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<p>In this study, a comprehensive virome analysis was conducted on a germplasm collection of autochthonous pear varieties (<i>Pyrus communis</i> L.) in the Apulia region of southern Italy. A total of 239 pear tree samples were collected and individually tested for specific viruses using reverse transcription PCR (RT-PCR) and reverse transcription quantitative PCR (RT-qPCR). The survey targeted five viruses, two viroids, and one phytoplasma included in the Italian voluntary certification protocol for plant propagation material. Both molecular approaches (RT-PCR and RT-qPCR) enabled the detection of apple stem pitting virus (ASPV, 45.1%), apple chlorotic leaf spot virus (ACLSV, 33.9%), apple stem grooving virus (ASGV, 0.4%), apple rubbery wood virus 1 (ARWV1, 2.5%), apple rubbery wood virus 2 (ARWV2, 11.7%), pear blister canker viroid (PBCVd, 5%), and ‘<i>Candidatus</i> Phytoplasma pyri’ (0.4%). No amplification was obtained for apple scar skin viroid (ASSVd). Based on these results, a pool of 25 nucleic acid extracts that tested negative for the targeted pathogens was further analyzed by high-throughput sequencing (HTS). Illumina-based HTS revealed the presence of citrus virus A (CiVA), which had not been included in the initial screening. Follow-up PCR testing confirmed CiVA infection in 18% of the pear tree samples. The complete genome of the local CiVA isolate, designated IT-Apulia, was sequenced, revealing RNA1 of 6,658 nucleotides and RNA2 of 2,721nucleotides. These sequences have been deposited in GenBank under accession numbers PV665642 and PV665643, respectively. This is the first report of the presence of CiVA in pear trees in Italy.</p>

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First report of citrus virus A (CiVA) in Italian pear trees and sanitary assessment of native pear varieties in Apulia (southern Italy)

  • 期刊论文

  • 发表日期：

    2025年11月19日

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  • Amani Ben Slimen,
  • Antonella Karam,
  • Pasquale Venerito,
  • Michele Digiaro

摘要

In this study, a comprehensive virome analysis was conducted on a germplasm collection of autochthonous pear varieties (Pyrus communis L.) in the Apulia region of southern Italy. A total of 239 pear tree samples were collected and individually tested for specific viruses using reverse transcription PCR (RT-PCR) and reverse transcription quantitative PCR (RT-qPCR). The survey targeted five viruses, two viroids, and one phytoplasma included in the Italian voluntary certification protocol for plant propagation material. Both molecular approaches (RT-PCR and RT-qPCR) enabled the detection of apple stem pitting virus (ASPV, 45.1%), apple chlorotic leaf spot virus (ACLSV, 33.9%), apple stem grooving virus (ASGV, 0.4%), apple rubbery wood virus 1 (ARWV1, 2.5%), apple rubbery wood virus 2 (ARWV2, 11.7%), pear blister canker viroid (PBCVd, 5%), and ‘Candidatus Phytoplasma pyri’ (0.4%). No amplification was obtained for apple scar skin viroid (ASSVd). Based on these results, a pool of 25 nucleic acid extracts that tested negative for the targeted pathogens was further analyzed by high-throughput sequencing (HTS). Illumina-based HTS revealed the presence of citrus virus A (CiVA), which had not been included in the initial screening. Follow-up PCR testing confirmed CiVA infection in 18% of the pear tree samples. The complete genome of the local CiVA isolate, designated IT-Apulia, was sequenced, revealing RNA1 of 6,658 nucleotides and RNA2 of 2,721nucleotides. These sequences have been deposited in GenBank under accession numbers PV665642 and PV665643, respectively. This is the first report of the presence of CiVA in pear trees in Italy.