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马铃薯帚顶病毒在美国马铃薯上的首次报道。

First Report of Potato mop-top virus on Potato from the United States.

作者信息

Lambert D H, Levy L, Mavrodieva V A, Johnson S B, Babcock M J, Vayda M E

机构信息

Department of Plant, Soil and Environmental Sciences, University of Maine, Orono 04469.

APHIS-PPQ-CPHST, National Plant Germplasm and Biotechnology Laboratory, Beltsville, MD 20705.

出版信息

Plant Dis. 2003 Jul;87(7):872. doi: 10.1094/PDIS.2003.87.7.872A.

Abstract

Potato mop-top virus (PMTV) is a tripartite pomovirus vectored by the powdery scab plasmodiophoromycete Spongospora subterranea pv. subterranea (1). PMTV occurs on potato (Solanum tuberosum) in Europe, the Andes, Asia, and Canada. Internal necrotic arc and fleck tuber symptoms ("spraing") may reduce commercial acceptance of some cultivars (3). PMTV symptoms were discovered in 'Shepody' tubers at the Aroostook Research Farm, Presque Isle, ME in May 2002 and subsequently in 'Russet Burbank' tubers in commercial storage from the 2001 Maine crop. Symptomatic tubers exhibited single or multiple concentric necrotic arcs that were partial or complete, but exhibited no distinct external symptoms. The presence of PMTV in eight 'Shepody' tubers was indicated by positive enzyme-linked immunosorbent assay (ELISA; Adgen, Ltd., Auchincruive, Ayr, Scotland) and confirmed by reverse transcription polymerase chain reaction (RT-PCR). 'Russet Burbank' potatoes were visually diagnosed, and the corresponding halves of 128 symptomatic tubers were forwarded to the University of Maine and APHIS (Beltsville, MD). Of these, ELISA readings in Maine were strongly positive (>3 × background) for 88, ambiguous (1.5-3 × background) for 13, and negative for 27. Subsamples from these three categories were positive by PCR in 17 of 17, 9 of 9, and 12 of 14 cases, respectively. A similar rating, positive or ambiguous, in ELISA testing was identical for all but one case at Beltsville. Confirmation of PMTV required PCR testing, resulting in a characteristic PCR product of 401 bp that was generated from the coat protein coding region on RNA 2 (2) using the primer pair PMTV 1 5'-GCAGCCGTCGAGAATAGATA-3' (RNA nucleotides 316-335) and PMTV 4 5'-GCGAGTTGATGTGCC ACATT-3' (complementary to RNA 2 nucleotides 716-697). An immunocapture RT-PCR using this primer set and the coating antibody from the Adgen ELISA kit was also successful in detecting PMTV. In separate reactions, a second product of 646 bp was generated from the triple gene block on RNA 3 (4) using the primer pair PMTV 5 5'-GGTGAACACGAGGACAAGGT-3' (RNA 3 nucleotides 1417-1436) and PMTV 7 5'-AACAGTCCGGTCTTGTGAAC-3' (complementary to RNA 3 nucleotides 2063-2044). The sequence of these products was 98 to 100% identical to PMTV published sequences. The discovery of this virus will result in adjustments to U.S. and Canadian seed potato certification standards and symptom characterization for common North American cultivars. References: (1) R. A. C. Jones and B. D. Harrison. Ann. Appl. Biol 63:1, 1969. (2) S. Kashiwazak et al. Virology 206:701, 1995. (3) M. Sandgren et al. Am. J. Potato Res. 79:205, 2002. (4) K. P. Scott et al. J. Gen. Virol.75:3561, 1994.

摘要

马铃薯帚顶病毒(PMTV)是一种三分体粉痂病毒,由粉状痂霉根肿菌马铃薯粉痂菌致病型(Spongospora subterranea pv. subterranea)传播(1)。PMTV在欧洲、安第斯地区、亚洲和加拿大的马铃薯(Solanum tuberosum)上均有发生。内部坏死弧和斑点块茎症状(“帚顶病”)可能会降低某些品种的商业接受度(3)。2002年5月,在缅因州普雷斯克岛的阿鲁斯图克研究农场的“希波蒂”(Shepody)块茎中发现了PMTV症状,随后在2001年缅因州作物的商业储存“褐皮伯班克”(Russet Burbank)块茎中也发现了该症状。有症状的块茎表现出单个或多个同心坏死弧,这些坏死弧部分或完整,但没有明显的外部症状。通过酶联免疫吸附测定(ELISA;Adgen有限公司,苏格兰艾尔郡奥金克鲁夫)检测,8个“希波蒂”块茎呈PMTV阳性,并通过逆转录聚合酶链反应(RT-PCR)得到确认。对“褐皮伯班克”马铃薯进行了目视诊断,128个有症状块茎的相应部分被送往缅因大学和美国动植物卫生检验局(马里兰州贝尔茨维尔)。其中,缅因州的ELISA读数显示,88个呈强阳性(>3×背景值),13个为模糊阳性(1.5 - 3×背景值),27个为阴性。从这三类样本中抽取的子样本通过PCR检测,在17个样本中有17个呈阳性,9个样本中有9个呈阳性,14个样本中有12个呈阳性。在贝尔茨维尔,除一个病例外,ELISA检测的阳性或模糊阳性结果与缅因州的结果完全相同。PMTV的确认需要进行PCR检测,使用引物对PMTV 1 5'-GCAGCCGTCGAGAATAGATA-3'(RNA核苷酸316 - 335)和PMTV 4 5'-GCGAGTTGATGTGCC ACATT-3'(与RNA 2核苷酸716 - 697互补),从RNA 2上的外壳蛋白编码区产生一个401 bp的特征性PCR产物(2)。使用该引物组和Adgen ELISA试剂盒中的包被抗体进行免疫捕获RT-PCR也成功检测到了PMTV。在单独的反应中,使用引物对PMTV 5 5'-GGTGAACACGAGGACAAGGT-3'(RNA 3核苷酸1417 - 1436)和PMTV 7 5'-AACAGTCCGGTCTTGTGAAC-3'(与RNA 3核苷酸2063 - 2044互补),从RNA 3上的三基因块产生了一个646 bp的第二个产物(4)。这些产物的序列与已发表的PMTV序列有98%至100%的同一性。这种病毒的发现将导致美国和加拿大种薯认证标准以及北美常见品种症状特征的调整。参考文献:(1)R. A. C. Jones和B. D. Harrison。《应用生物学年鉴》63:1,1969年。(2)S. Kashiwazak等人。《病毒学》206:701,1995年。(3)M. Sandgren等人。《美国马铃薯研究杂志》79:205,2002年。(4)K. P. Scott等人。《普通病毒学杂志》75:3561,1994年。

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