In-Plate Quantitative Characterization of Arabidopsis thaliana Susceptibility to the Fungal Vascular Pathogen Fusarium oxysporum.

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2020-09-01 DOI:10.1002/cppb.20113

Apolonio Ignacio Huerta, Christopher Kesten, Alexandra Lauren Menna, Gloria Sancho-Andrés, Clara Sanchez-Rodriguez

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引用次数: 6

Abstract

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.

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拟南芥对血管真菌病原菌尖孢镰刀菌敏感性的板内定量鉴定。

根维管病原体是世界上最具破坏性的植物病原体之一。然而，用于确定植物对这类病原体的易感性的方法是费力的，多变的，而且在大多数情况下是定性的。在这里，我们提出了一种快速、简单和强大的感染试验，用于鉴定拟南芥对真菌根病原体尖孢镰刀菌的抗性。该方法利用真菌根维管渗透和真菌诱导的根生长抑制来提供具有空间和时间分辨率的植物敏感性定量评估。这些植物敏感性指标与半自动数据分析管道相结合，提供植物对根维管病原体(如尖孢镰刀菌)敏感性的可重复评估。©2020 Wiley期刊有限责任公司基本方案1:拟南芥平板感染试验使用荧光标记的尖孢镰刀菌支持方案1:拟南芥萌发板的制备支持方案2:尖孢镰刀菌培养的制备基本方案2:尖孢镰刀菌植物感染试验的数据采集支持方案3:使用Fiji获取根生长抑制数据。

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Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.