Characterization of the Potential Probiotic Vibrio sp. V33 Antagonizing Vibrio Splendidus Based on Iron Competition
Characterization of the Potential Probiotic Vibrio sp. V33 Antagonizing Vibrio Splendidus Based on Iron Competition
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DOI：
10.30498/ijb.2019.85192
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发表时间：
2019-03
影响因子：
1.3
通讯作者：
Ningning Liu;T. Song;Shanshan Zhang;Huijie Liu;Xuelin Zhao;Y. Shao;Chenghua Li;Weiwei Zhang
中科院分区：
文献类型：
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作者：
Ningning Liu;T. Song;Shanshan Zhang;Huijie Liu;Xuelin Zhao;Y. Shao;Chenghua Li;Weiwei Zhang
背景：灿烂弧菌Vs是一种重要的水产养殖病原体，可以感染广泛的海洋生物宿主。在我们之前的研究中，一种拮抗细菌弧菌。鉴定出对致病分离株 V. splendidus Vs 的生长和毒力具有抑制作用的 V33。目的：我们从铁竞争的角度进一步探讨拮抗物质和拮抗作用。材料与方法：弧菌上清液中的主要拮抗物质。 V33 使用生物测定引导方法进行鉴定。 V. splendidus Vs 在弧菌无细胞上清液攻击下的反应。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和实时逆转录PCR测定V33。结果：弧菌产生的主要拮抗物质。 V33分子量低，易溶于水，是热稳定物质。同时，弧菌的铁吸收率。 V33 高于 V. splendidus Vs。在存在来自弧菌的无细胞上清液的情况下。 V33 中与 V. splendidus Vs 中铁吸收过程相关的两个功能基因 viuB 和 asbJ 的表达上调，而编码铁吸收阻遏物的furVs 则被抑制到 0.5 倍以下。编码磷酸丙酮酸水合酶的一个基因在 mRNA 水平上没有变化，但在蛋白质水平上上调。结论：我们的结果表明弧菌具有拮抗作用。致病性分离株V. splendidus Vs上的V33部分是由于弧菌更强的能力。 V33抢铁。这种来自弧菌的无细胞上清液。 V33为V. splendidus Vs创造了一个铁限制环境，这导致了与V. splendidus Vs铁吸收相关的基因表达谱的改变。
Background: Vibrio Splendidus Vs is an important aquaculture pathogen that can infect a broad host of marine organisms. In our previous study, an antagonistic bacterium Vibrio sp. V33 that possessed inhibitory effects on the growth and virulence of a pathogenic isolate V. splendidus Vs was identified. Objectives: Here, we further explored the antagonistic substances and antagonistic effects from the viewpoint of iron competition. Materials and Methods: The main antagonistic substances in the supernatants from Vibrio sp. V33 were identified using the bioassay-guided method. The response of V. splendidus Vs under the challenge of cell-free supernatant from Vibrio sp. V33 was determined via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and real-time reverse-transcription PCR. Results: The main antagonistic substances produced by Vibrio sp. V33 have low molecular weights, are water soluble, and are heat-stable substances. Meanwhile, the iron uptake rate of Vibrio sp. V33 was higher than that of V. splendidus Vs. In the presence of cell-free supernatant from Vibrio sp. V33, expressions of two functional genes, viuB and asbJ related to ferric uptake processes in V. splendidus Vs, were up-regulated, whereas furVs coding the ferric uptake repressor was suppressed below 0.5-fold. One gene coding phosphopyruvate hydratase does not change at mRNA level, but was up-regulated at protein level. Conclusions: Our results suggested that antagonistic effect of Vibrio sp. V33 on the pathogenic isolate V. splendidus Vs was partially due to the stronger ability of Vibrio sp. V33 to seize iron. This cell-free supernatant from Vibrio sp. V33 created an iron-limited milieu for V. splendidus Vs, which led to the changed expression profiles of genes that were related to iron uptake in V. splendidus Vs.