Genomic characterization of Calla lily chlorotic spot virus and design of broad-spectrum primers for detection of tospoviruses
Genomic characterization of Calla lily chlorotic spot virus and design of broad-spectrum primers for detection of tospoviruses
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DOI：
10.1111/j.1365-3059.2011.02484.x
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发表时间：
2012-02-01
期刊：
影响因子：
2.7
通讯作者：
Yeh, S. -D.
中科院分区：
文献类型：
--
作者：
Chen, T. -C.;Li, J-T.;Yeh, S. -D.
从台湾表现症状的马蹄莲植株上分离到的马蹄莲褪绿斑病毒，经S核糖核酸序列比对，初步确定为马蹄莲病毒属的一个新种。在这次调查中，测定了ITS L和M RNAs的全序列。L核糖核酸全长8911个核苷酸，编码2882个氨基酸(332 KDa)的病毒互补核糖核酸聚合酶。MRNA全长4704个核苷酸，编码309个氨基酸(35 KDa)的非结构NSM蛋白，编码1123个氨基酸(128 KDa)的Gn/GC糖蛋白前体。系统进化分析表明，CCSV与番茄带斑病毒有较近的亲缘关系，是从中国南部番茄上分离到的一种新的病毒株。从L和NSM基因高度保守区分别设计了两对简并引物gL2740/gL3920c和gM410/gM870c，经序列比对，成功地用逆转录-聚合酶链式反应检测了温室培养的12种Tospo病毒，其中包括6个正式种和6个暂定种。并对台湾地区大田瓜类样品中的甜瓜黄斑病毒和西瓜银斑病毒进行了检测。结果表明，这两对简并引物可用于大多数TospoVirus的快速检测和未知TospoVirus的探索。
Calla lily chlorotic spot virus (CCSV) collected from calla lily plants showing symptoms in Taiwan has been identified as a tentative species of the genus Tospovirus based on the comparison of S RNA sequences. In this investigation, the complete sequences of its L and M RNAs were determined. The L RNA contains 8911 nucleotides (nt) and encodes a putative RNA-dependent RNA polymerase of 2882 amino acids (aa) (332 kDa) in the viral complementary (vc) sense. The M RNA was shown to be 4704 nt in length, encoding a nonstructural NSm protein of 309 aa (35 kDa) in the viral sense and a Gn/Gc glycoprotein precursor (GP) of 1123 aa (128 kDa) in the vc sense. Phylogenetic analysis of the individual tospoviral proteins indicated that CCSV is closely related to Tomato zonate spot virus, a new tentative tospovirus isolated from tomato in southern China. Two degenerate primer pairs gL2740/gL3920c and gM410/gM870c, designed, after sequence comparison, from the highly conserved regions of the L and NSm genes, respectively, were successfully used to detect 12 greenhouse-cultured tospoviruses, including six formal species and six tentative species, by reverse transcription-polymerase chain reaction. They were also used to detect Melon yellow spot virus and Watermelon silver mottle virus from field cucurbit samples in Taiwan. The results indicate that the two degenerate primer pairs can be used for prompt detection of most tospoviruses and exploration of unknown tospovirus species.