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Virologica Sinica, 29 (3) : 155-161, 2014
Research Article
Development of a concentration method for detection of tobacco mosaic virus in irrigation water
1. State Key Laboratory of Crop Stress Biology for Arid Areas and Key Laboratory of Crop Pest Integrated Pest Management on the Loess Plateau of Ministry of Agriculture, College of Plant Protection, Northwest A&F University, Yangling 712100, China

2. Shaanxi Tobacco Research Institute, Xi'an 710061, China
 Correspondence: wuyf@nwsuaf.edu.cn
(1906.82KB)  
Abstract
Tobacco mosaic virus (TMV) causes significant yield loss in susceptible crops irrigated with contaminated water. However, detection of TMV in water is difficult owing to extremely low concentrations of the virus. Here, we developed a simple method for the detection and quantifi cation of TMV in irrigation water. TMV was reliably detected at concentrations as low as 10 viral copies/μL with real-time PCR. The sensitivity of detection was further improved using polyethylene glycol 6000 (PEG6000, MW 6000) to concentrate TMV from water samples. Among the 28 samples from Shaanxi Province examined with our method, 17 were tested positive after virus concentration. Infectivity of TMV in the original water sample as well as after concentration was confi rmed using PCR. The limiting concentration of TMV in water to re-infect plants was determined as 102 viral copies/mL. The method developed in this study offers a novel approach to detect TMV in irrigation water, and may provide an effective tool to control crop infection.
Received: 18 Jun 2014  Accepted: 21 Apr 2014  Published online: 1 Jun 2014
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