A yeast based screening method for Arabidopsis zearalenone-metabolism mutants

Juan Antonio Torres Acosta; Ulrike Werner; Herwig Bachmann; Franz Berthiller; Rainer Schuhmacher; Rudolf Krska; Marie Theres Hauser; Gerhard Adam

A yeast based screening method for Arabidopsis zearalenone-metabolism mutants

Juan Antonio Torres Acosta^*, Ulrike Werner, Herwig Bachmann, Franz Berthiller, Rainer Schuhmacher, Rudolf Krska, Marie Theres Hauser, Gerhard Adam

^*Corresponding author for this work

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Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Arabidopsis thaliana and other plants are able to rapidly metabolize the estrogenic Fusarium metabolite zearalenone (ZON) into ZON-glucoside and further unknown compounds. We present here a rapid and efficient method to screen for Arabidopsis mutants unable to metabolize ZON. Arabidopsis seedling are grown in 96 well microtiter plates and subsequently incubated with ZON and β-glucosidase. The remaining ZON in the supernatant is detected with an engineered estrogen-responsive yeast strain. Extracellular ZON-4- O-glucoside (ZON-Glc), which is not detected by the indicator strain, is converted back to ZON by the added almond β-glucosidase. Wildtype Arabidopsis seedlings inactivate 2 ppm ZON in three days, which is easily detected by the colorimetric readout. The assay should be useful to identify mutants affected in ZON metabolism.

Original language	English
Pages (from-to)	545-548
Number of pages	4
Journal	Cereal Research Communications
Volume	36
Issue number	SUPPL. 6
Publication status	Published - 2008

Keywords

Arabidopsis
Biotransformation
Yeast bioassay
Zearalenone

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title = "A yeast based screening method for Arabidopsis zearalenone-metabolism mutants",

abstract = "Arabidopsis thaliana and other plants are able to rapidly metabolize the estrogenic Fusarium metabolite zearalenone (ZON) into ZON-glucoside and further unknown compounds. We present here a rapid and efficient method to screen for Arabidopsis mutants unable to metabolize ZON. Arabidopsis seedling are grown in 96 well microtiter plates and subsequently incubated with ZON and β-glucosidase. The remaining ZON in the supernatant is detected with an engineered estrogen-responsive yeast strain. Extracellular ZON-4- O-glucoside (ZON-Glc), which is not detected by the indicator strain, is converted back to ZON by the added almond β-glucosidase. Wildtype Arabidopsis seedlings inactivate 2 ppm ZON in three days, which is easily detected by the colorimetric readout. The assay should be useful to identify mutants affected in ZON metabolism.",

keywords = "Arabidopsis, Biotransformation, Yeast bioassay, Zearalenone",

author = "Acosta, {Juan Antonio Torres} and Ulrike Werner and Herwig Bachmann and Franz Berthiller and Rainer Schuhmacher and Rudolf Krska and Hauser, {Marie Theres} and Gerhard Adam",

year = "2008",

language = "English",

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journal = "Cereal Research Communications",

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TY - JOUR

T1 - A yeast based screening method for Arabidopsis zearalenone-metabolism mutants

AU - Acosta, Juan Antonio Torres

AU - Werner, Ulrike

AU - Bachmann, Herwig

AU - Berthiller, Franz

AU - Schuhmacher, Rainer

AU - Krska, Rudolf

AU - Hauser, Marie Theres

AU - Adam, Gerhard

PY - 2008

Y1 - 2008

N2 - Arabidopsis thaliana and other plants are able to rapidly metabolize the estrogenic Fusarium metabolite zearalenone (ZON) into ZON-glucoside and further unknown compounds. We present here a rapid and efficient method to screen for Arabidopsis mutants unable to metabolize ZON. Arabidopsis seedling are grown in 96 well microtiter plates and subsequently incubated with ZON and β-glucosidase. The remaining ZON in the supernatant is detected with an engineered estrogen-responsive yeast strain. Extracellular ZON-4- O-glucoside (ZON-Glc), which is not detected by the indicator strain, is converted back to ZON by the added almond β-glucosidase. Wildtype Arabidopsis seedlings inactivate 2 ppm ZON in three days, which is easily detected by the colorimetric readout. The assay should be useful to identify mutants affected in ZON metabolism.

AB - Arabidopsis thaliana and other plants are able to rapidly metabolize the estrogenic Fusarium metabolite zearalenone (ZON) into ZON-glucoside and further unknown compounds. We present here a rapid and efficient method to screen for Arabidopsis mutants unable to metabolize ZON. Arabidopsis seedling are grown in 96 well microtiter plates and subsequently incubated with ZON and β-glucosidase. The remaining ZON in the supernatant is detected with an engineered estrogen-responsive yeast strain. Extracellular ZON-4- O-glucoside (ZON-Glc), which is not detected by the indicator strain, is converted back to ZON by the added almond β-glucosidase. Wildtype Arabidopsis seedlings inactivate 2 ppm ZON in three days, which is easily detected by the colorimetric readout. The assay should be useful to identify mutants affected in ZON metabolism.

KW - Arabidopsis

KW - Biotransformation

KW - Yeast bioassay

KW - Zearalenone

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M3 - Article

AN - SCOPUS:80052787591

SN - 0133-3720

VL - 36

SP - 545

EP - 548

JO - Cereal Research Communications

JF - Cereal Research Communications

IS - SUPPL. 6

ER -

A yeast based screening method for Arabidopsis zearalenone-metabolism mutants

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