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通过RNA测序对荧光假单胞菌对表没食子儿茶素没食子酸酯反应的转录组分析。

Transcriptomic analysis of the response of Pseudomonas fluorescens to epigallocatechin gallate by RNA-seq.

作者信息

Liu Xiaoxiang, Shen Bimiao, Du Peng, Wang Nan, Wang Jiaxue, Li Jianrong, Sun Aihua

机构信息

Faculty of Basic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, P.R. China.

Department of Laboratory Medicine, Hangzhou Medical College, Hangzhou, Zhejiang, P.R. China.

出版信息

PLoS One. 2017 May 17;12(5):e0177938. doi: 10.1371/journal.pone.0177938. eCollection 2017.

Abstract

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.

摘要

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,且被认为食用安全。然而,EGCG的潜在抗菌机制以及细菌对EGCG的反应尚不清楚。在本研究中,使用RNA测序技术对响应EGCG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1 mM EGCG处理和未处理的荧光假单胞菌分别产生了总共26365414和23287092条clean reads,并将这些clean reads与参考基因组进行比对。差异表达分析揭示了291个上调基因和134个下调基因,并且使用RT-qPCR对差异表达基因(DEG)进行了验证。EGCG处理显著上调了大多数参与铁摄取、抗氧化、DNA修复、外排系统、细胞包膜和细胞表面成分合成的DGE,而大多数与能量产生相关的基因则被下调。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处理显著改变了编码胞外功能sigma因子(PvdS、RpoE和AlgU)和双组分传感器组氨酸激酶(BaeS和RpfG)的特定基因的表达,这可能在调节荧光假单胞菌对EGCG的应激反应中起重要作用。本数据为EGCG的分子作用以及EGCG介导的荧光假单胞菌可能的交叉抗性提供了重要见解,这最终可能有助于绿茶多酚在食品保鲜中的最佳应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fceb/5435343/8ae2b1383361/pone.0177938.g001.jpg

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