Peterschmitt M, Granier M, Mekdoud R, Dalmon A, Gambin O, Vayssieres J F, Reynaud B
CIRAD, BP 5035, 34032 Montpellier Cedex 1, France.
SPV, 2 route Ligne Paradis, 97410 Saint Pierre, Réunion, France.
Plant Dis. 1999 Mar;83(3):303. doi: 10.1094/PDIS.1999.83.3.303B.
In September 1997, stunting, reduced leaf size, leaf curling, and yellow margins were observed on tomato plants on a farm on the south coast of Réunion, a French island belonging to the Mascarenes archipelago. To our knowledge, these symptoms appeared to be characteristic of a tomato yellow leaf curl virus (TYLCV) infection. Diseased plants gave positive reactions with a triple antibody sandwich-enzyme-linked immunosorbent assay (TAS-ELISA), using ADGEN antibodies specific for begomoviruses (1). The serological results were confirmed by polymerase chain reaction (PCR) with a pair of degenerate primers-MP16, 5'-CCTCTAGATAATATTAC(C/T)(G/T)(G/A)(A/T)(T/G)G(G/A)CC-3' and MP82, 5'-CGGAATTC(T/C)TGNAC(C/T)TT(G/A)CANGGNCC(T/C)T C(G/A)CA-3'-designed by Malla Padidam (ILTAB, San Diego, CA) to amplify a region of the A component of begomoviruses, between the intergenic conserved nonanucleotide sequence (TAATATTAC) and the first 5' quarter of the capsid protein gene. A 500-bp PCR product was obtained from a symptomatic plant but not from a healthy looking one. After cloning the PCR product in a pGEM-T Easy vector (Promega, Madison, WI) and sequencing it with plasmid-specific primers (SP6, T7), the sequence was compared with the sequences of the NCBI data base, with the use of BLAST. Nineteen sequences among those producing the highest scoring segment pairs were compared with each other and with the 500-bp PCR product from Réunion by the Clustal method of MegAlign (DNASTAR, London). The Réunion sequence (AJ010790) was at least 94% similar to sequences of TYLCV isolates from the Dominican Republic (AF024715), Cuba (AJ223505), and Israel (X15656, X76319 for the mild clone). Based on these results, it appeared that the analyzed tomato plant was infected by a geminivirus isolate belonging to the Israeli species of TYLCV. A preliminary survey was carried out from December 1997 to April 1998 in both outdoor and protected tomato crops. Infected plants were detected by TAS-ELISA in 52 of the 123 locations visited. Severe economic losses were observed: 14 locations with 60 to 100% yield reduction and 11 locations with 40 to 60% yield reduction. All the infected samples were collected in the leeward coast, which is the driest region of the island. Although Bemisia tabaci (Gennadius) has been recorded since 1938 in Réunion (2), it has been observed on tomato crops only since 1997 and population levels were low compared with those of Trialeurodes vaporariorum Westwood. During the first six months of 1998, B. tabaci was found on Euphorbia heterophylla L., Lantana camara L., Solanum melongena L., S. nigrum L., and Phaseolus vulgaris L. These host plants often occur near infected tomato crops. References: (1) S. Macintosh et al. Ann. Appl. Biol. 121:297, 1992. (2) L. Russell and J. Etienne. Proc. Entomol. Soc. Wash. 87:202, 1985.
1997年9月,在留尼汪岛(属于马斯克林群岛的一个法国岛屿)南海岸的一个农场里,番茄植株上出现了发育迟缓、叶片变小、叶片卷曲和叶缘发黄的症状。据我们所知,这些症状似乎是番茄黄化曲叶病毒(TYLCV)感染的特征。患病植株通过使用针对双生病毒的ADGEN抗体进行的三抗体夹心酶联免疫吸附测定(TAS - ELISA)呈现阳性反应(1)。聚合酶链反应(PCR)使用由Malla Padidam(加利福尼亚州圣地亚哥的ILTAB)设计的一对简并引物 - MP16,5'-CCTCTAGATAATATTAC(C/T)(G/T)(G/A)(A/T)(T/G)G(G/A)CC - 3'和MP82,5'-CGGAATTC(T/C)TGNAC(C/T)TT(G/A)CANGGNCC(T/C)T C(G/A)CA - 3',扩增双生病毒A组分的一个区域,该区域位于基因间保守的九核苷酸序列(TAATATTAC)和衣壳蛋白基因的第一个5'四分之一之间,从而证实了血清学结果。从一株有症状的植株中获得了一个500 bp的PCR产物,但从看起来健康的植株中未获得。将PCR产物克隆到pGEM - T Easy载体(Promega,威斯康星州麦迪逊)中,并用质粒特异性引物(SP6,T7)进行测序后,使用BLAST将该序列与NCBI数据库中的序列进行比较。通过MegAlign(DNASTAR,伦敦)的Clustal方法,将产生最高得分片段对的19个序列相互比较,并与留尼汪岛的500 bp PCR产物进行比较。留尼汪岛序列(AJ010790)与来自多米尼加共和国(AF024715)、古巴(AJ223505)和以色列(X15656,轻度克隆的X76319)的TYLCV分离株序列至少有94%的相似性。基于这些结果,似乎所分析的番茄植株被一种属于TYLCV以色列种的双生病毒分离株感染。1997年12月至1998年4月,在室外和保护地番茄作物中进行了初步调查。在所访问的123个地点中的52个地点,通过TAS - ELISA检测到了受感染的植株。观察到了严重的经济损失:有14个地点产量降低60%至100%,11个地点产量降低40%至60%。所有受感染的样本都采集自该岛最干燥的背风海岸。尽管自1938年以来留尼汪岛就有烟粉虱(Gennadius)的记录(2),但直到1997年才在番茄作物上观察到,并且与温室白粉虱(Trialeurodes vaporariorum Westwood)相比,其种群数量较低。在1998年的前六个月里,在异叶大戟、马缨丹、茄子、龙葵和菜豆上发现了烟粉虱。这些寄主植物经常出现在受感染的番茄作物附近。参考文献:(1)S. Macintosh等人,《应用生物学年鉴》121:297,1992。(2)L. Russell和J. Etienne,《华盛顿昆虫学会会刊》87:202,1985。