Molecular characterization of a distinct begomovirus species from Vernonia cinerea and its associated DNA-β using the bacteriophage Φ29 DNA polymerase
Molecular characterization of a distinct begomovirus species from Vernonia cinerea and its associated DNA-β using the bacteriophage Φ29 DNA polymerase
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DOI：
10.1007/s11262-010-0484-5
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发表时间：
2010-04
期刊：
影响因子：
1.6
通讯作者：
R. M. Packialakshmi;N. Srivastava;K. Girish;R. Usha
中科院分区：
文献类型：
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作者：
R. M. Packialakshmi;N. Srivastava;K. Girish;R. Usha
在马杜赖的农田周围收集具有黄脉症状的斑鸠菊。用简并引物从从病叶样品中纯化的总DNA中扩增出AV 1基因的一部分（550 bp），并进行克隆和测序。使用衍生自上述序列的特异性引物扩增具有菜豆花叶病毒DNA A（EMBL登录号AM 182232）的典型基因组结构的2，745个核苷酸。与其它菜豆病毒的序列比较表明，该病毒与中国一点红黄脉病毒（EmYVV-[Fz 1]）的同源性最高（82.4%），与其它已知菜豆病毒的同源性低于80%。因此，国际病毒分类委员会（ICTV）已将斑鸠菊黄脉病毒（VeYVV）确认为一种独特的菜豆花叶病毒属。常规PCR不能从病组织中扩增出DNA B或DNA β。然而，通过使用Φ29 DNA聚合酶的滚环扩增-限制性片段长度多态性方法（RCA-RFLP）获得了与该疾病相关的β DNA（1364 bp）（登录号FN 435836）。序列分析表明，VeYVV的DNA β与Sigesbeckia yellow vein广西β卫星（SibYVGxB-[CN：Gx 111：05]）的DNA β的同源性最高（56.8%），与其它双生子病毒的DNA β的同源性为56-53%。这是首次报道VeYVV的分子特征。印度的电影。完整的分子特征，系统发育分析，和推定的重组事件VeYVV的报告。
Vernonia cinereaplants with yellow vein symptoms were collected around crop fields in Madurai. A portion (550 bp) of the AV1 gene amplified using degenerate primers from the total DNA purified from diseased leaf sample was cloned and sequenced. Specific primers derived from the above sequence were used to amplify 2,745 nucleotides with the typical genome organization of begomoviral DNA A (EMBL Accession No. AM182232). Sequence comparison with other begomoviruses revealed the greatest identity (82.4%) withEmilia yellow vein virus(EmYVV-[Fz1]) from China and less than 80% with all other known begomoviruses. The International Committee on Taxonomy of Viruses (ICTV) has therefore recognizedVernonia yellow vein virus(VeYVV) as a distinct begomovirus species. Conventional PCR could not amplify the DNA B or DNA β from the diseased tissue. However, the β DNA (1364 bp) associated with the disease was obtained (Accession No. FN435836) by the rolling circle amplification–restriction fragment length polymorphism method (RCA-RFLP) using Φ29 DNA polymerase. Sequence analysis shows that DNA β of VeYVV has the highest identity (56.8%) with DNA β ofSigesbeckia yellow vein Guangxi betasatellite(SibYVGxB-[CN: Gx111:05]) and 56–53% with DNA β associated with other begomoviruses. This is the first report of the molecular characterization of VeYVV fromV. cinereain India. The complete molecular characterization, phylogenetic analysis, and putative recombination events in VeYVV are reported.