Characterization of a Novel Emaravirus Affecting Ash Species (Fraxinus spp.) in Europe

We identified a novel virus in diseased European ash (Fraxinus excelsior) and manna ash (F. ornus) trees exhibiting chlorotic ringspots, mottle and leaf deformation such as curling and shoestring symptoms. High-throughput sequencing (HTS, Illumina RNASeq) of tot...

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Autores principales:	Thomas R. Gaskin, Max Tischendorf, Ines Günther, Marius Rehanek, Carmen Büttner, Susanne von Bargen
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	forest and urban trees <i>Fraxinus excelsior</i> <i>F. ornus</i> leaf chloroses high-throughput sequencing novel emaravirus Plant ecology QK900-989
Acceso en línea:	https://doaj.org/article/a60c3aee9e58418faf2db63b9a019dd8
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spelling	oai:doaj.org-article:a60c3aee9e58418faf2db63b9a019dd82021-11-25T17:38:59ZCharacterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe10.3390/f121115741999-4907https://doaj.org/article/a60c3aee9e58418faf2db63b9a019dd82021-11-01T00:00:00Zhttps://www.mdpi.com/1999-4907/12/11/1574https://doaj.org/toc/1999-4907We identified a novel virus in diseased European ash (<i>Fraxinus excelsior)</i> and manna ash (<i>F. ornus</i>) trees exhibiting chlorotic ringspots, mottle and leaf deformation such as curling and shoestring symptoms. High-throughput sequencing (HTS, Illumina RNASeq) of total RNA isolated from diseased leaf material in combination with RT-PCR-based amplification techniques and Sanger sequencing determined five complete genome segments, each encoding a single open reading frame. Sequence analyses of RNA1–RNA5 revealed a genome organization typical for emaraviruses, i.e., (i) conserved and complementary terminal 5′ and 3′ termini of each genome segment (ii) proteins showing significant homologies to the RNA-dependent RNA polymerase (RdRP) encoded by RNA1, the glycoprotein precursor (GPP) encoded by RNA2, the viral nucleocapsid protein (N, RNA3), the movement protein (MP, RNA4), and a protein of 26 kDA (P26, RNA5) highly similar to proteins of unknown function encoded by other emaraviruses. Furthermore, we identified spherical particles (double-membrane bodies, DMB) of different sizes (70–80 nm in diameter) which are typical for emaraviruses exclusively in virus-infected leaf tissue exhibiting mottle and leaf deformation. Sequence comparison and phylogenetic analyses confirmed the identified novel virus as a new member of the genus <i>Emaravirus</i>. We established a species-specific RT-PCR detection protocol and could associate the observed disease symptoms with the infection of the novel emaravirus in <i>F. excelsior</i> and <i>F. ornus</i>. Therefore, we propose the name ash shoestring-associated emaravirus (ASaV). Investigation of ASaV-infected sample trees originating from different locations in Switzerland, Germany, Italy and Sweden provided a wide geographical distribution of the virus in affected ash species. To our knowledge, this is the first confirmation of an emaravirus affecting ash tree species with shoestring symptoms of leaves in Europe.Thomas R. GaskinMax TischendorfInes GüntherMarius RehanekCarmen BüttnerSusanne von BargenMDPI AGarticleforest and urban trees<i>Fraxinus excelsior</i><i>F. ornus</i>leaf chloroseshigh-throughput sequencingnovel emaravirusPlant ecologyQK900-989ENForests, Vol 12, Iss 1574, p 1574 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	forest and urban trees <i>Fraxinus excelsior</i> <i>F. ornus</i> leaf chloroses high-throughput sequencing novel emaravirus Plant ecology QK900-989
spellingShingle	forest and urban trees <i>Fraxinus excelsior</i> <i>F. ornus</i> leaf chloroses high-throughput sequencing novel emaravirus Plant ecology QK900-989 Thomas R. Gaskin Max Tischendorf Ines Günther Marius Rehanek Carmen Büttner Susanne von Bargen Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe
description	We identified a novel virus in diseased European ash (<i>Fraxinus excelsior)</i> and manna ash (<i>F. ornus</i>) trees exhibiting chlorotic ringspots, mottle and leaf deformation such as curling and shoestring symptoms. High-throughput sequencing (HTS, Illumina RNASeq) of total RNA isolated from diseased leaf material in combination with RT-PCR-based amplification techniques and Sanger sequencing determined five complete genome segments, each encoding a single open reading frame. Sequence analyses of RNA1–RNA5 revealed a genome organization typical for emaraviruses, i.e., (i) conserved and complementary terminal 5′ and 3′ termini of each genome segment (ii) proteins showing significant homologies to the RNA-dependent RNA polymerase (RdRP) encoded by RNA1, the glycoprotein precursor (GPP) encoded by RNA2, the viral nucleocapsid protein (N, RNA3), the movement protein (MP, RNA4), and a protein of 26 kDA (P26, RNA5) highly similar to proteins of unknown function encoded by other emaraviruses. Furthermore, we identified spherical particles (double-membrane bodies, DMB) of different sizes (70–80 nm in diameter) which are typical for emaraviruses exclusively in virus-infected leaf tissue exhibiting mottle and leaf deformation. Sequence comparison and phylogenetic analyses confirmed the identified novel virus as a new member of the genus <i>Emaravirus</i>. We established a species-specific RT-PCR detection protocol and could associate the observed disease symptoms with the infection of the novel emaravirus in <i>F. excelsior</i> and <i>F. ornus</i>. Therefore, we propose the name ash shoestring-associated emaravirus (ASaV). Investigation of ASaV-infected sample trees originating from different locations in Switzerland, Germany, Italy and Sweden provided a wide geographical distribution of the virus in affected ash species. To our knowledge, this is the first confirmation of an emaravirus affecting ash tree species with shoestring symptoms of leaves in Europe.
format	article
author	Thomas R. Gaskin Max Tischendorf Ines Günther Marius Rehanek Carmen Büttner Susanne von Bargen
author_facet	Thomas R. Gaskin Max Tischendorf Ines Günther Marius Rehanek Carmen Büttner Susanne von Bargen
author_sort	Thomas R. Gaskin
title	Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe
title_short	Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe
title_full	Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe
title_fullStr	Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe
title_full_unstemmed	Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe
title_sort	characterization of a novel emaravirus affecting ash species (<i>fraxinus</i> spp.) in europe
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/a60c3aee9e58418faf2db63b9a019dd8
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Characterization of a Novel Emaravirus Affecting Ash Species (<i>Fraxinus</i> spp.) in Europe

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