Development of PCR assays for the detection and differentiation of Fusarium sporotrichioides and Fusarium langsethiae.
Isolates of the type-A trichothecene producing Fusarium sporotrichioides and Fusarium langsethiae were grouped and differentiated in a phylogenetic tree using ITS sequence dissimilarity. An attempt was made to develop a PCR-based assay for the detection and differentiation of Fusarium sporotrichiodes from other Fusarium species using the 5'-region of the tri5 gene as a template. However, this assay was unable to differentiate, to a satisfactory level, between isolates of Fusarium sporotrichioides and Fusarium langsethiae, providing further genetic evidence for their close genetic relationship. A robust and repeatable PCR-assay was developed for the detection and differentiation of both species based on sequence determined from differentially amplified RAPD-PCR products. These assays were able to detect both species in samples of grain taken from the field.
Wilson
Anthony
A
Department of Metabolic Biology, John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK.
Simpson
Duncan
D
Chandler
Elizabeth
E
Jennings
Phil
P
Nicholson
Paul
P
D001483:Base Sequence; D017931:DNA Primers; D004271:DNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics; D005670:Fusarium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D005656:Fungal Proteins