# Characterization of a Novel Emaravirus Affecting Ash Species (_Fraximus_ spp.) in Europe
## 3 Results
\begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline
**RNA** & \multicolumn{1}{c}{**Scaffold**} & \multicolumn{1}{c}{**Size (n0)**} & \multicolumn{1}{c}{**\% Total**} & \multicolumn{1}{c}{**Mean**} & \multicolumn{1}{c}{**Query**} & \multicolumn{1}{c}{**E Value**} & \multicolumn{1}{c}{**Sequence**} \\
**Genomic** & **Number** & **Size (n0)** & **Reads** & **Corerage** & **Best Match in Blast (Accession Number)** & **Coverage (\%)** & **Coverage** & **Identify (\%)** \\ \hline \hline \multicolumn{10}{c}{_Frazimus exclusive_} \\ \hline RNA1 & S34 & 1455 & 0.37 & 6.54 & R4 RP, Actinida emaravirus 2 (GEE82886.1) & 99 & 0.0 & 73.76 \\  & S50 & 1195 & 0.31 & 7.40 & R4 RP, Actinida emaravirus 2 (GEE82886.1) & 99 & 0.0 & 87.12 \\  & S104 & 797 & 0.20 & 5.27 & R4 RP, Actinida emaravirus 2 (GEE82886.1) & 95 & \(1\times 10^{-102}\) & 63.53 \\  & S152 & 661 & 0.17 & 6.33 & R4 RP, Pseudocapearity mosaic emaravirus 2 (QRA3607.1) & 94 & \(3\times 10^{-116}\) & 85.58 \\  & S206 & 548 & 0.14 & 5.83 & R4 RP, Actinida emaravirus 2 (QRE2886.1) & 99 & \(6\times 10^{-106}\) & 75.41 \\  & S294 & 465 & 0.12 & 7.54 & R4 RP, Fig mosaic emaravirus (QBP2768.1) & 99 & \(1\times 10^{-106}\) & 87.66 \\ \hline RNA2 & S68 & 1043 & 0.27 & 18.90 & GPP, Pigopone steparity mosaic emaravirus 2 (Y.009268865.1) & 90 & \(5\times 10^{-128}\) & 56.51 \\  & S71 & 1024 & 0.26 & 18.45 & CPP, Pigopone steparity mosaic emaravirus 2 (Y.07,00268865.1) & 96 & \(7\times 10^{-153}\) & 61.49 \\ \hline RNA3 & S55 & 1177 & 0.30 & 24.46 & NC, Pen-associated emaravirus (QJX15716.1) & 80 & 0.0 & 99.05 \\ \hline RNA4 & S65 & 1070 & 0.27 & 16.79 & MP, Fig mosaic emaravirus (QRT2245.1) & 97 & 0.0 & 77.30 \\ \hline \hline \multicolumn{10}{c}{_Frazimus_} \\ \hline RNA1 & S10 & 7122 & 0.12 & 57.80 & R4 RP, Actinida emaravirus 2 (GEE82886.1) & 98 & 0.0 & 77.88 \\ \hline RNA2 & S146 & 1929 & 0.03 & 233.64 & GPP, Pigopone steparity mosaic emaravirus 2 (QY\({}_{00}\),009268865.1) & 95 & 0.0 & 59.90 \\ \hline RNA3 & S1644 & 1463 & 0.01 & 267.67 & NC, Pen-associated emaravirus (QJX15716.1) & 64 & 0.0 & 99.05 \\ \hline RNA4 & S81 & 1496 & 0.04 & 121.29 & MP, Fig mosaic emaravirus (QRT2245.1) & 72 & 0.0 & 77.29 \\ \hline RNA5 & S164 & 996 & 0.03 & 70.28 & P6, Actinida emaravirus 2 (GEE82891.1) & 60 & \(1\times 10^{-48}\) & 47.50 \\ \hline \hline \end{tabular}
\end{table} 
Table 2: A-Nembold scaffolds of the novel virus relating to emaravirus RNA segments identified by HTS in _Frazimus exclusive_ sample pool (\(n=2\); E55262, E55268) and _Frazimus arms_ sample pool (\(n=12\); E57321, E57322, E57322, E573228-E573326, E573328-E57330, E573328-E57330). Identified scaffolds are presented with size in nucleotides (nt), mean coverage and percentage of mapped reads. Best matches in black search including query coverage. E value and sequence identity are given for each scaffold. The sample E55270 from _F. excelsior_ was analyzed further to obtain the complete sequence of all five genome segments of the novel virus identified in _Frazinus_. The complete RNA1 was determined by assembling eight overlapping sequenced RT-PCR products, generated by eight specific primer sets (Table S1, Supplementary Information). The complete sequences of the RNA2-5 were determined by cloning and sequencing of full-length PCR products. No further RNA segments were amplified by this method, suggesting that the five RNAs represent the complete genome of the novel virus. The 234 3\({}^{\prime}\) terminal nucleotides of the RNA1 were confirmed by a tailing PCR. Likewise, the 13 nucleotides of the 3\({}^{\prime}\) terminal of RNA2 could be confirmed, which were determined by the PDAP213 primer sequence. At least two individual clones were analyzed for sequence confirmation. Genome Organization of the Novel Virus Identified in Frazinus spp. and Comparison with Members of the Genus Emaravirus  
The novel virus identified in _F. excelsior_ (E55270) contained five RNA genome segments, each encoding a single open reading frame (ORF).