侵染墨氏中华根瘤菌噬菌体主要衣壳基因g23的分离及系统发育分析。

微生物学报 Pub Date : 2017-02-04
Hao Yu, Junjie Liu, Guoquan Fan, Guanghua Wang
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引用次数: 0

摘要

目的:为研究侵染中华根瘤菌的噬菌体生态学提供科学依据,对侵染中华根瘤菌的噬菌体形态特征及其主要captain蛋白g23的系统发育状况进行研究。方法:采用双层平板法分离根瘤体,病原菌USDA1002T。用透射电镜研究了根瘤体的形态特征。同时提取根瘤体DNA,选择编码噬菌体主要衣壳蛋白的g23作为目的基因进行PCR扩增。结果:分离到3个根瘤体,均具有直径约81 ~ 86 nm的二十面体头和54 ~ 70 nm长的可收缩尾。国家生物技术信息中心(NCBI)网站的基本局部比对工具检索结果显示,本研究获得的g23氨基酸序列之间具有较高的同源性,但与t -even、pseudot -even、schizot -even和exot -even的同源性较低。系统发育分析表明,分离的g23序列与来自不同生态系统的克隆形成了一个独特的分支。结论:分离得到的根瘤菌属T4噬菌体新类群肌病毒科,与在不同环境下获得的g23无性系同源性较低。
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[Isolation and phylogenetic analysis of major capsid gene (g23) of bacteriophages infecting Sinorhizobium meliloti].

Objective: In order to provide scientific data for studying the ecology of phage infecting Sinorhizobium meliloti, we examined morphological characteristics of rhizobiophages and their phylogenetic status of the major captain protein g23.

Methods: Rhizobiophages were isolated by the double-layer plate method with host Sinorhizobium meliloti USDA1002T. The morphological characteristic of rhizobiophages were studied by transmission electron microscope. Meanwhile, rhizobiophage DNA was extracted, and the g23 that encodes the major capsid protein of bacteriophages was chosen as objective gene in PCR amplification.

Results: Three rhizobiophages were isolated, all had an icosahedral head with approximately 81 to 86 nm in diameter and a long contractile tail with 54 to 70 nm in length. Basic local alignment search tool searches in website of national center for biotechnology information (NCBI) revealed that the g23 amino acid sequences obtained in this study had high identity with each other, but had very lower identity with those from T-evens, PseudoT-evens, SchizoT-evens and ExoT-evens. Phylogenetic analysis showed that the isolated g23 sequences formed a unique clade with those clones obtained from different ecosystem.

Conclusion: All results indicated that the isolated rhizobiophages belong to family Myoviridae, a new group of T4 phages, which had lower identity with the g23 clones obtained in different environment.

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