Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 431482
Title Rapid identification of fusarium graminearum species complex using Rolling Circle Amplification (RCA)
Author(s) Davari, M.; Diepeningen, A.D. van; Babai-Ahari, A.; Arzanlou, M.; Javad Najafzadeh, M.; Lee, T.A.J. van der; Hoog, G.S. de
Source Journal of Microbiological Methods 89 (2012)1. - ISSN 0167-7012 - p. 63 - 70.
DOI https://doi.org/10.1016/j.mimet.2012.01.017
Department(s) Laboratory of Phytopathology
PRI BIOINT Moleculair Phytopathology
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) head blight pathogen - dna-sequence database - genealogical concordance - padlock probes - north-america - wheat - mycotoxins - cereals - barley - populations
Abstract Rolling Circle Amplification (RCA) of DNA is a sensitive and cost effective method for the rapid identification of pathogenic fungi without the need for sequencing. Amplification products can be visualized on 1% agarose gel to verify the specificity of probe-template binding or directly by adding fluorescent dyes. Fusarium Head Blight (FHB) is currently the world's largest threat to the production of cereal crops with the production of a range of mycotoxins as an additional risk. We designed sets of RCA padlock probes based on polymorphisms in the elongation factor 1-a (EF-1a) gene to detect the dominant FHB species, comprising lineages of the Fusarium graminearum species complex (FGSC). The method also enabled the identification of species of the Fusarium oxysporum (FOSC), the Fusarium incarnatum-equiseti (FIESC), and the Fusarium tricinctum (FTSC) species complexes, and used strains from the CBS culture collection as reference. Subsequently probes were applied to characterize isolates from wheat and wild grasses, and inoculated wheat kernels. The RCA assays successfully amplified DNA of the target fungi, both in environmental samples and in the contaminated wheat samples, while no cross reactivity was observed with uncontaminated wheat or related Fusarium species. As RCA does not require expensive instrumentation, the technique has a good potential for local and point of care screening for toxigenic Fusarium species in cereals
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