Reported here is the complete genome sequence of Mourilyan virus (MoV) that infects giant tiger (Penaeus monodon) and kuruma prawns (P. japonicas) in Australia. Its genome was determined using various PCR strategies based on the sequences of 3 randomly-amplified cDNA clones to its L and M RNA segments discovered in a library generated to determine the genome sequence of gill-associated ronivirus. The sequences of PCR products and clones obtained showed the MoV genome to comprise 4 ssRNA segments (L, M, S1 and S2), as confirmed by Northern blotting using RNA from naïve and MoV-infected prawns, and by Illumina sequence analysis of semi-purified MoV. BLASTn searches identified the MoV L, M and S1 RNA segments to be homologous to Wēnzhōu shrimp virus 1 (WzSV1) segments discovered recently in a P. monodon RNA-Seq library (SRR1745808). Mapping this read library to the MoV S2 RNA segment identified WzSV1 to also possess an equivalent segment. BLASTp searches identified the putative non-structural protein (NSs2; 393–394 aa) encoded in their S2 RNA segments to have no homologs in GenBank. Possibly due to NSs2 being encoded in a discrete RNA segment rather than in ambisense relative to the N protein as in the S RNA segments of other phenuiviruses, each of 6 MoV S1 RNA segment clones sequenced possessed a variable-length (≤ 645 nt) imperfect GA-repeat extending from the N protein stop codon to the more variable ∼90 nt segment terminal sequence. Read mapping of RNA-Seq library SRR1745808 showed the WzSV1 S1 RNA segment to possess a similar GA-repeat. However, paired-read variations hindered definitive assembly of a consensus sequence. All 4 MoV and WzSV1 RNA segments terminated with a 10 nt inverted repeat sequence (5′-ACACAAAGAC.) identical to the RNA segment termini of uukuviruses. Phylogenetic analyses of MoV/WzSV1 RNA-dependant RNA polymerase (L RNA), G1G2 precursor glycoprotein (M RNA) and nucleocapsid (N) protein (S1 RNA) sequences generally clustered them with as yet unassigned crustacean/diptera bunya-like viruses on branches positioned closely to others containing tick-transmitted phenuiviruses. As genome sequences of most phenuiviruses discovered recently have originated from meta-transcriptomics studies, the data presented here showing the MoV and WzSV1 genomes to comprise more than 3 RNA segments, like the plant tenuiviruses, suggests a need to investigate the genomes of these unassigned viruses more closely.

这里报道的是感染巨虎（Penaeus monodon）和大虾（P. japonicas） 在澳大利亚。它的基因组是使用各种 PCR 策略确定的，该策略基于 3 个随机扩增的 cDNA 克隆到其 L 和 M RNA 片段的序列，该序列是在为确定鳃相关罗尼病毒的基因组序列而生成的文库中发现的。获得的 PCR 产物和克隆的序列显示 MoV 基因组包含 4 个 ssRNA 片段（L、M、S1 和 S2），这通过使用来自未感染和 MoV 感染的对虾的 RNA 的 Northern印迹以及 Illumina 对半纯化的 MoV。BLASTn 搜索确定 MoV​​ L、M 和 S1 RNA 片段与最近在斑节对虾中发现的温州虾病毒 1 (WzSV1) 片段同源RNA-Seq 文库 (SRR1745808)。将此读取库映射到 MoV S2 RNA 片段，确定 WzSV1 也拥有等效片段。BLASTp 搜索确定了在其 S2 RNA 片段中编码的假定非结构蛋白（NSs2；393-394 aa）在 GenBank 中没有同源物。可能是由于 NSs2 是在一个离散的 RNA 片段中编码的，而不是像在其他苯类病毒的 S RNA 片段中那样相对于 N 蛋白在双义中编码，因此测序的 6 个 MoV S1 RNA 片段克隆中的每一个都具有可变长度（≤ 645 nt）不完美GA-重复从 N 蛋白终止密码子延伸到更可变的 ~90 nt 片段末端序列。RNA-Seq 文库 SRR1745808 的读取映射显示 WzSV1 S1 RNA 片段具有类似的 GA 重复。然而，配对读取变异阻碍了一致序列的最终组装。所有 4 个 MoV 和 WzSV1 RNA 片段都以与 uukuviruses 的 RNA 片段末端相同的 10 nt 反向重复序列 (5'-ACACAAAGAC.) 终止。MoV/WzSV1 RNA 依赖性 RNA 聚合酶 (L RNA)、G1G2 前体糖蛋白 (M RNA) 和核衣壳 (N) 蛋白 (S1 RNA) 序列的系统发育分析通常将它们与分支上尚未分配的甲壳类/双翅目类病毒聚集在一起与其他含有蜱传播的非病毒病毒的位置很接近。由于最近发现的大多数 phenuiviruses 的基因组序列源自元转录组学研究，此处提供的数据显示 MoV 和 WzSV1 基因组包含超过 3 个 RNA 片段，如植物细病毒，表明需要研究这些未分配病毒的基因组更紧密。