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m6A methyltransferase AflIme4 orchestrates mycelial growth, development and aflatoxin B1 biosynthesis in Aspergillus

Liuke Liang1, Xiaoyan Wang1, Shan Wei1

  • 1College of biological engineering, Henan University of Technology, Zhengzhou 450001, China.

Microbiological Research
|April 9, 2024
PubMed

Insights

A novel enzyme, AflIme4, regulates mRNA methylation in Aspergillus flavus, significantly reducing aflatoxin B1 (AFB1) production and impacting fungal growth. This discovery offers a potential target for controlling AFB1 contamination.

Area of Science:

  • Molecular Biology
  • Mycology
  • Food Safety

Background:

  • Aflatoxin B1 (AFB1) produced by Aspergillus flavus is a significant threat to agriculture, food safety, and human health.
  • Messenger RNA (mRNA) N6-methyladenosine (m6A) methylation is known to regulate A. flavus growth and AFB1 production, but its specific role requires further investigation.

Purpose of the Study:

  • To identify and characterize a novel m6A methyltransferase, AflIme4, in A. flavus.
  • To elucidate the function of AflIme4 in regulating fungal development and AFB1 biosynthesis.

Main Methods:

  • Identification and characterization of the AflIme4 gene and its protein product.
  • Generation of A. flavus ΔAflIme4 knockout strains.
  • Analysis of fungal growth, development, and AFB1 production.
  • RNA sequencing (RNA-Seq) and RT-qPCR for gene expression analysis.
  • Methylated RNA immunoprecipitation-qPCR (MeRIP-qPCR) to assess m6A modification levels and mRNA stability.

Main Results:

  • AflIme4 was localized to the cytoplasm and its absence significantly reduced overall mRNA methylation.
  • Knockout of AflIme4 impaired A. flavus growth, conidiation, hydrophobicity, sclerotium yield, and pathogenicity, while increasing sensitivity to oxidative and osmotic stress.
  • Crucially, AFB1 production was markedly inhibited in ΔAflIme4 strains.
  • RNA-Seq and MeRIP-qPCR revealed that AflIme4 enhances m6A methylation of genes in the AFB1 biosynthesis pathway, reducing their mRNA stability and consequently AFB1 production.

Conclusions:

  • AflIme4 functions as an m6A methyltransferase that attenuates mRNA stability of AFB1 biosynthesis genes, thereby inhibiting AFB1 production.
  • AflIme4 is a potential molecular target for developing strategies to control A. flavus growth and mitigate aflatoxin contamination in agriculture and food.
Keywords:
AflIme4Aflatoxin B(1)Aspergillus flavusM(6)A methylation modification

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