Characterization of Fusarium spp. isolates by PCR-RFLP analysis of the intergenic spacer region of the rRNA gene (rDNA)

6533b825fe1ef96bd1282971

RESEARCH PRODUCT

Characterization of Fusarium spp. isolates by PCR-RFLP analysis of the intergenic spacer region of the rRNA gene (rDNA)

R. Mateo Misericordia Jiménez A. Llorens María Teresa González-jaén Antonio F. Logrieco Francisco M. Valle-algarra M.j. Hinojo

subject

Fusarium Trichothecene Food Contamination Biology Polymerase Chain Reaction Microbiology Gas Chromatography-Mass Spectrometry Microbiology chemistry.chemical_compound Fusarium Species Specificity Vomitoxin DNA Ribosomal Spacer Fusarium oxysporum Fusarium culmorum Cluster Analysis DNA Fungal Mycological Typing Techniques Zearalenone Phylogeny food and beverages RNA Fungal DNA Restriction Enzymes General Medicine biology.organism_classification DNA Fingerprinting chemistry RNA Ribosomal Zearalenone Gibberella fujikuroi Restriction fragment length polymorphism Edible Grain Trichothecenes Polymorphism Restriction Fragment Length Food Science

description

In the present study, 44 Fusarium spp. isolates (5 Fusarium culmorum, 7 Fusarium graminearum, 1 Fusarium cerealis, 1 Fusarium poae, 26 Fusarium oxysporum, and 4 Gibberella fujikuroi species complex) were characterized morphologically, physiologically and genetically. All except one (Dutch Collection: CBS 620.72) were isolated from different hosts grown in various Spanish localizations. Morphological characterization was made according to macroscopic and microscopic aspects. Physiological characterization was based on their ability to produce zearalenone (ZEA) and type B trichothecenes (deoxynivalenol, nivalenol and 3-acetyldeoxynivalenol). ZEA was determined by liquid chromatography and trichothecenes by gas chromatography. Confirmation was carried out by liquid chromatography-ion trap-mass spectrometry (ZEA) or gas chromatography-mass spectrometry (trichothecenes). Molecular characterization of isolates was performed using an optimized, simple and low-cost method for isolation of DNA from filamentous fungi and polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) of the intergenic spacer region (IGS) of the rRNA gene (rDNA). The results indicate that F. graminearum, F. culmorum and F. cerealis isolates were high ZEA and type B trichothecene producers, the F. poae isolate produced very low level of nivalenol while F. oxysporum and the G. fujikuroi complex isolates did not show this ability. Restriction patterns of the IGS region did not show any relationship with the host, geographic origin of the isolate and mycotoxin-producing capacity. However, the haplotypes obtained with six restriction enzymes (CfoI, AluI, HapII, XhoI, EcoRI and PstI) permitted to discern the six assayed Fusarium species. Therefore, this is a rapid and suitable methodology that allows closely related strains to group and to estimate the genetic relationships between the groups.

year	journal	country	edition	language
2004-10-28	International Journal of Food Microbiology

https://doi.org/10.1016/j.ijfoodmicro.2005.09.005