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A multiple reverse transcription PCR assay for simultaneous detection of four main viruses in kiwifruit

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Abstract

Kiwifruit (Actinidia spp.) is an important horticultural crop in the world. Previous studies showed that a number of viruses were found in kiwifruit around the world. To detect the multiple viruses in kiwifruit quickly and effectively, a multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) method was developed to detect four viruses simultaneously in kiwifruit: Actinidia chlorotic ringspot-associated virus (AcCRaV), Actinidia virus 1(AcV-1), Actinidia virus A (AcVA), Citrus leaf blotch virus (CLBV), respectively. Four pairs of primers were designed according to the coat protein (CP) genes of the aforementioned viruses and then were confirmed by RT-PCR assay. Actin gene (ACT1) was used as an internal control to improve reliability of the multiplex RT-PCR. Determination of detection limit of the multiplex RT-PCR showed that this method could detect four viruses at the dilution of 10−4 cDNA. Furthermore, the developed multiplex RT-PCR could detect viruses in kiwifruit field samples successfully.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31772131, 31270290) and Applied Basic Research Fund of Sichuan Province (2019YJ0135) and the Fundamental Research Funds for the Central Universities (SCU2019D013). We are grateful to Dr. Qiguo Zhuang (Institute of Kiwifruit, Sichuan Provincial Academy of Natural Resource Science, Sichuan Province, China) for kindly collecting of kiwifruit samples.

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Authors and Affiliations

  1. Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, Sichuan, People’s Republic of China

    Qiding Peng, Long Qiu, Ting Yang, Jiachen Ning, Qianyi Xu & Dehui **

  2. Institute of Biotechnology and Germplasm Resources, Yunnan Academy of Agricultural Sciences, Yunnan Provincial Key Laboratory of Agricultural Biotechnology, Key Lab of Southwestern Crop Gene Resource and Germplasm Innovation, Ministry of Agriculture, Kunming, 650204, China

    Jiahong Dong

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  1. Qiding Peng
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  2. Long Qiu
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  3. Ting Yang
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  4. Jiachen Ning
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  5. Qianyi Xu
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  6. Jiahong Dong
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  7. Dehui **
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Correspondence to Jiahong Dong or Dehui **.

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Peng, Q., Qiu, L., Yang, T. et al. A multiple reverse transcription PCR assay for simultaneous detection of four main viruses in kiwifruit. Eur J Plant Pathol 156, 1207–1212 (2020). https://doi.org/10.1007/s10658-020-01950-w

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