Salem Nidá M, Golino Deborah A, Falk Bryce W, Rowhani Adib
Department of Plant Pathology, University of California, One Shields Avenue, Davis, CA 95616, USA.
Arch Virol. 2008;153(3):455-62. doi: 10.1007/s00705-007-0008-3. Epub 2008 Jan 3.
The three double-stranded (ds) RNAs were detected in Rosa multiflora plants showing rose spring dwarf (RSD) symptoms. Northern blot analysis revealed three dsRNAs in preparations of both dsRNA and total RNA from R. multiflora plants. The complete sequences of the dsRNAs (referred to as dsRNA 1, dsRNA 2 and dsRNA 3) were determined based on a combination of shotgun cloning of dsRNA cDNAs and reverse transcription-polymerase chain reaction (RT-PCR). The largest dsRNA (dsRNA 1) was 1,762 bp long with a single open reading frame (ORF) that encoded a putative polypeptide containing 479 amino acid residues with a molecular mass of 55.9 kDa. This polypeptide contains amino acid sequence motifs conserved in the RNA-dependent RNA polymerases (RdRp) of members of the family Partitiviridae. Both dsRNA 2 (1,475 bp) and dsRNA 3 (1,384 bp) contained single ORFs, encoding putative proteins of unknown function. The 5' untranslated regions (UTR) of all three segments shared regions of high sequence homology. Phylogenetic analysis using the RdRp sequences of the various partitiviruses revealed that the new sequences would constitute the genome of a virus in family Partitiviridae. This virus would cluster with Fragaria chiloensis cryptic virus and Raphanus sativus cryptic virus 2. We suggest that the three dsRNA segments constitute the genome of a novel cryptic virus infecting roses; we propose the name Rosa multiflora cryptic virus (RMCV). Detection primers were developed and used for RT-PCR detection of RMCV in rose plants.
在表现出蔷薇春季矮化（RSD）症状的多花蔷薇植株中检测到三种双链（ds）RNA。Northern杂交分析显示，多花蔷薇植株的dsRNA和总RNA制剂中均有三种dsRNA。基于dsRNA cDNA的鸟枪法克隆和逆转录-聚合酶链反应（RT-PCR）相结合的方法，确定了dsRNA（分别称为dsRNA 1、dsRNA 2和dsRNA 3）的完整序列。最大的dsRNA（dsRNA 1）长1762 bp，有一个单一的开放阅读框（ORF），编码一个推定的多肽，该多肽含有479个氨基酸残基，分子量为55.9 kDa。该多肽含有在双分病毒科成员的RNA依赖性RNA聚合酶（RdRp）中保守的氨基酸序列基序。dsRNA 2（1475 bp）和dsRNA 3（1384 bp）均含有单一的ORF，编码功能未知的推定蛋白。所有三个片段的5'非翻译区（UTR）具有高度序列同源性区域。使用各种双分病毒的RdRp序列进行的系统发育分析表明，新序列将构成双分病毒科中一种病毒的基因组。该病毒将与智利草莓隐性病毒和萝卜隐性病毒2聚类。我们认为，这三个dsRNA片段构成了一种感染蔷薇的新型隐性病毒的基因组；我们提议将其命名为多花蔷薇隐性病毒（RMCV）。开发了检测引物，并用于RT-PCR检测蔷薇植株中的RMCV。
