Perez-Egusquiza Z, Ward L I, Clover G R G, Fletcher J D
Plant Health and Environment Laboratory, MAF Biosecurity New Zealand, P.O. Box 2095, Auckland 1140, New Zealand.
Plant and Food Research, Private Bag 4704, Christchurch, New Zealand.
Plant Dis. 2009 Apr;93(4):427. doi: 10.1094/PDIS-93-4-0427B.
In New Zealand, sweet potato (Ipomoea batatas) is a crop of cultural importance and an important food source; it is grown mainly in the districts of Kaipara, Auckland, and the Bay of Plenty in the North Island. In January of 2008, virus symptoms that included chlorotic spots, ring spots, and mottling were observed on the leaves of commercial sweet potato crops (cvs. Beauregard, Owairaka Red, and Toka Toka Gold) growing in the three main production areas. A survey was done to determine the extent of virus infection in these crops. Fifty to one hundred leaves were collected randomly from each of 26 different fields. Leaves from each field were bulked into groups of 10, giving a total of 173 composite samples. All samples tested negative for Cucumber mosaic virus, C-6 virus, Sweet potato caulimo-like virus, Sweet potato chlorotic fleck virus, Sweet potato chlorotic stunt virus (SPCSV), Sweet potato latent virus, and Sweet potato mild specking virus by nitrocellulose membrane enzyme-linked immunosorbent assays (International Potato Center-CIP, Lima, Peru). Total nucleic acid was extracted from all 173 composite samples and used in real-time PCR assays specific for Sweet potato leaf curl virus (SPLCV) and real-time reverse transcription (RT)-PCR specific for SPCSV, Sweet potato feathery mottle virus (SPFMV), Sweet potato virus G (SPVG), and Sweet potato virus 2 (SPV2; synonym Sweet potato virus Y) (1). No samples were positive for SPLCV or SPCSV, but 107 and 138 samples tested positive for SPFMV and SPVG, respectively. SPFMV and SPVG have been reported previously in New Zealand (2,3). Sixty four samples from 16 different fields tested positive for SPV2. Of the 64 samples, 52 were also infected with SPVG and SPFMV, and 10 were co-infected with SPVG but not SPFMV; no samples were co-infected with SPV2 and SPFMV when SPVG was absent. From a representative SPV2 positive sample, the 70-bp amplicon obtained by the real-time RT-PCR primers was cloned and sequenced A BLAST search showed 100% nucleotide sequence identity with SPV2 (GenBank Accession Nos. AM050887 and AY178992). Subsequently, primers (V2-F1c: 5'-AGAACAGGACAAACTCAACC-3'; V2-R1: 5'-TAATCACCCTTCACACCTTC-3') were designed to amplify an approximately 434-bp fragment within the SPV2 coat protein gene. One-step RT-PCR was done on four of the SPV2 positive samples and amplicons of the expected size were sequenced directly (GenBank Accession No. FJ461774). Sequence comparison showed 99% nucleotide sequence identity with SPV2 (GenBank Accession Nos. AM050886, AM050887, AY178992, and EF577437). SPV2 is a member of the genus Potyvirus but the virus has not been fully characterized. It is known that single-potyvirus infections cause mild or no symptoms in sweet potato, and consequently, no significant yield reduction is observed generally. However, co-infection with other viruses such as SPCSV produces a synergistic effect and more severe disease symptoms (4). To our knowledge, this is the first report of SPV2 infecting sweet potato in New Zealand. References: (1) C. D. Kokinos and C. A. Clark. Plant Dis. 90:783, 2006. (2) M. N. Pearson et al. Australas. Plant Pathol. 35:217, 2006. (3) M. Rännäli et al. Plant Dis. 92:1313, 2008. (4) M. Untiveros et al. Plant Dis. 91:669, 2007.
在新西兰,甘薯(Ipomoea batatas)是具有重要文化意义的作物和重要的食物来源;它主要种植在北岛的凯帕拉、奥克兰以及丰盛湾地区。2008年1月,在三个主要产区种植的商业甘薯作物(品种Beauregard、Owairaka Red和Toka Toka Gold)的叶片上观察到包括褪绿斑点、环斑和斑驳在内的病毒症状。开展了一项调查以确定这些作物中病毒感染的程度。从26个不同田块中随机采集了50至100片叶子。每个田块的叶子被分成每组10片,共得到173个混合样本。通过硝酸纤维素膜酶联免疫吸附测定法(国际马铃薯中心 - CIP,秘鲁利马),所有样本对黄瓜花叶病毒、C - 6病毒、甘薯花椰菜花叶病毒样病毒、甘薯褪绿斑点病毒、甘薯褪绿矮化病毒(SPCSV)、甘薯潜隐病毒和甘薯轻度斑驳病毒检测均为阴性。从所有173个混合样本中提取总核酸,并用于针对甘薯卷叶病毒(SPLCV)的实时PCR测定以及针对SPCSV、甘薯羽状斑驳病毒(SPFMV)、甘薯病毒G(SPVG)和甘薯病毒2(SPV2;同义词甘薯病毒Y)的实时逆转录(RT) - PCR(1)。没有样本对SPLCV或SPCSV呈阳性,但分别有107和138个样本对SPFMV和SPVG检测呈阳性。SPFMV和SPVG此前已在新西兰报道过(2,3)。来自16个不同田块的64个样本对SPV2检测呈阳性。在这64个样本中,52个也感染了SPVG和SPFMV,10个与SPVG共同感染但未感染SPFMV;当不存在SPVG时,没有样本与SPV2和SPFMV共同感染。从一个代表性的SPV2阳性样本中,通过实时RT - PCR引物获得的70 bp扩增子被克隆并测序。BLAST搜索显示其与SPV2(GenBank登录号AM050887和AY178992)的核苷酸序列同一性为100%。随后,设计引物(V2 - F1c:5'-AGAACAGGACAAACTCAACC-3';V2 - R1:5'-TAATCACCCTTCACACCTTC-)以扩增SPV2外壳蛋白基因内约434 bp的片段。对4个SPV2阳性样本进行了一步RT - PCR,并对预期大小的扩增子直接测序(GenBank登录号FJ46177)。序列比较显示其与SPV2(GenBank登录号AM050886、AM050887、AY178992和EF577437)的核苷酸序列同一性为99%。SPV2是马铃薯Y病毒属的成员,但该病毒尚未得到充分鉴定。已知单马铃薯Y病毒感染在甘薯中引起轻微症状或无症状,因此,通常未观察到显著的产量降低。然而,与其他病毒如SPCSV共同感染会产生协同效应并导致更严重的病害症状(4)。据我们所知,这是SPV2在新西兰感染甘薯的首次报道。参考文献:(1)C.D. Kokinos和C.A. Clark。植物病害。90:783,2006。(2)M.N. Pearson等人。澳大利亚植物病理学。35:217,2006。(3)M. Rännäli等人。植物病害。92:1313,2008。(4)M. Untiveros等人。植物病害。91:669, 2007。