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. | Turkish Journal of Agriculture and Forestry Turk J Agric For (2014) 38: 471-477 © TÜBİTAK doi: Research Article Concentration and detection of tobacco etch virus from irrigation water using real-time PCR 1 1 1 1 2 1, Wei CHEN , Jin DAI , Huawei ZHANG , Honghong JIAO , Julong CHENG , Yunfeng WU * State Key Laboratory of Crop Stress Biology for Arid Areas, Northwest A&F University, Yangling, Shaanxi, . China 2 Shaanxi Tobacco Research Institute, Xi’an, . China 1 Received: Accepted: Published Online: Printed: 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 . 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. Key words: Tobacco etch virus, irrigation water, polyethylene glycol, quantitative real-time PC 1. Introduction Water plays a very important role in the activities of plants and animals (Novo, 2012). However, more than hundreds of thousands of .
