PCR detection of Fusarium oxysporum f.sp. gladioli race 1, causal agent of Gladiolus yellows disease, from infected corms

L.A.M. De Haan*, A. Numansen, E.J.A. Roebroeck, J. Van Doorn

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    26 Citations (Scopus)

    Abstract

    Fusarium oxysporum f.sp. gladioli (FOG) race 1 infects both large- and small-flowered Gladiolus cultivars. Race 2 isolates infect only small-flowered cultivars but can be present as epiphytes on large-flowered plants. When 160 arbitrary 10-mer oligonucleotide primers were tested on FOG by PCR to find RAPD markers specific for race 1, the RAPD primer G12 amplified two discriminating DNA fragments, AB (609 bp) and EF (1196 bp), in race 1 isolates only. Both fragments were cloned and sequenced. Two pairs of race 1-specific primers for multiplex PCR were designed. Tests of 112 E oxysporum isolates by PCR showed that, in almost all cases, race 1 isolates of vegetative compatibility group 0340 could be distinguished with these primers. Seven putative race 1 isolates did not react in multiplex PCR; hybridization studies with labelled AB and EF DNA fragments showed that these isolates belong to separate groups. A bioassay was developed to detect corms that were latently infected with FOG race 1. Gladiolus corms were homogenized and incubated for 5 days at 28°C in a semiselective medium to induce growth of Fusarium. Cultivated mycelium was isolated and subjected to the developed multiplex PCR after standard DNA isolation or disruption by microwave treatment.

    Original languageEnglish
    Pages (from-to)89-100
    Number of pages12
    JournalPlant Pathology
    Volume49
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2000

    Keywords

    • Bioassay
    • Fusarium oxysporum f.sp. gladioli
    • Gladiolus
    • Multiplex PCR
    • Races
    • RAPDs

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