4.7 Article

Identification and Characterization of Citrus Concave Gum-Associated Virus Infecting Citrus and Apple Trees by Serological, Molecular and High-Throughput Sequencing Approaches

期刊

PLANTS-BASEL
卷 10, 期 11, 页码 -

出版社

MDPI
DOI: 10.3390/plants10112390

关键词

apple rubbery wood virus 2; ARWV-2; CCGaV; coguvirus; concave gum disease; DAS-ELISA; DTBIA; next-generation sequencing; virus detection

资金

  1. Regione Campania [DGR 690/2016]
  2. European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant [734736]

向作者/读者索取更多资源

CCGaV is a negative-stranded RNA virus that can be detected in citrus and apple trees. The development of an anti-CCGaV polyclonal antiserum has allowed for sensitive and reliable detection of the virus in DAS-ELISA, revealing its relationships with different hosts.
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 (Citrus or Malus), 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.

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