Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
Molecular Phytobacteriology Laboratory, Infectious Disease Research Center, Korea Research Institute of Bioscience &Biotechnology (KRIBB), Daejeon, South Korea.
Nat Protoc. 2017 Jul;12(7):1359-1377. doi: 10.1038/nprot.2017.023. Epub 2017 Jun 15.
Airborne chemical signals emitted by bacteria influence the behavior of other bacteria and plants. We present an overview of in vitro methods for evaluating bacterial and plant responses to bacterial volatile compounds (BVCs). Three types of equipment have been used to physically separate the bacterial test strains from either other bacterial strains or plants (in our laboratory we use either Arabidopsis or tobacco plant seedlings): a Petri dish containing two compartments (BI Petri dish); two Petri dishes connected with tubing; and a microtiter-based assay. The optimized procedure for the BI Petri dish system is described in this protocol and can be widely used for elucidation of potential function in interactions between diverse microbes and those plant and chemical volatiles emitted by bacteria that are most likely to mediate bacterial or plant responses to BVCs. We also describe a procedure for metabolome-based BVC profiling via dynamic (i.e., continuous airflow) or static headspace sampling using solid-phase microextraction (SPME). Using both these procedures, bacteria-bacteria communications and bacteria-plant interactions mediated by BVCs can be rapidly investigated (within 1-4 weeks).
细菌释放的空气传播化学信号会影响其他细菌和植物的行为。我们概述了用于评估细菌和植物对细菌挥发性化合物（BVC）反应的体外方法。已经使用了三种设备将细菌测试菌株与其他细菌菌株或植物物理分离（在我们的实验室中，我们使用拟南芥或烟草植物幼苗）：一个包含两个隔室的培养皿（BI 培养皿）；两个通过管道连接的培养皿；和一个微量滴定板测定法。该协议中描述了 BI 培养皿系统的优化程序，该程序可广泛用于阐明不同微生物之间以及最有可能介导细菌或植物对 BVC 反应的细菌所释放的植物和化学挥发物之间相互作用的潜在功能。我们还描述了一种通过固相微萃取（SPME）使用动态（即连续气流）或静态顶空采样进行基于代谢组学的 BVC 分析的程序。使用这两种程序，可以快速研究 BVC 介导的细菌-细菌通讯和细菌-植物相互作用（在 1-4 周内）。
