Hafez E E, Abdelkhalek A A, El-Morsi A A, El-Shahaby O A
Molecular Plant Pathology Department, City for Scientific Research and Technology Applications (MuCSAT), 21934, Alexandria, Egypt.
Botany Department, Faculty of Science, Mansoura University, Mansoura, Egypt.
Plant Dis. 2012 Apr;96(4):594. doi: 10.1094/PDIS-10-11-0902.
Egyptian leek (Allium ampeloprasum), garlic (A. sativum), and onion (A. cepa) are key vegetables produced by small- and large-scale farmers in Egypt for national and international markets. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an economically important viral pathogen of bulb and seed onion crops in many onion-growing areas of the world (1,3). During February and March of 2011, symptoms of spindle-shaped, straw-colored, irregular lesions with occasional green islands were observed on onion, garlic, and Egyptian leek cultivated on large and small farms in Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiut governorates in Egypt. The presence of IYSV was confirmed by specific double antibody sandwich (DAS)-ELISA Flash Kits (Agdia Inc., Elkhart, IN) (2). A survey was carried out by collecting 100 plant samples (10 asymptomatic and 90 symptomatic) of each plant species from fields in the governorates of Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiute and testing the plants using DAS-ELISA. For onion and garlic, 45% of the symptomatic samples and 0% of the asymptomatic plants tested positive. For leek, 34% of the symptomatic samples tested positive and 0% of the asymptomatic samples. ELISA-positive samples were tested using a reverse transcription (RT)-PCR assay with primers specific to the S RNA of IYSV (forward primer 5'-TAAAACAAACATTCAAACAA-3' and reverse primer 5'-CTCTTAAACACATTTAACAAGCAC-3') (2). Amplicons of approximately 1,100 bp were obtained from all symptomatic samples that were ELISA positive, but none of the asymptomatic plants nor the sterile water control sample produced PCR amplicons. The amplicons were cloned (at least three clones per plant species) using the TOPO TA Cloning Kit (Invitrogen, Grand Island, NY), and sequenced. The Egyptian onion IYSV isolate (GenBank No. JN541273) had the greatest nucleotide sequence identity (86%) with the corresponding S RNA region of IYSV isolates from India (GenBank Nos. EU310290, EU310284, and EU310276). The Egyptian garlic IYSV isolate (GenBank No. JN541275) showed the strongest identity (93%) with that of a Sri Lankan IYSV isolate (GenBank No. GU901211). The Egyptian leek IYSV isolate (GenBank No. JN541274) exhibited 91% sequence identity with that of the same Sri Lankan isolate (No. GU901211). To our knowledge, this is the first report of IYSV infecting garlic and Egyptian leek in Egypt. IYSV infection of onion was reported previously from the agricultural farm of the Faculty of Agriculture, Cairo University, Giza (4), but to our knowledge, this is the first report of natural infection by the virus in commercial onion production in Egypt. Further surveys and monitoring of IYSV incidence and distribution in the entire Egyptian governorate are under investigation. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) A. Manal et al. Egypt. J. Virol. 3:49, 2006.
埃及韭葱(葱属宽叶韭)、大蒜(蒜)和洋葱(洋葱)是埃及的小型和大型农场为国内和国际市场生产的主要蔬菜。鸢尾黄斑病毒(IYSV;布尼亚病毒科番茄斑萎病毒属)是世界上许多洋葱种植地区的鳞茎和种子洋葱作物的一种具有经济重要性的病毒病原体(1,3)。2011年2月和3月期间,在埃及达卡利亚、加比亚、卡卢比亚、明亚、基纳和阿斯尤特省的大小农场种植的洋葱、大蒜和埃及韭葱上观察到纺锤形、稻草色、不规则病斑,偶尔带有绿色岛屿的症状。通过特异性双抗体夹心(DAS)-ELISA快速检测试剂盒(Agdia公司,美国印第安纳州埃尔克哈特)确认了IYSV的存在(2)。通过从达卡利亚、加比亚、卡卢比亚、明亚、基纳和阿斯尤特省的田间收集每种植物的100个植物样本(10个无症状样本和90个有症状样本)并使用DAS-ELISA检测这些植物,进行了一项调查。对于洋葱和大蒜,45%的有症状样本检测呈阳性,无症状植物的检测阳性率为0%。对于韭葱,34%的有症状样本检测呈阳性,无症状样本的检测阳性率为0%。使用针对IYSV的S RNA的特异性引物(正向引物5'-TAAAACAAACATTCAAACAA-3'和反向引物5'-CTCTTAAACACATTTAACAAGCAC-3')的逆转录(RT)-PCR检测对ELISA阳性样本进行检测(2)。从所有ELISA阳性的有症状样本中获得了约1100 bp的扩增子,但无症状植物和无菌水对照样本均未产生PCR扩增子。使用TOPO TA克隆试剂盒(Invitrogen公司,美国纽约州大岛)对扩增子进行克隆(每种植物至少三个克隆)并进行测序。埃及洋葱IYSV分离株(GenBank登录号JN541273)与来自印度的IYSV分离株(GenBank登录号EU310290、EU310284和EU310276)的相应S RNA区域具有最高的核苷酸序列同一性(86%)。埃及大蒜IYSV分离株(GenBank登录号JN541275)与斯里兰卡的IYSV分离株(GenBank登录号GU901211)具有最强的同一性(93%)。埃及韭葱IYSV分离株(GenBank登录号JN541274)与同一斯里兰卡分离株(登录号GU901211)表现出91%的序列同一性。据我们所知,这是IYSV在埃及感染大蒜和埃及韭葱的首次报道。此前曾有报道称开罗大学农业学院位于吉萨的农场的洋葱感染了IYSV(4),但据我们所知,这是该病毒在埃及商业洋葱生产中自然感染的首次报道。目前正在对埃及整个省份IYSV的发病率和分布进行进一步的调查和监测。参考文献:(1)D. H. Gent等人,《植物病害》88:446,2004年。(2)H. R. Pappu等人,《病毒学档案》151:1015,2006年。(3)H. R. Pappu等人,《病毒研究》141:219,2009年。(4)A. Manal等人,《埃及病毒学杂志》3:49,2006年。