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食品级不锈钢上的干燥初始转录组反应及适应性

Initial Transcriptomic Response and Adaption of to Desiccation on Food Grade Stainless Steel.

作者信息

Kragh Martin Laage, Truelstrup Hansen Lisbeth

机构信息

National Food Institute, Technical University of Denmark, Kongens Lyngby, Denmark.

出版信息

Front Microbiol. 2020 Jan 22;10:3132. doi: 10.3389/fmicb.2019.03132. eCollection 2019.

Abstract

The foodborne pathogen survives exposure to a variety of stresses including desiccation in the food industry. Strand-specific RNA sequencing was applied to analyze changes in the transcriptomes of two strains of (Lm 568 and Lm 08-5578) during desiccation [15°C, 43% relative humidity (RH)] on food grade stainless steel surfaces over 48 h to simulate a weekend with no food production. Both strains showed similar survival during desiccation with a 1.8-2 Log CFU/cm reduction after 48 h. Analysis of differentially expressed (DE) genes (>twofold, adjusted -value <0.05) revealed that the initial response to desiccation was established after 6 h and remained constant with few new genes being DE after 12, 24, and 48 h. A core of 81 up- and 73 down-regulated DE genes were identified as a shared, strain independent response to desiccation. Among common upregulated genes were energy and oxidative stress related genes e.g., (cytochrome aa) (pyruvate dehydrogenase complex) and (manganese transporter). Common downregulated genes related to anaerobic growth, proteolysis and the two component systems / and , which are involved in cold growth and flagellin production, respectively. Both strains upregulated additional genes involved in combatting oxidative stress and reactive oxygen species (ROS), including (superoxide dismutase), (catalase), (thiol peroxidase) and several thioredoxins including , and . Osmotic stress related genes were also upregulated in both strains, including (glycine betaine transporter) and several chaperones , , and . Significant strain differences were also detected with the food outbreak strain Lm 08-5578 differentially expressing 1.9 × more genes (726) compared to Lm 568 (410). Unique to Lm 08-5578 was a significant upregulation of the expression of the alternative transcription factor σ and its regulon. A number of long antisense transcripts (lasRNA) were upregulated during desiccation including , , , and , with the latter controlling flagellum biosynthesis and possibly the downregulation of motility genes observed in both strains. This exploration of the transcriptomes of desiccated provides further understanding of how this bacterium encounters and survives the stress faced when exposed to dry conditions in the food industry.

摘要

食源性病原体在食品工业中能够抵御包括干燥在内的多种应激。采用链特异性RNA测序分析两株单核细胞增生李斯特菌(Lm 568和Lm 08 - 5578)在食品级不锈钢表面干燥(15°C,43%相对湿度)48小时期间转录组的变化,以模拟无食品生产的周末。两株菌在干燥过程中存活情况相似,48小时后每平方厘米菌落形成单位(CFU/cm)减少1.8 - 2个对数。对差异表达(DE)基因(变化两倍以上,校正P值<0.05)的分析表明,对干燥的初始反应在6小时后确立,12、24和48小时后保持稳定,几乎没有新的基因差异表达。确定了81个上调和73个下调的核心DE基因,作为对干燥的共同、菌株非依赖性反应。上调的常见基因包括与能量和氧化应激相关的基因,如细胞色素aa3、丙酮酸脱氢酶复合体和锰转运蛋白。下调的常见基因与厌氧生长、蛋白水解以及分别参与冷生长和鞭毛蛋白产生的双组分系统ArcA/B和FlgM/N有关。两株菌均上调了参与对抗氧化应激和活性氧(ROS)的其他基因,包括超氧化物歧化酶、过氧化氢酶、硫醇过氧化物酶以及几种硫氧还蛋白,如TrxA、TrxC和TrxY。两株菌中与渗透应激相关的基因也上调,包括甘氨酸甜菜碱转运蛋白以及几种伴侣蛋白DnaK、GroEL和GroES。还检测到显著的菌株差异,与食源性暴发菌株Lm 08 - 5578相比,Lm 568差异表达的基因少1.9倍(410个对726个)。Lm 08 - 5578特有的是替代转录因子σ及其调控子的表达显著上调。干燥过程中一些长反义转录本(lasRNA)上调,包括Rli20、Rli21、Rli22和Rli23,后者控制鞭毛生物合成,并可能导致两株菌中观察到的运动基因下调。对干燥单核细胞增生李斯特菌转录组的探索进一步了解了该细菌在食品工业中暴露于干燥条件时如何应对并在这种应激下存活。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eddf/6987299/3f178fcdd0d7/fmicb-10-03132-g001.jpg

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