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ISHS Acta Horticulturae 482: International Symposium on Cut Flowers in the Tropics

IDENTIFICATION OF MOLECULAR MARKERS OF FUSARIUM OXYSPORUM F. SP.DIANTHI BY RAPD

Authors:   J.F. Hernandez, M.A. Posada, P. del Portillo, G. Arbeláez
Abstract:
The Random Amplified Polymorphic DNA (RAPD) technique was used to identify genetic markers useful in the development of a diagnostic method for Fusarium oxysporum f.sp. dianthi (F.o.d.), the etiologic agent of the carnation wilt disease. With the aim of identifying genetic fragments common to F.o.d, a total of 18 different F.o.d. strains isolated from different locations around the world, as well as 17 F. oxysporum isolates corresponding to formae speciales other than dianthi (non-dianthi F. oxysporum) were amplified using 15 different primers. Although, none of the primers used in this study allowed the identification of a single amplification band common to all the different F.o.d. isolates, primer OPA17 generated an RAPD banding pattern which allowed the identification of 4 different amplification groups within the F.o.d. isolates and discrimination between F.o.d. and non-dianthi F. oxysporum. No direct correlation was observed between the RAPD pattern and the race of an isolate, (previously determined by biological assays). Molecular hybridization analysis using OPA 17-amplified fragments as probes showed the feasibility of identifying molecular markers that could be used in the development of a PCR-based molecular diagnostic tool for Fusarium oxysporum f.sp. dianthi.

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