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Stable transformation of Erysiphe graminis an obligate biotrophic pathogen of barley

Nature Biotechnology volume 18, pages 205–207 (2000)Cite this article

Abstract

Barley powdery mildew, Erysiphe graminis f.sp. hordei, is an obligate biotrophic pathogen and as such cannot complete its life cycle without a living host. The inability to transform this fungus and manipulate its genome has constrained research towards understanding its life cycle and pathogenicity. Here we describe an in planta transformation system based on delivery of DNA using a gold-particle gun and selection using benomyl or bialaphos. Using this method, we consistently obtained stable transformants with efficiencies comparable to other filamentous fungi. Stable expression of the β-glucuronidase in E. graminis was demonstrated by co-transforming the uidA gene with the selectable markers.

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Figure 1: (A) Map of the transformation vectors used in this study.
Figure 2: Southern blot of DNA from uninfected barley leaves (1), leaves infected with nontransformed wild-type E. graminis (2), with E. graminis transformed with pBTmut (3–5) or pBT-BAR (6–8).
Figure 3: Barley leaves infected with E. graminis co-transformed with pBT-GUS and pBT-BAR.

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Acknowledgements

We wish to thank W. Harwood for the generous gift of BASTA-resistant barley and S. Foster for discussions. This work was funded by a BBSRC grant. P.D.S. is a Royal Society University Research Fellow.

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Authors and Affiliations

  1. Department of Plant Sciences, University of Oxford, South Parks Rd, Oxford, OX1 3RB, UK

    Pushpalata Chaure & Sarah J. Gurr

  2. Department of Biology, Imperial College of Science Technology and Medicine, Imperial College Rd, London, SW7 2AZ, UK

    Pietro Spanu

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  1. Pushpalata Chaure
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Correspondence to Pietro Spanu.

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Chaure, P., Gurr, S. & Spanu, P. Stable transformation of Erysiphe graminis an obligate biotrophic pathogen of barley. Nat Biotechnol 18, 205–207 (2000). https://doi.org/10.1038/72666

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