Cloning and heterologous expression of candidate DON-inactivating UDP-glucosyltranferases from rice and wheat in yeast.
Wolfgang Schweiger
wolfgang.schweiger@boku.ac.atDepartment of Applied Genetics and Cell Biology, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria. (Austria)
Barbara Steiner
Institute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. (Austria)
Apinun Limmongkon
Institute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. (Austria)
Kurt Brunner
Institute of Chemical Engineering, Vienna University of Technology, A-1060 Vienna, Austria. (Austria)
Marc Lemmens
Institute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. (Austria)
Franz Berthiller
Center for Analytical Chemistry, Depart- ment of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. (Austria)
Hermann Bürstmayr
Institute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. (Austria)
Gerhard Adam
Department of Applied Genetics and Cell Biology, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria. (Austria)
Abstrakt
Fusarium graminearum and related species causing Fusarium head blight of cereals and ear rot of maize produce the trichothecene toxin and virulence factor deoxynivalenol (DON). Plants can detoxify DON to a variable extent into deoxynivalenol-3-O-glucoside (D3G). We have previously reported the DON inactivat- ing glucosyltransferase (UGT) AtUGT73C5 from Arabidopsis thaliana (Poppenberger et al, 2003). Our goal was to identify UGT genes from monocotyledonous crop plants with this enzymatic activity. The two selected rice candidate genes with the highest sequence similarity with AtUGT73C5 were expressed in a toxin sensitive yeast strain but failed to protect against DON. A full length cDNA clone corresponding to a transcript derived fragment (TDF108) from wheat, which was reported to be specifically expressed in wheat genotypes contain- ing the quantitative trait locus Qfhs.ndsu-3BS for Fusarium spreading resistance (Steiner et al, 2009) was reconstructed. Only cDNAs with a few sequence deviations from TF108 could be cloned. However, toxin sensitive yeast strains expressing this wheat UGT cDNA did not show a resistant phenotype. The main diffi- culty in generating full length cDNAs for functional validation by heterologous expression in yeast is the enormous number of the UGT superfamily members in plants, with 107 UGT genes plus some pseudogenes in Arabidopsis thaliana and about 150 putative UGT genes in grasses. We conclude that neither sequence simi- larity nor inducibility are good predictors of substrate specificity.
Słowa kluczowe:
deoxynivalenol, Fusarium graminearum, phase II detoxification, rice, UDP-glucosyltranferase, wheatBibliografia
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Autorzy
Wolfgang Schweigerwolfgang.schweiger@boku.ac.at
Department of Applied Genetics and Cell Biology, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria. Austria
Autorzy
Barbara SteinerInstitute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. Austria
Autorzy
Apinun LimmongkonInstitute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. Austria
Autorzy
Kurt BrunnerInstitute of Chemical Engineering, Vienna University of Technology, A-1060 Vienna, Austria. Austria
Autorzy
Marc LemmensInstitute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. Austria
Autorzy
Franz BerthillerCenter for Analytical Chemistry, Depart- ment of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. Austria
Autorzy
Hermann BürstmayrInstitute of Biotechnology in Plant Production, Department of Agrobiotechnology, University of Natural Resources and Applied Life Sciences, A-3430 Tulln, Austria. Austria
Autorzy
Gerhard AdamDepartment of Applied Genetics and Cell Biology, University of Natural Resources and Applied Life Sciences, A-1190 Vienna, Austria. Austria
Statystyki
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