Reingold Victoria, Lachman Oded, Sela Noa, Luria Neta, Dombrovsky Aviv
Department of Plant Pathology, Agriculture Research Organization, The Volcani Center, Bet Dagan 50250, Israel.
Plant Dis. 2016 Jun;100(6):1176-1183. doi: 10.1094/PDIS-09-15-1040-RE. Epub 2016 Mar 21.
In spring 2014, unfamiliar watermelon disease symptoms were observed on 1,000 ha of watermelon plants (Citrullus lanatus Thunb.) growing in open fields in Jordan and Beit-She'an Valleys, Israel. These represented systemic wilt and yellowing of leaves with necrosis on leaves and stems, in some cases leading to plant dieback, fruit exocarp deterioration, and rotting of the fleshy mesocarp, leading to unmarketable fruit. Virus purification was carried out from watermelon exocarp and necrotic leaves, and transmission electron microscopy revealed viral particles with flexible filamentous morphology. The disease was transmitted by mechanical inoculation from symptomatic fruit and by the silverleaf whitefly Bemisia tabaci from symptomatic to healthy cucurbits. A reverse-transcription polymerase chain reaction (RT-PCR) test was conducted on purified virus preparation of Squash vein yellowing virus (SqVYV) using specific primers targeting the capsid protein gene revealing the expected amplicon size. The complete viral genome was sequenced and assembled by next-generation sequencing (NGS) Illumina MiSeq of small interfering RNA purified from symptomatic watermelon fruit, producing 92% genome coverage, and RT-PCR amplification and Sanger sequencing to close the genome gaps, validating the NGS sequence. The complete SqVYV-IL genome sequence shared 84% nucleotide sequence identity with the two complete genomes of SqVYV isolates from Florida, and 91% identity with the deduced amino acid sequence of the viral polyprotein.
2014年春季,在约旦和以色列贝特谢安山谷露天种植的1000公顷西瓜植株(西瓜)上观察到了不常见的西瓜病害症状。这些症状表现为系统性枯萎、叶片发黄,叶片和茎部出现坏死,在某些情况下导致植株枯死、果实外果皮变质以及果肉中果皮腐烂,致使果实无法上市销售。从西瓜外果皮和坏死叶片中进行了病毒纯化,透射电子显微镜显示病毒粒子呈柔韧的丝状形态。该病害可通过机械接种从有症状的果实传播,也可通过烟粉虱从有症状的葫芦科作物传播至健康的葫芦科作物。使用针对衣壳蛋白基因的特异性引物,对南瓜脉黄化病毒(SqVYV)的纯化病毒制剂进行了逆转录聚合酶链反应(RT-PCR)检测,结果显示出预期的扩增子大小。通过对从有症状的西瓜果实中纯化的小干扰RNA进行新一代测序(NGS)Illumina MiSeq,对完整的病毒基因组进行了测序和组装,基因组覆盖率达到92%,并通过RT-PCR扩增和桑格测序填补基因组缺口,验证了NGS序列。完整的SqVYV-IL基因组序列与来自佛罗里达州的SqVYV分离株的两个完整基因组的核苷酸序列同一性为84%,与病毒多聚蛋白的推导氨基酸序列的同一性为91%。