, and the high-throughput sequencing represents a good method to understand viral diversity.

摘要

肌动蛋白噬菌体是一类感染蓝藻的病毒,数量众多,在元素循环中发挥着重要作用。在此,我们使用高通量测序技术和新设计的引物对,针对编码肌动蛋白噬菌体gp23主要衣壳蛋白(MCP)的基因片段(长度约为145bp),研究了从四个海洋和三个湖泊地点采集的保留在0.2μm滤膜上的颗粒相关病毒群落以及沉积物样本(代表古代肌动蛋白噬菌体群落)。在所有样本中均检测到了多样的病毒群落。扩增子中长度为142bp、145bp和148bp的片段最为丰富,且大多数序列(>92%)属于肌动蛋白噬菌体。此外,不同的测序深度导致对病毒群落多样性的估计不同。从MCP基因深度测序获得的操作分类单元涵盖了从浅度测序获得的大部分单元,这表明深度测序比浅度测序更能完整地呈现肌动蛋白噬菌体群落的情况。我们的研究结果还揭示了海洋肌动蛋白噬菌体广泛的地理分布,即肌动蛋白噬菌体群落的差异越大,采样地点之间的距离就越大。总体而言,这项研究表明新设计的引物对可有效用于研究不同环境中肌动蛋白噬菌体的群落和多样性,高通量测序是了解病毒多样性的一种良好方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0e32/5943533/fbc51d004a6d/fmicb-09-00887-g001.jpg

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