Machuca Alvaro, Martinez Victor
FAVET-INBIOGEN, Faculty of Veterinary Sciences, University of Chile, Santiago, Chile.
PLoS One. 2016 Dec 29;11(12):e0168855. doi: 10.1371/journal.pone.0168855. eCollection 2016.
The intracellular facultative bacteria Piscirickettsia salmonis is one of the most important pathogens of the Chilean aquaculture. However, there is a lack of information regarding the whole genomic transcriptional response according to different extracellular environments. We used next generation sequencing (NGS) of RNA (RNA-seq) to study the whole transcriptome of an isolate of P. salmonis (FAVET-INBIOGEN) using a cell line culture and a modified cell-free liquid medium, with or without iron supplementation. This was done in order to obtain information about the factors there are involved in virulence and iron acquisition. First, the isolate was grown in the Sf21 cell line; then, the bacteria were cultured into a cell-free liquid medium supplemented or not with iron. We identified in the transcriptome, genes associated with type IV secretion systems, genes related to flagellar structure assembly, several proteases and sigma factors, and genes related to the development of drug resistance. Additionally, we identified for the first time several iron-metabolism associated genes including at least two iron uptake pathways (ferrous iron and ferric iron uptake) that are actually expressed in the different conditions analyzed. We further describe putative genes that are related with the use and storage of iron in the bacteria, which have not been previously described. Several sets of genes related to virulence were expressed in both the cell line and cell-free culture media (for example those related to flagellar structure; such as basal body, MS-ring, C-ring, proximal and distal rod, and filament), which may play roles in other basic processes rather than been restricted to virulence.
细胞内兼性细菌鲑鱼立克次氏体是智利水产养殖中最重要的病原体之一。然而,关于其在不同细胞外环境下的全基因组转录反应,目前仍缺乏相关信息。我们使用RNA的下一代测序(NGS)技术(RNA测序),通过细胞系培养和改良的无细胞液体培养基,在添加或不添加铁的情况下,研究了鲑鱼立克次氏体(FAVET-INBIOGEN)分离株的全转录组。这样做是为了获取有关参与毒力和铁摄取的因素的信息。首先,将该分离株在Sf21细胞系中培养;然后,将细菌接种到添加或不添加铁的无细胞液体培养基中。我们在转录组中鉴定出与IV型分泌系统相关的基因、与鞭毛结构组装相关的基因、几种蛋白酶和sigma因子,以及与耐药性发展相关的基因。此外,我们首次鉴定出几个与铁代谢相关的基因,包括至少两条在分析的不同条件下实际表达的铁摄取途径(亚铁摄取和铁离子摄取)。我们进一步描述了与细菌中铁的利用和储存相关的假定基因,这些基因以前未曾描述过。几组与毒力相关的基因在细胞系和无细胞培养基中均有表达(例如与鞭毛结构相关的基因;如基体、MS环、C环、近端和远端杆以及鞭毛丝),这些基因可能在其他基本过程中发挥作用,而不仅限于毒力。