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Transcriptomic profiling of microbe–microbe interactions reveals the specific response of the biocontrol strain P. fluorescens In5 to the phytopathogen Rhizoctonia solani

机译:微生物相互作用的转录组分析揭示了生防菌株P. fluorescens In5对植物病原菌Rhizoctonia solani的特异性反应

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BackgroundFew studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order to gain insights into the potential mechanism underlying the antagonism of the antimicrobial producing strain P. fluorescens In5 against the phytopathogens Rhizoctonia solani and Pythium aphanidermatum , global RNA sequencing was performed. MethodsDifferential gene expression profiling of P. fluorescens In5 in response to either R. solani or P. aphanidermatum was investigated using transcriptome sequencing (RNA-seq). Total RNA was isolated from single bacterial cultures of P. fluorescens In5 or bacterial cultures in dual-culture for 48?h with each pathogen in biological triplicates. RNA-seq libraries were constructed following a default Illumina stranded RNA protocol including rRNA depletion and were sequenced 2?×?100 bases on Illumina HiSeq generating approximately 10 million reads per sample. ResultsNo significant changes in global gene expression were recorded during dual-culture of P. fluorescens In5 with any of the two pathogens but rather each pathogen appeared to induce expression of a specific set of genes. A particularly strong transcriptional response to R. solani was observed and notably several genes possibly associated with secondary metabolite detoxification and metabolism were highly upregulated in response to the fungus. A total of 23 genes were significantly upregulated and seven genes were significantly downregulated with at least respectively a threefold change in expression level in response to R. solani compared to the no fungus control. In contrast, only one gene was significantly upregulated over threefold and three transcripts were significantly downregulated over threefold in response to P. aphanidermatum. Genes known to be involved in synthesis of secondary metabolites, e.g. non-ribosomal synthetases and hydrogen cyanide were not differentially expressed at the time points studied. ConclusionThis study demonstrates that genes possibly involved in metabolite detoxification are highly upregulated in P. fluorescens In5 when co-cultured with plant pathogens and in particular the fungus R. solani . This highlights the importance of studying microbe–microbe interactions to gain a better understanding of how different systems function in vitro and ultimately in natural systems where biocontrol agents can be used for the sustainable management of plant diseases.
机译:背景技术迄今为止,很少有研究报道生物防治细菌对植物病原体的转录反应。为了深入了解产生抗菌素的菌株P. fluorescens In5对抗植物病原菌Rhozoctonia solani和Pythium aphanidermatum的潜在潜在机制,进行了全局RNA测序。方法使用转录组测序(RNA-seq)研究萤光假单胞菌(P.fluorescens In5)对茄红假单胞菌(R. solani)或瓜果假单胞菌(P. aphanidermatum)的差异表达基因。从荧光假单胞菌In5的单一细菌培养物中分离总RNA,或在双重培养物中将细菌培养物分离48分钟,每种病原体均一式三份。按照默认的Illumina链RNA协议(包括rRNA耗竭)构建RNA-seq文库,并在Illumina HiSeq上对2?×?100碱基进行测序,每个样品产生大约1000万个读数。结果在荧光假单胞菌In5与两种病原体中的任何一种双重培养期间,未记录到全球基因表达的显着变化,但每种病原体似乎都诱导了一组特定基因的表达。观察到了对茄尼罗非鱼的特别强的转录反应,并且特别地,与真菌的反应高度上调了可能与次级代谢产物解毒和代谢相关的几个基因。与无真菌对照相比,响应于茄红假单胞菌,总共23个基因被显着上调,而七个基因被显着下调,至少分别对茄红假单胞菌的表达水平发生了三倍的变化。相比之下,只有一个基因被显着上调了三倍,而三个转录本被显着下调了三倍,以应对P. aphanidermatum。已知参与次级代谢产物合成的基因,例如在研究的时间点上,非核糖体合成酶和氰化氢没有差异表达。结论:这项研究表明,与植物病原体,尤其是真菌R. solani,共培养时,荧光代谢假单胞菌In5中可能与代谢物解毒有关的基因被上调。这凸显了研究微生物之间相互作用的重要性,以便更好地了解不同系统在体外以及最终在自然系统中的功能,在这些系统中生物防治剂可用于植物疾病的可持续管理。

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