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Rapid diagnosis of wheat head blight caused by Fusarium asiaticum using a loop-mediated isothermal amplification assay

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Abstract

Fusarium asiaticum is an important fungal pathogen that causes wheat head blight or scab in China. In this study, we describe a loop-mediated isothermal amplification assay that directly detects F. asiaticum in diseased wheat head tissues, thus allowing rapid diagnosis of wheat head blight caused by this pathogen. We selected the CYP51C gene sequence to design loop-mediated isothermal amplification (LAMP) primers specific for F. asiaticum. The target gene (CYP51C) was efficiently amplified in 60 min at 63 °C using the LAMP assay. The sensitivity of the assay was 100 pg·μL−1. Specificity was compared with those for other Fusarium spp. and other fungal species. The result was directly visualized by adding hydroxynaphthol blue prior to amplification; a color change (sky blue) was observed only in the presence of F. asiaticum, whereas all other isolates showed no color change. We successfully applied the LAMP assay to diagnose the suspected disease in wheat head samples collected in the field. This study provides a new tool for the diagnosis of Fusarium head blight caused by F. asiaticum.

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Acknowledgements

This research was supported by grants of the National High-Tech R&D Program of China (863 Program: 2012AA101501) to Xiaobo Zheng, and the China Agriculture Research System (CARS-004-PS14) to Yuanchao Wang.

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Authors and Affiliations

  1. Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China

    Miao Xu, Wenwu Ye, Dandan Zeng, Yuanchao Wang & Xiaobo Zheng

  2. Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing, 210095, China

    Miao Xu, Wenwu Ye, Dandan Zeng, Yuanchao Wang & Xiaobo Zheng

Authors
  1. Miao Xu
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  2. Wenwu Ye
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  3. Dandan Zeng
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  4. Yuanchao Wang
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  5. Xiaobo Zheng
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Correspondence to Xiaobo Zheng.

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Xu, M., Ye, W., Zeng, D. et al. Rapid diagnosis of wheat head blight caused by Fusarium asiaticum using a loop-mediated isothermal amplification assay. Australasian Plant Pathol. 46, 261–266 (2017). https://doi.org/10.1007/s13313-017-0487-y

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  • DOI: https://doi.org/10.1007/s13313-017-0487-y

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