Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Jan 23:5:100023.
doi: 10.1016/j.toxcx.2020.100023. eCollection 2020 Mar.

A mycotoxin transporter (4D) from a library of deoxynivalenol-tolerant microorganisms

Celia Jimenez-Sanchez  1 , Nina Wilson  1 , Nicole McMaster  1 , Dash Gantulga  1 , Benjamin G Freedman  2 , Ryan Senger  2 , David G Schmale 3rd  1
Affiliations

A mycotoxin transporter (4D) from a library of deoxynivalenol-tolerant microorganisms

Celia Jimenez-Sanchez et al. Toxicon X. .

Abstract

New strategies are needed to mitigate the mycotoxin deoxynivalenol (DON) in feed and food products. Microbial DNA fragments were generated from a library of DON-tolerant microorganisms. These fragments were screened in DON-sensitive yeast strains for their ability to modify or transport DON. Fragments were cloned into a PCR8/TOPO vector, and recombined into the yeast vector, pYES-DEST52. Resulting yeast transformants were screened in the presence of 100 ppm DON. Transformants that were able to grow in the presence of DON were plated on a selective medium, and the cloned microbial DNA fragments were sequenced. BLAST queries of one microbial DNA fragment (4D) showed a high degree of similarity to an ABC transporter. A series of screening and inhibition assays were conducted with a transport inhibitor (propanol), to test the hypothesis that 4D is a mycotoxin transporter. DON concentrations did not change for yeast transformants expressing 4D. The ability of yeast transformants expressing 4D to transport DON was inhibited by the addition of propanol. Moreover, yeast transformants expressing a known efflux pump (PDR5) showed similar trends in propanol transport inhibition compared to 4D. Future work should consider mycotoxin transporters such as 4D to the development of transgenic plants to limit DON accumulation in seeds.

Keywords: Deoxynivalenol; Microbial DNA fragment; Mycotoxin; Transporter; Yeast.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Schematic of sample collection, selection, library generation, screening assays, and sequencing to identify the mycotoxin transporter, 4D.
Fig. 2
Fig. 2
Growth of DON-sensitive yeast strains with or without microbial DNA fragments in the presence of 100 ppm DON. Data shown from six replicates. Error bars represent standard error of the mean.
Fig. 3
Fig. 3
Growth of DON-sensitive yeast strains with or without Pdr5 or 4D in the presence of induction medium, DON (40 ppm), propanol (1.5%), and DON (40 ppm) plus propanol (1.5%). Data shown from three replicates for experiment 1 (left) and experiment 2 (right). Error bars represent standard error of the mean.
Fig. 4
Fig. 4
Percentage of DON following screening assays of DON-sensitive yeast strains with or without Pdr5 or 4D in the presence of induction medium, DON (40 ppm), propanol (1.5%), and DON (40 ppm) plus propanol (1.5%). Data shown from three replicates for experiment 1 (left) and experiment 2 (right). Error bars represent standard error of the mean. No DON was found in the treatments without DON (media and propanol), and DON did not change significantly in the treatments containing DON (DON and DON + propanol).
See this image and copyright information in PMC

References

    1. Abolmaali S., Mitterbauer R., Spadiut O., Peruci M., Weindorfer H., Lucyshyn D., Ellersdorfer G., Lemmens M., Moll W.-D., Adam G. Engineered bakers yeast as a sensitive bioassay indicator organism for the trichothecene toxin deoxynivalenol. J. Microbiol. Methods. 2008;72:306–312. doi: 10.1016/j.mimet.2007.12.013. - DOI - PubMed
    1. Alexander N.J., McCormick S.P., Hohn T.M. TRI12, a trichothecene efflux pump from Fusarium sporotrichioides: gene isolation and expression in yeast. Mol. Gen. Genet. 1999;261:977–984. doi: 10.1007/s004380051046. - DOI - PubMed
    1. Binder E.M., Tan L.M., Chin L.J., Handl J., Richard J. vol. 137. 2007. pp. 265–282. (Worldwide occurrence of mycotoxins in commodities, feeds and feed ingredients. Animal Feed Science and Technology, Fusarium and their toxins: Mycology, occurrence, toxicity, control and economic impact). - DOI
    1. Freedman B.G. 2014. Degenerate Oligonucleotide Primed Amplification of Genomic DNA for Combinatorial Screening Libraries and Strain Enrichment.
    1. Fuchs E., Binder E., Heidler D., Krska R. Characterisation of metabolites after the microbial degradation of A- and B-trichothecenes by BBSH 797. Mycotoxin Res. 2000;16(1):66–69. doi: 10.1007/BF02942984. - DOI - PubMed