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      Non-aflatoxigenic Aspergillus flavus to prevent aflatoxin contamination in crops: advantages and limitations

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          Abstract

          Aspergillus flavus is a diverse assemblage of strains that include aflatoxin-producing and non-toxigenic strains with cosmopolitan distribution. The most promising strategy currently being used to reduce preharvest contamination of crops with aflatoxin is to introduce non-aflatoxin (biocontrol) A. flavus into the crop environment. Whether or not introduction of biocontrol strains into agricultural fields is enough to reduce aflatoxin contamination to levels required for acceptance of the contaminated food as fit for consumption is still unknown. There is no question that biocontrol strains are able to reduce the size of the populations of aflatoxin-producing strains but the available data suggests that at most only a four- to five-fold reduction in aflatoxin contamination is achieved. There are many challenges facing this strategy that are both short term and long term. First, the population biology of A. flavus is not well understood due in part to A. flavus’s diversity, its ability to form heterokaryotic reproductive forms, and its unknown ability to survive for prolonged periods after application. Second, biocontrol strains must be selected that are suitable for the environment, the type of crop, and the soil into which they will be introduced. Third, there is a need to guard against inadvertent introduction of A. flavus strains that could impose an additional burden on food safety and food quality, and fourth, with global warming and resultant changes in the soil nutrients and concomitant microbiome populations, the biocontrol strategy must be sufficiently flexible to adapt to such changes. Understanding genetic variation within strains of A. flavus is important for developing a robust biocontrol strategy and it is unlikely that a “one size fits all” strategy will work for preharvest aflatoxin reduction.

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          Most cited references51

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          Outbreak of an acute aflatoxicosis in Kenya in 2004: identification of the causal agent.

          Maize contaminated with aflatoxins has been implicated in deadly epidemics in Kenya three times since 1981, but the fungi contaminating the maize with aflatoxins have not been characterized. Here we associate the S strain of Aspergillus flavus with lethal aflatoxicoses that took more than 125 lives in 2004.
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            Characterization and population analysis of the mating-type genes in Aspergillus flavus and Aspergillus parasiticus.

            We characterize the mating-type genes in Aspergillus flavus,Aspergillus parasiticus and Petromyces alliaceus. A single MAT1-1 or MAT1-2 gene was detected in the genomes of A. flavus and A. parasiticus, which is consistent with a potential heterothallic organization of MAT genes in these species. In contrast, the only known, functionally homothallic species in Aspergillus section Flavi, P. alliaceus, has tightly linked (<2kb) MAT1-1 and MAT1-2 genes, typical of other self-fertile homothallic euascomycetes. This is the first example of linked MAT genes within a homothallic species of Aspergillus. We tested the null hypothesis of no significant difference in the frequency of MAT1-1 and MAT1-2 in A. flavus and A. parasiticus sampled from a single peanut field in Georgia. For each species, mating-type frequencies were determined for the total population samples and for samples that were clone-corrected based on vegetative compatibility groups (VCGs) and aflatoxin gene cluster haplotypes. There was no significant difference in the frequency of the two mating types for A. flavus and A. parasiticus in either VCG or haplotype clone-corrected samples. The existence of both mating-type genes in equal proportions in A. flavus and A. parasiticus populations, coupled with their expression at the mRNA level and the high amino acid sequence identity of MAT1-1 (77%) and MAT1-2 (83%) with corresponding homologs in P. alliaceus, indicates the potential functionality of these genes and the possible existence of a sexual state in these agriculturally important species.
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              Aflatoxin biosynthesis cluster gene cypA is required for G aflatoxin formation.

              Aspergillus flavus isolates produce only aflatoxins B1 and B2, while Aspergillus parasiticus and Aspergillus nomius produce aflatoxins B1, B2, G1, and G2. Sequence comparison of the aflatoxin biosynthesis pathway gene cluster upstream from the polyketide synthase gene, pksA, revealed that A. flavus isolates are missing portions of genes (cypA and norB) predicted to encode, respectively, a cytochrome P450 monooxygenase and an aryl alcohol dehydrogenase. Insertional disruption of cypA in A. parasiticus yielded transformants that lack the ability to produce G aflatoxins but not B aflatoxins. The enzyme encoded by cypA has highest amino acid identity to Gibberella zeae Tri4 (38%), a P450 monooxygenase previously shown to be involved in trichodiene epoxidation. The substrate for CypA may be an intermediate formed by oxidative cleavage of the A ring of O-methylsterigmatocystin by OrdA, the P450 monooxygenase required for formation of aflatoxins B1 and B2.

                Author and article information

                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                10 February 2014
                2014
                : 5
                : 50
                Affiliations
                Southern Regional Research Center, United States Department of Agriculture – Agricultural Research Service New Orleans, LA, USA
                Author notes

                Edited by: Mahendra Kumar Rai, Sant Gadge Baba Amravati University at Amravati, India

                Reviewed by: Ebrahim Hadavi, Islamic Azad University, Iran; Zhao Chen, Clemson University, USA

                *Correspondence: Kenneth C. Ehrlich, Southern Regional Research Center, United States Department of Agriculture – Agricultural Research Service, New Orleans, LA 70124, USA e-mail: ken.ehrlich@ 123456ars.usda.gov

                This article was submitted to Food Microbiology, a section of the journal Frontiers in Microbiology.

                Article
                10.3389/fmicb.2014.00050
                3918586
                24575088
                3a390629-6fe1-4ae9-b6fe-21af5f515032
                Copyright © 2014 Ehrlich.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 18 December 2013
                : 23 January 2014
                Page count
                Figures: 2, Tables: 2, Equations: 0, References: 76, Pages: 9, Words: 0
                Categories
                Microbiology
                Review Article

                Microbiology & Virology
                recombination,biocontrol,aspergillus flavus,maize,aflatoxin,population diversity,cottonseed,food safety

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