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Fusarium verticillioides chitin synthases CHS5 and CHS7 are required for normal growth and pathogenicity

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Abstract

Fusarium verticillioides is both an endophyte and a pathogen of maize and is a health threat in many areas of the world because it can contaminate maize with fumonisins, a toxic secondary metabolite. We identified eight putative chitin synthase (CHS) genes in F. verticillioides genomic sequence, and phylogenetic evidence shows that they group into seven established CHS gene classes. We targeted two CHSs (CHS5 and CHS7) for deletion analysis and found that both are required for normal hyphal growth and maximal disease of maize seedlings and ears. CHS5 and CHS7 encode a putative class V and class VII fungal chitin synthase, respectively; they are located adjacent to each other and are divergently transcribed. Fluorescent microscopy found that both CHS deficient strains produce balloon-shaped hyphae, while growth assays indicated that they were more sensitive to cell wall stressing compounds (e.g., the antifungal compound Nikkomycin Z) than wild type. Pathogenicity assays on maize seedlings and ears indicated that both strains were significantly reduced in their ability to cause disease. Our results demonstrate that both CHS5 and CHS7 are necessary for proper hyphal growth and pathogenicity of F. verticillioides on maize.

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Acknowledgments

We would like to acknowledge Amber M. Anderson, Nathan Deppe, and Debbie Shane for technical assistance. The mention of firm names or trade products does not imply that they are endorsed or recommended by the US Department of Agriculture over the firms or similar products not mentioned.

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  1. Bacterial Foodborne Pathogens and Mycology Research Unit, Agricultural Research Service, U.S. Department of Agriculture, 1815 North University Street, Peoria, IL, 61604-3999, USA

    Troy M. Larson, David F. Kendra, Mark Busman & Daren W. Brown

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  1. Troy M. Larson
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Correspondence to Troy M. Larson.

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Communicated by B. Cormack.

Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable.

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Supplemental Fig. 1

Alignment of conserved region 1 (Con1) found in the catalytic site of the eight putative F.verticillioides CHSs. Identical amino acids are shaded. Roman numerals above the alignment define the three sub domains (I–III) required for proper chitin synthase activity and classification. (TIFF 587 kb)

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Larson, T.M., Kendra, D.F., Busman, M. et al. Fusarium verticillioides chitin synthases CHS5 and CHS7 are required for normal growth and pathogenicity. Curr Genet 57, 177–189 (2011). https://doi.org/10.1007/s00294-011-0334-6

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