Identification of aflatoxin biosynthesis genes by genetic complementation in an Aspergillus flavus mutant lacking the aflatoxin gene cluster.

Applied and Environmental Microbiology
R PrietoC P Woloshuk

Abstract

Aspergillus flavus mutant strain 649, which has a genomic DNA deletion of at least 120 kb covering the aflatoxin biosynthesis cluster, was transformed with a series of overlapping cosmids that contained DNA harboring the cluster of genes. The mutant phenotype of strain 649 was rescued by transformation with a combination of cosmid clones 5E6, 8B9, and 13B9, indicating that the cluster of genes involved in aflatoxin biosynthesis resides in the 90 kb of A. flavus genomic DNA carried by these clones. Transformants 5E6 and 20B11 and transformants 5E6 and 8B9 accumulated intermediate metabolites of the aflatoxin pathway, which were identified as averufanin and/or averufin, respectively. These data suggest that avf1, which is involved in the conversion of averufin to versiconal hemiacetal acetate, was present in the cosmid 13B9. Deletion analysis of 13B9 located the gene on a 7-kb DNA fragment of the cosmid. Transformants containing cosmid 8B9 converted exogenously supplied O-methylsterigmatocystin to aflatoxin, indicating that the oxidoreductase gene (ord1), which mediates the conversion of O-methylsterigmatocystin to aflatoxin, is carried by this cosmid. The analysis of transformants containing deletions of 8B9 led to the localizat...Continue Reading

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Citations

Jul 1, 1999·FEMS Microbiology Letters·M J Sweeney, A D Dobson
Jun 4, 2005·Applied and Environmental Microbiology·Ying WenKimiko Yabe
Apr 9, 1998·FEMS Microbiology Letters·C P Woloshuk, R Prieto
May 11, 2010·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·M DonnerP J Cotty
Mar 12, 2004·Annual Review of Phytopathology·G A Payne, M P Brown
Oct 2, 2014·The Journal of Microbiology·Sheikh Imranudin Sheikh-AliMd Enamul Hoque
Mar 10, 2004·Applied and Environmental Microbiology·Jiujiang YuJoan W Bennett
May 18, 1999·Fungal Genetics and Biology : FG & B·M P BrownG A Payne
Oct 31, 2000·Applied and Environmental Microbiology·P K ChangT E Cleveland
Oct 6, 1998·Applied and Environmental Microbiology·D M MeyersG A Payne
Nov 18, 2008·Fungal Genetics and Biology : FG & B·D Ryan Georgianna, Gary A Payne
Feb 20, 2002·Chemical Reviews·Robert E. Minto, Craig A. Townsend

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