Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i>
Green Sichuan pepper (<i>Zanthoxylum armatum</i>) is an important economic fruit crop, which is widely planted in the southwest region of China. Recently, a serious disease, namely flower yellowing disease (FYD), broke out, and the virus of green Sichuan pepper nepovirus (GSPNeV) was ide...
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MDPI AG
2022-05-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/11/9/1242 |
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| author | Zhenfei Dong Xiaoli Zhao Junjie Liu Binhui Zhan Shifang Li |
| author_facet | Zhenfei Dong Xiaoli Zhao Junjie Liu Binhui Zhan Shifang Li |
| author_sort | Zhenfei Dong |
| collection | DOAJ |
| description | Green Sichuan pepper (<i>Zanthoxylum armatum</i>) is an important economic fruit crop, which is widely planted in the southwest region of China. Recently, a serious disease, namely flower yellowing disease (FYD), broke out, and the virus of green Sichuan pepper nepovirus (GSPNeV) was identified to be highly correlated with the viral symptoms. Meanwhile, green Sichuan pepper idaeovirus (GSPIV) and green Sichuan pepper enamovirus (GSPEV) were also common viruses infecting green pepper. In our research, specific primers were designed according to the reported sequences of the three viruses, and a multiplex reverse transcription-polymerase chain reaction (RT-PCR) method for the simultaneous detection of GSPNeV, GSPIV, and GSPEV was established. The annealing temperature, extension time, and cycle number affecting the multiplex RT-PCR reaction were adjusted and optimized. Sensitivity analysis showed that the system could detect the three viruses simultaneously from the complementary deoxyribonucleic acid (cDNA) samples diluted by 10<sup>−3</sup>. The results of the ten samples detected by the multiplex RT-PCR system were consistent with the results of a single PCR, indicating that the method can be successfully used for rapid detection of field samples. |
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| id | doaj.art-3261d2c79d77412182217ca9d74f1e93 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T03:47:39Z |
| publishDate | 2022-05-01 |
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| spelling | doaj.art-3261d2c79d77412182217ca9d74f1e932023-11-23T09:03:33ZengMDPI AGPlants2223-77472022-05-01119124210.3390/plants11091242Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i>Zhenfei Dong0Xiaoli Zhao1Junjie Liu2Binhui Zhan3Shifang Li4State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaDepartment of Fruit Science, College of Horticulture, China Agricultural University, Beijing 100107, ChinaDepartment of Fruit Science, College of Horticulture, China Agricultural University, Beijing 100107, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, ChinaGreen Sichuan pepper (<i>Zanthoxylum armatum</i>) is an important economic fruit crop, which is widely planted in the southwest region of China. Recently, a serious disease, namely flower yellowing disease (FYD), broke out, and the virus of green Sichuan pepper nepovirus (GSPNeV) was identified to be highly correlated with the viral symptoms. Meanwhile, green Sichuan pepper idaeovirus (GSPIV) and green Sichuan pepper enamovirus (GSPEV) were also common viruses infecting green pepper. In our research, specific primers were designed according to the reported sequences of the three viruses, and a multiplex reverse transcription-polymerase chain reaction (RT-PCR) method for the simultaneous detection of GSPNeV, GSPIV, and GSPEV was established. The annealing temperature, extension time, and cycle number affecting the multiplex RT-PCR reaction were adjusted and optimized. Sensitivity analysis showed that the system could detect the three viruses simultaneously from the complementary deoxyribonucleic acid (cDNA) samples diluted by 10<sup>−3</sup>. The results of the ten samples detected by the multiplex RT-PCR system were consistent with the results of a single PCR, indicating that the method can be successfully used for rapid detection of field samples.https://www.mdpi.com/2223-7747/11/9/1242multiplex RT-PCR<i>Zanthoxylum armatum</i>GSPNeVGSPIVGSPEVITS2 |
| spellingShingle | Zhenfei Dong Xiaoli Zhao Junjie Liu Binhui Zhan Shifang Li Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i> Plants multiplex RT-PCR <i>Zanthoxylum armatum</i> GSPNeV GSPIV GSPEV ITS2 |
| title | Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i> |
| title_full | Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i> |
| title_fullStr | Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i> |
| title_full_unstemmed | Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i> |
| title_short | Detection and Simultaneous Differentiation of Three Co-infected Viruses in <i>Zanthoxylum armatum</i> |
| title_sort | detection and simultaneous differentiation of three co infected viruses in i zanthoxylum armatum i |
| topic | multiplex RT-PCR <i>Zanthoxylum armatum</i> GSPNeV GSPIV GSPEV ITS2 |
| url | https://www.mdpi.com/2223-7747/11/9/1242 |
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