Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches
Citrus concave gum-associated virus (CCGaV) is a negative-stranded RNA virus, first reported a few years ago in citrus trees from Italy. It has been reported in apple trees in the USA and in Brazil, suggesting a wider host range and geographic distribution. Here, an anti-CCGaV polyclonal antiserum t...
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| Format: | Article |
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MDPI AG
2021-11-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/10/11/2390 |
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| author | Maria Minutolo Maria Cinque Michela Chiumenti Francesco Di Serio Daniela Alioto Beatriz Navarro |
| author_facet | Maria Minutolo Maria Cinque Michela Chiumenti Francesco Di Serio Daniela Alioto Beatriz Navarro |
| author_sort | Maria Minutolo |
| collection | DOAJ |
| description | Citrus concave gum-associated virus (CCGaV) is a negative-stranded RNA virus, first reported a few years ago in citrus trees from Italy. It has been reported in apple trees in the USA and in Brazil, suggesting a wider host range and geographic distribution. Here, an anti-CCGaV polyclonal antiserum to specifically detect the virus has been developed and used in a standard double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) that has been validated as a sensitive and reliable method to detect this virus both in citrus and apple trees. In contrast, when the same antiserum was used in direct tissue-blot immunoassay, CCGaV was efficiently detected in citrus but not in apple. Using this antiserum, the first apple trees infected by CCGaV were identified in Italy and the presence of CCGaV in several apple cultivars in southern Italy was confirmed by field surveys. High-throughput sequencing (HTS) allowed for the assembling of the complete genome of one CCGaV Italian apple isolate (CE-c3). Phylogenetic analysis of Italian CCGaV isolates from apple and citrus and those available in the database showed close relationships between the isolates from the same genus (<i>Citrus</i> or <i>Malus</i>), regardless their geographical origin. This finding was further confirmed by the identification of amino acid signatures specific of isolates infecting citrus or apple hosts. Analysis of HTS reads also revealed that the CE-c3 Italian apple tree, besides CCGaV, was simultaneously infected by several viruses and one viroid, including apple rubbery wood virus 2 which is reported for the first time in Italy. The complete or almost complete genomic sequences of the coinfecting agents were determined. |
| first_indexed | 2024-03-10T05:07:36Z |
| format | Article |
| id | doaj.art-149cc19b99854e39a9f7d68b40695445 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T05:07:36Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-149cc19b99854e39a9f7d68b406954452023-11-23T01:04:53ZengMDPI AGPlants2223-77472021-11-011011239010.3390/plants10112390Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing ApproachesMaria Minutolo0Maria Cinque1Michela Chiumenti2Francesco Di Serio3Daniela Alioto4Beatriz Navarro5Dipartimento di Agraria, Università degli Studi di Napoli Federico II, 80055 Portici, ItalyDipartimento di Agraria, Università degli Studi di Napoli Federico II, 80055 Portici, ItalyIstituto per la Protezione Sostenibile delle Piante, Consiglio Nazionale delle Ricerche, 70126 Bari, ItalyIstituto per la Protezione Sostenibile delle Piante, Consiglio Nazionale delle Ricerche, 70126 Bari, ItalyDipartimento di Agraria, Università degli Studi di Napoli Federico II, 80055 Portici, ItalyIstituto per la Protezione Sostenibile delle Piante, Consiglio Nazionale delle Ricerche, 70126 Bari, ItalyCitrus concave gum-associated virus (CCGaV) is a negative-stranded RNA virus, first reported a few years ago in citrus trees from Italy. It has been reported in apple trees in the USA and in Brazil, suggesting a wider host range and geographic distribution. Here, an anti-CCGaV polyclonal antiserum to specifically detect the virus has been developed and used in a standard double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) that has been validated as a sensitive and reliable method to detect this virus both in citrus and apple trees. In contrast, when the same antiserum was used in direct tissue-blot immunoassay, CCGaV was efficiently detected in citrus but not in apple. Using this antiserum, the first apple trees infected by CCGaV were identified in Italy and the presence of CCGaV in several apple cultivars in southern Italy was confirmed by field surveys. High-throughput sequencing (HTS) allowed for the assembling of the complete genome of one CCGaV Italian apple isolate (CE-c3). Phylogenetic analysis of Italian CCGaV isolates from apple and citrus and those available in the database showed close relationships between the isolates from the same genus (<i>Citrus</i> or <i>Malus</i>), regardless their geographical origin. This finding was further confirmed by the identification of amino acid signatures specific of isolates infecting citrus or apple hosts. Analysis of HTS reads also revealed that the CE-c3 Italian apple tree, besides CCGaV, was simultaneously infected by several viruses and one viroid, including apple rubbery wood virus 2 which is reported for the first time in Italy. The complete or almost complete genomic sequences of the coinfecting agents were determined.https://www.mdpi.com/2223-7747/10/11/2390apple rubbery wood virus 2ARWV-2CCGaVcoguvirusconcave gum diseaseDAS-ELISA |
| spellingShingle | Maria Minutolo Maria Cinque Michela Chiumenti Francesco Di Serio Daniela Alioto Beatriz Navarro Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches Plants apple rubbery wood virus 2 ARWV-2 CCGaV coguvirus concave gum disease DAS-ELISA |
| title | Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches |
| title_full | Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches |
| title_fullStr | Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches |
| title_full_unstemmed | Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches |
| title_short | Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches |
| title_sort | identification and characterization of citrus concave gum associated virus infecting citrus and apple trees by serological molecular and high throughput sequencing approaches |
| topic | apple rubbery wood virus 2 ARWV-2 CCGaV coguvirus concave gum disease DAS-ELISA |
| url | https://www.mdpi.com/2223-7747/10/11/2390 |
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