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南美洲野生马铃薯( Solanum acaule)上种传樱桃卷叶病毒的首次报道

First Report of Seedborne Cherry leaf roll virus in Wild Potato, Solanum acaule, from South America.

作者信息

Crosslin J M, Eastwell K C, Davitt C M, Abad J A

机构信息

USDA-ARS Vegetable and Forage Crops Research Unit, Prosser, WA 99350.

Washington State University, Prosser, 99350.

出版信息

Plant Dis. 2010 Jun;94(6):782. doi: 10.1094/PDIS-94-6-0782C.

Abstract

A virus, designated JCM-79, was isolated from wild potato (Solanum acaule Bitt.) plants grown from true seed received at USDA-APHIS Potato Quarantine Program from Peru. JCM-79 was mechanically transmissible to Nicotiana clevelandii and N. tabacum cv. Samsun NN. Symptoms in the original S. acaule were general chlorosis and spreading necrotic lesions. Symptoms in N. tabacum and N. clevelandii included necrotic ringspots on inoculated leaves and oak-leaf patterns or necrotic spots, respectively, on upper leaves. Cultivated potatoes (S. tuberosum) infected with JCM-79 by grafting from N. clevelandii were symptomless but virus was detected by back-inoculation to N. clevelandii. Viral nucleoproteins were purified by differential centrifugation and sucrose density gradient fractionation from N. clevelandii and N. tabacum. Transmission electron microscopy of nucleoproteins revealed isometric particles approximately 25 nm in diameter. Two RNA species of approximately 8,000 and 6,500 nucleotides were obtained from nucleoproteins digested with sodium dodecyl sulfate and Proteinase K. The above characteristics suggested JCM-79 was a nepovirus or nepovirus-like in nature. Reverse transcription (RT)-PCR tests for Cherry rasp leaf virus, genus Cheravirus, which was reported from potato (3), were negative. An approximately 1,600-bp cDNA clone was obtained from RNA of JCM-79 by oligo dT primed reverse transcription and second strand cDNA synthesis. Sequence analysis (GenBank No. GU321989) revealed the closest homology (82%) to nucleotides 327 to 1801 of Accession No. S84125 Cherry leaf roll virus (CLRV), genus Nepovirus. Subsequent RT-PCR tests with CLRV-specific primers (4) resulted in amplification of a 417-bp product from nucleic acid extracts of infected N. clevelandii and N. tabacum. The amplified product from N. clevelandii was cloned and three clones were sequenced in both directions. The consensus sequence (GenBank No. GU321988) showed approximately 90% homology to the 3' untranslated region of isolates of CLRV including those from birch, walnut, and sweet cherry (GenBank Nos. S84124, Z34265, and AJ877128, respectively). JCM-79 was also detected in extracts of infected plants by ELISA using CLRV-cherry reagents (Bioreba AG, Reinach, Switzerland). These results indicate JCM-79 represents a new variant of CLRV. To our knowledge, this is the first report of CLRV naturally infecting S. acaule. S. acaule is common in the Andean regions of South America and has been used for crosses with S. tuberosum because of its pathogen resistance (1). The fact that JCM-79 is seed transmitted in S. acaule suggests that this virus could be a threat to potato-breeding programs. Another nepo-like virus with properties similar to JCM-79, designated Potato virus U (PVU), was reported from South America, but PVU was not serologically related to CLRV (2). References: (1) K. Hosaka and D. M. Spooner. Theor. Appl. Genet. 84:851, 1992. (2) R. A. C. Jones et al. Phytopathology 73:195, 1983. (3) J. R. Thompson et al. Arch. Virol. 149:2141, 2004. (4) B. Werner et al. Eur. J. For. Pathol. 27:309, 1997.

摘要

从美国农业部动植物卫生检验局马铃薯检疫项目从秘鲁收到的实生种子培育出的野生马铃薯(Solanum acaule Bitt.)植株中分离出一种病毒,命名为JCM - 79。JCM - 79可通过机械接种传播给克利夫兰烟草(Nicotiana clevelandii)和烟草(N. tabacum)品种Samsun NN。原始的S. acaule植株症状为普遍黄化和扩展的坏死斑。烟草和克利夫兰烟草的症状分别为接种叶片上的坏死环斑以及上部叶片上的橡树叶状图案或坏死斑。通过从克利夫兰烟草嫁接感染JCM - 79的栽培马铃薯(S. tuberosum)无症状,但通过回接至克利夫兰烟草检测到病毒。通过差速离心和蔗糖密度梯度分级从克利夫兰烟草和烟草中纯化病毒核蛋白。核蛋白的透射电子显微镜观察显示直径约25 nm的等轴粒子。用十二烷基硫酸钠和蛋白酶K消化核蛋白后得到两种分别约8000和6500个核苷酸的RNA种类。上述特征表明JCM - 79本质上是一种线虫传多面体病毒或类线虫传多面体病毒。对马铃薯中报道的樱桃锉叶病毒属(Cheravirus)樱桃锉叶病毒进行的逆转录(RT)-PCR检测呈阴性。通过寡聚dT引发的逆转录和第二链cDNA合成从JCM - 79的RNA中获得了一个约1600 bp的cDNA克隆。序列分析(GenBank登录号GU321989)显示与登录号S84125樱桃叶卷病毒(CLRV)属线虫传多面体病毒的核苷酸327至1801的同源性最高(82%)。随后用CLRV特异性引物进行的RT - PCR检测从感染的克利夫兰烟草和烟草的核酸提取物中扩增出一个417 bp的产物。从克利夫兰烟草扩增出的产物被克隆,对三个克隆进行双向测序。共有序列(GenBank登录号GU321988)与包括来自桦树、核桃和甜樱桃的CLRV分离株(分别为GenBank登录号S84124、Z34265和AJ877128)的3'非翻译区显示出约90%的同源性。使用CLRV - 樱桃试剂(瑞士雷纳赫市Bioreba AG公司)通过ELISA在感染植物的提取物中也检测到了JCM - 79。这些结果表明JCM - 79代表CLRV的一个新变种。据我们所知,这是CLRV自然感染S. acaule的首次报道。S. acaule在南美洲安第斯地区很常见,因其抗病性已被用于与S. tuberosum杂交(1)。JCM - 79在S. acaule中通过种子传播这一事实表明该病毒可能对马铃薯育种项目构成威胁。另一种与JCM - 79特性相似的类线虫传多面体病毒,命名为马铃薯U病毒(PVU),已在南美洲报道,但PVU与CLRV无血清学关系(2)。参考文献:(1)K. Hosaka和D. M. Spooner。《理论与应用遗传学》84:851,1992。(2)R. A. C. Jones等人。《植物病理学》73:195,1983。(3)J. R. Thompson等人。《病毒学档案》149:2141,2004。(4)B. Werner等人。《欧洲森林病理学杂志》27:309,1997。

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