Department of Biological Sciences, Virginia Tech, Blacksburg, VA, United States of America.
PLoS One. 2019 Jun 14;14(6):e0218220. doi: 10.1371/journal.pone.0218220. eCollection 2019.
Bacterial endospores can survive harsh environmental conditions and long-term dormancy in the absence of nutrients, but can rapidly germinate under favorable conditions. In the present study, we employed transposon sequencing (Tn-seq) to identify genes with previously uncharacterized roles in spore germination. Identified genes that encoded spore inner membrane proteins were chosen for study of defined mutants, which exhibited delayed germination in several assays in response to varying germinants. Significantly slowed release of DPA indicated that mutants were affected in Stage I of germination. Several mutants exhibited phenotypic traits consistent with failure of a GerA germinant receptor-mediated response, while others appeared to have a more general loss of response to varied germinants. Use of a gerA-lacZ transcriptional fusion and quantitative western blotting of GerAC allowed mutants to be classified based upon normal or decreased gerA transcription and normal or reduced GerA accumulation. Fourteen genes were identified to have newly described roles within Bacillus spore germination. A more complete understanding of this process can contribute to the development of better spore decontamination procedures.
细菌芽孢能够在缺乏营养物质的情况下，在恶劣的环境条件下生存并长期休眠，但在有利条件下可以迅速发芽。在本研究中，我们采用转座子测序（Tn-seq）来鉴定在芽孢发芽中具有先前未被描述作用的基因。选择编码芽孢内膜蛋白的基因进行了明确突变体的研究，这些突变体在几种检测中对不同的发芽剂表现出延迟发芽。DPA 的显著释放减缓表明突变体在发芽的第一阶段受到影响。一些突变体表现出与 GerA 发芽剂受体介导的反应失败一致的表型特征，而其他突变体似乎对不同的发芽剂的反应更为普遍丧失。使用 gerA-lacZ 转录融合和定量 Western 印迹分析 GerAC，突变体可以根据正常或降低的 gerA 转录和正常或减少的 GerA 积累进行分类。确定了 14 个基因在芽孢发芽过程中具有新描述的作用。更全面地了解这一过程有助于开发更好的芽孢消毒程序。
