Quiñones M, Fonseca D, Martinez Y, Accotto G P
Departamento de Fitopatología, Centro Nacional de Sanidad Agropecuaria (CENSA), Apdo. 10, La Habana, Cuba.
Istituto di Fitovirologia Applicata, CNR, Strada Delle Cacce 73, 10135 Torino, Italy.
Plant Dis. 2002 Jan;86(1):73. doi: 10.1094/PDIS.2002.86.1.73A.
The begomovirus Tomato yellow leaf curl virus (TYLCV) is one of the major threats to tomato production in tropical and subtropical regions worldwide. TYLCV was found in Cuba in 1994 and later became the most serious constraint to tomato production (2). During a field survey in 2001, pepper plants (Capsicum annuum) were observed in a greenhouse in Camagüey Province, showing mild interveinal yellowing and curling of leaves. Total nucleic acids were extracted from these plants and from pepper samples collected in previous years that showed similar symptoms. Polymerase chain reaction (PCR) was performed on extracts using a primer pair (TY-1/TY-2) (1) specific for the capsid protein (CP) gene of begomoviruses and a second primer pair (IR2353+: CTGAATGTTTGGATGGAAATGTGC; IR255-:GCTCGTAAGTTTCCT CAACGGAC) designed to amplify the part of the genome encompassing the intergenic region (IR) of the Cuban isolate of TYLCV-IS (2). With these primer pairs, amplicons of the expected size were obtained from five samples (one collected in 1995 in Havana Province, two in 1999 in Sancti Spiritus, and two in 2001 in Camagüey.) The CP fragment was digested with RsaI, while the IR amplicon was digested with AvaII and EcoRI. In all cases the patterns obtained corresponded to digestion patterns for identical PCR fragments obtained from TYLCV-infected tomatoes. The IR amplicon sequence from one sample showed ≈99% identity with the corresponding region of the TYLCV-IS isolated from tomato in Cuba. To our knowledge, this is the first report of TYLCV-IS infection in peppers in Cuba. References: (1) G. P. Accotto et al. Eur. J. Plant. Pathol. 106:179, 2000. (2) Y. Martínez et al. J. Phytopathol.144:277, 1996.
双生病毒番茄黄化曲叶病毒（TYLCV）是全球热带和亚热带地区番茄生产面临的主要威胁之一。1994年在古巴发现了TYLCV，后来它成为番茄生产最严重的制约因素（2）。在2001年的一次田间调查中，在卡马圭省的一个温室里观察到辣椒植株（辣椒属），叶片出现轻度脉间黄化和卷曲。从这些植株以及前几年收集的表现出类似症状的辣椒样本中提取了总核酸。使用一对针对双生病毒衣壳蛋白（CP）基因的引物（TY-1/TY-2）（1）和第二对引物（IR2353+：CTGAATGTTTGGATGGAAATGTGC；IR255-：GCTCGTAAGTTTCCT CAACGGAC）对提取物进行聚合酶链反应（PCR），第二对引物设计用于扩增包含古巴TYLCV-IS分离株基因间隔区（IR）的基因组部分（2）。使用这些引物对，从五个样本（一个于1995年在哈瓦那省采集，两个于1999年在圣斯皮里图斯采集，两个于2001年在卡马圭采集）中获得了预期大小的扩增子。CP片段用RsaI酶切，而IR扩增子用AvaII和EcoRI酶切。在所有情况下获得的图谱都与从感染TYLCV的番茄中获得的相同PCR片段的酶切图谱一致。来自一个样本的IR扩增子序列与从古巴番茄中分离的TYLCV-IS的相应区域显示出约99%的同一性。据我们所知，这是古巴辣椒中TYLCV-IS感染的首次报道。参考文献：（1）G.P.Accotto等人，《欧洲植物病理学杂志》106：179，2000年。（2）Y.Martínez等人，《植物病理学杂志》144：277，1996年。
