Identificacion de marcadores genéticos del agente causal del marchitamiento del clavel fusarium oxysporum f.sp. dianthi mediante amplificacion arbitraria de fragmentos polimórficos de adn
Identification of genetic markers of fusarium oxysporum f.sp. dianthi , the carnation curling agent, using the random amplified polymorphic dna (rapd) technique
Keywords:
RAPD, Fusarium oxysporum, polimorfismos, clavel (es)RAPD, Fusarium oxysporum, polymorphism, carnation (en)
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La técnica de Amplificación Arbitraria de Fragmentos Polimórficos de ADN (RAPD) fue utilizada para identificar marcadores genéticos útiles para el desarrollo de un método diagnóstico para Fusarium oxysporum f.sp. dianthi, el agente etiológico de la enfermedad del marchitamiento del clavel. Con el fin de identificar fragmentos genéticos característicos de este patógeno, un total de 18 aislados diferentes, provenientes de diferentes lugares del mundo y 17 cepas de F. oxysporum de otras formas especiales fueron amplificadas utilizando 15 iniciadores diferentes. Aunque ninguno de los iniciadores empleados en este estudio amplificó una banda común a todas las formas especiales dianthi, el iniciador OPA 17 mostró un patrón de RAPD que permitió la identificación de cuatro grupos polimórficos dentro de este grupo taxonómico. Este mismo iniciador, permitió la discriminación entre aislados de Fusarium oxysporum f.sp. dianthi y cepas de F. oxysporum de otras formas especiales. No se observó una correlación directa entre el patrón de RAPD y las razas reportadas para F. oxysporum f.sp. dianthi, previamente determinadas mediante ensayos biológicos por otros grupos de investigadores. Los análisis de hibridación molecular con fragmentos escogidos de estos patrones de RAPD, permitieron el reconocimiento selectivo de los cuatro grupos descritos. Los fragmentos genómicos identificados, son candidatos para el desarrollo de un sistema diagnóstico por PCR para este patógeno del clavel.
The Random Amplified Polymorphic DNA (RAPD) technique was used to analyse genetic markers useful in developing a method to diagnosis Fusarium oxysporum f.sp. dianthi infections. To identify characteristic genetic fragments of this pathogen, 18 strains were isolated from locations around the world and 17 special forms of F. oxysporum (non-.dianthi F. oxysporum) were amplified using 15 different primers. None of these primers allowed the identification of a single amplification band common to the Fusarium oxysporum f.sp. dianthi samples. However, the OPA 17 primer generated an RAPD pattern that allowed the identification of four amplification groups within this taxonomic group. The same marker permitted to distinguish between the Fusarium oxysporum f.sp. dianthi and the special forms of F. oxysporum. No direct correlation was observed between the RAPD pattern and the race of the samples established in previous works. Molecular hybridization analysis, using the OPA 17 amplified fragments as probes, showed the feasibility of identifying molecular markers that can be used in developing a PCR method for diagnosis of Fusarium oxysporum f.sp. dianthi infections.
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