浙江农业学报 ›› 2023, Vol. 35 ›› Issue (1): 50-57.DOI: 10.3969/j.issn.1004-1524.2023.01.06
赵赛赛1(), 张皓杰1, 蔡秀磊1, 王丽荣2, 申小冉2, 曹志1, 单虎1()
收稿日期:
2022-05-24
出版日期:
2023-01-25
发布日期:
2023-02-21
通讯作者:
*单虎,E-mail: hushan67@163.com
作者简介:
赵赛赛(1995—),女,山东青岛人,硕士,研究方向为预防兽医学。E-mail: 18754297793@163.com
基金资助:
ZHAO Saisai1(), ZHANG Haojie1, CAI Xiulei1, WANG Lirong2, SHEN Xiaoran2, CAO Zhi1, SHAN Hu1()
Received:
2022-05-24
Online:
2023-01-25
Published:
2023-02-21
摘要:
本实验室从水产养殖池防污附钢片的微生物黏膜中分离到一株菌DL3,为了明确该菌株的活性及作为有益菌进行应用的可能,通过细菌分离培养、16S rRNA测序和系统发育分析对其做了初步鉴定,并采用牛津杯法、平板法分别测定了菌株的抑菌活性和群体感应淬灭活性。基于菌株DL3 16S rRNA基因序列的系统发育分析表明,该菌株与解肽假交替单胞菌(Pseudoalteromonas peptidolytica )NBRC 101021T的相似性最高,亲缘关系最近,为99.64%;抑菌试验显示,该菌株对鳗弧菌(Vibrio anguillarum)和铜绿假单胞菌(Pseudomonas aeruginosa)的抑制作用最强,对金黄色葡萄球菌(Staphylococcus aureus)、大肠埃希菌(Escherichi coli)、沙门氏菌(Salmonella)、链球菌(Streptococcus)、志贺氏杆菌(Shigella)、河弧菌(V. fluvialis)、东方弧菌(V. orientalis)、哈维氏弧菌(V. harveyi)和短小芽孢杆菌(Bacillus pumilus)的生长表现出不同程度的抑制作用,且在培养72 h时抑菌活性最强;平板法检测结果表明,菌株DL3具有群体感应淬灭活性,对3-oxo-C6-HSL、C10-HSL、C12-HSL、3-oxo-C14-HSL等信号分子具有较强的淬灭效果。本研究成功分离到假交替单胞菌DL3,并明确了其拮抗病原菌的活性,为后续深入开展菌株DL3及其活性产物的抑菌机制研究、探索海洋微生物在动物细菌性病害防治中的应用奠定了基础。
中图分类号:
赵赛赛, 张皓杰, 蔡秀磊, 王丽荣, 申小冉, 曹志, 单虎. 一株假交替单胞菌DL3的抑菌与群体感应淬灭活性研究[J]. 浙江农业学报, 2023, 35(1): 50-57.
ZHAO Saisai, ZHANG Haojie, CAI Xiulei, WANG Lirong, SHEN Xiaoran, CAO Zhi, SHAN Hu. Study on antibacterial and quorum quenching activity of a Pseudoalteromonas sp. DL3[J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 50-57.
菌种名称 Strain Name | 菌株号 Strain No. | 数量 Number |
---|---|---|
Escherichia coli | E1 | 1 |
E. coli | E2 | 1 |
E. coli | E3 | 1 |
E. coli | E4 | 1 |
Staphylococcus aureus | PMJ4-1 | 1 |
S. aureus | PMJ10-1 | 1 |
S. aureus | PMJ30-2 | 1 |
S. aureus | TTB1-1 | 1 |
S. aureus | TTB2-1 | 1 |
S. aureus | QTB1-1 | 1 |
Vibrio anguillarum | VIB72 | 1 |
V. fluvialis | VIB292 | 1 |
V. harveyi | VIB295 | 1 |
V. orientalis | VIB303 | 1 |
Bacillus pumilus | FYB01 | 1 |
Shigella | T3 | 1 |
Pseudomonas aeruginosa | PA2 | 1 |
Pseudomonas aeruginosa | PA1 | 1 |
Salmonella | QS1 | 1 |
Streptococcus | SS-1 | 1 |
Streptococcus | SS-2 | 1 |
Streptococcus | SS-7 | 1 |
Streptococcus | SS-9 | 1 |
总计Total | 23 |
表1 二十三株临床病原菌名称及菌株号
Table 1 Names and numbers of 23 clinical pathogen strains
菌种名称 Strain Name | 菌株号 Strain No. | 数量 Number |
---|---|---|
Escherichia coli | E1 | 1 |
E. coli | E2 | 1 |
E. coli | E3 | 1 |
E. coli | E4 | 1 |
Staphylococcus aureus | PMJ4-1 | 1 |
S. aureus | PMJ10-1 | 1 |
S. aureus | PMJ30-2 | 1 |
S. aureus | TTB1-1 | 1 |
S. aureus | TTB2-1 | 1 |
S. aureus | QTB1-1 | 1 |
Vibrio anguillarum | VIB72 | 1 |
V. fluvialis | VIB292 | 1 |
V. harveyi | VIB295 | 1 |
V. orientalis | VIB303 | 1 |
Bacillus pumilus | FYB01 | 1 |
Shigella | T3 | 1 |
Pseudomonas aeruginosa | PA2 | 1 |
Pseudomonas aeruginosa | PA1 | 1 |
Salmonella | QS1 | 1 |
Streptococcus | SS-1 | 1 |
Streptococcus | SS-2 | 1 |
Streptococcus | SS-7 | 1 |
Streptococcus | SS-9 | 1 |
总计Total | 23 |
图2 菌株DL3的16S rRNA基因序列N-J系统发育树 “●”表示该节点同ML、MP方法结果一致;“○”表示该节点同ML法结果一致。
Fig.2 Phylogenetic tree of strain DL3 based on 16S rRNA gene sequences by neighbour-joining method “●” indicated that the corresponding nodes are also recovered in tree generated with the ML and MP algorithms; “○” indicated that the corresponding nodes were also recovered in tree generated with the ML algorithm.
图3 不同培养时间对菌株DL3生物量、蛋白质浓度与抑菌活性的影响 A,不同培养时间对菌株DL3生物量的影响;B,不同培养时间对菌株DL3蛋白质浓度的影响;C,不同培养时间对菌株DL3抑菌活性的影响。“**”表示差异极显著(P<0.01),“ns”表示差异不显著(P>0.05)。
Fig.3 Effects of different culture time on biomass, protein concentration and antibacterial activity of strain DL3 A, Effect of culture time on the biomass of strain DL3; B, Effect of culture time on the protein concentration of strain DL3; C. Effect of culture time on the antibacterial activity of strain DL3. “**” meant significant difference (P<0.01), “ns” meant no significant difference (P>0.05).
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