Simultaneous detection ofFusarium asiaticumandFusarium graminearumin wheat seeds using a real-time PCR method

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Abstract

Aims:

To develop a PCR-based method for quantitative detection of Fusarium asiaticum (Fa) and Fusarium graminearum (Fg) in wheat seeds.

Methods and Results:

Based on the sequences of the cyp51A gene, two primer pairs FaF + FaR and FgF + FgR were developed for the species-specific detection of Fa and Fg, respectively. To simultaneously detect these two phylogenetic species, a pair of primers FgaF + FgaR was developed based on the first and the second introns of β-tubulin gene. This primer pair amplified a 228-bp fragment only from Fa and Fg isolates, but not from 22 other Fusarium spp. and 13 other fungal species. A real-time PCR with this primer pair was able to quantify minute amounts of Fa and Fg DNA in wheat seeds rapidly.

Conclusions:

PCR primers designed based on the sequence of cyp51A or intron region of β-tubulin gene could allow differentiation of genetically related fungal species.

Significance and Impact of the Study:

The sensitive and quantitative detection method can be readily used in epidemiological studies and in assessing risk of Fusarium mycotoxin contamination in wheat samples.

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