Liu H-Y, Sears J L, Bandla M, Harness A M, Kulemeka B
USDA-ARS, Salinas, CA 93905.
Agdia, Inc., Elkhart, IN 46514.
Plant Dis. 2003 Dec;87(12):1538. doi: 10.1094/PDIS.2003.87.12.1538A.
Calibrachoa mottle virus (CbMV), a tentative carmovirus, was first isolated and reported by Liu et al. (1) from infected Calibrachoa plants. During the spring of 2003, petunia samples from Florida and California sent to testing services at Agdia, Inc (Elkhart IN) tested positive for CbMV by enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay (ImmunoStrips). These samples also tested positive by carmovirus group-specific polymerase chain reaction (PCR) primers and by immunocapture PCR (2). RNA extracted from these samples with the RNeasy Plant Kit (Qiagen Inc., Valencia, CA) hybridized with a digoxigenin labeled probe derived from purified CbMV viral RNA. All plant samples that tested positive for CbMV were symptomless except one symptomatic sample that also tested positive for Tobacco mosaic virus. From samples that tested positive for CbMV only, mechanical inoculations were made to Chenopodium quinoa at a USDA-ARS greenhouse in Salinas, CA. Representative single, local lesions were used to inoculate additional C. quinoa plants. The resulting local lesions from these inoculations were freeze-dried and further used as virus inoculum (CbMV petunia). Similar inoculum was made with CbMV isolated from Calibrachoa plants (CbMV calibrachoa). Virus-free Petunia hybrida cultivars Surfinia 'Baby Pink' and Surfinia 'Violet' (Jackson and Perkins Inc., Somis, CA) were mechanically inoculated with CbMV petunia and CbMV calibrachoa. Four weeks postinoculation, all plants were tested using ELISA for the presence of CbMV. In greenhouse conditions, 14.3% of 'Baby Pink' plants were positive for CbMV petunia, whereas none were positive for CbMV calibrachoa. 'Violet' plants were 64.3 and 33.3% positive for CbMV petunia and CbMV calibrachoa, respectively. None of the positive plants expressed virus-like symptoms. Virus particles resembling those of CbMV were observed from infected petunia plants with transmission electron microscopy in leaf-dip preparations. To our knowledge, this is the first report of CbMV infecting petunia. Commercial reproduction of petunia plants and maintenance of genetic mother stock are usually by vegetative propagation. CbMV can be transmitted mechanically and is readily propagated along with its host. To produce healthy petunia plants, virus-free mother stock should be used, which requires regular screening of mother stock for CbMV. Reference: (1) H.-Y. Liu et al. Plant Dis. 87:167, 2003. (2) A. M. Harness et al. (Abstr.) Phytopathology 92:S34, 2002.
小花矮牵牛斑驳病毒(CbMV),一种暂定的香石竹斑驳病毒属病毒,最早由Liu等人(1)从受感染的小花矮牵牛植株中分离并报道。2003年春季,来自佛罗里达州和加利福尼亚州的矮牵牛样本被送往位于印第安纳州埃尔克哈特的Agdia公司的检测服务部门,通过酶联免疫吸附测定(ELISA)和侧向流动免疫测定(免疫试纸条)检测,结果显示CbMV呈阳性。这些样本通过香石竹斑驳病毒属特异性聚合酶链反应(PCR)引物和免疫捕获PCR检测也呈阳性(2)。用RNeasy植物试剂盒(Qiagen公司,加利福尼亚州瓦伦西亚)从这些样本中提取的RNA与源自纯化的CbMV病毒RNA的地高辛标记探针杂交。所有检测CbMV呈阳性的植物样本均无症状,只有一个有症状的样本同时检测出烟草花叶病毒呈阳性。从仅检测CbMV呈阳性的样本中,在加利福尼亚州萨利纳斯的美国农业部农业研究局温室中对藜麦进行了机械接种。用代表性的单个局部病斑接种更多的藜麦植株。这些接种产生的局部病斑经冷冻干燥后进一步用作病毒接种物(CbMV矮牵牛)。用从小花矮牵牛植株中分离出的CbMV制备了类似的接种物(CbMV小花矮牵牛)。用CbMV矮牵牛和CbMV小花矮牵牛对无病毒的矮牵牛品种“冲浪者粉宝宝”和“冲浪者紫罗兰”(杰克逊和珀金斯公司,加利福尼亚州索米斯)进行机械接种。接种四周后,使用ELISA检测所有植株是否存在CbMV。在温室条件下,“粉宝宝”植株中有14.3%检测CbMV矮牵牛呈阳性,而检测CbMV小花矮牵牛呈阳性的植株无一株。“紫罗兰”植株中检测CbMV矮牵牛和CbMV小花矮牵牛呈阳性的分别为64.3%和33.3%。所有阳性植株均未表现出病毒样症状。在叶浸制标本中,通过透射电子显微镜观察到受感染的矮牵牛植株中有类似CbMV的病毒颗粒。据我们所知,这是CbMV感染矮牵牛的首次报道。矮牵牛植株的商业繁殖和遗传母本的保存通常通过营养繁殖。CbMV可通过机械传播,并很容易与其寄主一起繁殖。为了生产健康的矮牵牛植株,应使用无病毒的母本,这需要定期对母本进行CbMV筛查。参考文献:(1)H.-Y. Liu等人,《植物病害》87:167,200