A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region

Citrus are affected by many viruses and viroids, some globally widespread and some restricted to particular countries or areas. In this study, we simulated the use of high throughput sequencing (HTS) and the bioinformatic analysis of small interfering RNAs (siRNA) as a pre-screening method to guide...

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Main Authors: Grazia Licciardello, Rosario Ferraro, Giuseppe Scuderi, Marcella Russo, Antonino F. Catara
Format: Article
Language:English
Published: MDPI AG 2021-04-01
Series:Agriculture
Subjects:
Online Access:https://www.mdpi.com/2077-0472/11/5/400
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author Grazia Licciardello
Rosario Ferraro
Giuseppe Scuderi
Marcella Russo
Antonino F. Catara
author_facet Grazia Licciardello
Rosario Ferraro
Giuseppe Scuderi
Marcella Russo
Antonino F. Catara
author_sort Grazia Licciardello
collection DOAJ
description Citrus are affected by many viruses and viroids, some globally widespread and some restricted to particular countries or areas. In this study, we simulated the use of high throughput sequencing (HTS) and the bioinformatic analysis of small interfering RNAs (siRNA) as a pre-screening method to guide bioindexing and molecular detection to enhance the surveillance survey of some key or emerging citrus viruses, such as non-European citrus tristeza virus isolates (non-EU CTV), citrus tatter leaf virus, citrus leprosis virus, citrus yellow mosaic virus, and citrus bark cracking viroid, present in the EPPO lists, and the citrus yellow vein clearing virus. The HTS’s ability to detect other citrus viroids was also evaluated. The results demonstrate that HTS provides a comprehensive phytosanitary status of citrus samples either in single and multiple infections of viruses and viroids. It also provides effective information on citrus tristeza virus mixed infections despite not being able to identify the non-EU variants of the virus. Bioindexing checks each single virus infection but does not differentiate viroids on the Etrog citron indicator and is time-consuming. Molecular assays are valuable as confirmation tests of viruses and viroids but many pairs of primers are needed for a full screening and new or non-target pathogens remain undetected. In addition, the genomes of two isolates of the citrus yellow vein clearing virus and the citrus tatter leaf virus, detected in a sample from China, are described.
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spelling doaj.art-24d08ee46dae4219926317f723f6af6b2023-11-21T17:26:16ZengMDPI AGAgriculture2077-04722021-04-0111540010.3390/agriculture11050400A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO RegionGrazia Licciardello0Rosario Ferraro1Giuseppe Scuderi2Marcella Russo3Antonino F. Catara4Consiglio per la Ricerca in Agricoltura e L’analisi dell’Economia Agraria-Centro di Ricerca Olivicoltura, Frutticoltura e Agrumicoltura (CREA), C. da Li Rocchi, Via Settimio Severo 83, 87036 Rende (CS), ItalyAgrobiotech Soc. Coop. z.i. Blocco Palma I, Via V. Lancia 57, 95121 Catania, ItalyAgrobiotech Soc. Coop. z.i. Blocco Palma I, Via V. Lancia 57, 95121 Catania, ItalyAgrobiotech Soc. Coop. z.i. Blocco Palma I, Via V. Lancia 57, 95121 Catania, ItalyFormerly, Department of Phytosanitary Science and Technologies, University of Catania, 95123 Catania, ItalyCitrus are affected by many viruses and viroids, some globally widespread and some restricted to particular countries or areas. In this study, we simulated the use of high throughput sequencing (HTS) and the bioinformatic analysis of small interfering RNAs (siRNA) as a pre-screening method to guide bioindexing and molecular detection to enhance the surveillance survey of some key or emerging citrus viruses, such as non-European citrus tristeza virus isolates (non-EU CTV), citrus tatter leaf virus, citrus leprosis virus, citrus yellow mosaic virus, and citrus bark cracking viroid, present in the EPPO lists, and the citrus yellow vein clearing virus. The HTS’s ability to detect other citrus viroids was also evaluated. The results demonstrate that HTS provides a comprehensive phytosanitary status of citrus samples either in single and multiple infections of viruses and viroids. It also provides effective information on citrus tristeza virus mixed infections despite not being able to identify the non-EU variants of the virus. Bioindexing checks each single virus infection but does not differentiate viroids on the Etrog citron indicator and is time-consuming. Molecular assays are valuable as confirmation tests of viruses and viroids but many pairs of primers are needed for a full screening and new or non-target pathogens remain undetected. In addition, the genomes of two isolates of the citrus yellow vein clearing virus and the citrus tatter leaf virus, detected in a sample from China, are described.https://www.mdpi.com/2077-0472/11/5/400citrus tristeza viruscitrus yellow vein clearing viruscitrus tatter leaf virusnon-EU virusesgenotypegenome coverage
spellingShingle Grazia Licciardello
Rosario Ferraro
Giuseppe Scuderi
Marcella Russo
Antonino F. Catara
A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region
Agriculture
citrus tristeza virus
citrus yellow vein clearing virus
citrus tatter leaf virus
non-EU viruses
genotype
genome coverage
title A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region
title_full A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region
title_fullStr A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region
title_full_unstemmed A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region
title_short A Simulation of the Use of High Throughput Sequencing as Pre-Screening Assay to Enhance the Surveillance of Citrus Viruses and Viroids in the EPPO Region
title_sort simulation of the use of high throughput sequencing as pre screening assay to enhance the surveillance of citrus viruses and viroids in the eppo region
topic citrus tristeza virus
citrus yellow vein clearing virus
citrus tatter leaf virus
non-EU viruses
genotype
genome coverage
url https://www.mdpi.com/2077-0472/11/5/400
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