In October 2019, tomato (Solanum lycopersicum L.) plants showing chlorosis and brown necrosis in apical leaflets and rugose surface in fruits were observed in a greenhouse in Vicar, Almería, Spain. A total of 0.5% of the tomato plants in the greenhouse (1,38 ha) showed these symptoms. The presence of tomato brown rugose fruit virus (ToBRFV) was suspected. A total of 5 symptomatic and 2 symptomless leaf samples were collected and analyzed by double-antibody sandwich (DAS)-ELISA with antibodies for ToBRFV, tobacco mosaic virus (TMV) and tomato mosaic virus (ToMV) (Loewe Biochemica, Germany). Symptomatic samples tested positive by DAS-ELISA only for ToBRFV. Therefore, one sample was selected and analyzed by reverse transcription (RT)-PCR with specific primers ToBRFV-F5722/ToBRFV-R6179 for ToBRFV (Panno et al. 2019a) which amplified a 458 bp fragment of the coat protein gene. The sequence obtained by Sanger sequencing from the amplicon showed 99.7% nt identity with ToBRFV isolate from United Kingdom (Acc. No. MN182533) and was deposited in the GenBank database under the accession number MT211630. Further surveys were performed in Vicar and El Egido (Almería, Spain) on plants showing viral-like symptoms. A total of 50 tomato and two pepper leaf samples from 28 greenhouses were collected and analyzed by DAS-ELISA for ToBRFV, TMV, ToMV, pepino mosaic virus (PepMV) , tomato spotted wilt virus (TSWV) and tomato yellow leaf curl virus (TYLCV) with respective antibodies (Loewe Biochemica, Germany). Five tomato plants (four from Vicar and one from El Ejido) tested positive for ToBRFV. PepMV, ToMV and TYLCV were also detected in one, three and five tomato plants, respectively. The ELISA positive results for ToBRFV were confirmed by end point RT-PCR with primer pairs ToBRFV-F5722/ToBRFV-R6179 and ToBRFV-F/ToBRFV-R (Alkouni et al., 2019) and by real-time RT-PCR with Taqman probe ToB-probe and specific primers ToB5520F/ToB5598R (Panno et al., 2019b). Tomato seeds used for plantation in these greenhouses were also analyzed, but ToBRFV was not detected. Eradication measures have been undertaken to prevent the virus spread and to control this outbreak. Official seed analysis by DAS-ELISA and real time RT-PCR (ISF, 2019) are being conducted on the tomato and pepper imported seeds to prevent the appearance of new sources of ToBRFV inoculum in Spain. References: Alkowni, R., et al. 2019. J. Plant Pathol. 101: 719. doi: 10.1007/s42161-019-00240-7. ISF, 2019. International Seed Federation. Version 1.3, September 2019. Available at https://www.worldseed.org/wp-content/uploads/2019/09/Tomato-ToBRFV_2019.09.pdf Panno, S., et al. 2019a. Plant Dis. 103: 1443. https://doi.org/10.1094/PDIS-12-18-2254-PDN Panno, S. et al. 2019b. PeerJ 7:e7928 DOI 10.7717/peerj.7928.

中文翻译：

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西班牙番茄中番茄棕色皱纹水果病毒的首次报道。

2019年10月，在西班牙阿尔梅里亚Vicar的一个温室中观察到了番茄（Solanum lycopersicum L.）植株，在其根尖小叶和果实的皱纹表面均显示出萎黄和棕色坏死。大棚中总计0.5％的番茄植株（1.38公顷）表现出这些症状。怀疑存在番茄棕色皱纹果病毒（ToBRFV）。总共收集了5个有症状和2个无症状叶片样品，并通过双抗体夹心（DAS）-ELISA分析了ToBRFV，烟草花叶病毒（TMV）和番茄花叶病毒（ToMV）的抗体（德国Loewe Biochemica）。有症状的样品仅通过DAS-ELISA检测为ToBRFV阳性。因此，选择了一个样品，并通过针对ToBRFV的特异性引物ToBRFV-F5722 / ToBRFV-R6179进行逆转录（RT）-PCR分析（Panno等。2019a）扩增了外壳蛋白基因的458 bp片段。通过Sanger测序从扩增子获得的序列与来自英国的ToBRFV分离株（Acc。MN182533）表现出99.7％的核苷酸同一性，并以登录号MT211630保藏在GenBank数据库中。在Vicar和El Egido（西班牙阿尔梅里亚）对表现出病毒样症状的植物进行了进一步调查。收集了来自28个温室的50个番茄和两个辣椒叶样品，并通过DAS-ELISA分析了ToBRFV，TMV，ToMV，pepino花叶病毒（PepMV），番茄斑萎病毒（TSWV）和番茄黄叶卷曲病毒（TYLCV） ）和相应的抗体（Loewe Biochemica，德国）。五株番茄植株（四株来自Vicar，一株来自El Ejido）的ToBRFV检测呈阳性。还发现了PepMV，ToMV和TYLCV，三个和五个西红柿。通过使用引物对ToBRFV-F5722 / ToBRFV-R6179和ToBRFV-F / ToBRFV-R进行终点RT-PCR（Alkouni等，2019）以及通过Taqman实时RT-PCR证实了ToBRFV的ELISA阳性结果探针ToB-探针和特异性引物ToB5520F / ToB5598R（Panno等，2019b）。还分析了这些温室中用于种植的番茄种子，但未检测到ToBRFV。已经采取了消灭措施，以防止病毒传播并控制这种暴发。目前正通过DAS-ELISA和实时RT-PCR（ISF，2019）对进口的番茄和辣椒种子进行官方种子分析，以防止西班牙出现ToBRFV接种物的新来源。参考：Alkowni，R.等。2019.植物病理学杂志。101：719。doi：10.1007 / s42161-019-00240-7。ISF，2019。国际种子联合会。版本1.3,2019年9月。可在https://www.worldseed.org/wp-content/uploads/2019/09/Tomato-ToBRFV_2019.09.pdf Panno，S.，et al。2019年 植物病 103：1443。https：//doi.org/10.1094/PDIS-12-18-2254-PDN Panno，S.等。2019b。PeerJ 7：e7928 DOI 10.7717 / peerj.7928。