Perez-Egusquiza Z, Liefting L W, Ward L I
Plant Health and Environment Laboratory, Ministry for Primary Industries, P.O. Box 2095, Auckland 1140, New Zealand.
Plant Dis. 2013 Aug;97(8):1122. doi: 10.1094/PDIS-10-12-0981-PDN.
Rose is the top selling cut flower in New Zealand and is the most popular garden plant in the world. Several virus-like diseases have been described in roses, but the causal agents for many remain unknown. Most of the described viruses infecting rose belong to the genera Ilarvirus and Nepovirus. Only recently, a number of new viruses have been or are in the process of being characterized (1,2,3,4). In January 2011, 10 rose samples showing virus-like symptoms were collected from the Wanganui region on the North Island of New Zealand. Total nucleic acid was extracted from these samples using an InviMag Plant DNA Mini Kit (Invitek GmbH, Berlin, Germany) and a KingFisher mL workstation (Thermo Scientific, Waltham, MA). PCR and reverse transcription (RT)-PCR was conducted using specific primers for Arabis mosaic virus (ArMV), Cherry leaf roll virus, Prunus necrotic ringspot virus (PNRSV), Rosa rugosa leaf distortion virus, Rose spring dwarf associated virus, Rose yellow leaf virus, Rose yellow mosaic virus, Rose yellow vein virus (RYVV), and Strawberry latent ringspot virus. Samples were also tested using generic primers for carlavirus, potexvirus, potyvirus, tombusvirus, and phytoplasmas. Two samples (cvs. Pauls Himalayan Musk and Bloomfield) were positive for ArMV, four samples (cvs. Leda, Rosa Mundi, Charles de Mills, and Indica Major) were positive for PNRSV, and two samples (cvs. Leda and Zephirine Drouhin) were positive for RYVV. Samples were negative for all other tested viruses and phytoplasmas. RYVV was detected using two sets of primers (D. Mollov, personal communication) designed to amplify fragments of estimated sizes of 797 bp and 684 bp of the movement protein (MP) and coat protein (CP) genes of RYVV, respectively. RYVV amplicons were sequenced directly (GenBank Accession Nos. JX887423 to JX887426). A BLASTn search of the MP and CP fragments showed the highest nucleotide identity of 98% and 96 to 97%, respectively, with the type isolate of RYVV (JX028536). RYVV has been reported as the causal agent of a vein yellowing disease in rose (2). Symptoms observed in the 'Leda' sample infected with PNRSV and RYVV (vein yellowing and chlorotic mottle in the apex of leaves) were not typical of PNRSV, so they may be caused by RYVV. Symptoms in samples of cv. Zephirine Drouhin (curling of leaves and mottle), observed in both RYVV-positive and -negative samples, may not be associated with RYVV infection. This suggests that vein yellowing may be influenced by cultivar. RYVV has been reported in several rose cultivars, but only in the United States (2). To the best of our knowledge, this is the first report of RYVV infecting rose in New Zealand, where it is likely that the virus has been present for some time. The virus may have a much wider geographical distribution than that reported as the virus was only recently characterized (3). References: (1) B. Lockhart et al. Page 31 in: Program and Abstracts of The 12th International Symposium on Virus Diseases of Ornamental Plants, 2008. (2) D. Mollov et al. Phytopathology 99:S87, 2009. (3) D. Mollov et al. Arch Virol. 158:877, 2012. (4) N. Salem et al. Plant Dis. 92:508, 2008.
玫瑰是新西兰销量最高的切花，也是全球最受欢迎的园林植物。在玫瑰中已描述了几种类似病毒的病害，但许多病害的病原体仍不明确。大多数已描述的感染玫瑰的病毒属于等轴不稳环斑病毒属和线虫传多面体病毒属。直到最近，才有一些新病毒已被鉴定或正在被鉴定（参考文献1、2、3、4）。2011年1月，从新西兰北岛旺阿努伊地区采集了10个表现出类似病毒症状的玫瑰样本。使用InviMag植物DNA微量提取试剂盒（德国柏林Invitek公司）和KingFisher mL工作站（美国马萨诸塞州沃尔瑟姆赛默飞世尔科技公司）从这些样本中提取总核酸。使用针对南芥菜花叶病毒（ArMV）、樱桃卷叶病毒、李坏死环斑病毒（PNRSV）、玫瑰皱叶变形病毒、玫瑰春季矮化相关病毒、玫瑰黄叶病毒、玫瑰黄花叶病毒、玫瑰黄脉病毒（RYVV）和草莓潜隐环斑病毒的特异性引物进行PCR和逆转录（RT）-PCR。还使用了针对香石竹潜隐病毒属、马铃薯X病毒属、马铃薯Y病毒属、番茄丛矮病毒属和植原体的通用引物对样本进行检测。两个样本（品种为Pauls Himalayan Musk和Bloomfield）ArMV检测呈阳性，四个样本（品种为Leda、Rosa Mundi、Charles de Mills和Indica Major）PNRSV检测呈阳性，两个样本（品种为Leda和Zephirine Drouhin）RYVV检测呈阳性。所有其他检测的病毒和植原体样本均为阴性。使用两组引物（D. Mollov，个人交流）检测RYVV，这两组引物分别设计用于扩增RYVV运动蛋白（MP）和外壳蛋白（CP）基因估计大小为797 bp和684 bp的片段。对RYVV扩增子进行直接测序（GenBank登录号：JX887423至JX887426）。对MP和CP片段进行BLASTn搜索显示，与RYVV模式分离株（JX028536）的核苷酸同一性最高分别为98%和96%至97%。RYVV已被报道为玫瑰叶脉黄化病的病原体（参考文献2）。在感染PNRSV和RYVV的“Leda”样本中观察到的症状（叶脉黄化和叶尖褪绿斑驳）并非PNRSV的典型症状，因此可能是由RYVV引起的。在RYVV阳性和阴性样本中均观察到Zephirine Drouhin品种样本的症状（叶片卷曲和斑驳），可能与RYVV感染无关。这表明叶脉黄化可能受品种影响。RYVV已在多个玫瑰品种中被报道，但仅在美国（参考文献2）。据我们所知，这是RYVV在新西兰感染玫瑰的首次报道，在新西兰该病毒可能已存在一段时间。该病毒的地理分布可能比报道的范围更广，因为该病毒直到最近才被鉴定（参考文献3）。参考文献：（1）B. Lockhart等人。载于《第12届国际观赏植物病毒病研讨会会议议程和摘要》，2008年，第31页。（2）D. Mollov等人。《植物病理学》99：S87，2009年。（3）D. Mollov等人。《病毒学档案》158：877，2012年。（4）N. Salem等人。《植物病害》92：508，2008年。
