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Method for monitoring deletions in the aflatoxin biosynthesis gene cluster of A spergillus flavus with multiplex PCR
Author(s) -
Callicott K.A.,
Cotty P.J.
Publication year - 2015
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12337
Subject(s) - aspergillus flavus , aflatoxin , biology , multiplex polymerase chain reaction , multiplex , indel , gene cluster , genetics , aspergillus , gene , population , mycotoxin , microbiology and biotechnology , polymerase chain reaction , genotype , single nucleotide polymorphism , demography , sociology
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 ( CAP s). CAP markers are amplified in four multiplex PCR s, 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. Significance and Impact of the Study 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.