College of Food Science, Sichuan Agricultural University, Ya'an, 625014, China; Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, 510070, China.
College of Food Science, Sichuan Agricultural University, Ya'an, 625014, China.
Food Microbiol. 2023 Oct;115:104323. doi: 10.1016/j.fm.2023.104323. Epub 2023 Jun 14.
Lawn-harvest method uses a solid medium (e.g., tryptic soy agar, TSA) to produce bacterial lawns and is widely accepted for the culture of microorganisms in microbial studies of low-moisture foods (LMFs, foods with water activity less than 0.85). It produces desiccation-tolerant cells with higher D-values in LMFs; however, little is known about the molecular mechanisms underlying bacterial resistance. Salmonella enterica Enteritidis PT 30 (S. Enteritidis), the most pertinent pathogen in LMFs, was cultured in TSA and tryptic soy broth (TSB). Cells were harvested and inoculated on filter papers to assess their performance under a relative humidity of 32 ± 2%. Transcriptome analysis of cultured cells during long-term desiccation (24, 72, and 168 h) was conducted in TruSeq PE Cluster Kit (Illumina) by paired-end methods. Lawn-cultured S. Enteritidis cells have stronger survivability (only decreased by 0.78 ± 0.12 log after 130 d of storage) and heat tolerance (higher D/β value) than those from the broth method. More desiccation genes of lawn-cultured cells were significantly upregulated from growth to long-term desiccation. Differentially expressed genes were the most enriched in the ribosome and sulfur metabolism pathways in the lawn- and broth-cultured groups. This study tracked the transcriptomic differences between two cultured groups in response to long-term desiccation stress and revealed some molecular mechanisms underlying their different suitability in microbial studies of LMFs.
草皮采集法使用固体培养基(如胰蛋白酶大豆琼脂、TSA)来产生细菌草皮,被广泛应用于低水分食品(水分活度小于 0.85 的食品)微生物研究中的微生物培养。它在低水分食品中产生具有更高 D 值的耐干燥细胞;然而,关于细菌抗性的分子机制知之甚少。肠炎沙门氏菌 Enteritidis PT 30(S. Enteritidis)是低水分食品中最相关的病原体,在 TSA 和胰蛋白酶大豆肉汤(TSB)中进行培养。细胞被收获并接种在滤纸上,以评估它们在相对湿度为 32 ± 2%下的性能。通过使用 TruSeq PE Cluster Kit(Illumina)进行的配对末端方法,对长期干燥(24、72 和 168 小时)过程中的培养细胞进行了转录组分析。与肉汤法相比,草皮培养的 S. Enteritidis 细胞具有更强的生存能力(在 130 天的储存后仅下降了 0.78 ± 0.12 个对数)和耐热性(更高的 D/β 值)。与生长相比,草皮培养细胞的更多干燥基因在长期干燥过程中显著上调。差异表达基因在草皮和肉汤培养组中最富集于核糖体和硫代谢途径。本研究跟踪了两个培养组在应对长期干燥应激时的转录组差异,并揭示了它们在低水分食品微生物研究中不同适用性的一些分子机制。