Differential gene expression in individual papilla-resistant and powdery mildew-infected barley epidermal cells

T. Gjetting, Timothy L. W. Carver, Leif Skøt, Michael Foged Lyngkjær

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Resistance and susceptibility in barley to the powdery mildew fungus (Blumeria graminis f. sp. hordei) is determined at the single-cell level. Even in genetically compatible interactions, attacked plant epidermal cells defend themselves against attempted fungal penetration by localized responses leading to papilla deposition and reinforcement of their cell wall. This conveys a race-nonspecific form of resistance. However, this defense is not complete, and a proportion of penetration attempts succeed in infection. The resultant mixture of infected and uninfected leaf cells makes it impossible to relate powdery mildew-induced gene expression in whole leaves or even dissected epidermal tissues to resistance or susceptibility. A method for generating transcript profiles from individual barley epidermal cells was established and proven useful for analyzing resistant and successfully infected cells separately. Contents of single epidermal cells (resistant, infected, and unattacked controls) were collected, and after cDNA synthesis and PCR amplification, the resulting sample was hybridized to dot-blots spotted with genes, including some previously reported to be induced upon pathogen attack. Transcripts of several genes, (e.g., PR1a, encoding a pathogenesis related protein, and GLP4, encoding a germin-like protein) accumulated specifically in resistant cells, while GRP94, encoding a molecular chaperone, accumulated in infected cells. Thus, the single-cell method allows discrimination of transcript profiles from resistant and infected cells. The method will be useful for microarray expression profiling for simultaneous analysis of many genes.
    Original languageEnglish
    JournalMolecular Plant - Microbe Interactions
    Volume17
    Issue number7
    Pages (from-to)729-738
    ISSN0894-0282
    DOIs
    Publication statusPublished - 2004

    Keywords

    • single-cell analysis

    Fingerprint Dive into the research topics of 'Differential gene expression in individual papilla-resistant and powdery mildew-infected barley epidermal cells'. Together they form a unique fingerprint.

    Cite this