Evans C K, Bag S, Frank E, Reeve J, Ransom C, Drost D, Pappu H R
Utah State University, Biology Department, Logan.
Department of Plant Pathology, Washington State University, Pullman.
Plant Dis. 2009 Jun;93(6):670. doi: 10.1094/PDIS-93-6-0670C.
Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is a serious virus pathogen in onion bulb and seed crops in the United States and several parts of the world (1). The virus is exclusively transmitted by onion thrips (Thrips tabaci). Besides onion and other susceptible crops such as garlic, leek, chives, and several ornamentals, weeds could be serving as potential reservoir sources of virus inoculum. There are reports of several weeds found naturally infected with IYSV (1,2,4). However, there is no report of IYSV infection of a grass species. Leaves of green foxtail (Setaria viridis (L.) Beauv.) were collected from two naturally occurring plants approximately 30 m apart in a weed trial conducted in commercial onions grown in Box Elder County, UT on 24 September 2008. Notes of IYSV symptoms on green foxtail were made only on the two grass plants sampled. Density of green foxtail in the weed trial was low and was not recorded. Leaves on both plants displayed a range of symptoms that included streaking, purpling, and chlorotic and necrotic lesions along leaf margins oriented along the axis of longitudinal venation. Samples were positive for IYSV by double-antibody sandwich-ELISA with a commercially available kit (Agdia Inc., Elkhart, IN). ELISA values of the grass samples were 2.64 and 2.23 for each plant sampled. Negative and positive control readings were 0.24 and 4.33, respectively. All absorbance readings were made at 405 nm. To provide a contrast of the grass data in context to the onion field where the weed trial was located, final visual assessments of onions in the field were made on 4 September 2009. Approximately 300 onion plants were assessed for incidence and severity of disease. Incidence of the disease among onions was 100% and the severity of iris yellow spot on leaves was 20 lesions per leaf. The average ELISA value over 30 individual onions arbitrarily sampled from the field on the same day was 3.50, and the ELISA values among the samples ranged from 1.37 to 4.38. The negative and positive controls were 0.19 and 4.40, respectively. To further verify the presence of IYSV in the grass specimen, reverse transcription-PCR was performed on total nucleic acid extracts obtained from the symptomatic parts of the leaves. Primers specific to the nucleocapsid (N) gene coded by the small (S)-RNA of IYSV were used (3). The forward and reverse primer pairs, 5'-TCAGAAATCGAGAAACTT-3' and 5'-CACCAATGTCTTCAACAATCTT-3', respectively, amplify a 751-nt fragment of the N gene (3). An amplicon of expected size was obtained, cloned, and sequenced. The nucleotide sequence analysis and comparison with known IYSV S-RNA sequences showed that the amplicon from foxtail (GenBank Accession No. FJ652594) samples had the highest nucleotide sequence identity (98%) with the corresponding region of an IYSV isolate from Jefferson County, OR (GenBank Accession No. DQ233479). To our knowledge, this is the first report of natural infection of a grass species by IYSV and the first report of a Tospovirus infecting a grass species. The data suggests grasses may serve as a new host reservoir for IYSV. The increasing number of weed hosts of IYSV warrants further study on the role of these weeds as hosts for onion thrips and in IYSV epidemiology. References: (1) D. Gent et al. Plant Dis. 90:1468, 2006. (2) C. Nischwitz et al. Plant Dis. 91:1518, 2007. (3) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (4) R. Sampangi et al. Plant Dis. 91:1683, 2007.
鸢尾黄斑病毒(IYSV;布尼亚病毒科番茄斑萎病毒属)是美国及世界其他一些地区洋葱鳞茎和种子作物中的一种严重病毒病原体(1)。该病毒仅由葱蓟马(西花蓟马)传播。除了洋葱和其他易感作物,如大蒜、韭菜、细香葱以及几种观赏植物外,杂草可能是病毒接种体的潜在储存源。有报道称发现几种杂草自然感染了IYSV(1,2,4)。然而,尚无关于禾本科植物感染IYSV的报道。2008年9月24日,在犹他州博克斯埃尔德县种植的商业洋葱田进行的杂草试验中,从相距约30米的两株自然生长的绿色狗尾草(Setaria viridis (L.) Beauv.)植株上采集了叶片。仅对所采集的这两株禾本科植物记录了绿色狗尾草上IYSV的症状。杂草试验中绿色狗尾草的密度较低,未作记录。两株植物的叶片均表现出一系列症状,包括沿着纵向叶脉轴线的叶片边缘出现条纹、发紫以及褪绿和坏死病斑。使用市售试剂盒(Agdia公司,印第安纳州埃尔克哈特)通过双抗体夹心ELISA检测,样本对IYSV呈阳性。所采集的每株禾本科植物样本的ELISA值分别为2.64和2.23。阴性和阳性对照读数分别为0.24和4.33。所有吸光度读数均在405nm处测定。为了将禾本科植物的数据与进行杂草试验的洋葱田情况进行对比,于2009年9月4日对田间洋葱进行了最终目视评估。对约300株洋葱植株的发病率和病情严重程度进行了评估。洋葱的发病率为100%,叶片上鸢尾黄斑的严重程度为每片叶20个病斑。同一天从田间随机抽取的30株单个洋葱的平均ELISA值为3.50,样本中的ELISA值范围为1.37至4.38。阴性和阳性对照分别为0.19和4.40。为进一步验证禾本科植物样本中IYSV的存在,对从叶片症状部位获得的总核酸提取物进行了逆转录PCR。使用了针对IYSV小(S)RNA编码的核衣壳(N)基因特异的引物(3)。正向和反向引物对分别为5'-TCAGAAATCGAGAAACTT-3'和5'-CACCAATGTCTTCAACAATCTT-3',可扩增N基因的一个751 nt片段(3)。获得了预期大小的扩增子,进行了克隆和测序。核苷酸序列分析以及与已知IYSV S-RNA序列的比较表明,狗尾草(GenBank登录号FJ652594)样本的扩增子与俄勒冈州杰斐逊县的一个IYSV分离株的相应区域具有最高的核苷酸序列同一性(98%)(GenBank登录号DQ233479)。据我们所知,这是关于禾本科植物自然感染IYSV的首次报道,也是番茄斑萎病毒属病毒感染禾本科植物的首次报道。数据表明禾本科植物可能是IYSV的新宿主储存库。IYSV杂草宿主数量的增加值得进一步研究这些杂草作为葱蓟马宿主以及在IYSV流行病学中的作用。参考文献:(1)D. Gent等人,《植物病害》90:1468,2006年。(2)C. Nischwitz等人,《植物病害》91:1518,2007年。(3)H. R. Pappu等人,《病毒学档案》151:1015,2006年。(4)R. Sampangi等人,《植物病害》91:1683,2007年。