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A multiplex RT-PCR for simultaneous detection and identification of five viruses and two viroids infecting chrysanthemum

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Abstract

Pathogens causing significant economic losses in chrysanthemum include tomato aspermy virus (TAV), chrysanthemum virus B (CVB), cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), potato virus Y (PVY), chrysanthemum stunt viroid (CSVd) and chrysanthemum chlorotic mottle viroid (CChMVd). A multiplex reverse transcription polymerase chain reaction (RT-PCR) method, using specific primer sets for each virus or viroid, was developed for simultaneous detection and differentiation of TAV, CVB, CMV, TMV, PVY, CChMVd, and CSVd. The RT-PCR method was validated by testing chrysanthemum samples collected from different regions of China. In this study, CVB, TAV, TMV, PVY, CSVd, CMV, and CChMVd were detected, respectively, in 24.7 %, 17.5 %, 4.4 %, 4.4 %, 2.9 %, 2.5 %, and 1.5 % of the samples tested. These results indicate that CVB and TAV (24.7 % and 17.5 %) are common, whereas CMV, TMV, CChMVd, CSVd, and PVY (all below 5 %) are less frequently encountered. This new multiplex RT-PCR method has potential to be used routinely in large-scale virus and viroid surveys.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 31171990 and No. 31372105).

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    Authors and Affiliations

    1. Department of Biological Science, College of Life Sciences, Henan Normal University, Construction East Road No. 46, Muye District, **nxiang, 453007, People’s Republic of China

      **ting Zhao & Mingjun Li

    2. Bei**g Entry-Exit Inspection and Quarantine Bureau, Tianshuiyuan Street No. 6, Chaoyang District, Bei**g, 100026, People’s Republic of China

      **ngliang Liu

    3. State Key Laboratory of Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Yuanmingyuan West Road No. 2, Haidian District, Bei**g, 100193, People’s Republic of China

      Beibei Ge

    4. Department of Ornamental Horticulture, College of Agronomy and Biotechnology, China Agricultural University, Yuanmingyuan West Road No. 2, Haidian District, Bei**g, 100193, People’s Republic of China

      **ngliang Liu & Bo Hong

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    1. **ting Zhao
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    2. **ngliang Liu
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    Correspondence to Bo Hong.

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    X. Zhao and X. Liu contributed equally to this work.

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    Zhao, X., Liu, X., Ge, B. et al. A multiplex RT-PCR for simultaneous detection and identification of five viruses and two viroids infecting chrysanthemum. Arch Virol 160, 1145–1152 (2015). https://doi.org/10.1007/s00705-015-2360-z

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    • DOI: https://doi.org/10.1007/s00705-015-2360-z

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