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首页> 外文期刊>Applied Microbiology >RNA Sequencing Analysis of the Broad-Host-Range Strain Sinorhizobium fredii NGR234 Identifies a Large Set of Genes Linked to Quorum Sensing-Dependent Regulation in the Background of a traI and ngrI Deletion Mutant
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RNA Sequencing Analysis of the Broad-Host-Range Strain Sinorhizobium fredii NGR234 Identifies a Large Set of Genes Linked to Quorum Sensing-Dependent Regulation in the Background of a traI and ngrI Deletion Mutant

机译：广泛的宿主菌株Srinohizobium fredii NGR234的RNA测序分析确定了traI和ngrI缺失突变体背景下与仲裁感应依赖的调控相关的大量基因。

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The alphaproteobacterium Sinorhizobium fredii NGR234 has an exceptionally wide host range, as it forms nitrogen-fixing nodules with more legumes than any other known microsymbiont. Within its 6.9-Mbp genome, it encodes two N -acyl-homoserine-lactone synthase genes (i.e., traI and ngrI ) involved in the biosynthesis of two distinct autoinducer I-type molecules. Here, we report on the construction of an NGR234-Δ traI and an NGR234-Δ ngrI mutant and their genome-wide transcriptome analysis. A high-resolution RNA sequencing (RNA-seq) analysis of early-stationary-phase cultures in the NGR234-Δ traI background suggested that up to 316 genes were differentially expressed in the NGR234-Δ traI mutant versus the parent strain. Similarly, in the background of NGR234-Δ ngrI 466 differentially regulated genes were identified. Accordingly, a common set of 186 genes was regulated by the TraI/R and NgrI/R regulon. Coregulated genes included 42 flagellar biosynthesis genes and 22 genes linked to exopolysaccharide (EPS) biosynthesis. Among the genes and open reading frames (ORFs) that were differentially regulated in NGR234-Δ traI were those linked to replication of the pNGR234 a symbiotic plasmid and cytochrome c oxidases. Biotin and pyrroloquinoline quinone biosynthesis genes were differentially expressed in the NGR234-Δ ngrI mutant as well as the entire cluster of 21 genes linked to assembly of the NGR234 type III secretion system (T3SS-II). Further, we also discovered that genes responsible for rhizopine catabolism in NGR234 were strongly repressed in the presence of high levels of N -acyl-homoserine-lactones. Together with nodulation assays, the RNA-seq-based findings suggested that quorum sensing (QS)-dependent gene regulation appears to be of higher relevance during nonsymbiotic growth rather than for life within root nodules.
机译：阿尔法变形杆菌Sinorhizobium fredii NGR234具有异常广泛的宿主范围，因为它形成了固氮结节，比任何其他已知的微共生菌都有更多的豆类。在其6.9-Mbp基因组中，它编码两个N-酰基-高丝氨酸-内酯合酶基因（即traI和ngrI），参与两个不同的自诱导I型分子的生物合成。在这里，我们报告了NGR234-ΔtraI和NGR234-ΔngrI突变体的构建及其全基因组转录组分析。对NGR234-ΔtraI背景中的早期平稳期培养物的高分辨率RNA测序（RNA-seq）分析表明，NGR234-ΔtraI突变体与亲本菌株相比，多达316个基因差异表达。类似地，在NGR234-ΔngrI 466的背景中，鉴定到差异调节的基因。因此，TraI / R和NgrI / R调节子调节一组共186个基因。共调节基因包括42个鞭毛生物合成基因和22个与胞外多糖（EPS）生物合成相关的基因。在NGR234-ΔtraI中受到差异调节的基因和开放阅读框（ORF）中，有与共生质粒pNGR234复制和细胞色素C氧化酶相关的基因。生物素和吡咯并喹啉醌生物合成基因在NGR234-ΔngrI突变体以及与NGR234 III型分泌系统（T3SS-II）组装相关的21个基因的整个簇中差异表达。此外，我们还发现，在高水平的N-酰基-高丝氨酸-内酯存在下，NGR234中负责rhozopine分解代谢的基因被强烈抑制。与结节化测定一起，基于RNA-seq的发现表明，在非共生生长过程中，群体感应（QS）依赖性基因调控似乎具有更高的相关性，而不是根瘤中的生命。

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