Huerta Apolonio Ignacio, Kesten Christopher, Menna Alexandra Lauren, Sancho-Andrés Gloria, Sanchez-Rodriguez Clara
Department of Biology, ETH Zurich, Zurich, Switzerland.
Curr Protoc Plant Biol. 2020 Sep;5(3):e20113. doi: 10.1002/cppb.20113.
Root vascular pathogens are some of the world's most devastating plant pathogens. However, the methods used to determine plant susceptibility to this class of pathogen are laborious, variable, and in most cases qualitative. Here we present a rapid, simple, and robust infection assay for the characterization of Arabidopsis thaliana resistance to the fungal root pathogen Fusarium oxysporum. The method utilizes fungal root vascular penetrations and fungal-induced root growth inhibition to deliver a quantitative assessment of plant susceptibility with spatial and temporal resolution. These plant susceptibility indicators are paired with a semiautomated data analysis pipeline to deliver a reproducible assessment of plant susceptibility to root vascular pathogens such as F. oxysporum. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Arabidopsis thaliana plate infection assay using fluorescently labeled Fusarium oxysporum Support Protocol 1: Preparation of A. thaliana germination plates Support Protocol 2: Preparation of the F. oxysporum culture Basic Protocol 2: Data acquisition of F. oxysporum plant infection assay Support Protocol 3: Acquiring root growth inhibition data using Fiji.
根部维管束病原体是世界上最具破坏力的一些植物病原体。然而,用于确定植物对这类病原体易感性的方法既费力、结果多变,而且在大多数情况下是定性的。在此,我们提出一种快速、简单且可靠的感染检测方法,用于表征拟南芥对真菌根部病原体尖孢镰刀菌的抗性。该方法利用真菌对根部维管束的穿透以及真菌诱导的根部生长抑制,以时空分辨率对植物易感性进行定量评估。这些植物易感性指标与一个半自动化的数据分析流程相结合,以对植物对诸如尖孢镰刀菌等根部维管束病原体的易感性进行可重复的评估。© 2020威利期刊公司。基本方案1:使用荧光标记的尖孢镰刀菌进行拟南芥平板感染检测 支持方案1:拟南芥萌发平板的制备 支持方案2:尖孢镰刀菌培养物的制备 基本方案2:尖孢镰刀菌植物感染检测的数据采集 支持方案3:使用Fiji获取根部生长抑制数据