title: Utility of the polymerase chain reaction-restriction fragment length polymorphisms of the intergenic spacer region of the rDNA for characterizing Gibberella fujikuroi isolates.
abstract: In the present report, a total of thirty-one isolates of Gibberella fujikuroi (Sawada) Wollenw. species complex of Fusarium (section Liseola) morphologically classified as F. moniliforme according to the taxonomy of Nelson, Toussoun and Marasas (1983) were analyzed for their ability to produce fumonisin B1 and fumonisin B2 by an optimized liquid chromatographic method. They were isolated from three hosts (Zea mays, Musa sapientum and Pinus pinea). The results indicate that M. sapientum is a preferential host for G. fujikuroi isolates with low or null capacity for producing fumonisins, while isolates from Z. mays and P. pinea are generally high fumonisin producers. The molecular characterization of isolates was carried out in parallel using an optimized, simple and low-cost method for isolating DNA from filamentous fungi and polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) of the rDNA intergenic spacer (IGS) region. The haplotypes obtained with Hha I enzyme and combinations of Hha I, EcoR I, Alu I, Pst I and Xho I enzymes provided very characteristic groupings of G. fujikuroi isolates as a function of host type and fumonisin B1 and B2 producing capacity. IGS region restriction patterns showed no relationship to isolate geographical origin. This is the first report on this method's capacity to detect polymorphism permitting discrimination between G. fujikuroi isolates from different hosts and with different toxigenic profiles.
authors.affiliation: Departamento de Microbiología y Ecología, Facultad de Biología, Universidad de Valencia, Burjassot, Valencia, Spain.
authors.forename: María J
authors.initials: MJ
authors.lastname: Hinojo
authors.forename: Amparo
authors.initials: A
authors.lastname: Llorens
authors.forename: Rufino
authors.initials: R
authors.lastname: Mateo
authors.forename: Belén
authors.initials: B
authors.lastname: Patiño
authors.forename: María Teresa
authors.initials: MT
authors.lastname: González-Jaén
authors.forename: Misericordia
authors.initials: M
authors.lastname: Jiménez
mesh_terms: D002845:Chromatography; D016172:DNA Fingerprinting; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D021903:DNA, Ribosomal Spacer / Q000235:genetics* / Q000302:isolation & purification; D037341:Fumonisins / Q000032:analysis / Q000302:isolation & purification; D005874:Gibberella / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006239:Haplotypes; D028521:Musa / Q000382:microbiology; D016533:Mycological Typing Techniques; D010802:Phylogeny; D028223:Pinus / Q000382:microbiology; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D003313:Zea mays / Q000382:microbiology
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D037341:Fumonisins; C056934:fumonisin B2; C056933:fumonisin B1; D004262:DNA Restriction Enzymes