RESUMEN
Fusarium head blight (FHB) is a disease of small-grain cereals and corn caused by a complex of fungal species of the genus Fusarium. The disease reduces the yield and quality of seeds and results in the accumulation of various mycotoxins which cause a variety of toxic effects on humans and livestock. Beauvericin (BEA) and enniatins (ENs) are a group of toxins with antimicrobial, insecticidal and phytotoxic activities produced mainly by F. avenaceum, F. poae and F. tricinctum. In this study, primer sets were designed that were targeted to esyn1 gene homologs encoding multifunctional enzyme enniatin synthetase. Primers used in multiplex PCR amplified products from the FHB species reported to produce (ENs) and/or BEA. The use of the marker developed on asymptomatic wheat seed samples originating from Northern and Southern Poland demonstrated that all samples were positive for the presence of potential enniatin-producing Fusarium species.
Asunto(s)
Grano Comestible/microbiología , Contaminación de Alimentos/análisis , Fusarium/genética , Péptido Sintasas/genética , Reacción en Cadena de la Polimerasa/métodos , Cartilla de ADN , ADN de Hongos/análisis , Depsipéptidos , Grano Comestible/química , Genes FúngicosRESUMEN
A polymerase chain reaction assay was developed for detection of Fusarium sporotrichioides, a plant pathogen in many parts of the world. Based on small nucleotide differences in ITS2 (Internal Transcribed Spacer) rDNA of our local isolate of F. sporotrichioides (Accession No. AY510069) and other isolates found in NCBI/GeneBank database, species specific primer FspITS2K was selected. Primer pair FspITS2K and P28SL amplified a fragment of 288 bp containing a portion of ITS2 and 28S rDNA of all the F. sporotrichioides isolates tested, originated from different hosts and regions of the world but did not amplify any other species of Fusarium and plant's DNA. To use the PCR assay in seed health testing, a protocol was setup for the rapid and effective preparations of fungal DNA from wheat seeds. The method developed may be useful for the rapid detection and identification of F. sporotrichioides both from culture and from plant tissue.