Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species
The banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus that infects <i>Musa</i> spp. and has a very wide geographical distribution. The current BanMMV indexing process for an accession requires the testing of no less than four pla...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-01-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/11/2/226 |
| _version_ | 1797490980810653696 |
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| author | Marwa Hanafi Wei Rong Lucie Tamisier Chadi Berhal Nicolas Roux Sebastien Massart |
| author_facet | Marwa Hanafi Wei Rong Lucie Tamisier Chadi Berhal Nicolas Roux Sebastien Massart |
| author_sort | Marwa Hanafi |
| collection | DOAJ |
| description | The banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus that infects <i>Musa</i> spp. and has a very wide geographical distribution. The current BanMMV indexing process for an accession requires the testing of no less than four plants cultivated in a greenhouse for at least 6 months and causes a significant delay for the distribution of the germplasm. We evaluated the sensitivity of different protocols for BanMMV detection from in vitro plants to accelerate the testing process. We first used corm tissues from 137 in vitro plants and obtained a diagnostic sensitivity (DSE) of only 61% when testing four plants per accession. After thermotherapy was carried out to eliminate BanMMV infection, the meristem was recovered and further grown in vitro. The same protocol was evaluated in parallel on the corm tissue surrounding the meristem, as a rapid screening to evaluate virus therapy success, and was compared to the results obtained following the standard protocol. The obtained results showed 28% false negatives when conducting testing from corm tissues, making this protocol unsuitable in routine processes. Furthermore, RT-PCR and high-throughput sequencing (HTS) tests were applied on tissues from the base (<i>n</i> = 39) and the leaves (<i>n</i> = 36). For RT-PCR, the average DSE per sample reached 65% from either the base or leaves. HTS was applied on 36 samples and yielded 100% diagnostic specificity (DSP) and 100% DSE, whatever the sampled tissue, allowing the identification of a new Betaflexiviridae species infecting <i>Musa</i>. These results suggest that a reliable diagnostic of BanMMV from in vitro plants using RT-PCR or HTS technologies might represent an efficient alternative for testing after greenhouse cultivation. |
| first_indexed | 2024-03-10T00:40:51Z |
| format | Article |
| id | doaj.art-03721fee47c8477db026295dbb58973a |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T00:40:51Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-03721fee47c8477db026295dbb58973a2023-11-23T15:07:53ZengMDPI AGPlants2223-77472022-01-0111222610.3390/plants11020226Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae SpeciesMarwa Hanafi0Wei Rong1Lucie Tamisier2Chadi Berhal3Nicolas Roux4Sebastien Massart5Integrated and Urban Plant Pathology Laboratory, Gembloux Agro-Bio Tech, University of Liège, 2, Passage des Déportés, 5030 Gembloux, BelgiumIntegrated and Urban Plant Pathology Laboratory, Gembloux Agro-Bio Tech, University of Liège, 2, Passage des Déportés, 5030 Gembloux, BelgiumIntegrated and Urban Plant Pathology Laboratory, Gembloux Agro-Bio Tech, University of Liège, 2, Passage des Déportés, 5030 Gembloux, BelgiumIntegrated and Urban Plant Pathology Laboratory, Gembloux Agro-Bio Tech, University of Liège, 2, Passage des Déportés, 5030 Gembloux, BelgiumBioversity International, Parc Scientifique Agropolis II, 34397 Montpellier, FranceIntegrated and Urban Plant Pathology Laboratory, Gembloux Agro-Bio Tech, University of Liège, 2, Passage des Déportés, 5030 Gembloux, BelgiumThe banana mild mosaic virus (BanMMV) (Betaflexiviridae, Quinvirinae, unassigned species) is a filamentous virus that infects <i>Musa</i> spp. and has a very wide geographical distribution. The current BanMMV indexing process for an accession requires the testing of no less than four plants cultivated in a greenhouse for at least 6 months and causes a significant delay for the distribution of the germplasm. We evaluated the sensitivity of different protocols for BanMMV detection from in vitro plants to accelerate the testing process. We first used corm tissues from 137 in vitro plants and obtained a diagnostic sensitivity (DSE) of only 61% when testing four plants per accession. After thermotherapy was carried out to eliminate BanMMV infection, the meristem was recovered and further grown in vitro. The same protocol was evaluated in parallel on the corm tissue surrounding the meristem, as a rapid screening to evaluate virus therapy success, and was compared to the results obtained following the standard protocol. The obtained results showed 28% false negatives when conducting testing from corm tissues, making this protocol unsuitable in routine processes. Furthermore, RT-PCR and high-throughput sequencing (HTS) tests were applied on tissues from the base (<i>n</i> = 39) and the leaves (<i>n</i> = 36). For RT-PCR, the average DSE per sample reached 65% from either the base or leaves. HTS was applied on 36 samples and yielded 100% diagnostic specificity (DSP) and 100% DSE, whatever the sampled tissue, allowing the identification of a new Betaflexiviridae species infecting <i>Musa</i>. These results suggest that a reliable diagnostic of BanMMV from in vitro plants using RT-PCR or HTS technologies might represent an efficient alternative for testing after greenhouse cultivation.https://www.mdpi.com/2223-7747/11/2/226high-throughput sequencing(IC)-RT-PCRdiagnostic performanceBanMMVin vitro plantsmeristem culture |
| spellingShingle | Marwa Hanafi Wei Rong Lucie Tamisier Chadi Berhal Nicolas Roux Sebastien Massart Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species Plants high-throughput sequencing (IC)-RT-PCR diagnostic performance BanMMV in vitro plants meristem culture |
| title | Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species |
| title_full | Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species |
| title_fullStr | Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species |
| title_full_unstemmed | Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species |
| title_short | Detection of Banana Mild Mosaic Virus in <i>Musa</i> In Vitro Plants: High-Throughput Sequencing Presents Higher Diagnostic Sensitivity Than (IC)-RT-PCR and Identifies a New Betaflexiviridae Species |
| title_sort | detection of banana mild mosaic virus in i musa i in vitro plants high throughput sequencing presents higher diagnostic sensitivity than ic rt pcr and identifies a new betaflexiviridae species |
| topic | high-throughput sequencing (IC)-RT-PCR diagnostic performance BanMMV in vitro plants meristem culture |
| url | https://www.mdpi.com/2223-7747/11/2/226 |
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