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Multiplex RT–PCR Assay for the Simultaneous Detection and Identification of Five Sugarcane Viruses

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Abstract

Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), Sugarcane streak mosaic virus (SCSMV), Sugarcane yellow leaf virus (SCYLV), and Sugarcane bacilliform virus (SCBV) are the major viruses infecting sugarcane worldwide. This study proposed a multiplex reverse transcription–polymerase chain reaction (RT–PCR) assay for the simultaneous detection and identification of these sugarcane viruses. Results showed that five primer pairs amplified their respective target viruses, indicating the assay specificity. The sensitivity of the assay was similar to or 10–100-folds lower than that of singleplex RT–PCR for detecting SCMV or the four other viruses, respectively. When detecting the five viruses in field sugarcane samples, the detection results of multiplex RT–PCR were approximately identical to those of singleplex RT–PCR, thereby indicating the relative reliability of the former. The proposed multiplex RT–PCR can simultaneously detect SCMV, SrMV, SCSMV, SCYLV, and SCBV in sugarcane, making it a suitable, efficient, and practical detection method for sugarcane viruses.

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Abbreviations

PCR:

Polymerase chain reaction

RT:

Reverse transcription

SCBV:

Sugarcane bacilliform virus

SCMV:

Sugarcane mosaic virus

SCSMV:

Sugarcane streak mosaic virus

SCYLV:

Sugarcane yellow leaf virus

SrMV:

Sorghum mosaic virus

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Acknowledgements

This study was funded by the National Key Research and Development Program of China (2019YFD1000503), Central Public-interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural Sciences (No. 19CXTD-33), National Natural Science Foundation of China (No. 31771865), and Sugar Crop Research System (CARS-170301).

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Authors and Affiliations

  1. Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, No. 4, College Road, Longhua District, Haikou, 571101, Hainan, China

    Xiao-Yan Feng, Wen-Zhi Wang, Lin-Bo Shen, Jun-Gang Wang, Guo-Ru Xiong, Cui-Lian Feng, Ting-Ting Zhao & Shu-Zhen Zhang

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  1. Xiao-Yan Feng
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  2. Wen-Zhi Wang
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  3. Lin-Bo Shen
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  4. Jun-Gang Wang
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  5. Guo-Ru Xiong
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  6. Cui-Lian Feng
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  7. Ting-Ting Zhao
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  8. Shu-Zhen Zhang
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Contributions

SZZ conceived and devised the experiments. XYF performed the experiments, analyzed the results, and wrote the manuscript. WZW, LBS, JGW, GRX, CLF, and TTZ revised the manuscript. All authors have read and approved the manuscript.

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Correspondence to Shu-Zhen Zhang.

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Feng, XY., Wang, WZ., Shen, LB. et al. Multiplex RT–PCR Assay for the Simultaneous Detection and Identification of Five Sugarcane Viruses. Sugar Tech 22, 662–670 (2020). https://doi.org/10.1007/s12355-020-00805-2

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