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Plant Molecular Biology

, Volume 55, Issue 1, pp 1–15 | Cite as

Identification of powdery mildew-induced barley genes by cDNA-AFLP: functional assessment of an early expressed MAP kinase

  • Christina Eckey
  • Michael Korell
  • Katja Leib
  • Dagmar Biedenkopf
  • Carin Jansen
  • Gregor Langen
  • Karl-Heinz Kogel
Article

Abstract

Gene expression analysis by cDNA-AFLP in barley (Hordeum vulgare L.) after powdery mildew (Blumeria graminis f.sp. hordei, Bgh) inoculation revealed 615 (3.7%) of 16 500 screened cDNA fragments being differentially regulated 4 and/or 12 h after inoculation. Of these transcript derived fragments (TDFs), 120 were sequenced, and for 28 out of 29 tested, induction was confirmed via RT-PCR. Most TDFs did not show any homology to sequences with known functions, others showed homology to genes involved in primary and secondary metabolism, pathogen response, redox regulation, and signal transduction. TDFs with homology to a MAP kinase (PWMK1), a WRKY transcription factor, a heparanase, an immunophilin, a cytochrome P450, and a receptor-like protein kinase were isolated as full length cDNAs. Knockdown by RNA interference via biolistic delivery of sequence specific double stranded RNA to leaf segments tagged two of these genes as possible candidates being causally involved in the outcome of the barley-Bgh interaction. Knockdown of the receptor-like protein kinase and the WRKY transcription factor increased resistance to the fungus, while knockdown of PWMK1only led to a slightly enhanced susceptibility of epidermal cells to Bgh. This suggests that the receptor-like protein kinase and the WRKY protein are candidates for negative regulators of powdery mildew resistance. Based on expression analyses, PWMK1appears to be more generally involved in stress response.

barley cDNA-AFLP functional analysis MAPK powdery mildew RNA interference transformation assay WRKY transcription factor 

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Supplementary material

APp.DO00000275_1.doc (202 kb)
Supplementary material

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Christina Eckey
    • 1
  • Michael Korell
    • 1
  • Katja Leib
    • 1
  • Dagmar Biedenkopf
    • 1
  • Carin Jansen
    • 1
  • Gregor Langen
    • 1
  • Karl-Heinz Kogel
    • 1
  1. 1.Interdisciplinary Research Centre for Environmental Sciences, Institute of Phytopathology and Applied ZoologyJustus-Liebig-UniversityHeinrich-Buff-Ring 26Germany

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