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A DNA-Based Procedure for In Planta Detection of Fusarium oxysporum f. sp. phaseoli

March 2002 , Volume 92 , Number  3
Pages  237 - 244

Fernando M. Alves-Santos , Brisa Ramos , M. Asunción García-Sánchez , Arturo P. Eslava , and José María Díaz-Mínguez

Area de Genética, Centro Hispano-Luso de Investigaciones Agrarias, Universidad de Salamanca, Avda. Campo Charro s/n 37007 Salamanca, Spain

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Accepted for publication 25 October 2001.

We have characterized strains of Fusarium oxysporum from common bean fields in Spain that were nonpathogenic on common bean, as well as F. oxysporum strains (F. oxysporum f. sp. phaseoli) pathogenic to common bean by random amplified polymorphic DNA (RAPD) analysis. We identified a RAPD marker (RAPD 4.12) specific for the highly virulent pathogenic strains of the seven races of F. oxysporum f. sp. phaseoli. Sequence analysis of RAPD 4.12 allowed the design of oligonucleotides that amplify a 609-bp sequence characterized amplified region (SCAR) marker (SCAR-B310A280). Under controlled environmental and greenhouse conditions, detection of the pathogen by polymerase chain reaction was 100% successful in root samples of infected but still symptomless plants and in stem samples of plants with disease severity of ≥4 in the Centro Internacional de Agricultura Tropical (CIAT; Cali, Colombia) scale. The diagnostic procedure can be completed in 5 h and allows the detection of all known races of the pathogen in plant samples at early stages of the disease with no visible symptoms.

© 2002 The American Phytopathological Society