Abstract
Irrigation water can be polluted by the tobacco etch virus (TEV), which causes serious economic loss in tobacco. However, it is difficult to monitor the sanitary status of irrigation water because TEV presents at extremely low concentrations. This study designed a procedure for concentrating TEV from a standard water sample using polyethylene glycol and detecting it using SYBR Green-based quantitative real-time polymerase chain reaction (qRT-PCR). The concentration factors were optimized through orthogonal tests and the highest recovery efficiency of the TEV genome from a standard water sample was 92.05%. The sensitivity was evaluated using nine 10-fold dilution series of TEV plasmids, and the detection limit of the qRT-PCR was about 10 viral copies/µL. In the infectivity test, TEV was first detected in the roots of NC89 at 7 days after treatment and in the upper leaves at 14 days after treatment. Field diagnosis results showed that 56 of the 180 samples tested positive for TEV, which indicated that this method may be suitable for concentrating and detecting TEV in irrigation water.
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