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Article

Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a

1
Department of Entomology, Agricultural Research Organization—the Volcani Center, 68 HaMaccabim Road, P.O. Box 15159, Rishon LeZion 7505101, Israel
2
The Shmunis School of Molecular Cell Biology & Biotechnology, Faculty of Life Science, Tel Aviv University, Tel Aviv 69978, Israel
3
Department of Plant Pathology and Weed Research, Agricultural Research Organization—the Volcani Center, 68 HaMaccabim Road, P.O. Box 15159, Rishon LeZion 7505101, Israel
*
Authors to whom correspondence should be addressed.
Academic Editors: Ozgur Batuman and Aviv Dombrovsky
Plants 2021, 10(6), 1256; https://doi.org/10.3390/plants10061256
Received: 11 May 2021 / Revised: 14 June 2021 / Accepted: 16 June 2021 / Published: 21 June 2021
(This article belongs to the Special Issue Tobamoviruses and Interacting Viruses in Modern Agriculture)
CRISPR/Cas12a-based detection is a novel approach for the efficient, sequence-specific identification of viruses. Here we adopt the use of CRISPR/Cas12a to identify the tomato brown rugose fruit virus (ToBRFV), a new and emerging tobamovirus which is causing substantial damage to the global tomato industry. Specific CRISPR RNAs (crRNAs) were designed to detect either ToBRFV or the closely related tomato mosaic virus (ToMV). This technology enabled the differential detection of ToBRFV and ToMV. Sensitivity assays revealed that viruses can be detected from 15–30 ng of RT-PCR product, and that specific detection could be achieved from a mix of ToMV and ToBRFV. In addition, we show that this method can enable the identification of ToBRFV in samples collected from commercial greenhouses. These results demonstrate a new method for species-specific detection of tobamoviruses. A future combination of this approach with isothermal amplification could provide a platform for efficient and user-friendly ways to distinguish between closely related strains and resistance-breaking pathogens. View Full-Text
Keywords: CRISPR/Cas12a; Solanum lycopersicum; Tobamovirus; ToBRFV; tomato brown rugose fruit virus; tomato mosaic virus; ToMV CRISPR/Cas12a; Solanum lycopersicum; Tobamovirus; ToBRFV; tomato brown rugose fruit virus; tomato mosaic virus; ToMV
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MDPI and ACS Style

Alon, D.M.; Hak, H.; Bornstein, M.; Pines, G.; Spiegelman, Z. Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a. Plants 2021, 10, 1256. https://doi.org/10.3390/plants10061256

AMA Style

Alon DM, Hak H, Bornstein M, Pines G, Spiegelman Z. Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a. Plants. 2021; 10(6):1256. https://doi.org/10.3390/plants10061256

Chicago/Turabian Style

Alon, Dan M., Hagit Hak, Menachem Bornstein, Gur Pines, and Ziv Spiegelman. 2021. "Differential Detection of the Tobamoviruses Tomato Mosaic Virus (ToMV) and Tomato Brown Rugose Fruit Virus (ToBRFV) Using CRISPR-Cas12a" Plants 10, no. 6: 1256. https://doi.org/10.3390/plants10061256

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