Abstract

Aspergillus flavus and Fusarium verticillioides infect maize kernels and contaminate them with the mycotoxins aflatoxin, and fumonisin, respectively. Genetic resistance in maize to these fungi and to mycotoxin contamination has been difficult to achieve due to lack of identified resistance genes. The objective of this study was to identify new candidate resistance genes by characterizing their temporal expression in response to infection and comparing expression of these genes with genes known to be associated with plant defense. Fungal colonization and transcriptional changes in kernels inoculated with each fungus were monitored at 4, 12, 24, 48, and 72 h post inoculation (hpi). Maize kernels responded by differential gene expression to each fungus within 4 hpi, before the fungi could be observed visually, but more genes were differentially expressed between 48 and 72 hpi, when fungal colonization was more extensive. Two-way hierarchal clustering analysis grouped the temporal expression profiles of the 5,863 differentially expressed maize genes over all time points into 12 clusters. Many clusters were enriched for genes previously associated with defense responses to either A. flavus or F. verticillioides. Also within these expression clusters were genes that lacked either annotation or assignment to functional categories. This study provided a comprehensive analysis of gene expression of each A. flavus and F. verticillioides during infection of maize kernels, it identified genes expressed early and late in the infection process, and it provided a grouping of genes of unknown function with similarly expressed defense related genes that could inform selection of new genes as targets in breeding strategies.

Highlights

  • Aspergillus flavus is an opportunistic fungal pathogen that can grow either as a saprophyte in the soil or as a pathogen of many plant species

  • Maize kernels developing in the field were inoculated with either A. flavus or F. verticillioides (Figure 1a) and colonization

  • Extensive colonization by A. flavus and F. verticillioides was observed at 48 hpi in the aleurone and the outermost layer of the endosperm (Figures 1d,e)

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Summary

Introduction

Aspergillus flavus is an opportunistic fungal pathogen that can grow either as a saprophyte in the soil or as a pathogen of many plant species. Hosts of A. flavus include maize kernels, peanuts, cottonseeds and tree nuts (Payne, 1992; Payne and Yu, 2010; Scheidegger and Payne, 2003; St. Leger et al, 2000). Unlike A. flavus, Fusarium verticillioides can colonize maize seeds as an endophyte or necrotroph. Often related to plant stress, F. verticillioides can cause seedling blight, ear rot, and stalk rot (Bacon et al, 1992; Munkvold, 2003; Pei-Bao et al, 2010).

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