Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events

A RT–PCR assay developed to amplify the full coat protein (CP) gene of apple stem pitting virus (ASPV) was evaluated using 180 Greek apple and pear samples and showed a broad detection range. This method was used to investigate the presence of ASPV in quince in Greece and showed a high incidence of...

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Main Authors: Matthaios M. Mathioudakis, Varvara I. Maliogka, Thierry Candresse, Osmar Nickel, Thor Vinicius Martins Fajardo, Daria Budzyńska, Beata Hasiów-Jaroszewska, Nikolaos I. Katis
Format: Article
Language:English
Published: MDPI AG 2021-05-01
Series:Plants
Subjects:
Online Access:https://www.mdpi.com/2223-7747/10/5/917
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spelling doaj-68b00a22bf944a2da5af577115b1507c2021-05-31T23:07:23ZengMDPI AGPlants2223-77472021-05-011091791710.3390/plants10050917Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination EventsMatthaios M. Mathioudakis0Varvara I. Maliogka1Thierry Candresse2Osmar Nickel3Thor Vinicius Martins Fajardo4Daria Budzyńska5Beata Hasiów-Jaroszewska6Nikolaos I. Katis7Plant Pathology Laboratory, Institute of Olive Tree, Subtropical Plants & Viticulture, ELGO-DIMITRA, Karamanlis Ave. 167, Gr-73134 Chania, GreeceLaboratory of Plant Pathology, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, Gr-54124 Thessaloniki, GreeceINRAE, UMR BFP, University of Bordeaux, CS20032, CEDEX, 33882 Villenave d’Ornon, FranceEmbrapa Uva e Vinho, 95.701-008, Bento Gonçalves, Rio Grande do Sul 96200, BrazilEmbrapa Uva e Vinho, 95.701-008, Bento Gonçalves, Rio Grande do Sul 96200, BrazilDepartment of Virology and Bacteriology, Institute of Plant Protection-National Research Institute, ul. Wł. Węgorka 20, 60-318 Poznań, PolandDepartment of Virology and Bacteriology, Institute of Plant Protection-National Research Institute, ul. Wł. Węgorka 20, 60-318 Poznań, PolandLaboratory of Plant Pathology, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, Gr-54124 Thessaloniki, GreeceA RT–PCR assay developed to amplify the full coat protein (CP) gene of apple stem pitting virus (ASPV) was evaluated using 180 Greek apple and pear samples and showed a broad detection range. This method was used to investigate the presence of ASPV in quince in Greece and showed a high incidence of 52%. The sequences of 14 isolates from various hosts with a distinct RFLP profile were determined. ASPV population genetics and the factors driving ASPV evolution were analyzed using the Greek ASPV sequences, novel sequences from Brazilian apple trees and Chinese botanical <i>Pyrus</i> species, and homologous sequences retrieved from GenBank. Fourteen variant types of Greek, Brazilian and botanical isolates, which differ in CP gene length and presence of indels, were identified. In addition, these analyses showed high intra- and inter-group variation among isolates from different countries and hosts, indicating the significant variability present in ASPV. Recombination events were detected in four isolates originating from Greek pear and quince and two from Brazilian apples. In a phylogenetic analysis, there was a tendency for isolates to cluster together based on CP gene length, the isolation host, and the detection method applied. Although there was no strict clustering based on geographical origin, most isolates from a given country tended to regroup in specific clusters. Interestingly, it was found that the phylogeny was correlated to the type, position, and pattern of indels, which represent hallmarks of specific lineages and indicate their possible role in virus diversification, rather than the CP size itself. Evidence of recombination between isolates from botanical and cultivated species and the clustering of isolates from botanical species and isolates from cultivated species suggest the existence of a possible undetermined transmission mechanism allowing the exchange of ASPV isolates between the cultivated and wild/ornamental hosts.https://www.mdpi.com/2223-7747/10/5/917ASPVCP-specific RT–PCRpome fruitsbotanical pearsvariantsCP variability
collection DOAJ
language English
format Article
sources DOAJ
author Matthaios M. Mathioudakis
Varvara I. Maliogka
Thierry Candresse
Osmar Nickel
Thor Vinicius Martins Fajardo
Daria Budzyńska
Beata Hasiów-Jaroszewska
Nikolaos I. Katis
spellingShingle Matthaios M. Mathioudakis
Varvara I. Maliogka
Thierry Candresse
Osmar Nickel
Thor Vinicius Martins Fajardo
Daria Budzyńska
Beata Hasiów-Jaroszewska
Nikolaos I. Katis
Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events
Plants
ASPV
CP-specific RT–PCR
pome fruits
botanical pears
variants
CP variability
author_facet Matthaios M. Mathioudakis
Varvara I. Maliogka
Thierry Candresse
Osmar Nickel
Thor Vinicius Martins Fajardo
Daria Budzyńska
Beata Hasiów-Jaroszewska
Nikolaos I. Katis
author_sort Matthaios M. Mathioudakis
title Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events
title_short Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events
title_full Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events
title_fullStr Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events
title_full_unstemmed Molecular Characterization of the Coat Protein Gene of Greek Apple Stem Pitting Virus Isolates: Evolution through Deletions, Insertions, and Recombination Events
title_sort molecular characterization of the coat protein gene of greek apple stem pitting virus isolates: evolution through deletions, insertions, and recombination events
publisher MDPI AG
series Plants
issn 2223-7747
publishDate 2021-05-01
description A RT–PCR assay developed to amplify the full coat protein (CP) gene of apple stem pitting virus (ASPV) was evaluated using 180 Greek apple and pear samples and showed a broad detection range. This method was used to investigate the presence of ASPV in quince in Greece and showed a high incidence of 52%. The sequences of 14 isolates from various hosts with a distinct RFLP profile were determined. ASPV population genetics and the factors driving ASPV evolution were analyzed using the Greek ASPV sequences, novel sequences from Brazilian apple trees and Chinese botanical <i>Pyrus</i> species, and homologous sequences retrieved from GenBank. Fourteen variant types of Greek, Brazilian and botanical isolates, which differ in CP gene length and presence of indels, were identified. In addition, these analyses showed high intra- and inter-group variation among isolates from different countries and hosts, indicating the significant variability present in ASPV. Recombination events were detected in four isolates originating from Greek pear and quince and two from Brazilian apples. In a phylogenetic analysis, there was a tendency for isolates to cluster together based on CP gene length, the isolation host, and the detection method applied. Although there was no strict clustering based on geographical origin, most isolates from a given country tended to regroup in specific clusters. Interestingly, it was found that the phylogeny was correlated to the type, position, and pattern of indels, which represent hallmarks of specific lineages and indicate their possible role in virus diversification, rather than the CP size itself. Evidence of recombination between isolates from botanical and cultivated species and the clustering of isolates from botanical species and isolates from cultivated species suggest the existence of a possible undetermined transmission mechanism allowing the exchange of ASPV isolates between the cultivated and wild/ornamental hosts.
topic ASPV
CP-specific RT–PCR
pome fruits
botanical pears
variants
CP variability
url https://www.mdpi.com/2223-7747/10/5/917
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