Acevedo V, Rodrigues J C V, de Jensen C E, Webster C G, Adkins S, Wessel-Beaver L
University of Puerto Rico (UPR), Río Piedras Agricultural Experiment Station, Virology Laboratory, San Juan, PR 00926.
UPR, Fortuna Experiment Station, Juana Díaz, PR 00795.
Plant Dis. 2013 Nov;97(11):1516. doi: 10.1094/PDIS-03-13-0322-PDN.
In 2005, symptoms of watermelon vine decline (WVD) were observed on a 200-acre watermelon farm in Santa Isabel, on the south central coast of Puerto Rico. WVD symptoms included leaf curling, mosaics, and internode necrosis. In early growth stages of WVD, reduced vigor and plant stunting occurred. At flowering, symptoms progressed to necrosis and wilting of vines. A 2006 to 2007 survey demonstrated that fungal pathogens were not associated with the presence of WVD symptoms (3,4). By 2006, other watermelon fields were also affected. Field trials in 2007 and 2008 with insect-proof cages and insecticides suggested a role of whiteflies (Bemisia tabaci) in the transmission of a virus (3,4). Here, we report that watermelon and pumpkin plants were successfully infected in Puerto Rico by mechanical inoculation and through B. tabaci transmission assays, similarly to transmissions previously conducted in Florida with Squash vein yellowing virus (SqVYV) (1). In addition, plants of Cucurbita moschata exhibited vein clearing symptoms typical of SqVYV after mechanical inoculation with extracts from watermelon plants with WVD symptoms. In 2011, eight watermelon samples from plants exhibiting WVD syndrome were collected in Guánica, Santa Isabel, Juana Díaz, and Mayagüez, and two Momordica charantia samples were collected from Mayagüez. RNA was extracted from all 10 original samples, as well as from plants that were used in mechanical and vector transmission assays, using RNeasy Plant Mini Kit (Qiagen, Valencia, California), and all samples were found positive for SqVYV by reverse transcription-PCR, using previously described primers and methods (1,2). In all cases, a single ~1-kb PCR fragment was revealed, and PCR fragments from four samples were selected for direct sequencing. All sequences showed high levels (>99%) of nucleotide identity with SqVYV sequences from Florida (JF897989, JF897985, and JF897984). Sequences of the SqVYV CP gene from Puerto Rico were deposited in GenBank under accession numbers KC713961 to KC713964. To our knowledge, this is the first report of SqVYV in Puerto Rico associated with WVD syndrome in cucurbits, and thus has implications for management of viral diseases of watermelon in the Caribbean. This is also the first detection of SqVYV outside of the continental United States in both watermelon and a wild species, M. charantia (bitter gourd). References: (1) S. Adkins et al. Phytopathology 97:145, 2007. (2) S. Adkins et al. Plant Dis. 92:1119, 2008. (3) C. Estévez de Jensen et al. Phytopathology 98:S52, 2008. (4) L. Polanco-Florián. El marchitamiento súbito de la sandía [Citrullus lanatus (Thumb.) Matsum & Nakai]. M.S. thesis, University of Puerto Rico, Mayagüez, PR, 2009.
2005年,在波多黎各中南部海岸圣伊莎贝尔的一个200英亩的西瓜农场里,观察到西瓜藤蔓衰退(WVD)的症状。WVD症状包括叶片卷曲、花叶以及节间坏死。在WVD的早期生长阶段,植株活力下降且生长受阻。到开花期,症状发展为藤蔓坏死和枯萎。2006年至2007年的一项调查表明,真菌病原体与WVD症状的出现无关(3,4)。到2006年,其他西瓜田也受到了影响。2007年和2008年在防虫网罩和使用杀虫剂的田间试验表明,烟粉虱(Bemisia tabaci)在一种病毒的传播中起到了作用(3,4)。在此,我们报告,与之前在佛罗里达州用南瓜脉黄化病毒(SqVYV)进行的传播试验类似,在波多黎各通过机械接种和烟粉虱传播试验成功感染了西瓜和南瓜植株(1)。此外,用有WVD症状的西瓜植株提取物对南瓜(Cucurbita moschata)进行机械接种后,南瓜植株表现出典型的SqVYV脉明症状。2011年,在瓜尼卡、圣伊莎贝尔、胡安娜·迪亚斯和马亚圭斯采集了8个表现出WVD综合征的西瓜样本,在马亚圭斯采集了2个苦瓜(Momordica charantia)样本。使用RNeasy植物小提试剂盒(Qiagen公司,加利福尼亚州瓦伦西亚)从所有10个原始样本以及用于机械接种和媒介传播试验的植株中提取RNA,通过逆转录 - PCR,使用先前描述的引物和方法(1,2),所有样本均被检测出SqVYV呈阳性。在所有情况下,均显示出一个约1千碱基的PCR片段,从四个样本中选取PCR片段进行直接测序。所有序列与来自佛罗里达州的SqVYV序列(JF897989、JF897985和JF897984)显示出高水平(>99%)的核苷酸同一性。来自波多黎各的SqVYV CP基因序列已存入GenBank,登录号为KC713961至KC713964。据我们所知,这是波多黎各首次报道SqVYV与葫芦科作物的WVD综合征相关,因此对加勒比地区西瓜病毒病的管理具有重要意义。这也是在美国大陆以外首次在西瓜和野生种苦瓜(M. charantia)中检测到SqVYV。参考文献:(1)S. Adkins等人,《植物病理学》97:145,2007年。(2)S. Adkins等人,《植物病害》92:1119,2008年。(3)C. Estévez de Jensen等人,《植物病理学》98:S52,2008年。(4)L. Polanco - Florián,《西瓜的突然枯萎病[Citrullus lanatus (Thumb.) Matsum & Nakai]》,硕士论文,波多黎各大学,马亚圭斯,PR,2009年。