Applied Microbiology and Biotechnology

, Volume 102, Issue 12, pp 5209–5220 | Cite as

Aspergillus flavus GPI-anchored protein-encoding ecm33 has a role in growth, development, aflatoxin biosynthesis, and maize infection

  • Perng-Kuang Chang
  • Qi Zhang
  • Leslie Scharfenstein
  • Brian Mack
  • Akira Yoshimi
  • Ken Miyazawa
  • Keietsu Abe
Applied genetics and molecular biotechnology


Many glycosylphosphatidylinositol-anchored proteins (GPI-APs) of fungi are membrane enzymes, organization components, and extracellular matrix adhesins. We analyzed eight Aspergillus flavus transcriptome sets for the GPI-AP gene family and identified AFLA_040110, AFLA_063860, and AFLA_113120 to be among the top 5 highly expressed genes of the 36 family genes analyzed. Disruption of the former two genes did not drastically affect A. flavus growth and development. In contrast, disruption of AFLA_113120, an orthologue of Saccharomyces cerevisiae ECM33, caused a significant decrease in vegetative growth and conidiation, promoted sclerotial production, and altered conidial pigmentation. The A. flavus ecm33 null mutant, compared with the wild type and the complemented strain, produced predominantly aflatoxin B2 but accumulated comparable amounts of cyclopiazonic acid. It showed decreased sensitivity to Congo red at low concentrations (25–50 μg/mL) but had increased sensitivity to calcofluor white at high concentrations (250–500 μg/mL). Analyses of cell wall carbohydrates indicated that the α-glucan content was decreased significantly (p < 0.05), but the contents of chitin and ß-glucan were increased in the mutant strain. In a maize colonization study, the mutant was shown to be impaired in its infectivity and produced 3- to 4-fold lower amounts of conidia than the wild type and the complemented strain. A. flavus Ecm33 is required for proper cell wall composition and plays an important role in normal fungal growth and development, aflatoxin biosynthesis, and seed colonization.


Aspergillus flavus Glycosylphosphatidylinisotol Cell wall integrity Glucan Chitin Calcofluor white 


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain studies with human participants or animals.

Supplementary material

253_2018_9012_MOESM1_ESM.pdf (825 kb)
ESM 1 (PDF 824 kb)


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Copyright information

© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2018

Authors and Affiliations

  • Perng-Kuang Chang
    • 1
  • Qi Zhang
    • 2
  • Leslie Scharfenstein
    • 1
  • Brian Mack
    • 1
  • Akira Yoshimi
    • 3
  • Ken Miyazawa
    • 4
  • Keietsu Abe
    • 3
    • 4
  1. 1.Southern Regional Research Center, Agricultural Research ServiceUS Department of AgricultureNew OrleansUSA
  2. 2.Oil Crops Research InstituteChinese Academy of Agricultural SciencesWuhanChina
  3. 3.ABE-Project, New Industry Creation Hatchery CenterTohoku UniversitySendaiJapan
  4. 4.Laboratory of Applied Microbiology, Department of Microbial Biotechnology, Graduate School of Agricultural SciencesTohoku UniversitySendaiJapan

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