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

2009/03/01 by Y. Yin, Yanni Yin, X. Liu +2
Agricultural and Biological Sciences · Biochemistry, Genetics and Molecular Biology · #Fungal and yeast genetics research #Mycotoxins in Agriculture and Food #Plant Pathogens and Fungal Diseases

paper · pdf · doi:10.1111/j.1472-765x.2009.02595.x

openalex publication_date 2009/03/01 · openalex created_date 2016/06/24 · openalex updated_date 2026/07/29

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 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. 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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