Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco
Abstract Background Chilli veinal mottle virus (ChiVMV), which belongs to the genus Potyvirus of the family Potyviridae, mainly infects solanaceous plants and has caused serious economic losses in Asia and Africa. Tobacco plants infected with ChiVMV suffered from punctate necrosis of leaves, leaf de...
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| Format: | Article |
| Language: | English |
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BMC
2020-03-01
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| Series: | Virology Journal |
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| Online Access: | http://link.springer.com/article/10.1186/s12985-020-01299-w |
| _version_ | 1818953356323848192 |
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| author | Yubing Jiao Chuantao Xu Jialun Li Yong Gu Chun Xia Qiang Xie Yunbo Xie Mengnan An Zihao Xia Yuanhua Wu |
| author_facet | Yubing Jiao Chuantao Xu Jialun Li Yong Gu Chun Xia Qiang Xie Yunbo Xie Mengnan An Zihao Xia Yuanhua Wu |
| author_sort | Yubing Jiao |
| collection | DOAJ |
| description | Abstract Background Chilli veinal mottle virus (ChiVMV), which belongs to the genus Potyvirus of the family Potyviridae, mainly infects solanaceous plants and has caused serious economic losses in Asia and Africa. Tobacco plants infected with ChiVMV suffered from punctate necrosis of leaves, leaf deformation, systemic necrosis of leaves and stems, and eventually plant death. However, ChiVMV infection could not usually be identified given the lack of rapid and efficient detection assays in tobacco plants. Therefore, an isolate of tobacco-infecting ChiVMV (ChiVMV-LZ) was obtained, and a novel isothermal amplification and detection technique, reverse transcription-recombinase polymerase amplification (RT-RPA), was established to detect ChiVMV in tobacco plants. Methods In this study, the full-length genome of ChiVMV-LZ was obtained using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) assays. The genome sequence of ChiVMV-LZ was characterized by sequence alignment and phylogenetic analysis. Then, a RT-RPA assay was established for rapid and sensitive detection of ChiVMV-LZ in tobacco. Additionally, the established RT-RPA assay was compared to the RT-PCR assay in aspect of sensitivity and application in field-collected tobacco samples. Results ChiVMV-LZ was isolated from diseased tobacco in Luzhou, Sichuan, China. The tobacco plants inoculated with ChiVMV-LZ showed typical symptoms of yellow and round spots on the leaves, and curled and folded leaf margin, similar to those observed on naturally ChiVMV-infected tobacco in the field. The full-length genomic sequence of ChiVMV-LZ was determined to be 9742 nucleotides. Sequence alignment and phylogenetic analysis showed that ChiVMV-LZ was most closely related to ChiVMV-Yp8 isolated from pepper plants in Sichuan province while distantly related to ChiVMV-YN from tobacco in Yunnan province, indicating a possibly geographical differentiation of ChiVMV isolates. Additionally, a RT-RPA assay was established for rapid detection of ChiVMV in tobacco. The RT-RPA has no cross-reaction with other related tobacco viruses and is about 10-fold more sensitive than conventional RT-PCR method. Conclusion The characterization of ChiVMV-LZ infecting tobacco was determined, and the established RT-RPA assay provides a reliable and effective method for rapid detection of ChiVMV in tobacco. |
| first_indexed | 2024-12-20T10:04:58Z |
| format | Article |
| id | doaj.art-a24310f19c6d43c78cc95aaf1be29775 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-20T10:04:58Z |
| publishDate | 2020-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-a24310f19c6d43c78cc95aaf1be297752022-12-21T19:44:15ZengBMCVirology Journal1743-422X2020-03-011711910.1186/s12985-020-01299-wCharacterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobaccoYubing Jiao0Chuantao Xu1Jialun Li2Yong Gu3Chun Xia4Qiang Xie5Yunbo Xie6Mengnan An7Zihao Xia8Yuanhua Wu9College of Plant Protection, Shenyang Agricultural UniversityCollege of Plant Protection, Shenyang Agricultural UniversityCollege of Plant Protection, Shenyang Agricultural UniversityLuzhou City Company of Sichuan Tobacco CompanyLuzhou City Company of Sichuan Tobacco CompanyLuzhou City Company of Sichuan Tobacco CompanySichuan Province Company of China Tobacco CorporationCollege of Plant Protection, Shenyang Agricultural UniversityCollege of Plant Protection, Shenyang Agricultural UniversityCollege of Plant Protection, Shenyang Agricultural UniversityAbstract Background Chilli veinal mottle virus (ChiVMV), which belongs to the genus Potyvirus of the family Potyviridae, mainly infects solanaceous plants and has caused serious economic losses in Asia and Africa. Tobacco plants infected with ChiVMV suffered from punctate necrosis of leaves, leaf deformation, systemic necrosis of leaves and stems, and eventually plant death. However, ChiVMV infection could not usually be identified given the lack of rapid and efficient detection assays in tobacco plants. Therefore, an isolate of tobacco-infecting ChiVMV (ChiVMV-LZ) was obtained, and a novel isothermal amplification and detection technique, reverse transcription-recombinase polymerase amplification (RT-RPA), was established to detect ChiVMV in tobacco plants. Methods In this study, the full-length genome of ChiVMV-LZ was obtained using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) assays. The genome sequence of ChiVMV-LZ was characterized by sequence alignment and phylogenetic analysis. Then, a RT-RPA assay was established for rapid and sensitive detection of ChiVMV-LZ in tobacco. Additionally, the established RT-RPA assay was compared to the RT-PCR assay in aspect of sensitivity and application in field-collected tobacco samples. Results ChiVMV-LZ was isolated from diseased tobacco in Luzhou, Sichuan, China. The tobacco plants inoculated with ChiVMV-LZ showed typical symptoms of yellow and round spots on the leaves, and curled and folded leaf margin, similar to those observed on naturally ChiVMV-infected tobacco in the field. The full-length genomic sequence of ChiVMV-LZ was determined to be 9742 nucleotides. Sequence alignment and phylogenetic analysis showed that ChiVMV-LZ was most closely related to ChiVMV-Yp8 isolated from pepper plants in Sichuan province while distantly related to ChiVMV-YN from tobacco in Yunnan province, indicating a possibly geographical differentiation of ChiVMV isolates. Additionally, a RT-RPA assay was established for rapid detection of ChiVMV in tobacco. The RT-RPA has no cross-reaction with other related tobacco viruses and is about 10-fold more sensitive than conventional RT-PCR method. Conclusion The characterization of ChiVMV-LZ infecting tobacco was determined, and the established RT-RPA assay provides a reliable and effective method for rapid detection of ChiVMV in tobacco.http://link.springer.com/article/10.1186/s12985-020-01299-wChilli veinal mottle virus (ChiVMV)CharacterizationRT-RPARapid detectionNicotinana tabacum |
| spellingShingle | Yubing Jiao Chuantao Xu Jialun Li Yong Gu Chun Xia Qiang Xie Yunbo Xie Mengnan An Zihao Xia Yuanhua Wu Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco Virology Journal Chilli veinal mottle virus (ChiVMV) Characterization RT-RPA Rapid detection Nicotinana tabacum |
| title | Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco |
| title_full | Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco |
| title_fullStr | Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco |
| title_full_unstemmed | Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco |
| title_short | Characterization and a RT-RPA assay for rapid detection of Chilli Veinal mottle virus (ChiVMV) in tobacco |
| title_sort | characterization and a rt rpa assay for rapid detection of chilli veinal mottle virus chivmv in tobacco |
| topic | Chilli veinal mottle virus (ChiVMV) Characterization RT-RPA Rapid detection Nicotinana tabacum |
| url | http://link.springer.com/article/10.1186/s12985-020-01299-w |
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