A Novel Strain of Pepper Leafroll Virus Infecting Common Bean and Soybean in Ecuador

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HomePlant DiseaseVol. 103, No. 1A Novel Strain of Pepper Leafroll Virus Infecting Common Bean and Soybean in Ecuador PreviousNext DISEASE NOTES OPENOpen Access licenseA Novel Strain of Pepper Leafroll Virus Infecting Common Bean and Soybean in EcuadorE. Fiallo-Olivé, D. T. Chirinos, R. Castro, and J. Navas-CastilloE. Fiallo-Olivé†Corresponding author: E. Fiallo-Olivé; E-mail: E-mail Address: [email protected]http://orcid.org/0000-0002-2460-1199, D. T. Chirinos, R. Castro, and J. Navas-CastilloAffiliationsAuthors and Affiliations E. Fiallo-Olivé † , Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora", Consejo Superior de Investigaciones Científicas – Universidad de Málaga (IHSM-CSIC-UMA), 29750 Algarrobo-Costa, Málaga, Spain D. T. Chirinos , Facultad de Ingeniería Agronómica, Universidad Técnica de Manabí, Manabí, Ecuador R. Castro , Universidad Agraria del Ecuador, Guayaquil, Ecuador J. Navas-Castillo , IHSM-CSIC-UMA, 29750 Algarrobo-Costa, Málaga, Spain. Published Online:6 Nov 2018https://doi.org/10.1094/PDIS-06-18-1076-PDNAboutSections ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Symptoms resembling those caused by begomoviruses (Geminiviridae: Begomovirus) have been observed in common bean and other leguminous crops in Ecuador in the last few years. The only begomovirus species found infecting legumes (common bean and cowpea) in the country to date is cabbage leaf curl virus (CabLCV) (Fiallo-Olivé et al. 2018). During May to June 2017, a survey of leguminous plants was conducted in the Ecuadorian localities of Santa Elena (Santa Elena province), Milagro (Guayas province), and Pallatanga (Chimborazo province). Twenty-four plants showing leaf deformation and/or mosaic symptoms were sampled, including common bean (Phaseolus vulgaris, n = 21), soybean (Glycine max, n = 2), and pigeon pea (Cajanus cajan, n = 1). Most common bean plants were infested with whiteflies (Hemiptera: Aleyrodidae) that were also collected. Total DNA was extracted from leaf samples and used as template in polymerase chain reaction (PCR) using specific primers for CabLCV (MA2540 [5′-GATGTTGAAGCATCTGCAAACATTC-3′] and MA2541 [5′-CTAGGAACATCAGGGCTTCTCAAC-3′]) (Fiallo-Olivé et al. 2018). All samples were negative for the presence of CabLCV. DNA extracts were then used as templates in rolling-circle amplification (RCA) using φ29 DNA polymerase (TempliPhi kit, GE Healthcare). Digestion of RCA products with a set of restriction enzymes yielded identical restriction patterns for six samples (five common bean and one soybean), suggesting the presence of a bipartite begomovirus. BamHI and NcoI fragments of approximately 2.6 kbp from a common bean plant were cloned in pBluescript II SK(+), and two selected clones were sequenced (Macrogen, Seoul, South Korea). BLAST analysis showed that the clones corresponded to begomoviral DNA-A (BamHI, 2,580 nt, GenBank accession no. MH481901) and DNA-B (NcoI, 2,561 nt, MH481902). Pairwise identity scores with isolates selected after BLAST analysis were calculated with SDT (Muhire et al. 2014). Cloned DNA-A and DNA-B showed the highest identity (93 and 92%, respectively) with pepper leafroll virus (PepLRV) (KC769819 and KC769820), a bipartite begomovirus identified infecting pepper, common bean, and tomato in Peru (Martínez-Ayala et al. 2014). To confirm the identity of the begomovirus present in the rest of the samples, PCR was performed with specific primers whose design was based on the cloned PepLRV DNA-A (MA2602 [5′-GTTAATGGACAATGAGGGTATGTG-3′] and MA2603 [5′-CACGAGGAGAATAATTGGCGTTAC-3′]) using Taq DNA polymerase (BIOTAQ, Bioline) and thermocycler conditions consisting of 35 cycles with 45 s annealing steps at 57°C. DNA fragments of the expected size (452 bp) were amplified from the six samples, and direct sequencing showed them to be identical to the cloned PepLRV DNA-A, supporting the presence of this virus. Based on the species/strain demarcation threshold for the viruses in the genus Begomovirus (Brown et al. 2015), the isolate characterized here belongs to a new strain of PepLRV. DNA was extracted from individual whiteflies (n = 42) using a Chelex-based method, and PCR was carried out using mitochondrial cytochrome oxidase I gene primers, C1-J-2195 (5′-TTGATTTTTTGGTCATCCAGAAGT-3′) and Btab-uni-PrimerR (5′-CTTAAATTTACTGCACTTTCTGCCAYATTAG-3′). Direct sequencing of PCR products showed that seven sequences (one deposited in GenBank with accession no. MH481903) were identical to numerous sequences of Bemisia tabaci Middle East-Asia Minor 1 (MEAM1, formerly B biotype), known to be an efficient vector of begomoviruses. To our knowledge, this is the first report of PepLRV in Ecuador and of soybean as a host for the virus, which had been identified only in Peru to date. Also, this is the first molecular evidence of the presence of B. tabaci MEAM1 in the country.References:Brown, J. K., et al. 2015. Arch. Virol. 160:1593. https://doi.org/10.1007/s00705-015-2398-y Crossref, ISI, Google ScholarFiallo-Olivé, E., et al. 2018. Plant Dis.doi.org/10.1094/PDIS-05-18-0817-PDN. Google ScholarMartínez-Ayala, A., et al. 2014. Ann. Appl. Biol. 164:62. https://doi.org/10.1111/aab.12074 Crossref, ISI, Google ScholarMuhire, B., et al. 2014. PLoS One 9:e108277. https://doi.org/10.1371/journal.pone.0108277 Crossref, ISI, Google ScholarFunding: This work was supported by grants from Consejo Superior de Investigaciones Científicas (CSIC, Spain; 2016SU0020) and Ministerio de Economía, Industria y Competitividad (MINECO, Spain; AGL2016-75819-C2-2-R) co-financed by the European Regional Development Fund. E. Fiallo-Olivé was recipient of a "Juan de la Cierva-Incorporación" contract (MINECO, Spain).DetailsFiguresLiterature CitedRelated Vol. 103, No. 1 January 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 Metrics Article History Issue Date: 4 Jan 2019Published: 6 Nov 2018First Look: 16 Aug 2018Accepted: 14 Aug 2018 Page: 167 Information© 2019 The American Phytopathological SocietyFundingConsejo Superior de Investigaciones CientíficasGrant/Award Number: 2016SU0020Ministerio de Economía, Industria y CompetitividadGrant/Award Number: AGL2016-75819-C2-2-RCited byBemisia tabaci (MEAM1) (silverleaf whitefly)CABI Compendium, Vol. CABI CompendiumPepper leafroll virusCABI Compendium, Vol. CABI CompendiumAn Annotated List of Legume-Infecting Viruses in the Light of Metagenomics10 July 2021 | Plants, Vol. 10, No. 7Biological and molecular characterization of bean bushy stunt virus, a novel bipartite begomovirus infecting common bean in northwestern Argentina28 February 2021 | Archives of Virology, Vol. 166, No. 5Population Dynamics of Whiteflies and Associated Viruses in South America: Research Progress and Perspectives28 November 2020 | Insects, Vol. 11, No. 12Glycine max (Soybean)6 June 2020Phaseolus vulgaris (Common bean/French bean/Snap bean)6 June 2020