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波兰大蒜上葱黄矮化病毒、大蒜普通潜隐病毒和红葱潜隐病毒的首次报道

First Report of Onion yellow dwarf virus, Garlic common latent virus and Shallot latent virus on Garlic in Poland.

作者信息

Chodorska M, Paduch-Cichal E, Kalinowska E, Szyndel M S

机构信息

Department of Plant Pathology, Faculty of Horticulture, Biotechnology and Landscape Architecture, Warsaw University of Life Sciences (WULS-SGGW), Nowoursynowska 166 ST, 02-787 Warsaw, Poland.

出版信息

Plant Dis. 2014 Jun;98(6):858. doi: 10.1094/PDIS-11-13-1160-PDN.

Abstract

Garlic (Allium sativum L.) is vegetatively propagated and can be affected by a virus complex (1) consisting of two potyviruses, Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV), and two carlaviruses, Garlic common latent virus (GCLV) and Shallot latent virus (SLV) (2). OYDV, GCLV, and SLV are economically important viral pathogens of bulb garlic crops in many garlic-growing areas of the world. A general mosaic and yellowing of leaves of four garlic cultivars (Blanko, Harnaś, Jarus, and Mega) was observed in 11 garlic-production fields in the Lodz, Mazowieckie, Małopolska, and Pomorskie regions of Poland in July 2012. ELISA was carried out with extracts from 29 collected garlic leaf samples to detect OYDV, GCLV, and SLV using commercial antiserum (DSMZ, Braunschweig, Germany). Results indicated that 6 samples (20.7%) were infected with OYDV, 25 samples (86.2%) were infected with GCLV, and 23 samples (79.3%) were infected with SLV. The presence of these viruses in garlic leaf samples was confirmed by reverse transcription (RT)-PCR using total RNA extracted using the Spectrum Plant Total RNA kit (Sigma-Aldrich, Munich, Germany) and primers, designed in this study, specific to the whole coat protein gene of OYDV (OYDVF 5'-TAGGGTTGGATTATGATTTCTCGA-3' and OYDVR 5'-TAGTGGTACACCACATTTCGT-3'), GCLV (GCLVF 5'-TTATAGGGACGGCACAAAATCAATCA-3' and GCLVR 5'-AATAGCACTCCTAGAACAACCATT-3') and SLV (SLVF 5'-AATYATTTACAATCGTCCAGCTA-3' and SLVR 5'-ATAATATCAATCAAATMCACACAATT-3'). Amplicons of the expected size were obtained for each virus. The amplified products were purified and sequenced in both directions. Sequence information of the CP genes of 9 OYDV, 12 GCLV, and 7 SLV isolates has been submitted to NCBI-GenBank with accession numbers KF862683 to KF862710. Sequence analysis showed that the coat protein gene of OYDV shared 86% identity with the coat protein gene of OYDV isolate MS/SW1 from Australia (GenBank Accession No. HQ258894). Comparison of the coat protein gene sequences of Polish GCLV isolates with those available in GenBank showed 85 to 91% sequence identities. Multiple sequence alignment revealed 84% nucleotide identity between the Polish isolate of SLV and an SLV isolate from Chinese garlic (AF314147) formerly referred to as Garlic latent virus (3). To the best of our knowledge, this is the first report of OYDV, GCLV, and SLV in garlic plants in Poland. The accurate identification of viruses present in garlic plants will help to use the appropriate strategies to reduce viral incidence in garlic-growing areas. References: (1) J. Chen et al. Arch Virol 146:1841, 2001. (2) A. M. G. King et al. Virus Taxonomy: Ninth Report of the International Committee on Taxonomy of Viruses. Elsevier Academic Press, San Diego, CA, 2011. (3) T. Tsuneyoshi et al. Arch. Virol. 143:1093, 1998.

摘要

大蒜(Allium sativum L.)通过营养繁殖,可能受到一种病毒复合体(1)的影响,该复合体由两种马铃薯Y病毒,即洋葱黄矮病毒(OYDV)和韭菜黄条病毒(LYSV),以及两种香石竹潜隐病毒,即大蒜普通潜隐病毒(GCLV)和葱潜隐病毒(SLV)组成(2)。OYDV、GCLV和SLV是世界上许多大蒜种植区中鳞茎大蒜作物的重要经济病毒性病原体。2012年7月,在波兰罗兹、马佐夫舍、小波兰和滨海地区的11个大蒜生产田中,观察到四个大蒜品种(布兰科、哈纳斯克、雅鲁斯和梅加)的叶片普遍出现花叶和黄化现象。使用商业抗血清(德国不伦瑞克的DSMZ)对29份采集的大蒜叶片样本提取物进行酶联免疫吸附测定(ELISA),以检测OYDV、GCLV和SLV。结果表明,6个样本(20.7%)感染了OYDV,25个样本(86.2%)感染了GCLV,23个样本(79.3%)感染了SLV。使用Spectrum植物总RNA试剂盒(德国慕尼黑的Sigma-Aldrich公司)提取总RNA,并使用本研究设计的针对OYDV全衣壳蛋白基因(OYDVF 5'-TAGGGTTGGATTATGATTTCTCGA-3'和OYDVR 5'-TAGTGGTACACCACATTTCGT-3')、GCLV(GCLVF 5'-TTATAGGGACGGCACAAAATCAATCA-3'和GCLVR 5'-AATAGCACTCCTAGAACAACCATT-3')和SLV(SLVF 5'-AATYATTTACAATCGTCCAGCTA-3'和SLVR 5'-ATAATATCAATCAAATMCACACAATT-3')的引物,通过逆转录(RT)-PCR确认大蒜叶片样本中这些病毒的存在。每个病毒均获得了预期大小的扩增子。对扩增产物进行双向纯化和测序。9个OYDV、12个GCLV和7个SLV分离株的CP基因序列信息已提交至NCBI-GenBank,登录号为KF862683至KF862710。序列分析表明,OYDV的衣壳蛋白基因与来自澳大利亚的OYDV分离株MS/SW1(GenBank登录号HQ258894)的衣壳蛋白基因具有86%的同一性。将波兰GCLV分离株的衣壳蛋白基因序列与GenBank中可用的序列进行比较,显示序列同一性为85%至91%。多重序列比对显示,波兰SLV分离株与来自中国大蒜的SLV分离株(AF314147,以前称为大蒜潜隐病毒)(3)之间的核苷酸同一性为84%。据我们所知,这是波兰大蒜植株中OYDV、GCLV和SLV的首次报道。准确鉴定大蒜植株中存在的病毒将有助于采用适当策略降低大蒜种植区的病毒发生率。参考文献:(1)J. Chen等人,《病毒学档案》146:1841,2001年。(2)A.M.G. King等人,《病毒分类学:国际病毒分类委员会第九次报告》。爱思唯尔学术出版社,加利福尼亚州圣地亚哥,2011年。(3)T. Tsuneyoshi等人,《病毒学档案》143:1093,1998年。

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