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


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 beta-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 beta-tubulin gene could allow differentiation of genetically related fungal species.


CONCLUSIONS
The sensitive and quantitative detection method can be readily used in epidemiological studies and in assessing risk of Fusarium mycotoxin contamination in wheat samples.
Institute of Biotechnology, Zhejiang University, Hangzhou, China.
Y
Y
Yin
X
X
Liu
Z
Z
Ma
D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D005670:Fusarium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016533:Mycological Typing Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012639:Seeds / Q000382:microbiology; D014908:Triticum / Q000382:microbiology*; D014404:Tubulin / Q000235:genetics
D017931:DNA Primers; D005656:Fungal Proteins; D014404:Tubulin; D003577:Cytochrome P-450 Enzyme System