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番茄黄化曲叶病毒Sar从地中海盆地的传播:在加那利群岛和摩洛哥的存在情况。

Spread of Tomato yellow leaf curl virus Sar from the Mediterranean Basin: Presence in the Canary Islands and Morocco.

作者信息

Monci F, Navas-Castillo J, Cenis J L, Lacasa A, Benazoun A, Moriones E

机构信息

CSIC, 29750 Algarrobo, Costa, Málaga, Spain.

CIDA, 30150 La Alberca, Murcia, Spain.

出版信息

Plant Dis. 2000 Apr;84(4):490. doi: 10.1094/PDIS.2000.84.4.490D.

Abstract

Severe outbreaks of tomato yellow leaf curl disease occurred during summer and autumn 1999 in tomato (Lycopersicon esculentum Mill.) crops in the Vecindario Region of Gran Canaria (Canary Islands, Spain) and Agadir (southwestern Atlantic coast of Morocco). Symptoms of the disease included upward curling of leaflet margins, reduction of leaflet area, and yellowing of young leaves, as well as stunting and flower abortion. High populations of whiteflies, Bemisia tabaci Gen., were present on tomatoes in Agadir, and analysis of adult individuals by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) identified them as the biotype Q. Samples were collected from symptomatic tomato plants: 5 plants from Gran Canaria and 22 from three areas in Agadir, (7 from Agadir/1, 12 from Agadir/2, and 3 from Agadir/3) in the Koudya Region. Samples were analyzed for Tomato yellow leaf curl virus (TYLCV) Sar or Is (genus Begomovirus, family Geminiviridae) infection by squash blot hybridization under high stringency conditions with digoxigenin-labeled DNA probes specific to TYLCV-Sar or -Is, as described previously (1,3). The TYLCV-Sar probe hybridized to the five samples from Gran Canaria, and the TYLCV-Is probe hybridized to the 22 samples from Agadir. The TYLCV-Sar probe also hybridized to the three samples from Agadir/3. Primer pairs MA-14/MA-15 and MA-30/MA-31, designed for specific amplification of the intergenic region (IR) of TYLCV-Sar or -Is reported from Spain, respectively (1), were used in PCR to amplify one sample each from Gran Canaria, Agadir/1, and Agadir/3. A fragment of the expected size was obtained from the samples from Gran Canaria and Agadir/3 using MA14/MA15 (342 bp) and from the two samples from Agadir using MA30/MA31 (357 bp). PCR products were directly sequenced (GenBank Accession nos. AF215819 to AF215822). The nucleotide sequences of the IR fragments amplified from the Gran Canaria and Agadir/3 sample using MA-14/MA-15 indicated their closest relationship (99.0 and 96.7% identity, respectively) was to the corresponding region of a TYLCV-Sar isolate reported from Spain (GenBank Accession no. L27708). The nucleotide sequences of the IR fragments amplified from the Agadir/1 and Agadir/3 samples using MA-30/MA-31 indicated their closest relationship (98.1% identity) was to the corresponding region of the TYLCV-Is isolate reported from Spain (GenBank Accession no. AF071228). Based on the hybridization and sequence data, we conclude that the symptomatic plants from Gran Canaria were infected by TYLCV-Sar, those from Agadir/1 and Agadir/2 were infected by TYLCV-Is, and those from Agadir/3 had mixed infections with TYLCV-Is and TYLCV-Sar. The presence of TYLCV-Is in Morocco has been described recently (2). However, this is the first report of TYLCV-Sar in the Canary Islands and Morocco and extends its geographic range beyond the Iberian Peninsula and Italy. References: (1) J. Navas-Castillo et al. Plant Dis. 83:29, 1999. (2) M. Peterschmitt et al. Plant Dis. 83:1074, 1999. (3) S. Sánchez-Campos et al. Phytopathology 89:1038, 1999.

摘要

1999年夏秋季节,西班牙加那利群岛大加那利岛的韦辛达里奥地区以及摩洛哥阿加迪尔(摩洛哥西南大西洋沿岸)的番茄(Lycopersicon esculentum Mill.)作物上爆发了严重的番茄黄化曲叶病。该病症状包括小叶边缘向上卷曲、小叶面积减小、幼叶黄化,以及植株矮化和落花。阿加迪尔的番茄上有大量烟粉虱(Bemisia tabaci Gen.),通过随机扩增多态性DNA聚合酶链反应(RAPD-PCR)对成虫个体进行分析,确定它们为Q生物型。从有症状的番茄植株上采集样本:来自大加那利岛的5株以及来自阿加迪尔库迪亚地区三个区域(阿加迪尔/1区7株、阿加迪尔/2区12株、阿加迪尔/3区3株)的22株。如先前所述(1,3),在高严谨条件下,使用地高辛标记的针对番茄黄化曲叶病毒(TYLCV)Sar或Is(双生病毒属,双生病毒科)的DNA探针,通过压片杂交分析样本是否感染TYLCV-Sar或-Is。TYLCV-Sar探针与来自大加那利岛的5个样本杂交,TYLCV-Is探针与来自阿加迪尔的22个样本杂交。TYLCV-Sar探针也与来自阿加迪尔/3区的3个样本杂交。分别设计用于特异性扩增西班牙报道的TYLCV-Sar或-Is基因间隔区(IR)的引物对MA-14/MA-15和MA-30/MA-31(1),用于PCR扩增来自大加那利岛、阿加迪尔/1区和阿加迪尔/3区的各一个样本。使用MA14/MA15从大加那利岛和阿加迪尔/3区的样本中获得了预期大小的片段(342 bp),使用MA30/MA31从阿加迪尔的两个样本中获得了预期大小的片段(357 bp)。对PCR产物进行直接测序(GenBank登录号:AF215819至AF215822)。使用MA-14/MA-15从大加那利岛和阿加迪尔/3区样本中扩增的IR片段的核苷酸序列表明,它们与西班牙报道的TYLCV-Sar分离株的相应区域关系最为密切(分别为99.0%和96.7%的同一性)(GenBank登录号:L27708)。使用MA-30/MA-31从阿加迪尔/1区和阿加迪尔/3区样本中扩增的IR片段的核苷酸序列表明,它们与西班牙报道的TYLCV-Is分离株的相应区域关系最为密切(98.1%的同一性)(GenBank登录号:AF071228)。根据杂交和序列数据,我们得出结论,大加那利岛有症状的植株感染了TYLCV-Sar,阿加迪尔/1区和阿加迪尔/2区的植株感染了TYLCV-Is,阿加迪尔/3区的植株同时感染了TYLCV-Is和TYLCV-Sar。最近已报道摩洛哥存在TYLCV-Is(2)。然而,这是加那利群岛和摩洛哥首次报道TYLCV-Sar,其地理分布范围扩展至伊比利亚半岛和意大利之外。参考文献:(1)J. Navas-Castillo等人,《植物病害》83:29,1999年。(2)M. Peterschmitt等人,《植物病害》83:1074,1999年。(3)S. Sánchez-Campos等人,《植物病理学》89:1038,1999年。

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