Aflatoxin formation and gene expression in response to carbon source media shift in Aspergillus parasiticus

Abstract

Aflatoxins are toxic and carcinogenic polyketide metabolites produced by fungal species, including Aspergillus flavus and A. parasiticus. The biosynthesis of aflatoxins is modulated by many environmental factors, including the availability of a carbon source. The gene expression profile of A. parasiticus was evaluated during a shift from a medium with low concentration of simple sugars, yeast extract (YE), to a similar medium with sucrose, yeast extract sucrose (YES). Gene expression and aflatoxins (B₁, B₂, G₁, and G₂) were quantified from fungal mycelia harvested pre- and post-shifting. When compared with YE media, YES caused temporary reduction of the aflatoxin levels detected at 3-h post-shifting and they remained low well past 12 h post-shift. Aflatoxin levels did not exceed the levels in YE until 24 h post-shift, at which time point a tenfold increase was observed over YE. Microarray analysis comparing the RNA samples from the 48-h YE culture to the YES samples identified a total of 2120 genes that were expressed across all experiments, including most of the aflatoxin biosynthesis genes. One-way analysis of variance (ANOVA) identified 56 genes that were expressed with significant variation across all time points. Three genes responsible for converting norsolorinic acid to averantin were identified among these significantly expressed genes. The potential involvement of these genes in the regulation of aflatoxin biosynthesis is discussed.

Keywords:

• Aspergillus
• norsolorinic acid
• averantin
• aflatoxins
• carbon medium
• microarray

Acknowledgements

The authors wish to thank Jennifer Tonos and Bobbie Johnson for their assistance in aflatoxin detection; and Janell Becker for her secretarial help. Work was approved for publication as Journal Article No. J-11069 of the Mississippi Agricultural and Forestry Experiment Station, Mississippi State University.