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旱金莲作为樱桃卷叶病毒在阿姆斯特丹岛的天然寄主的首次报道。

First Report of Nasturtium as a Natural Host of Cherry leaf roll virus on Amsterdam Island.

作者信息

Marais A, Faure C, Candresse T, Hullé M

机构信息

UMR GDPP, IBVM, INRA, Université Bordeaux 2, BP81, 33883 Villenave d'Ornon Cedex, France.

UMR BiO3P, INRA, Domaine de la Motte au Vicomte, BP 35327, 35653 Le Rheu Cedex, France.

出版信息

Plant Dis. 2010 Apr;94(4):477. doi: 10.1094/PDIS-94-4-0477B.

Abstract

Cherry leaf roll virus (CLRV) is a well-known virus belonging to the genus Nepovirus, but unlike most members of this genus, it is not known to be transmitted by nematodes but only through seeds and pollen. Since its first description in 1955 on Prunus avium L. in England (1), CLRV has been shown to have a worldwide distribution and a wide natural host range. During a survey of plant viruses in the French sub-Antarctic islands, samples from nasturtium plants (Tropaeolum majus), an introduced plant species, showing symptoms of leaf mosaic, deformation, and veinal necrosis were collected on Amsterdam Island. Upon mechanical transmission with sap extracts, necrotic ringspot and oak-leaf symptoms typical of Nepovirus infection were observed on the leaves of inoculated Nicotiana clevelandii and N. tabacum plants. Inoculation of healthy nasturtium plants resulted in mosaic and pin-point necrosis symptoms. Electron microscopy on negatively stained sap extracts revealed the presence of icosahedral virions, 28 to 30 nm in diameter, in the symptomatic Nicotiana leaves. Amplification by reverse transcription (RT)-PCR with a polyvalent test, which identifies viruses belonging to the family Comoviridae (2), yielded the expected 248-bp fragment. Sequencing of the cloned amplicon showed 80% nucleotide and 90% amino acid identity with a part of the RNA dependent RNA polymerase (RdRp) of CLRV (CAE83562). To confirm the presence of CLRV, an approximate 4.6-kbp cDNA fragment was PCR amplified from double-stranded RNAs purifed from infected Nicotiana plants using the sense primer 5'-GTGGGACTGCCATGCACCTACTC-3' and an oligo-T as antisense primer. This PCR product (GenBank Accession No. GU167974) spans the region between the VPg gene and the polyA tail at the 3' end of the genome and thus provides approximately 2.8 kb of new internal sequence information on RNA1 of CLRV. The presence of CLRV in the initial nasturtium samples was confirmed with a CLRV-specific RT-PCR assay that amplifies the 3' non-coding region of the CLRV genome (3). Sequence of the amplified fragment showed it to be identical to the corresponding part of the 3' non-coding region of 4.6-kbp clone obtained from the CLRV isolate mechanically transmitted to the N. tabacum and N. clevelandii plants. Experimental infection of nasturtium by CLRV has been reported (4), but to the best of our knowledge these results represent the first report of natural infection of T. majus by CLRV. Given its seed transmissible character in many hosts, CLRV likely was introduced in infected seeds of T. majus imported to the remote sub-Antarctic Amsterdam Island. References: (1) R. Cropley. Ann. Appl. Biol. 49:524, 1961. (2) V. Maliogka et al. J. Phytopathol. 152:404, 2004. (3) K. Rebenstorf et al. J. Virol. 80:2453, 2006. (4) K. Schmelzer. Phytopathol. Z. 55:317, 1966.

摘要

樱桃叶卷病毒(CLRV)是一种隶属于线虫传多面体病毒属的知名病毒,但与该属的大多数成员不同,它并非通过线虫传播,而是仅通过种子和花粉传播。自1955年在英国首次在欧洲甜樱桃(Prunus avium L.)上被描述以来(1),CLRV已被证明在全球范围内分布,且具有广泛的自然宿主范围。在对法国亚南极岛屿的植物病毒进行调查期间,在阿姆斯特丹岛上采集了来自引入植物种旱金莲(Tropaeolum majus)的样本,这些样本表现出叶片花叶、变形和叶脉坏死的症状。用汁液提取物进行机械接种后,在接种的克利夫兰烟草(Nicotiana clevelandii)和烟草(N. tabacum)植株的叶片上观察到了典型的线虫传多面体病毒感染的坏死环斑和橡树叶症状。对接种健康的旱金莲植株导致了花叶和针尖状坏死症状。对负染的汁液提取物进行电子显微镜观察,发现在有症状的烟草叶片中存在直径为28至30纳米的二十面体病毒粒子。使用一种能鉴定属于豇豆花叶病毒科的病毒的多价检测方法,通过逆转录(RT)-PCR进行扩增(2),得到了预期的248碱基对片段。对克隆扩增子的测序显示,其与CLRV的RNA依赖RNA聚合酶(RdRp)的一部分具有80%的核苷酸同一性和90%的氨基酸同一性(CAE83562)。为了确认CLRV的存在,使用正义引物5'-GTGGGACTGCCATGCACCTACTC-3'和寡聚T作为反义引物,从感染烟草植株中纯化的双链RNA中PCR扩增出一个约4.6千碱基对的cDNA片段。该PCR产物(GenBank登录号GU167974)跨越了基因组3'端VPg基因和多聚A尾之间的区域,因此提供了关于CLRV RNA1的约2.8千碱基对的新内部序列信息。通过一种扩增CLRV基因组3'非编码区的CLRV特异性RT-PCR检测方法,确认了最初的旱金莲样本中存在CLRV(3)。扩增片段的序列显示,它与从机械接种到烟草和克利夫兰烟草植株的CLRV分离株获得的4.6千碱基对克隆的3'非编码区的相应部分相同。已有关于CLRV对旱金莲进行实验性感染的报道(4),但据我们所知,这些结果代表了CLRV对旱金莲自然感染的首次报道。鉴于其在许多宿主中具有种子传播特性,CLRV很可能是通过进口到偏远的亚南极阿姆斯特丹岛的感染旱金莲种子引入的。参考文献:(1)R. Cropley. Ann. Appl. Biol. 49:524, 1961.(2)V. Maliogka等人. J. Phytopathol. 152:404, 2004.(3)K. Rebenstorf等人. J. Virol. 80:2453, 2006.(4)K. Schmelzer. Phytopathol. Z. 55:317, 1966.

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