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Genome analysis of Bacillus subtilis JCL16 and the synergistic relationship among its metabolites reveal its potential for biocontrol of Nocardia seriolae

文献类型： 外文期刊

作者： Wang, Xiaohua 1 ; Onchari, M. M. 1 ; Yang, Xueting 1 ; Xu, Lin 1 ; Yin, Xiulian 1 ; Wan, Faxiang 1 ; Chen, Yuewen 1 ; Guan, Ming 1 ; Li, Bin 1 ; Luo, Chuping 1 ;

作者机构： 1.Huaiyin Inst Technol, Jiangsu Prov Key Construct Lab Probiot Preparat, Huaian 223003, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Vet Med, Key Lab Vet Biol Engn & Technol, Minist Agr, Nanjing 210014, Peoples R China

关键词： Largemouth bass; Nocardiosis; Aquatic probiotics; Antibacterial metabolites; Synergistic inhibition

期刊名称：BIOLOGICAL CONTROL （ 影响因子：3.857； 五年影响因子：4.141 ）

ISSN： 1049-9644

年卷期： 2022 年 167 卷

页码：

收录情况： SCI

摘要： Nocardia seriolae is one of the major causative agents of fish nocardiosis, responsible for large-scale fish die-offs and economic losses in the aquaculture industry worldwide. Therefore, research efforts towards developing efficacious alternatives to aquatic antibiotics, such as vaccines, immunomodulatory additives including aquatic probiotics, to control this disease are of high importance. Recently, we isolated a soil-derived strain of Bacillus subtilis JCL16, and the 21-day challenge tests results showed that the addition of B. subtilis JCL16 culture to the basal control diet significantly increased the survival rate of largemouth bass with nocardiosis from 21.67% to 68.3%. To reveal the putative functional factors, we determined the genome sequence of B. subtilis JCL16 using PacBio RS II and Illumina HiSeq 4000 platform, and predicted its secondary metabolites using antiSMASH, which showed that it contains 11 functional gene clusters, three of which, srf, bac and sbo, have previously been verified to have antagonistic bacterial functions. Subsequently, mutants Delta srf, Delta bac, Delta sbo were constructed using homologous recombination techniques, and HPLC results further elucidated that the three gene clusters are responsible for the synthesis of surfactin, bacilysin and subtilosin A, respectively. To our surprise, a significant reduction in the inhibition of N. seriolae occurred in all three mutants compared to the wild type, so we spec-ulated that surfactin, bacilysin and subtilosin A might have adopted a synergistic strategy. Next, we prepared their pure products and further confirmed our inference by Oxford Cup and MIC assays. In summary, the good biocontrol effect of B. subtilis JCL16 against fish Nocardia is likely due to the synergistic strategy among surfactin, bacilysin and subtilosin A. Here we propose a synergistic mechanism among them, which provides some insight into the powerful biocontrol mechanisms of Bacillus.

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