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A rapid assay for chloroplast-encoded triazine resistance in higher plants
Authors:Wing Y. Cheung  Jean-Charles Côté  Diane L. Benoit  Benoit S. Landry
Affiliation:(1) a Division of Pioneer, Allelix Crop Technologies, 12111 Mississauga Road, L7G 4S7 Georgetown, Ontario, Canada;(2) Agriculture Canada Research Station, 430 Boulevard Gouin, J3B 3E6 St-Jean-sur-Richelieu, Québec, Canada
Abstract:We have designed a simple and rapid assay for chloroplast-based triazine resistance in higher plants using PCR amplification of thepsbA gene coupled toMaeI digestion of the amplified product to distinguish triazine resistant from sensitive biotypes. Our assay is universal and avoids the need of lengthy procedures of previously published assays, which either required spraying of seedlings in a controlled environment, quantification of chlorophyll fluorescence of leaf discs after incubation in triazine solution, DNA sequencing of thepsbA gene, or Southern-blot analysis. Our diagnostic system is qualitative, reliable, fast and simple. More than 100 seedlings taken directly from the field can be analyzed in one day. This system has a direct application towards a more rational use of herbicides in production fields. It also represents a valuable tool to monitor spreading of resistant biotypes through time and space and can serve as a model system applicable to other gene monitoring needs.
Keywords:Atrazineor triazine resistance  DNA markers  PCR  DNA diagnostic  STS   psbA
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