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Dongo A, Leflon M, Simier P & Delavault P (2011) Development of a high-throughput real-time quantitative PCR method to detect and quantify contaminating seeds of Phelipanche ramosa and Orobanche cumana in crop seed lots. Weed Research 52, 34–41. Summary The objective of this study was to develop a simple and robust quantitative PCR method for the detection of seeds from two parasitic plants, Phelipanche ramosa and Orobanche cumana, as well as their closely related species, in seed harvests of oilseed rape and sunflower. The method was based on the design of primers/probe sets specific to both parasitic plants and targeting internal transcribed spacer sequences for quantitative real-time PCR (TaqMan). Together with the proposed DNA extraction protocol, this diagnostic method allows rapid, high-throughput and accurate assessment of contamination with broomrape seeds, without the requirement of tedious purification steps and identification under a binocular microscope. The TaqMan assay is highly specific, because it did not detect any possible plant contaminants that are likely to be present in harvested crop seeds. The results of this assay can be expressed in terms of the number of parasite seeds per kilogram of crop seeds, a metric that could be utilised to help decisions regarding crop seed lot utilisation and commercialisation. |
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