Assessment of Wheat Resistance to Fusarium graminearum by Automated Image Analysis of Detached Leaves Assay   

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Original research article

A brief version of this protocol appeared in:
Plant Physiology
Dec 2015


Fusarium head blight (FHB) caused by Fusarium pathogens is a globally important cereal disease. To study Fusarium pathogenicity and host disease resistance, robust methods for disease assessment and quantification are needed. Here we describe the procedure of a detached leaves assay emphasizing the image analysis. The protocol provides the different steps of a rapid, automatic and quantitative image analysis to evaluate leaf area infected by Fusarium graminearum.

Keywords: Detached leaf assay, Disease assessment, Fusarium graminearum, Image analysis, Fiji, Pathogenicity, Wheat


Evaluation of wheat FHB resistance at the whole plant level is estimated by visual scoring at flowering stage which is laborious, time consuming and requires space. Therefore in vitro methods that expedites disease assessment for FHB resistance at early plant stage have been developed, such as seed germination assay (Browne, 2009), coleoptiles assay (Shin et al., 2014), detached leaf assay (Browne and Cooke, 2004) and seedling assay (Li et al., 2010). Detached leaves assay is commonly used to assess host responses to Fusarium and was successful in identifying components of FHB resistance (Browne and Cooke, 2004). In such assays pathogen establishment is visually assessed which is time-consuming and limits accurate measurement. The method described recently by Perochon et al. (2015) and detailed here resolved these two limitations by using an automatic method that quantifies leaf area infected by image analysis based on particle size.

Copyright: © 2016 The Authors; exclusive licensee Bio-protocol LLC.
How to cite:  Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
  1. Perochon, A. and Doohan, F. M. (2016). Assessment of Wheat Resistance to Fusarium graminearum by Automated Image Analysis of Detached Leaves Assay. Bio-protocol 6(24): e2065. DOI: 10.21769/BioProtoc.2065.
  2. Perochon, A., Jianguang, J., Kahla, A., Arunachalam, C., Scofield, S. R., Bowden, S., Wallington, E. and Doohan, F. M. (2015). TaFROG encodes a Pooideae orphan protein that interacts with SnRK1 and enhances resistance to the mycotoxigenic fungus Fusarium graminearum. Plant Physiol 169(4): 2895-2906.

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