Mallik I, Gudmestad N C
Department of Plant Pathology, North Dakota State University, Fargo 58108.
Plant Dis. 2015 Jan;99(1):164. doi: 10.1094/PDIS-08-14-0819-PDN.
Potato mop top virus (PMTV) is considered the type member of the genus Pomovirus. PMTV is an important pathogen of potato vectored by the plasmodiophorid Spongospora subterranea f. sp. subterranea (Sss), which causes powdery scab of potato (1). Sss and PMTV are usually associated with cool and humid environments. PMTV-infected potato tubers generally exhibit internal hollow necrotic spots or concentric rings, and the virus is known to cause significant economic losses in Northern Europe, North and South America, and Asia (4). PMTV in the United States was first reported in Maine (2). Potato (Solanum tuberosum L.) tubers cv. FL2048 and cv. Atlantic were sent to our laboratory from fields in Saguache County in Colorado and in San Juan County in New Mexico, respectively, during the spring of 2013. The tubers from both locations had multiple, internal, concentric, necrotic arcs and circles. Internal tissue with necrotic lesion from six symptomatic tubers from each location were crushed in liquid nitrogen followed by ribonucleic acid extraction using a Total RNA Isolation kit (Promega Corp., Madison, WI). These extracts were tested by reverse transcription (RT)-PCR using three different sets of previously published primers for molecular detection of PMTV. The primer set H360/C819 targeting the coat protein (CP) on RNA 3 of PMTV yielded an amplicon (H360-CO and H360-NM) of 460 bp (4). The second set of primers, pmtF4/pmtR4 (5), amplified a 417-bp product (PMTF-CO and PMTF-NM) in RNA 2, and the third set, PMTV-P9/PMTV-M9 (3), designed to amplify the region encoding an 8-KD cysteine-rich protein in RNA 3 of PMTV, yielded a 507-bp amplicon (PMTV9-CO and PMTV9-NM). The amplicons generated from RT-PCR using all three sets were cloned (PGEMT-easy) and sequenced. Since the sequences from symptomatic tuber extracts from each location were identical to their respective primer sets, a consensus sequence from each primer set was submitted to National Center for Biotechnology Information (NCBI) GenBank. Sequences obtained from the H360/C819 primer set (GenBank Accession Nos. KM207013 and KM207014 for H360-CO and H360-NM, respectively) were 100% identical to the corresponding CP regions of PMTV isolates from North Dakota (HM776172). Sequences from the pmtF4/pmtR4 primer set (KM207015 and KM207016 for PMTF-CO and PMTF-NM, respectively) were 100% identical to the corresponding protein in RNA2 of PMTV isolates from North Dakota (GenBank HM776171), and sequences from the PMTV-P9/PMTV-M9 primer set (KM207017 and KM207018 for PMTV9-CO and PMTV9-NM respectively) were 99% identical to the corresponding protein in RNA3 of PMTV isolates (AY187010). The 100-99% homology of the sequences from this study to the corresponding PMTV sequences published in NCBI confirmed the occurrence of symptoms in the tubers from both Colorado and New Mexico due to PMTV. None of the symptomatic tubers tested positive for Tobacco rattle virus, Tomato spotted wilt virus, Alfalfa mosaic virus, Potato leafroll virus, or the necrotic strains of Potato virus Y by RT-PCR. To our knowledge, this is the first report of PMTV in potato in states of Colorado and New Mexico. References: (1) R. A. C. Jones and B. D. Harrsion. Ann. Appl. Biol. 63:1, 1969. (2) D. H. Lambert et al. Plant Dis. 87:872, 2003. (3) T. Nakayama et al. Am. J. Pot. Res. 87:218, 2010. (4) J. Santala et al. Ann. Appl. Biol. Online publication. DOI: 10.1111/j.1744-7348.2010.00423.x (5) H. Xu et al. Plant Dis. 88:363, 2004.
马铃薯帚顶病毒(PMTV)被认为是帚顶病毒属的典型成员。PMTV是马铃薯的一种重要病原体,由根肿菌纲的马铃薯粉痂菌(Spongospora subterranea f. sp. subterranea,Sss)传播,该菌可引发马铃薯粉痂病(1)。Sss和PMTV通常与凉爽湿润的环境有关。受PMTV感染的马铃薯块茎通常会出现内部中空坏死斑或同心环,已知该病毒在北欧、南北美洲和亚洲造成了重大经济损失(4)。美国首次报道PMTV是在缅因州(2)。2013年春季,马铃薯(Solanum tuberosum L.)品种FL2048和品种Atlantic的块茎分别从科罗拉多州萨瓜什县和新墨西哥州圣胡安县的田间被送到我们实验室。这两个地点的块茎都有多个内部同心坏死弧和圆圈。将来自每个地点的六个有症状块茎的带有坏死病变的内部组织在液氮中研磨,然后使用总RNA分离试剂盒(Promega公司,威斯康星州麦迪逊市)提取核糖核酸。使用先前公布的三组不同引物通过逆转录(RT)-PCR对这些提取物进行检测,以分子检测PMTV。靶向PMTV RNA 3上外壳蛋白(CP)的引物对H360/C819产生了一个460 bp的扩增子(H360-CO和H360-NM)(4)。第二组引物pmtF4/pmtR4(5)在RNA 2中扩增出一个417 bp的产物(PMTF-CO和PMTF-NM),第三组引物PMTV-P9/PMTV-M9(3)设计用于扩增PMTV RNA 3中编码一种富含半胱氨酸的8 KD蛋白的区域,产生了一个507 bp的扩增子(PMTV9-CO和PMTV9-NM)。使用所有三组引物通过RT-PCR产生的扩增子被克隆(PGEMT-easy)并测序。由于来自每个地点有症状块茎提取物的序列与其各自的引物对相同,因此将每个引物对的一致序列提交给了美国国立生物技术信息中心(NCBI)基因库。从引物对H360/C819获得的序列(H360-CO和H360-NM的GenBank登录号分别为KM207013和KM207014)与来自北达科他州的PMTV分离株(HM776172)的相应CP区域100%相同。来自引物对pmtF4/pmtR4的序列(PMTF-CO和PMTF-NM的KM207015和KM207016)与来自北达科他州的PMTV分离株(GenBank HM776171)RNA2中的相应蛋白100%相同,来自引物对PMTV-P9/PMTV-M9的序列(PMTV9-CO和PMTV9-NM的KM207017和KM207018)与PMTV分离株(AY187010)RNA3中的相应蛋白99%相同。本研究中序列与NCBI上公布的相应PMTV序列的100 - 99%同源性证实了科罗拉多州和新墨西哥州的块茎中出现的症状是由PMTV引起的。通过RT-PCR检测,所有有症状的块茎均未检测出烟草脆裂病毒、番茄斑萎病毒、苜蓿花叶病毒、马铃薯卷叶病毒或马铃薯Y病毒坏死株呈阳性。据我们所知,这是科罗拉多州和新墨西哥州马铃薯中首次报道PMTV。参考文献:(1)R. A. C. Jones和B. D. Harrsion。《应用生物学年鉴》63:1,1969。(2)D. H. Lambert等人。《植物病害》87:872,2003。(3)T. Nakayama等人。《美国马铃薯研究杂志》87:218,2010。(4)J. Santala等人。《应用生物学年鉴》在线发表。DOI: 10.1111/j.1744 - 7348.2010.00423.x(5)H. Xu等人。《植物病害》88:363,2004。