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Ustilago maydis Seedling Infection Assay and Scoring.

Methods in molecular biology (Clifton, N.J.) · 1 Jan 2026 · 10.1007/978-1-0716-5364-7_3

Abstract

This protocol outlines a reproducible method for assessing the virulence of Ustilago maydis strains in Zea mays seedlings through targeted syringe inoculation of the leaf whorl. Using seedlings at the V3-V4 growth stage ensures optimal susceptibility and developmental uniformity. The method can be used with both solopathogenic strains and compatible mating-type combinations, enabling comparative analyses of infection efficiency and symptom development. Fungal cultures are prepared under controlled conditions to maintain virulence, and post-infection humidity is regulated to enhance colonization success. Symptom progression is monitored over 10-12 days and quantified using an ordinal disease scoring system. This assay provides a robust tool for evaluating effector mutants, analyzing host-pathogen interactions, and comparing strain virulence under standardized conditions.

Plant phenotyping relevance

トウモロコシ苗の感染症状を標準化して評価・スコア化する再現可能な表現型測定プロトコルが中心であり、単なる病原体検出ではない。

abstractThis protocol outlines a reproducible method for assessing the virulence of Ustilago maydis strains in Zea mays seedlings through targeted syringe inoculation of the leaf whorl.
abstractSymptom progression is monitored over 10-12 days and quantified using an ordinal disease scoring system.

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