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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Autores principales:	Maria Minutolo, Maria Cinque, Michela Chiumenti, Francesco Di Serio, Daniela Alioto, Beatriz Navarro
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	apple rubbery wood virus 2 ARWV-2 CCGaV coguvirus concave gum disease DAS-ELISA Botany QK1-989
Acceso en línea:	https://doaj.org/article/149cc19b99854e39a9f7d68b40695445
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spelling	oai:doaj.org-article:149cc19b99854e39a9f7d68b406954452021-11-25T18:46:13ZIdentification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches10.3390/plants101123902223-7747https://doaj.org/article/149cc19b99854e39a9f7d68b406954452021-11-01T00:00:00Zhttps://www.mdpi.com/2223-7747/10/11/2390https://doaj.org/toc/2223-7747Citrus 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.Maria MinutoloMaria CinqueMichela ChiumentiFrancesco Di SerioDaniela AliotoBeatriz NavarroMDPI AGarticleapple rubbery wood virus 2ARWV-2CCGaVcoguvirusconcave gum diseaseDAS-ELISABotanyQK1-989ENPlants, Vol 10, Iss 2390, p 2390 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	apple rubbery wood virus 2 ARWV-2 CCGaV coguvirus concave gum disease DAS-ELISA Botany QK1-989
spellingShingle	apple rubbery wood virus 2 ARWV-2 CCGaV coguvirus concave gum disease DAS-ELISA Botany QK1-989 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
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.
format	article
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
title	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_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_sort	identification and characterization of citrus concave gum-associated virus infecting citrus and apple trees by serological, molecular and high-throughput sequencing approaches
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/149cc19b99854e39a9f7d68b40695445
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Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches

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