Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses
Mycoviruses are viruses that specifically infect and replicate in fungi. Several mycoviruses have been previously reported in <i>Pleurotus ostreatus</i>, including the oyster mushroom spherical virus (OMSV), oyster mushroom isometric virus (OMIV), <i>Pleurotus ostreatus</i> s...
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MDPI AG
2022-11-01
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| Series: | Current Issues in Molecular Biology |
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| Online Access: | https://www.mdpi.com/1467-3045/44/11/392 |
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| author | Xiaoyan Zhang Haijing Hu Yifan Wang Junjie Yan Yu Liu Jianrui Wang Xianhao Cheng |
| author_facet | Xiaoyan Zhang Haijing Hu Yifan Wang Junjie Yan Yu Liu Jianrui Wang Xianhao Cheng |
| author_sort | Xiaoyan Zhang |
| collection | DOAJ |
| description | Mycoviruses are viruses that specifically infect and replicate in fungi. Several mycoviruses have been previously reported in <i>Pleurotus ostreatus</i>, including the oyster mushroom spherical virus (OMSV), oyster mushroom isometric virus (OMIV), <i>Pleurotus ostreatus</i> spherical virus (POSV), and <i>Pleurotus ostreatus</i> virus 1 (PoV1). This study was designed to develop a multiplex RT-PCR for simultaneous detection and differentiation of the four <i>P. ostreatus</i> mycoviruses. Four pairs of primers were designed from conserved regions based on the reported sequences and the multiplex RT-PCR products were 672 bp for OMSV, 540 bp for OMIV, 310 bp for POSV, and 200 bp for PoV1. The optimal annealing temperature of the multiplex RT-PCR was 62 °C and the detection limits of the plasmids were 100 fg for OMSV and OMIV and 1 pg for POSV and PoV1. This technique was successfully applied for the detection of OMSV, OMIV, and POSV from different <i>P. ostreatus</i> strains and the plasmid containing the PoV1 sequence. This methodology can serve as a powerful diagnostic tool for the survey of the incidence and epidemiology of the four <i>P. ostreatus</i> mycoviruses, further contributing to the prevention and treatment of mycoviral diseases in <i>P. ostreatus</i>. |
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| issn | 1467-3037 1467-3045 |
| language | English |
| last_indexed | 2024-03-09T18:24:41Z |
| publishDate | 2022-11-01 |
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| series | Current Issues in Molecular Biology |
| spelling | doaj.art-a448a07885304202b37f8469308e85262023-11-24T08:01:13ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452022-11-0144115778578710.3390/cimb44110392Multiplex Detection of <i>Pleurotus ostreatus</i> MycovirusesXiaoyan Zhang0Haijing Hu1Yifan Wang2Junjie Yan3Yu Liu4Jianrui Wang5Xianhao Cheng6Shandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaShandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaShandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaShandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaShandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaShandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaShandong Key Laboratory of Edible Mushroom Technology, School of Agriculture, Ludong University, Yantai 264025, ChinaMycoviruses are viruses that specifically infect and replicate in fungi. Several mycoviruses have been previously reported in <i>Pleurotus ostreatus</i>, including the oyster mushroom spherical virus (OMSV), oyster mushroom isometric virus (OMIV), <i>Pleurotus ostreatus</i> spherical virus (POSV), and <i>Pleurotus ostreatus</i> virus 1 (PoV1). This study was designed to develop a multiplex RT-PCR for simultaneous detection and differentiation of the four <i>P. ostreatus</i> mycoviruses. Four pairs of primers were designed from conserved regions based on the reported sequences and the multiplex RT-PCR products were 672 bp for OMSV, 540 bp for OMIV, 310 bp for POSV, and 200 bp for PoV1. The optimal annealing temperature of the multiplex RT-PCR was 62 °C and the detection limits of the plasmids were 100 fg for OMSV and OMIV and 1 pg for POSV and PoV1. This technique was successfully applied for the detection of OMSV, OMIV, and POSV from different <i>P. ostreatus</i> strains and the plasmid containing the PoV1 sequence. This methodology can serve as a powerful diagnostic tool for the survey of the incidence and epidemiology of the four <i>P. ostreatus</i> mycoviruses, further contributing to the prevention and treatment of mycoviral diseases in <i>P. ostreatus</i>.https://www.mdpi.com/1467-3045/44/11/392<i>Pleurotus ostreatus</i>multiplex PCRoyster mushroom spherical virusoyster mushroom isometric virus<i>Pleurotus ostreatus</i> spherical virus<i>Pleurotus ostreatus</i> virus 1 |
| spellingShingle | Xiaoyan Zhang Haijing Hu Yifan Wang Junjie Yan Yu Liu Jianrui Wang Xianhao Cheng Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses Current Issues in Molecular Biology <i>Pleurotus ostreatus</i> multiplex PCR oyster mushroom spherical virus oyster mushroom isometric virus <i>Pleurotus ostreatus</i> spherical virus <i>Pleurotus ostreatus</i> virus 1 |
| title | Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses |
| title_full | Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses |
| title_fullStr | Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses |
| title_full_unstemmed | Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses |
| title_short | Multiplex Detection of <i>Pleurotus ostreatus</i> Mycoviruses |
| title_sort | multiplex detection of i pleurotus ostreatus i mycoviruses |
| topic | <i>Pleurotus ostreatus</i> multiplex PCR oyster mushroom spherical virus oyster mushroom isometric virus <i>Pleurotus ostreatus</i> spherical virus <i>Pleurotus ostreatus</i> virus 1 |
| url | https://www.mdpi.com/1467-3045/44/11/392 |
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