title: Method for monitoring deletions in the aflatoxin biosynthesis gene cluster of Aspergillus flavus with multiplex PCR.
abstract: The report presents a rapid, inexpensive and simple method for monitoring indels with influence on aflatoxin biosynthesis within Aspergillus flavus populations. PCR primers were developed for 32 markers spaced approximately every 5 kb from 20 kb proximal to the aflatoxin biosynthesis gene cluster to the telomere repeat. This region includes gene clusters required for biosynthesis of aflatoxins and cyclopiazonic acid; the resulting data were named cluster amplification patterns (CAPs). CAP markers are amplified in four multiplex PCRs, greatly reducing the cost and time to monitor indels within this region across populations. The method also provides a practical tool for characterizing intraspecific variability in A. flavus not captured with other methods.
  
  
  CONCLUSIONS
  Aflatoxins, potent naturally-occurring carcinogens, cause significant agricultural problems. The most effective method for preventing contamination of crops with aflatoxins is through use of atoxigenic strains of Aspergillus flavus to alter the population structure of this species and reduce incidences of aflatoxin producers. Cluster amplification pattern (CAP) is a rapid multiplex PCR method for identifying and monitoring indels associated with atoxigenicity in A. flavus. Compared to previous techniques, the reported method allows for increased resolution, reduced cost, and greater speed in monitoring the stability of atoxigenic strains, incidences of indel mediated atoxigenicity and the structure of A. flavus populations.
authors.affiliation: Food and Feed Safety Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Tucson, AZ, USA.
authors.forename: K A
authors.initials: KA
authors.lastname: Callicott
authors.forename: P J
authors.initials: PJ
authors.lastname: Cotty
mesh_terms: D000348:Aflatoxins / Q000096:biosynthesis*; D001231:Aspergillus flavus / Q000235:genetics*; D017353:Gene Deletion; D005800:Genes, Fungal; D054643:INDEL Mutation; D005810:Multigene Family; D060885:Multiplex Polymerase Chain Reaction; D016133:Polymerase Chain Reaction
chemical_list: D000348:Aflatoxins
keywords: Aspergillus flavus; aflatoxin; biocontrol; cyclopiazonic acid; multiplex PCR; subtelomere stability