北京大学-researchgate.docVIP

一点红萝卜中一种新的隐潜病毒的基因组全序列分析 李力强1，田清花2，陈集双1，2，3，* （1.浙江大学 生命科学学院，杭州 310058；2.浙江理工大学 生物工程研究所，杭州 310018；3.南京工业大学，资源生物研究所） 我们从浙江本地的一点红萝卜品种中分离到两条双链RNA序列，分别命名为dsR1 和 dsR2。单引物扩增获得cDNA克隆测序分析它们的基因组结构特征显示：两条序列的5′非编码区（untranslated regions，UTR）高度保守，并含有一段完全相同的序列 5′-AGAAUUU-3′；3′ 端非编码区也有相同序列5′-UAAGAC-3′。自含有dsR1 和 dsR2的一点红萝卜中提取病毒粒子，然后从病毒粒子样品中提取核酸RT-PCR均能特异性检测到dsR1 和 dsR2，表明dsR1 和 dsR2来自同一病毒。BLASTP 检索显示dsR1最大开放阅读框编码的蛋白和双分病毒科病毒编码的依赖RNA的复制酶相似。利用双分病毒的复制酶和RasV3 dsR1编码的蛋白序列比对后，系统发生分析表明dsR1和双分病毒聚类到一起. 综上所述, RasV3 dsR1和dsR2是萝卜中的一种新的双分病毒的基因组，我们把这种病毒命名为 Raphanus sativus virus 3. 双链RNA；萝卜；RDRP；双链RNA 病毒 Complete nucleotide sequences of two double-stranded RNAs from Raphanus sativus cv. Yidianhong demonstrate the incidence of a novel cryptic virus Li Liqiang 1, Tian Qinghua 2，Chen Jishuang 1,2,3,* (1. College of Resources and Environmental Sciences, Northwest AF University, Yangling 712100, 2.State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Nanjing, 210008) Abstract: Two double-stranded RNAs were extracted from leaf tissues of Raphanus sativus cv. Yidianhong grown in Eastern China. Their full sequences, namely dsR1 and dsR2, were determined by a single primer amplification technique. Comparisons of 5′ untranslated regions (UTR) and 3′ UTR between dsR1 and dsR2 showed that they were highly conserved at 5′ UTR and an identical region 5′-AGAAUUU-3′ was discovered as the evidence of the same plant virus. 5′-UAAGAC-3′ was found as their identical region in 3′ UTR. BLASTP search in Genbank showed that dsR1 encoded putative protein was much similar with RNA dependent RNA polymerases (RdRps) of plant-infecting dsRNA viruses belonging to the family Partitiviridae. Phylogenetic analysis showed that dsR1 encoded putative protein clustered with the RdRps of members of the family Partitiviridae. RT-PCR detection indicated that both RasR7 and RasR8 existed in the virus-like particles. Therefore, it was suggested that dsR1 and dsR2 constitute the