Sensitive detection and discrimination method for studying multiple infections of five major plant viruses infecting ornamental plants in nursery environments. (23rd December 2014)

• Record Type: Journal Article
• Authors: Dobhal, S.; Arif, M.; Olson, J.; Mendoza‐Yerbafría, A.; Aguilar‐Moreno, S.; Perez‐Garcia, M.; Ochoa‐Corona, F. M.
• Is Part Of: Annals of applied biology. Volume 166:Number 2(2015)
• Page Start: 286
• Page End: 296
• Publication Date: 2014-12-23
• Subjects: Crop science -- Periodicals; Plants, Protection of -- Periodicals; Crops -- Ecology -- Periodicals; 630
• DOI: 10.1111/aab.12182
• Languages: English
• ISSNs: 0003-4746

Abstract

<abstract abstract-type="main" id="aab12182-abs-0001"> <title>Abstract</title> <p id="aab12182-para-0001"> <italic>Tobacco mosaic virus</italic> (TMV), <italic>Hosta virus X</italic> (HVX), <italic>Cucumber mosaic virus</italic> (CMV), <italic>Tomato spotted wilt virus</italic> (TSWV) and <italic>Impatiens necrotic spot virus</italic> (INSV) are a few of the major viruses that infect ornamental and nursery plants. These viruses cause significant losses that impact growers and the ornamental industry. Often, a single ornamental plant is co‐infected by more than one virus, which makes identification and discrimination of these viruses a difficult task, thus creating delays and limiting regulatory measures for effective quarantine. The aim of this study is to develop a sensitive, rapid, economic, and reliable multiplex Reverse Transcription PCR (RT‐PCR) for simultaneous detection and discrimination of these five viruses. Specific PCR primers were designed using the consensus sequences of corresponding capsid protein (CP) genes of HVX and CMV, the nucleocapsid protein (NP) genes of TSWV and INSV, and the movement protein (MP) and CP genes of TMV. The primers were validated <italic>in vitro</italic> using single and multiplex RT‐PCR assays. The detection limit of each primer set in multiplex RT‐PCR was 100 fg (TMV), 1 fg (HVX), 10 fg (CMV), 10 pg (TSWV) and 10 pg (INSV). Forty‐six infected nursery samples collected from different locations in the USA were screened for virus infections using this multiplex RT‐PCR. The multiplex RT‐PCR has a potential for its application in routine diagnostics, quarantine, and epidemiological studies. The developed method is reliable, sensitive, and economic for testing a wide range of ornamental and nursery plants for detection of these viruses.</p> </abstract>