Bourigault Yvann, Chane Andrea, Barbey Corinne, Jafra Sylwia, Czajkowski Robert, Latour Xavier
Laboratory of Microbiology Signals and Microenvironment (LMSM EA 4312), University of Rouen Normandy, 55 rue Saint-Germain, F-27000 Evreux, France.
Research Federations NORVEGE Fed4277 & NORSEVE, Normandy University, F-76821 Mont-Saint-Aignan, France.
Microorganisms. 2021 Feb 1;9(2):295. doi: 10.3390/microorganisms9020295.
Promoter-probe vectors carrying fluorescent protein-reporter genes are powerful tools used to study microbial ecology, epidemiology, and etiology. In addition, they provide direct visual evidence of molecular interactions related to cell physiology and metabolism. Knowledge and advances carried out thanks to the construction of soft-rot biosensors, often inoculated in potato , are discussed in this review. Under epifluorescence and confocal laser scanning microscopies, and -tagged strains managed to monitor in situ bacterial viability, microcolony and biofilm formation, and colonization of infected plant organs, as well as disease symptoms, such as cell-wall lysis and their suppression by biocontrol antagonists. The use of dual-colored reporters encoding the first fluorophore expressed from a constitutive promoter as a cell tag, while a second was used as a regulator-based reporter system, was also used to simultaneously visualize bacterial spread and activity. This revealed the chronology of events leading to tuber maceration and quorum-sensing communication, in addition to the disruption of the latter by biocontrol agents. The promising potential of these fluorescent biosensors should make it possible to apprehend other activities, such as subcellular localization of key proteins involved in bacterial virulence , in the near future.
携带荧光蛋白报告基因的启动子探针载体是用于研究微生物生态学、流行病学和病因学的强大工具。此外,它们还为与细胞生理和代谢相关的分子相互作用提供了直接的视觉证据。本综述讨论了由于构建软腐生物传感器(通常接种于马铃薯中)而取得的知识和进展。在落射荧光显微镜和共聚焦激光扫描显微镜下,携带荧光标签的菌株能够原位监测细菌活力、微菌落和生物膜形成、感染植物器官的定殖以及疾病症状,如细胞壁裂解及其被生物防治拮抗剂抑制的情况。使用双色报告基因,其中第一个荧光团由组成型启动子表达作为细胞标签,而第二个用作基于调控因子的报告系统,也用于同时可视化细菌传播和活性。这揭示了导致块茎浸软和群体感应通讯的事件顺序,以及生物防治剂对后者的破坏。这些荧光生物传感器的潜在前景有望在不久的将来实现对其他活动的理解,例如参与细菌毒力的关键蛋白的亚细胞定位。