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Characterization of novel mutants with an altered gibberellin spectrum in comparison to different wild-type strains of Fusarium fujikuroi

  • Applied genetics and molecular biotechnology
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Abstract

The rice pathogen Fusarium fujikuroi is known for producing a wide range of secondary metabolites such as pigments, mycotoxins, and a group of phytohormones, the gibberellic acids (GAs). Bioactive forms of these diterpenes are responsible for hyperelongation of rice stems, yellowish chlorotic leaves, and reduced grain formation during the bakanae disease leading to severely decreased crop yields. GAs are also successfully applied in agriculture and horticulture as plant growth regulators to enhance crop yields, fruit size, and to induce earlier flowering. In this study, six F. fujikuroi wild-type and mutant strains differing in GA yields and the spectrum of produced GAs were cultivated in high-quality lab fermenters for optimal temperature and pH control and compared regarding their growth, GA production, and GA gene expression levels. Comparative analysis of the six strains revealed that strain 6314/ΔDES/ΔPPT1, holding mutations in two GA biosynthetic genes and an additional deletion of the 4'-phosphopantetheinyl transferase gene PPT1, exhibits the highest total GA amount. Expression studies of two GA biosynthesis genes, CPS/KS and DES, showed a constantly high expression level for both genes under production conditions (nitrogen limitation) in all strains. By cultivating these genetically engineered mutant strains, we were able to produce not only mixtures of different bioactive GAs (GA3, GA4, and GA7) but also pure GA4 or GA7. In addition, we show that the GA yields are not only determined by different production rates, but also by different decomposition rates of the end products GA3, GA4, and GA7 explaining the varying GA levels of genetically almost identical mutant strains.

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Acknowledgments

We thank Marcus Straeten for cloning plasmid pΔppt1 during his diploma thesis as well as Niklas Danne-Rasche for generating strain 6314/ΔDES/ΔPPT1 during his bachelor thesis. Furthermore, we thank Professor Dr. Lewis Mander from the Australian National University, Canberra, Australia, as well as Professor Dr. Peter Hedden of the Rothamsted Research, Harpenden, Hertfordshire, UK, for providing GAs. We also thank Professor Dr. Arthur Ram from the University Leiden for helping to optimize fermentation parameters for Fusarium fujikuroi. This work was funded by the German Federation of Industrial Research Associations (AiF, project IGF16001N).

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Authors and Affiliations

  1. Institute of Plant Biology and Biotechnology, Westfälische Wilhelms Universiät Münster, Schlossplatz 8, 48143, Münster, Germany

    Sabine Albermann & Bettina Tudzynski

  2. Institute for Interfacial Engineering, University of Stuttgart, Nobelstraße 12, 70569, Stuttgart, Germany

    Tino Elter, Andreas Teubner & Thomas Hirth

  3. Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Nobelstr. 12, 70569, Stuttgart, Germany

    Wolfgang Krischke

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  1. Sabine Albermann
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  2. Tino Elter
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  3. Andreas Teubner
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  4. Wolfgang Krischke
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  5. Thomas Hirth
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  6. Bettina Tudzynski
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Correspondence to Bettina Tudzynski.

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Tino Elter and Sabine Albermann contributed equally to the work.

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Albermann, S., Elter, T., Teubner, A. et al. Characterization of novel mutants with an altered gibberellin spectrum in comparison to different wild-type strains of Fusarium fujikuroi . Appl Microbiol Biotechnol 97, 7779–7790 (2013). https://doi.org/10.1007/s00253-013-4917-7

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