Bacillomycin D突变体在生物膜形成中的转录组分析

发布时间：2018-05-29 23:07

本文选题：解淀粉芽孢杆菌SQR + bacillomycin　；参考：《南京农业大学学报》2017年05期

【摘要】：[目的]植物促生细菌的根际生物膜形成过程是其功能发挥的关键因素,本研究利用转录组技术遴选参与这一过程的相关基因。[方法]利用转录组技术分析解淀粉芽孢杆菌SQR9的突变体SQR9M1和野生型SQR9生物膜形成24 h时基因的表达差异,并用定量Real-time PCR验证;通过电感耦合等离子质谱(ICP-MS)的方法比较突变体SQR9M1和野生型SQR9的胞内铁离子浓度的差异;细胞培养板结合定量PCR监测外源添加铁离子后对突变体SQR9M1生物膜形成表型和相关基因表达的影响。[结果]突变体SQR9M1的生物膜形成能力在培养前期(24 h)显著低于野生型SQR9。与野生型相比,突变体生物膜细胞中共有1 261个基因表达发生改变,其中574个基因上调(P0.01),687个基因下调(P0.01)。功能明确的变化基因约占全部差异基因数量的70%,变化最大的基因功能类群是无机离子转运和代谢。突变体SQR9M1的生物膜形成延迟现象与其铁离子转运能力受损有关,外源添加铁离子能诱导野生型SQR9和突变体SQR9M1生物膜的形成和相关基因的表达。[结论]突变体SQR9M1铁离子ABC转运蛋白Fhu BGC、Feu ABC和Yfm CDEF相关基因显著下调,其胞内铁离子浓度在生长前期显著低于野生型,外源添加铁离子后能恢复突变体的生物膜表型。
[Abstract]:[objective] the process of biofilm formation in rhizosphere of plant-promoting bacteria is the key factor for its function. In this study, transcriptome technique was used to select the genes involved in this process. [methods] the difference of gene expression between mutant SQR9M1 of Bacillus amylolyticus SQR9 and wild-type SQR9 biofilm at 24 h was analyzed by transcriptional technique and verified by quantitative Real-time PCR. Inductively coupled plasma mass spectrometry (ICP-MS) was used to compare the difference of intracellular iron concentration between the mutant SQR9M1 and the wild-type SQR9. Cell culture plates combined with quantitative PCR were used to detect the effects of exogenous iron ions on the biofilm formation phenotype and related gene expression of mutant SQR9M1. [results] the biofilm forming ability of mutant SQR9M1 was significantly lower than that of wild type SQR9 at 24 h. Compared with wild type, there were 1 261 gene expression changes in the mutant biofilm cells, of which 574 genes up-regulated the expression of P0.01G and 687 genes down-regulated P0.01G. About 70% of the total differentially expressed genes were functionally altered, and the most varied functional groups were inorganic ion transport and metabolism. The delayed biofilm formation of mutant SQR9M1 is related to the loss of its iron ion transport ability. Exogenous iron ions can induce the formation of wild type SQR9 and mutant SQR9M1 biofilm and the expression of related genes. [conclusion] the SQR9M1 iron ion ABC transporter Fhu BGCfFeu ABC and Yfm CDEF related genes were down-regulated, and the intracellular iron ion concentration was significantly lower in the early stage of growth than in the wild type. Exogenous iron ions could restore the biofilm phenotype of the mutant.
【作者单位】：南京农业大学资源与环境科学学院/江苏省固体有机废弃物资源化高技术研究重点实验室;江苏省耕地质量与农业环境保护站;
【基金】：国家自然科学基金项目(31501833) 江苏省自然科学基金项目(20150658)
【分类号】：S182

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