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Revisado por pares
First Report of Tomato Chlorosis Virus Infecting Cucumber in Brazil
2019; American Phytopathological Society; Volume: 104; Issue: 2 Linguagem: Inglês
10.1094/pdis-07-19-1490-pdn
ISSN1943-7692
AutoresVinicius Henrique Bello, Eduardo Silva Gorayeb, Luís Fernando Maranho Watanabe, Bruno Rossitto De Marchi, Marcos Roberto Ribeiro‐Júnior, Eduardo Vicentin, F. B. da Silva, Renate Krause‐Sakate,
Tópico(s)Plant Pathogenic Bacteria Studies
ResumoHomePlant DiseaseVol. 104, No. 2First Report of Tomato Chlorosis Virus Infecting Cucumber in Brazil PreviousNext DISEASE NOTES OPENOpen Access licenseFirst Report of Tomato Chlorosis Virus Infecting Cucumber in BrazilV. H. Bello, E. S. Gorayeb, L. F. M. Watanabe, B. R. De Marchi, M. R. Ribeiro-Junior, E. Vicentin, F. B. da Silva, and R. Krause-SakateV. H. Bello†Corresponding author: V. H. Bello; E-mail Address: vhbello@hotmail.comhttp://orcid.org/0000-0002-2869-8119Faculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, E. S. Gorayebhttp://orcid.org/0000-0001-8594-6070Faculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, L. F. M. WatanabeFaculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, B. R. De Marchihttp://orcid.org/0000-0002-4401-2594Faculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, M. R. Ribeiro-Juniorhttp://orcid.org/0000-0002-5024-1801Faculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, E. VicentinFaculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, F. B. da SilvaFaculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil, and R. Krause-SakateFaculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil AffiliationsAuthors and Affiliations V. H. Bello † E. S. Gorayeb L. F. M. Watanabe B. R. De Marchi M. R. Ribeiro-Junior E. Vicentin F. B. da Silva R. Krause-Sakate Faculdade de Ciências Agronômicas, UNESP, Botucatu, SP, Brazil Published Online:30 Nov 2019https://doi.org/10.1094/PDIS-07-19-1490-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Cucumber plants (Cucumis sativus L.) exhibiting chlorosis and mottling symptoms on young leaves were found in a commercial greenhouse, in the county of Oleo, State of São Paulo, Brazil, in February 2019. The crop was highly infested by whiteflies. To identify the whitefly species in the greenhouse, total DNA was extracted from 10 whiteflies according to the Chelex protocol (Walsh et al. 1991). The DNA samples were subjected to initial PCR amplification of a mitochondrial cytochrome oxidase subunit 1 fragment using the primer pair 2196Bt and C012/Bt-sh2 (Mugerwa et al. 2018), followed by amplicon restriction fragment length polymorphism (RFLP) analysis of all samples and amplicon Sanger sequencing of one sample. The whiteflies were identified as Bemisia tabaci Mediterranean species (MED, also known as Q biotype) based on the RFLP analysis and BLASTn search of the amplicon sequence (GenBank accession no. MN177614). To identify the agent that might have caused the disease, total RNA was extracted from five symptomatic leaves and 15 asymptomatic fruits using the PureLink Viral RNA/DNA Kit (Invitrogen). The RNA was pooled and used for construction of a cDNA library using the Complete ScriptSeq Kit (Epicenter, Illumina) and transcriptome sequencing with the Illumina HiSeq2500 platform (Roossinck et al. 2015). A total of 33,774,040 raw reads (average length 50 bp) were generated by high-throughput sequencing (HTS). The raw reads were trimmed and de novo assembled using CLC Genomics Workbench software version 9.0.3. The 1,441 obtained contigs (average length 350 bp) were subjected to a BLASTn using Geneious software version 9.1.5. The contigs then were blasted against the NCBI virus genome database. This analysis revealed the complete nucleotide sequence of two contigs of 8,573 bp (GenBank accession no. MN172419) and 8,223 bp (GenBank accession no. MN172420) in length, showing 99.8 and 99.6% sequence identity with tomato chlorosis virus (ToCV) RNA1 (JQ952600) and RNA2 (JQ952601), respectively. No other viruses were detected. Subsequently, individual RNA samples were tested for ToCV. Reverse transcription PCR (RT-PCR) was performed with a newly designed primer pair, p22-7537 (5′-ATAGGTACAATTTACCACCG-3′)/p22-8371-R (5′-GAATGTCAACAAAACACCTACT-3′), which was anticipated to result in an amplicon of 834 bp from RNA1 of ToCV. One leaf sample and three fruits tested positive by the RT-PCR assay. In order to validate the results, the amplicon from one sample was Sanger sequenced. The amplicon (GenBank accession no. MN164488) showed 100% nucleotide sequence identity with the HTS-derived sequence and 99.6% with a tomato ToCV isolate (MF415521) from Brazil. To fulfill Koch's postulates, adults of B. tabaci MED were transferred to ToCV-infected cucumber leaves for 72 h for virus acquisition, followed by inoculation access (50 B. tabaci per plant) to 10 tomato (cv. Santa Clara) and 10 cucumber (cv. Aodai) plants. Forty-five days postinoculation, ToCV was detected by RT-PCR in one tomato and three cucumber plants. The tomato plant infected with the ToCV-cucumber isolate developed typical ToCV symptoms, whereas the cucumber plants developed chlorosis and mottling in young leaves. ToCV is one of the most common whitefly-transmitted viruses of solanaceous plants in Brazil (Macedo et al. 2019), and its impact depends on vector prevalence and abundance (Fariña et al. 2019). These findings may raise a great concern for cucumber growers in Brazil, because B. tabaci MED can efficiently transmit ToCV (Bello et al. 2019) and is often found on cucumber crops grown in greenhouses (Moraes et al. 2018). Additionally, cucumbers may serve as inoculum sources of ToCV for other economically important solanaceous crops. To the best of our knowledge, this is the first report of ToCV infecting cucumber plants.The author(s) declare no conflict of interest.References:Bello, V. H., et al. 2019. Phytoparasitica 47:293. https://doi.org/10.1007/s12600-019-00729-y Crossref, ISI, Google ScholarFariña, A. E., et al. 2019. Plant Dis. 103:1132. https://doi.org/10.1094/PDIS-11-18-1941-RE Link, ISI, Google ScholarMacedo, M. A., et al. 2019. Plant Pathol. 68:72. https://doi.org/10.1111/ppa.12920 Crossref, ISI, Google ScholarMoraes, L. A., et al. 2018. Sci. Rep. 8:14589. https://doi.org/10.1038/s41598-018-32913-1 Crossref, Google ScholarMugerwa, H., et al. 2018. Sci. Rep. 8:2734. https://doi.org/10.1038/s41598-018-20956-3 Crossref, Google ScholarRoossinck, M. J., et al. 2015. Phytopathology 105:716. https://doi.org/10.1094/PHYTO-12-14-0356-RVW Link, ISI, Google ScholarWalsh, P. S., et al. 1991. Biotechniques 10:506. ISI, Google ScholarThe author(s) declare no conflict of interest.Funding: This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES), Finance Code 001; Fundação de Amparo à Pesquisa do Estado de São Paulo (processes number 2017/21588-7 and 2018/18274-3); and Conselho Nacional de Desenvolvimento Científico e Tecnológico (405684/2018-5 and 303411/2018-0).DetailsFiguresLiterature CitedRelated Vol. 104, No. 2 February 2020SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionSymptom of maize ear rot caused by Fusarium sporotrichioides (B. B. Wang et al.). Photo credit: C. X. Duan. Systemic symptoms of alfalfa mosaic virus (AMV) isolate CaM on leaves of potato (X. Z. Nie et al.). Photo credit: X. Z. Nie. Metrics Downloaded 2,817 times Article History Issue Date: 31 Jan 2020Published: 30 Nov 2019First Look: 23 Sep 2019Accepted: 17 Sep 2019 Page: 603 Information© 2020 The American Phytopathological SocietyFundingFundação de Amparo à Pesquisa do Estado de São PauloGrant/Award Number: 2017/21588-7Grant/Award Number: 2018/18274-3Conselho Nacional de Desenvolvimento Científico e TecnológicoGrant/Award Number: 303411/2018-0Grant/Award Number: 405684/2018-5KeywordsToCVcucumberMEDBemisia tabaciThe author(s) declare no conflict of interest.Cited ByPerformance and preference of Bemisia tabaci on cucumbers: Understanding the recent outbreaks of Mediterranean cryptic species in BrazilCrop Protection, Vol. 143Population Dynamics of Whiteflies and Associated Viruses in South America: Research Progress and Perspectives28 November 2020 | Insects, Vol. 11, No. 12Outbreaks of Bemisia tabaci Mediterranean species in vegetable crops in São Paulo and Paraná States, Brazil28 January 2020 | Bulletin of Entomological Research, Vol. 7
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