title: Identification and characterization of three Vibrio alginolyticus non-coding RNAs involved in adhesion, chemotaxis, and motility processes.
abstract: The capability of Vibrio alginolyticus to adhere to fish mucus is a key virulence factor of the bacteria. Our previous research showed that stress conditions, such as Cu(2+), Pb(2+), Hg(2+), and low pH, can reduce this adhesion ability. Non-coding (nc) RNAs play a crucial role in regulating bacterial gene expression, affecting the bacteria's pathogenicity. To investigate the mechanism(s) underlying the decline in adhesion ability caused by stressors, we combined high-throughput sequencing with computational techniques to detect stressed ncRNA dynamics. These approaches yielded three commonly altered ncRNAs that are predicted to regulate the bacterial chemotaxis pathway, which plays a key role in the adhesion process of bacteria. We hypothesized they play a key role in the adhesion process of V. alginolyticus. In this study, we validated the effects of these three ncRNAs on their predicted target genes and their role in the V. alginolyticus adhesion process with RNA interference (i), quantitative real-time polymerase chain reaction (qPCR), northern blot, capillary assay, and in vitro adhesion assays. The expression of these ncRNAs and their predicted target genes were confirmed by qPCR and northern blot, which reinforced the reliability of the sequencing data and the target prediction. Overexpression of these ncRNAs was capable of reducing the chemotactic and adhesion ability of V. alginolyticus, and the expression levels of their target genes were also significantly reduced. Our results indicated that these three ncRNAs: (1) are able to regulate the bacterial chemotaxis pathway, and (2) play a key role in the adhesion process of V. alginolyticus.
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Lixing
authors.initials: L
authors.lastname: Huang
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Jiao
authors.initials: J
authors.lastname: Hu
authors.affiliation: College of Ocean and Earth Sciences, Xiamen University Xiamen, China.
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Su
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Yingxue
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Wendi
authors.initials: W
authors.lastname: Kong
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Xiaojin
authors.initials: X
authors.lastname: Xu
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Mao
authors.initials: M
authors.lastname: Lin
authors.affiliation: Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University Xiamen, China.
authors.forename: Qingpi
authors.initials: Q
authors.lastname: Yan
mesh_terms: D001422:Bacterial Adhesion; D015152:Blotting, Northern; D002633:Chemotaxis; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D020868:Gene Silencing; D008124:Locomotion; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D022661:RNA, Untranslated / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D044149:Vibrio alginolyticus / Q000235:genetics / Q000502:physiology*
chemical_list: D012329:RNA, Bacterial; D022661:RNA, Untranslated
keywords: RNAi; Vibrio alginolyticus; adhesion; chemotaxis pathway; ncRNA