Tan H D, Li S Y, Du X F, Seno M
Dalian Institute of Biotechnology, Liaoning Academy of Agricultural Sciences, Dalian 116023, People's Republic of China.
Department of Medical and Bioengineering Science Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan.
Plant Dis. 2010 Nov;94(11):1378. doi: 10.1094/PDIS-07-10-0549.
From the spring of 2003 to the summer of 2006, sweet cherry (Prunus avium) trees in orchards near Lvshun City, in the northeast People's Republic of China, had symptoms suggestive of those caused by Cucumber mosaic virus (CMV; genus Cucumovirus, family Bromoviridae). Symptoms included chlorotic patches or mottling on leaves that were also deformed (4). In April 2006, 20 symptomatic leaves sampled from 10 trees in each of four orchards were assayed for CMV with a CMV-specific antiserum (Agdia Inc., Elkhart, IN) in a double-antibody sandwich-ELISA. Of the 80 symptomatic leaf samples, 27 tested positive for the presence of CMV. CMV was detected in all four orchards, within which incidence varied between 0.5 and 4%. Viral nucleoproteins were purified by differential centrifugation and sucrose density gradient fractionation from symptomatic leaves. Transmission electron microscopy of nucleoproteins revealed isometric particles approximately 30 nm in diameter, which is also typical of CMV. Total RNA was also extracted from 100 mg of symptomatic tissue following a Trizol-based protocol (1). A reverse transcriptase-PCR assay with nucleocapsid gene-specific primers was then used (forward primer 5'-ATGGCGACGTCCTCGTTCA-3'; reverse primer 5'-CATCGTTCCCTTCAAAATAG-3') (3). A PCR product of approximately 633 bp was obtained. The PCR product was cloned and sequenced. The sequence (GenBank Accession No. HM996559) had 95% identity with the RNA-1 sequence from CMV 'Fny' strain in GenBank (Accession No. D00356.1). The People's Republic of China is one of the major producers of sweet cherry in Asia and the spread of CMV in China may cause significant economic losses. Thus, virus-infected material should not be used for propagation and surveys should be undertaken to determine if the aphid vectors capable of transmitting CMV are present (2).To our knowledge, this is the first report of CMV occurring in sweet cherry orchards in the People's Republic of China. References: (1) P. Chomczynski and K. Mackey. Biotechniques 19:942, 1995. (2) F. E. Gildow et al. Phytopathology 98:1233, 2008. (3) T. M. Rizzo and P. Palukaitis. J. Gen. Virol. 70:1, 1989. (4) J. Shang et al. Z. Naturforsch. C 65:73, 2010.
2003年春至2006年夏,在中国东北旅顺市附近果园中的甜樱桃(Prunus avium)树出现了类似黄瓜花叶病毒(CMV;黄瓜花叶病毒属,雀麦花叶病毒科)所致的症状。症状包括叶片上出现褪绿斑或斑驳,叶片也会变形(4)。2006年4月,从四个果园中各10棵树上采集了20片有症状的叶片,用CMV特异性抗血清(Agdia公司,美国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法检测CMV。在80片有症状的叶片样本中,27片检测出CMV呈阳性。在所有四个果园中均检测到CMV,其发病率在0.5%至4%之间。通过差速离心和蔗糖密度梯度分级从有症状的叶片中纯化病毒核蛋白。核蛋白的透射电子显微镜观察显示直径约为30 nm的等轴颗粒,这也是CMV的典型特征。还按照基于Trizol的方案从100 mg有症状组织中提取了总RNA(1)。然后使用针对核衣壳基因的特异性引物进行逆转录聚合酶链反应测定(正向引物5'-ATGGCGACGTCCTCGTTCA-3';反向引物5'-CATCGTTCCCTTCAAAATAG-3')(3)。获得了一个约633 bp的聚合酶链反应产物。该聚合酶链反应产物被克隆并测序。该序列(GenBank登录号HM996559)与GenBank中CMV“Fny”株的RNA-1序列(登录号D00356.1)有95%的同一性。中国是亚洲甜樱桃的主要生产国之一,CMV在中国的传播可能会造成重大经济损失。因此,不应使用受病毒感染的材料进行繁殖,并且应进行调查以确定是否存在能够传播CMV的蚜虫传播媒介(2)。据我们所知,这是CMV在中国甜樱桃果园中出现的首次报道。参考文献:(1)P. Chomczynski和K. Mackey。生物技术19:942,1995。(2)F. E. Gildow等人。植物病理学98:1233,2008。(3)T. M. Rizzo和P. Palukaitis。普通病毒学杂志70:1,1989。(4)J. Shang等人。自然科学研究C辑65:73,2010。