Mullis S W, Langston D B, Gitaitis R D, Sherwood J L, Csinos A C, Riley D G, Sparks A N, Torrance R L, Cook M J
Department of Plant Pathology, University of Georgia, P.O. Box 748, Tifton 31793.
Department of Entomology, University of Georgia, Coastal Plain Experiment Station, Tifton 31793.
Plant Dis. 2004 Nov;88(11):1285. doi: 10.1094/PDIS.2004.88.11.1285B.
Vidalia onion is an important crop in Georgia's agriculture with worldwide recognition as a specialty vegetable. Vidalia onions are shortday, Granex-type sweet onions grown within a specific area of southeastern Georgia. Tomato spotted wilt virus (TSWV) has been endemic to Georgia crops for the past decade, but has gone undetected in Vidalia onions. Tobacco thrips (Frankliniella fusca) and Western flower thrips (Frankliniella occidentalis) are the primary vectors for TSWV in this region, and a number of plant species serve as reproductive reservoirs for the vector or virus. Iris yellow spot virus (IYSV), an emerging tospovirus that is potentially a devastating pathogen of onion, has been reported in many locations in the western United States (2,4). Thrips tabaci is the known vector for IYSV, but it is unknown if noncrop plants play a role in its epidemiology in Georgia. During October 2003, a small (n = 12) sampling of onions with chlorosis and dieback of unknown etiology from the Vidalia region was screened for a variety of viruses, and TSWV and IYSV infections were serologically detected. Since that time, leaf and bulb tissues from 4,424 onion samples were screened for TSWV and IYSV using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) with commercial kits (Agdia Inc., Elkhart, IN). Samples were collected from 53 locations in the Vidalia region during the growing season between November 2003 and March 2004. Plants exhibiting stress, such as tip dieback, necrotic lesions, chlorosis or environmental damage were selected. Of these, 306 were positive for TSWV and 396 were positive for IYSV using positive threshold absorbance of three times the average plus two standard deviations of healthy negative onion controls. Positive serological findings of the onion tissues were verified by immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) for TSWV (3) and RT-PCR for IYSV (1). In both instances, a region of the viral nucleocapsid (N) gene was amplified. The PCR products were analyzed with gel electrophoresis with an ethidium bromide stain in 0.8% agarose. Eighty-six percent (n = 263) of the TSWV ELISA-positive samples exhibited the expected 774-bp product and 55 percent (n = 217) of the IYSV ELISA-positive samples exhibited the expected 962-bp product. The reduced success of the IYSV verification could be attributed to the age and deteriorated condition of the samples at the time of amplification. Thrips tabaci were obtained from onion seedbeds and cull piles within the early sampling (n = 84) and screened for TSWV by the use of an indirect-ELISA to the nonstructural (NSs) protein of TSWV. Of the thrips sampled, 25 were positive in ELISA. While the incidence of IYSV and TSWV in the Vidalia onion crop has been documented, more research is needed to illuminate their potential danger to Vidalia onions. References: (1) I. Cortês et al. Phytopathology 88:1276, 1998. (2) L. J. du Toit et al. Plant Dis. 88:222, 2004. (3) R. K. Jain et al. Plant Dis. 82:900, 1998. (4) J. W. Moyer et al. (Abstr.) Phytopathology 93(suppl.):S115, 2003.
维达利亚洋葱是佐治亚州农业中的一种重要作物,作为特色蔬菜在全球范围内享有盛誉。维达利亚洋葱是短日照、格兰ex型甜洋葱,生长在佐治亚州东南部的特定区域。在过去十年中,番茄斑萎病毒(TSWV)在佐治亚州的作物中一直存在,但在维达利亚洋葱中未被发现。烟草蓟马(Frankliniella fusca)和西花蓟马(Frankliniella occidentalis)是该地区TSWV的主要传播媒介,许多植物物种是该传播媒介或病毒的繁殖宿主。鸢尾黄斑病毒(IYSV)是一种新出现的番茄斑萎病毒属病毒,可能是洋葱的一种毁灭性病原体,已在美国西部的许多地方被报道(2,4)。烟蓟马是IYSV已知的传播媒介,但非作物植物在佐治亚州其流行病学中是否起作用尚不清楚。2003年10月,对来自维达利亚地区12株病因不明的出现黄化和枯死的洋葱进行了小规模抽样,检测了多种病毒,并通过血清学方法检测到了TSWV和IYSV感染。从那时起,使用商业试剂盒(Agdia公司,印第安纳州埃尔克哈特),通过双抗体夹心酶联免疫吸附测定(DAS-ELISA)对4424份洋葱样品的叶片和鳞茎组织进行了TSWV和IYSV检测。在2003年11月至2004年3月的生长季节,从维达利亚地区的53个地点采集了样品。选择表现出胁迫症状的植株进行检测,如叶尖枯死、坏死斑、黄化或环境损伤。其中,以健康阴性洋葱对照平均吸光度的三倍加两个标准差作为阳性阈值吸光度,306份样品TSWV呈阳性,396份样品IYSV呈阳性。洋葱组织血清学检测阳性结果通过针对TSWV的免疫捕获逆转录聚合酶链反应(IC-RT-PCR)(3)和针对IYSV 的逆转录聚合酶链反应(RT-PCR)(1)进行验证。在这两种情况下,均扩增了病毒核衣壳(N)基因的一个区域。PCR产物用含溴化乙锭的0.8%琼脂糖凝胶电泳进行分析。86%(n = 263)的TSWV ELISA阳性样品呈现预期的774 bp产物,55%(n = 217)的IYSV ELISA阳性样品呈现预期的962 bp产物。IYSV验证成功率较低可能归因于扩增时样品的老化和变质状况。在早期抽样(n = 84)中从洋葱苗床和淘汰堆中采集烟蓟马,并通过针对TSWV非结构(NSs)蛋白的间接ELISA检测TSWV。在抽样的蓟马中,25份ELISA呈阳性。虽然已记录了维达利亚洋葱作物中IYSV和TSWV的发病率,但仍需要更多研究来阐明它们对维达利亚洋葱的潜在危害。参考文献:(1)I. Cortês等人,《植物病理学》88:1276,1998年。(2)L. J. du Toit等人,《植物病害》88:222,2004年。(3)R. K. Jain等人,《植物病害》82:900,1998年。(4)J. W. Moyer等人(摘要),《植物病理学》93(增刊):S115,2003年。