Development of TaqMan assays for the quantitative detection of Fusarium avenaceum/Fusarium tricinctum and Fusarium poae esyn1 genotypes from cereal grain.
Fungi of the genus Fusarium are important plant pathogens and contaminants of cereal grains producing different types of mycotoxins. Enniatins are a group of mycotoxins with ionophoric properties frequently detected in North European grains. Within the Fusarium complex responsible for grain infection, Fusarium avenaceum, Fusarium poae and Fusarium tricinctum are the most potential enniatins producers. This study presents the development of two quantitative TaqMan MGB (Minor Groove Binder) assays for the specific quantification of F. avenaceum/F. tricinctum and F. poae esyn1 genotypes, respectively. Two sets of genotype-specific primers/probes were designed on the basis of esyn1 gene homologues encoding multifunctional enzyme enniatin synthetase. The specificity of the assays developed has been tested successfully on 111 Fusarium isolates from different geographical origins. The detection limits for F. avenaceum/F. tricinctum esyn1 genotype and F. poae genotype were 19 and 0.3 pg, respectively. The application of the assays developed on asymptomatic wheat grain samples revealed significant positive correlations between the enniatins levels and the amount of F. avenaceum/F. tricinctum esyn1 genotype (R=0.61) and F. poae esyn1 genotype (R=0.42).
Department of Diagnostics and Plant Pathophysiology, University of Warmia and Mazury, Olsztyn, Poland. tomaszkulik76@gmail.com
Tomasz
T
Kulik
Marika
M
Jestoi
Adam
A
Okorski
D017931:DNA Primers / Q000235:genetics; D005506:Food Contamination; D005656:Fungal Proteins / Q000235:genetics; D005670:Fusarium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005838:Genotype; D008969:Molecular Sequence Data; D016533:Mycological Typing Techniques / Q000379:methods*; D010453:Peptide Synthases / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism; D014908:Triticum / Q000382:microbiology*
D017931:DNA Primers; D005656:Fungal Proteins; D019914:Taq Polymerase; D010453:Peptide Synthases; C023122:enniatin B synthetase