title: Fusarium diversity in soil using a specific molecular approach and a cultural approach.
abstract: Fusarium species are ubiquitous in soil. They cause plant and human diseases and can produce mycotoxins. Surveys of Fusarium species diversity in environmental samples usually rely on laborious culture-based methods. In the present study, we have developed a molecular method to analyze Fusarium diversity directly from soil DNA. We designed primers targeting the translation elongation factor 1-alpha (EF-1α) gene and demonstrated their specificity toward Fusarium using a large collection of fungi. We used the specific primers to construct a clone library from three contrasting soils. Sequence analysis confirmed the specificity of the assay, with 750 clones identified as Fusarium and distributed among eight species or species complexes. The Fusarium oxysporum species complex (FOSC) was the most abundant one in the three soils, followed by the Fusarium solani species complex (FSSC). We then compared our molecular approach results with those obtained by isolating Fusarium colonies on two culture media and identifying species by sequencing part of the EF-1α gene. The 750 isolates were distributed into eight species or species complexes, with the same dominant species as with the cloning method. Sequence diversity was much higher in the clone library than in the isolate collection. The molecular approach proved to be a valuable tool to assess Fusarium diversity in environmental samples. Combined with high throughput sequencing, it will allow for in-depth analysis of large numbers of samples.
authors: INRA, UMR1347 Agroécologie, BP 86510, F-21000 Dijon, France. Electronic address: veronique.edel@dijon.inra.fr.
authors: Véronique
authors: V
authors: Edel-Hermann
authors: INRA, UMR1347 Agroécologie, BP 86510, F-21000 Dijon, France.
authors: Nadine
authors: N
authors: Gautheron
authors: INRA, UMR1347 Agroécologie, BP 86510, F-21000 Dijon, France.
authors: Arnaud
authors: A
authors: Mounier
authors: INRA, UMR1347 Agroécologie, BP 86510, F-21000 Dijon, France.
authors: Christian
authors: C
authors: Steinberg
mesh_terms: D017931:DNA Primers; D004271:DNA, Fungal; D005670:Fusarium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D014644:Genetic Variation; D058889:Molecular Typing; D016533:Mycological Typing Techniques; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D020648:Peptide Elongation Factor 1
keywords: Detection; Environment; Fusarium spp.; Genus-specific primers; Molecular diversity; Soil DNA