Transcriptomic analysis of the response of Pseudomonas fluorescens to epigallocatechin gallate by RNA-seq.
Transcriptomic analysis of the response of Pseudomonas fluorescens to epigallocatechin gallate by RNA-seq.
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通过 RNA-seq 转录组分析荧光假单胞菌对表没食子儿茶素没食子酸酯的反应
DOI：
10.1371/journal.pone.0177938
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Sun A
中科院分区：
文献类型：
--
作者：
Liu X;Shen B;Du P;Wang N;Wang J;Li J;Sun A
表没食子儿茶素没食子酸酯（EGCG）是绿茶多酚的主要成分，绿茶多酚被广泛用作食品防腐剂，被认为是安全的。然而，EGCG的潜在抗菌机制和细菌对EGCG的反应尚不清楚。在本研究中，利用RNA-seq技术对一种对EGCG有反应的典型腐败细菌荧光假单胞菌进行了全基因组转录分析。用或不加1 mM EGCG处理的P. fluorescens共产生26,365,414和23,287,092个clean reads，这些clean reads与参考基因组对齐。差异表达分析发现291个上调基因和134个下调基因，并通过RT-qPCR验证差异表达基因（DEGs）。EGCG处理后，与铁摄取、抗氧化、DNA修复、外排系统、细胞包膜和细胞表面成分合成相关的大部分基因显著上调，而与能量产生相关的大部分基因下调。这些转录组变化可能是荧光假单胞菌对铁限制和氧化应激以及EGCG引起的DNA和包膜损伤的适应性反应。EGCG处理显著改变了胞质外功能sigma因子（PvdS、RpoE和AlgU）和双组份组氨酸激酶（BaeS和RpfG）的表达，可能在调控荧光藻对EGCG的应激反应中起重要作用。本研究为进一步了解EGCG的分子作用以及EGCG介导的对荧光杆菌的交叉抗性提供了重要的见解，这可能最终有助于绿茶多酚在食品保鲜中的最佳应用。
Epigallocatechin gallate (EGCG) is a main constituent of green tea polyphenols that are widely used as food preservatives and are considered to be safe for consumption. However, the underlying antimicrobial mechanism of EGCG and the bacterial response to EGCG are not clearly understood. In the present study, a genome-wide transcriptional analysis of a typical spoilage bacterium, Pseudomonas fluorescens that responded to EGCG was performed using RNA-seq technology. A total of 26,365,414 and 23,287,092 clean reads were generated from P. fluorescens treated with or without 1 mM EGCG and the clean reads were aligned to the reference genome. Differential expression analysis revealed 291 upregulated genes and 134 downregulated genes and the differentially expressed genes (DEGs) were verified using RT-qPCR. Most of the DGEs involved in iron uptake, antioxidation, DNA repair, efflux system, cell envelope and cell-surface component synthesis were significantly upregulated by EGCG treatment, while most genes associated with energy production were downregulated. These transcriptomic changes are likely to be adaptive responses of P. fluorescens to iron limitation and oxidative stress, as well as DNA and envelope damage caused by EGCG. The expression of specific genes encoding the extra-cytoplasmic function sigma factor (PvdS, RpoE and AlgU) and the two-component sensor histidine kinase (BaeS and RpfG) were markedly changed by EGCG treatment, which may play important roles in regulating the stress responses of P. fluorescens to EGCG. The present data provides important insights into the molecular action of EGCG and the possible cross-resistance mediated by EGCG on P. fluorescens, which may ultimately contribute to the optimal application of green tea polyphenols in food preservation.