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Simultaneous detection of Fusarium asiaticum and Fusarium graminearum in wheat seeds using a real‐time PCR method
Author(s) -
Yin Y.,
Liu X.,
Ma Z.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02595.x
Subject(s) - fusarium , biology , primer (cosmetics) , mycotoxin , polymerase chain reaction , intron , gene , phylogenetic tree , genetics , botany , chemistry , organic chemistry
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.