Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus

[EN] A multiplex PCR method has been developed to detect, differentiate, and confirm the morphological identification of three root infecting Olpidium spp.: O. bornovanus, O. brassicae, and O. virulentus. Of the 132 root samples examined, 101 samples were infected by Olpidium spp.. Based on the morphology of resting spores, the presence of O. bornovanus was confirmed in 20.5 % of the samples, whereas species identity could not be determined for the remaining samples because they failed to reproduce sexually. With multiplex PCR, it was possible to determine the Olpidium identity of all the infected samples, even when resting spores were not formed. This method was also effective for detecting Olpidium spp. in water samples. In addition, the specificity and sensitivity of multiplex PCR were evaluated. The multiplex PCR method was validated with samples of 9 different crops from 11 countries of America, Europe, and Africa. Jose Angel Herrera Vasquez was supported by a predoctoral fellowship from the Agencia Espanola de Cooperacion Internacional Para el Desarrollo (AECID, Spain, 2004-2007 period) and the Instituto Para la Formacion y Aprovechamiento de Recursos Humanos and the Secretaria Nacional de Ciencia y Tecnologia (IFARHU-SENACYT, Panama, 2007-2010 period). We thank the Direccion Nacional de Sanidad Vegetal-Ministerio de Desarrollo Agropecuario (DNSV-MIDA, Panama) for their technical assistance. We also thank Dr. M. L Herrero (Bioforsk, Plant Health and Plant Protection Division, Norway) and Dr. J. C. Tello (Plant Production Department, University of Almeria, Spain) for providing the fungal isolates used in the specificity assays, and Dr. R. Beltran for providing the soil samples from Brazil. This work was supported by grants AGL 2005-06682-C03-01 from the Comision Interministerial de Ciencia y Tecnologia CICYT, Spain. Herrera-Vásquez, JA.; Cebrián, MDC.; Alfaro Fernández, AO.; Córdoba-Sellés, MDC.; Jordá, C. (2009). Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus. Mycological Research. 113:602-610. https://doi.org/10.1016/j.mycres.2009.01.007