Polston J E, Bois D, Ano G, Poliakoff F, Urbino C
University of Florida, GCREC, 5007 60th St. E., Bradenton 34203.
INRA-URPV, Pointe à Pitre, Guadeloupe.
Plant Dis. 1998 Jan;82(1):126. doi: 10.1094/PDIS.1998.82.1.126B.
Viruslike symptoms were observed in epidemic proportions in tomato, Lycopersicon esculentum Mill., in Guadeloupe and Martinique each year since 1992 and 1993, respectively, and in Puerto Rico since 1994 (1,3). Many tomato fields in Guadeloupe and Martinique had more than 70% of plants expressing symptoms of chlorotic mottling, leaf distortion, leaf rolling, and stunting in 1995 and 1996. The B biotype of Bemisia tabaci (aka B. argentifolii) was associated with all these epidemics. Ninety-three samples of tomato were collected from multiple locations from each island (65 samples from Guadeloupe, 11 from Martinique, and 17 from Puerto Rico) and assayed for the presence of geminivirus by polymerase chain reaction (PCR) with broad spectrum primers, PAL1v1978 and PAR1c496 for the A component, and PBL1v2040 and PCRc154 for the B component (4). Most samples tested positive for geminivirus (98% from Guadeloupe, 100% from Martinique, and 82% from Puerto Rico). Restriction analyses of amplified A component fragment with SacI, EcoRI, and AluI, and amplified B component fragment with EcoRI, AluI, VspI, PstI, and HaeIII were conducted on 34 samples (25 from Guadeloupe, six from Martinique, and three from Puerto Rico). All samples produced very similar restriction patterns, suggesting that they were infected by the same virus. One to three clones of amplified fragments of A and B components, obtained from one plant sample from each location, were at least 98.7% identical in sequence to each other and were 89.6% and 75.2 to 75.7% identical to equivalent regions in potato yellow mosaic virus (PYMV) A and B DNA, respectively (GenBank accession nos. D00940 and D00941). This isolate of PYMV was collected from potato in Venezuela in 1985 (2). Infectious full-length genomic clones of A and B component DNA from Guadeloupe were derived from the same tissue as the PCR-generated clones, and nucleotide sequences were found to be 99.1 and 99% identical to the PCR-generated fragments of A and B DNA, respectively. Nucleotide sequences of these full-length clones were 92.6 and 89.4% identical to full-length sequences of PYMV A and B DNA, respectively. The DNA sequence identity of the common regions of the A and B components between the Guadeloupe virus and PYMV was 95.1 and 91.6%, respectively. There was a nucleotide identity of 93% in the first 125 nucleotides of the coat protein gene. The virus found in Guadeloupe, Martinique, and Puerto Rico appears to be a strain of PYMV reported from Venezuela. This strain of PYMV is at least partially responsible for the epidemics in tomato in Guadeloupe, Martinique, and Puerto Rico. The similarity among geminivirus sequences at distant locations (Puerto Rico to Martinique is approximately 600 km) is unexpected and could be due to recent introductions. References: (1) J. K. Brown et al. Plant Dis. 79:1250, 1995. (2) R. H. A. Coutts et al. J. Gen. Virol. 72:1515, 1991. (3) B. Hostachy et al. Phytoma 456:24, 1993. (4) M. R. Rojas et al. Plant Dis. 77:340, 1993.
自1992年和1993年起,瓜德罗普岛和马提尼克岛的番茄(Lycopersicon esculentum Mill.)每年都会出现呈流行态势的类病毒症状,自1994年起波多黎各也出现了此类症状(1,3)。1995年和1996年,瓜德罗普岛和马提尼克岛的许多番茄田中有超过70%的植株表现出褪绿斑驳、叶片扭曲、卷叶和生长受阻的症状。烟粉虱B型生物型(又名银叶粉虱)与所有这些疫情有关。从每个岛屿的多个地点采集了93份番茄样本(瓜德罗普岛65份、马提尼克岛11份、波多黎各17份),并使用广谱引物通过聚合酶链反应(PCR)检测双生病毒的存在,用于A组分的引物为PAL1v1978和PAR1c496,用于B组分的引物为PBL1v2040和PCRc154(4)。大多数样本检测出双生病毒呈阳性(瓜德罗普岛98%、马提尼克岛100%、波多黎各82%)。对34份样本(瓜德罗普岛25份、马提尼克岛6份、波多黎各3份)进行了用SacI、EcoRI和AluI对扩增的A组分片段以及用EcoRI、AluI、VspI、PstI和HaeIII对扩增的B组分片段的限制性分析。所有样本产生了非常相似的限制性图谱,表明它们感染的是同一种病毒。从每个地点的一个植物样本中获得的A和B组分扩增片段的一至三个克隆,彼此之间的序列至少98.7%相同,并且分别与马铃薯黄花叶病毒(PYMV)A和B DNA的等效区域89.6%以及75.2%至75.7%相同(GenBank登录号:D00940和D00941)。该PYMV分离株于1985年从委内瑞拉的马铃薯中采集(2)。来自瓜德罗普岛的A和B组分DNA的感染性全长基因组克隆与PCR产生的克隆来自同一组织,并且发现核苷酸序列分别与A和B DNA的PCR产生的片段99.1%和99%相同。这些全长克隆的核苷酸序列分别与PYMV A和B DNA的全长序列92.6%和89.4%相同。瓜德罗普岛病毒与PYMV之间A和B组分共同区域的DNA序列同一性分别为95.1%和91.6%。外壳蛋白基因的前125个核苷酸中有93%的核苷酸同一性。在瓜德罗普岛、马提尼克岛和波多黎各发现的病毒似乎是从委内瑞拉报道的PYMV的一个株系。这种PYMV株系至少部分导致了瓜德罗普岛、马提尼克岛和波多黎各番茄的疫情。远距离地点(波多黎各到马提尼克岛约600公里)双生病毒序列之间的相似性出乎意料,可能是由于近期引入所致。参考文献:(1) J. K. Brown等人,《植物病害》79:1250, 1995。(2) R. H. A. Coutts等人,《普通病毒学杂志》72:1515, 1991。(3) B. Hostachy等人,《植物病理学》456:24, 1993。(4) M. R. Rojas等人,《植物病害》77:340, 1993。