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[1]谭筱醴 乔晋楠 杨丹妮 张义阳 李文馨 乔代蓉 曹毅**.一株烈性青枯菌噬菌体φRSm2C的分离纯化及生物学特征[J].应用与环境生物学报,2021,27(04):1-14.[doi:10.19675/j.cnki.1006-687x.2020.07067]
 TAN Xiaoli,QIAO Jinnan,YANG Danni,et al. Isolation, purification and biological characterization of a virulent Ralstonia phage RSm2C[J].Chinese Journal of Applied & Environmental Biology,2021,27(04):1-14.[doi:10.19675/j.cnki.1006-687x.2020.07067]
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一株烈性青枯菌噬菌体φRSm2C的分离纯化及生物学特征()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
27卷
期数:
2021年04期
页码:
1-14
栏目:
研究论文
出版日期:
2021-08-25

文章信息/Info

Title:
 Isolation, purification and biological characterization of a virulent Ralstonia phage RSm2C
作者:
谭筱醴 乔晋楠 杨丹妮 张义阳 李文馨 乔代蓉 曹毅**
四川大学生命科学学院微生物与代谢工程四川省重点实验室 成都 610065
Author(s):
TAN Xiaoli QIAO Jinnan YANG Danni ZHANG Yiyang LI WenxinQIAO Dairong CAO Yi**
Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, China
关键词:
植物病害青枯雷尔氏菌生物防治烈性噬菌体生物学特性
Keywords:
plant diseases Ralstonia solanacearum biological control virulent phage biological characteristics
DOI:
10.19675/j.cnki.1006-687x.2020.07067
摘要:
生物防治由于其高效无害的优势成为植物细菌性病害防治方面的研究热点。噬菌体有极强的宿主特异性,且来源广泛,易于获得,应用其防治青枯病具有良好的发展前景。通过双层平板法从青枯病爆发的田块土壤中分离噬菌体,经单斑纯化后,由透射电子显微镜进行形态学鉴定,并以平板稀释法测定其生物学特征。以致病性青枯菌为宿主,从发生辣椒青枯病的田间土壤中成功分离出一株噬菌体φRSm2C,依据形态学鉴定其为一株肌尾噬菌体,颗粒大小为128.27 ± 5.66nm。生物学特征测定显示φRSm2C的最佳MOI为1,潜伏期约为90 min,暴发期约为180 min,裂解量为112;φRSm2C能够裂解试验所用8株青枯菌中的3株,并且能同时裂解演化型I和演化型II的青枯菌;φRSm2C热稳定性、pH稳定性和有机试剂耐受性均较好,但对紫外线较为敏感;φRSm2C在60 ℃以下活性稳定,在pH 6 - 9范围内活性稳定,对乙醇较为敏感。本研究表明φRSm2C是一株烈性青枯菌肌尾噬菌体,能裂解多种青枯菌,且其稳定性较好,有作为生物农药防治植物青枯病的潜能。(图11 表2 参21)
Abstract:
Biological control has become a research hotspot in the field of bacterial wilt control due to its advantages of high efficiency and harmlessness. Phage has strong host specificity with extensive sources and it’s easy to obtain. Therefore, it has a great prospect of development in the prevention and treatment of bacterial wilt. The bacteriophages was isolated from the field soil of chili infected with bacterial wilt disease by double-layer plate method. After purification by single plaque, the morphological characteristics of the phage were observed using transmission electron microscope (TEM), and the biological characteristics were determined by plate dilution method. In this study, a Ralstonia phage φRSm2C was successfully isolated, and was classified in Myoviridae by morphological characteristics with phage particle size (128.27 ± 5.66) nm. It was determined that the phage had the best MOI(Multiplicity of Infection) of 1 for its host Rs. 1.356, the latent period was about 90 minutes, the burst period was about 180 minutes, and the burst size was about 112. It was found that the phage can i nfect 3/8 R. solanacearum and simultaneously cleave the R. solanacearum of phylotype I and phylotype II. Through the stability test, it was found that φRSm2C has good thermal stability, pH stability, and tolerance to organic reagents, but is sensitive to UV: the phage is stable at temperatures below 60 ℃, also stable in the range of pH 6 – 9, but intolerant of ethanol. This study shows that φRSm2C is a virulent Ralstonia phage which can lyse a variety of R. solanacearum, and has good stability. Thus it has the potential to control plant bacterial wilt as a biological pesticide.

备注/Memo

备注/Memo:
收稿日期 Received: 2020-07-31 接受日期 Accepted: 2020-09-30
四川省科技厅项目(2018GZ0375,2018TJPT0004)、国家微生物资源平台项目(NIMR-2019-8-1)和成都市科技局项目(2018-YF05-00738-SN,2017-GH02-00071-HZ)资助
**通讯作者 Corresponding author (E-mail: caoyi_01@163.com)
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更新日期/Last Update: 2020-10-05