一株枯草芽孢杆菌噬菌体Bac-S的生物学特性及全基因组序列分析  

Biological characterization and genome sequence of a bacteriophage strain Bac-S infecting Bacillus subtilis

作　　者：赵欣卓 王鑫淼 王丽艳 吴楠 李喜梅 于镇华[2] 荆瑞勇 刘俊杰[2] ZHAO Xinzhuo;WANG Xinmiao;WANG Liyan;WU Nan;LI Ximei;YU Zhenhua;JING Ruiyong;LIU Junjie(Heilongjiang Provincial Key Laboratory of Environmental Microbiology and Recycling of Argo-waste in Cold Region,College of Life Science and Biotechnology,Heilongjiang Bayi Agricultural University,Daqing 163319,Heilongjiang,China;Key Laboratory of Mollisols Agroecology,Northeast Institute of Geography and Agroecology,Chinese Academy of Sciences,Harbin 150081,Heilongjiang,China;Key Laboratory of Low-carbon Green Agriculture in Northeast Plain,Ministry of Agriculture and Rural Affairs,Daqing 163319,Heilongjiang,China)

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,黑龙江省寒区环境微生物与农业废弃物资源化利用重点实验室,黑龙江大庆163319 [2]中国科学院东北地理与农业生态研究所,黑土区农业生态重点实验室,黑龙江哈尔滨150081 [3]农业农村部东北平原农业绿色低碳重点实验室,黑龙江大庆163319

出　　处：《微生物学报》2024年第9期3379-3392,共14页Acta Microbiologica Sinica

基　　金：国家重点研发计划(2021YFD1500400);国家自然科学基金(31870477,31300425);黑龙江省自然科学基金(C2017045)。

摘　　要：【目的】枯草芽孢杆菌(Bacillus subtilis)施用于土壤可促进植物生长,但针对土壤中枯草芽孢杆菌噬菌体的研究鲜有报道,因此从土壤中分离出枯草芽孢杆菌噬菌体,并深入研究其生物学特性及基因组特征,不仅有助于枯草芽孢杆菌的实际应用,还能丰富枯草芽孢杆菌噬菌体相关生物信息库。【方法】以枯草芽孢杆菌SM13为宿主,从土壤中分离培养出一株噬菌体Bac-S,对其进行生物学特性测定、全基因组序列特征、基因功能注释和系统发育分析。【结果】透射电镜显示,噬菌体Bac-S头部直径约为43 nm,尾部极短无法测量;该噬菌体的最佳感染复数(multiplicity of infection,MOI)为0.1;一步生长曲线显示,潜伏期约为10 min,裂解量为30 PFU/cell;宿主谱广泛,能实现跨属侵染;测序结果显示其基因组长度为150019 bp,G+C含量为42.6%;该噬菌体共有237个开放阅读框(open reading frames,ORFs),BLASTn比对显示该噬菌体与NCBI数据库中其他芽孢杆菌噬菌体相似性较高。【结论】从土壤中分离出了一株具有潜伏期短、宿主谱广泛、耐受高温、不耐紫外光照射的特征枯草芽孢杆菌噬菌体,拓宽了对枯草芽孢杆菌噬菌体的认知。[Objective]The application of Bacillus subtilis in soil can promote plant growth,while few studies have reported the bacteriophages infecting B.subtilis.Therefore,the isolation,biological characterization,and genome sequencing of bacteriophages infecting B.subtilis from soil will contribute to the application of B.subtilis and enrich the biological information of bacteriophages infecting B.subtilis.[Methods]B.subtilis SM13 was used as the host to isolate a bacteriophage strain Bac-S from soil.Biological characterization,whole genome sequencing,gene function annotation,and phylogenetic analysis were performed for this bacteriophage strain.[Results]Transmission electron microscopy showed that Bac-S had a head with the diameter of about 43 nm and a tail too short to be measured.The optimal multiplicity of infection of Bac-S was 0.1.The one-step growth curve showed that Bac-S had an incubation period of about 10 min and a burst size of 30 PFU/cell.Bac-S had a wide host spectrum and can infect hosts of different genera.The sequencing results showed that the genome of Bac-S was 150019 bp,with the G+C content of 42.6%and 237 open reading frames(ORFs).The BLASTn comparison with the sequences in the NCBI database showed that Bac-S was similar to other bacteriophages infecting B.subtilis.[Conclusion]A bacteriophage strain infecting B.subtilis with a short incubation period,a wide host spectrum,tolerance to high temperature,and intolerance to UV light was isolated from soil.The biological and genetic characterization of this bacteriophage enriches our knowledge about the bacteriophages infecting B.subtilis.

关 键 词：枯草芽孢杆菌 噬菌体 生物学特性 基因组 

分 类 号：Q933[生物学—微生物学]