Use of molecular markers to compare Fusarium verticillioides pathogenic strains isolated from plants and humans.
Fusarium verticillioides is a pathogen of agriculturally important crops, especially maize. It is considered one of the most important pathogens responsible for fumonisin contamination of food products, which causes severe, chronic, and acute intoxication in humans and animals. Moreover, it is recognized as a cause of localized infections in immunocompetent patients and disseminated infections among severely immunosuppressed patients. Several molecular tools have been used to analyze the intraspecific variability of fungi. The objective of this study was to use molecular markers to compare pathogenic isolates of F. verticillioides and isolates of the same species obtained from clinical samples of patients with Fusarium mycoses. The molecular markers that we used were inter-simple sequence repeat markers (primers GTG5 and GACA4), intron splice site primer (primer EI1), random amplified polymorphic DNA marker (primer OPW-6), and restriction fragment length polymorphism-internal transcribed spacer (ITS) from rDNA. From the data obtained, clusters were generated based on the UPGMA clustering method. The amplification products obtained using primers ITS4 and ITS5 and loci ITS1-5.8-ITS2 of the rDNA yielded fragments of approximately 600 bp for all the isolates. Digestion of the ITS region fragment using restriction enzymes such as EcoRI, DraI, BshI, AluI, HaeIII, HinfI, MspI, and PstI did not permit differentiation among pathogenic and clinical isolates. The inter-simple sequence repeat, intron splice site primer, and random amplified polymorphic DNA markers presented high genetic homogeneity among clinical isolates in contrast to the high variability found among the phytopathogenic isolates of F. verticillioides.
Laboratório de Genética Molecular de Fungos, Departamento de Micologia, Universidade Federal de Pernambuco, Recife, PE, Brasil suzycchang@yahoo.com.br.
S C
SC
Chang
D P C
DP
Macêdo
C M
CM
Souza-Motta
N T
NT
Oliveira
D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics*; D060585:Fusariosis / Q000235:genetics / Q000382:microbiology; D005670:Fusarium / Q000235:genetics* / Q000472:pathogenicity; D005819:Genetic Markers; D014644:Genetic Variation; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*; D003313:Zea mays / Q000382:microbiology*
D004271:DNA, Fungal; D004275:DNA, Ribosomal; D005819:Genetic Markers