Characterization and Genome Structure of Virulent Phage EspM4VN to Control Enterobacter sp. M4 Isolated From Plant Soft Rot
Characterization and Genome Structure of Virulent Phage EspM4VN to Control Enterobacter sp. M4 Isolated From Plant Soft Rot
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DOI：
10.3389/fmicb.2020.00885
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发表时间：
2020-06
影响因子：
5.2
通讯作者：
N. C. Thanh;Yuko Nagayoshi;Yasuhiro Fujino;K. Iiyama;N. Furuya;Y. Hiromasa;T. Iwamoto;Katsumi Doi
中科院分区：
文献类型：
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作者：
N. C. Thanh;Yuko Nagayoshi;Yasuhiro Fujino;K. Iiyama;N. Furuya;Y. Hiromasa;T. Iwamoto;Katsumi Doi
肠杆菌属。从植物软腐病中分离到M4等细菌菌株。从土壤中分离的EspM4VN等毒力噬菌体是植物病害生物防治的趋势。该噬菌体有一个二十面体的头部(直径100 nm)、一个颈部和一个可收缩的鞘(长100 nm，宽18 nm)。它属于阿克曼病毒科，与志贺氏菌噬菌体Ag3和迪克雅噬菌体JA15和XF4相似。本文报道EspM4VN在10℃~50℃、pH 4~10范围内稳定，在70℃、pH 3、12下失活。该噬菌体仅在肠杆菌表面形成清晰的空斑。供试菌株中有M4。一步生长曲线显示，潜伏期为20min，上升期为10min，平均每个吸收细胞释放122个噬菌体颗粒。我们发现该噬菌体的基因组大小为160,766bp，它注释了219个开放阅读框。EspM4VN的基因组结构与沙门氏菌噬菌体SKML-39、迪克雅噬菌体Coodle、PP35、JA15和石灰石以及志贺氏菌噬菌体Ag3有很高的相似性。噬菌体EspM4VN有5个tRNA物种、4个尾尖蛋白和1个胸苷合成酶。基于结构蛋白和酶的系统发育分析表明，EspM4VN属于阿克曼病毒科Aglimvirinae亚科AgtreVirus属的一个成员。
Enterobacter sp. M4 and other bacterial strains were isolated from plant soft rot disease. Virulent phages such as EspM4VN isolated from soil are trending biological controls for plant disease. This phage has an icosahedral head (100 nm in diameter), a neck, and a contractile sheath (100 nm long and 18 nm wide). It belongs to the Ackermannviridae family and resembles Shigella phage Ag3 and Dickeya phages JA15 and XF4. We report herein that EspM4VN was stable from 10°C to 50°C and pH 4 to 10 but deactivated at 70°C and pH 3 and 12. This phage formed clear plaques only on Enterobacter sp. M4 among tested bacterial strains. A one-step growth curve showed that the latent phase was 20 min, rise period was 10 min, and an average of 122 phage particles were released from each absorbed cell. We found the phage’s genome size was 160,766 bp and that it annotated 219 open reading frames. The genome organization of EspM4VN has high similarity with the Salmonella phage SKML-39; Dickeya phages Coodle, PP35, JA15, and Limestone; and Shigella phage Ag3. The phage EspM4VN has five tRNA species, four tail-spike proteins, and a thymidylate synthase. Phylogenetic analysis based on structural proteins and enzymes indicated that EspM4VN was identified as a member of the genus Agtrevirus, subfamily Aglimvirinae, family Ackermannviridae.