ena-STUDY-HUMBOLDT UNIVERSITY BERLIN-30-07-2021-11:20:01:994-731
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 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) showing significant homologies to proteins of unknown function encoded by other emaraviruses. Further, 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 Emaravirus. We established a species-specific RT-PCR detection protocol and could associate the observed disease symptoms with the infection of the novel emaravirus in F. excelsior and F. ornus. Therefore, we propose the name ash shoestring-associated emaravirus (ASaV). Investigated ASaV-infected sample trees originated from different locations in Switzerland, Germany, Italy and Sweden proving 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.
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 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) showing significant homologies to proteins of unknown function encoded by other emaraviruses. Further, 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 Emaravirus. We established a species-specific RT-PCR detection protocol and could associate the observed disease symptoms with the infection of the novel emaravirus in F. excelsior and F. ornus. Therefore, we propose the name ash shoestring-associated emaravirus (ASaV). Investigated ASaV-infected sample trees originated from different locations in Switzerland, Germany, Italy and Sweden proving 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.