HomePlant DiseaseVol. 107, No. 7First Report of Passion Fruit Green Spot Virus in Yellow Passion Fruit (Passiflora edulis f. flavicarpa) in Casanare, Colombia PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Passion Fruit Green Spot Virus in Yellow Passion Fruit (Passiflora edulis f. flavicarpa) in Casanare, ColombiaAvijit Roy, Guillermo Leon M., Schyler Nunziata, Chellappan Padmanabhan, Yazmín Rivera, Ronald H. Brlansky, and John S. HartungAvijit Roy†Corresponding author: A. Roy; E-mail Address: [email protected]https://orcid.org/0000-0002-2268-9713USDA, Agricultural Research Service, Molecular Plant Pathology Laboratory, Beltsville, MD 20705, U.S.A., Guillermo Leon M.AGROSAVIA; Centro de Investigación La Libertad; Villavicencio, Meta, Colombia, Schyler NunziataUSDA, Animal Plant Health Inspection Service; Plant Protection and Quarantine, Science and Technology, Plant Pathogen Confirmatory Diagnostics Laboratory, Laurel, MD 20708, U.S.A., Chellappan PadmanabhanUSDA, Animal Plant Health Inspection Service; Plant Protection and Quarantine, Science and Technology, Plant Pathogen Confirmatory Diagnostics Laboratory, Laurel, MD 20708, U.S.A., Yazmín RiveraUSDA, Animal Plant Health Inspection Service; Plant Protection and Quarantine, Science and Technology, Plant Pathogen Confirmatory Diagnostics Laboratory, Laurel, MD 20708, U.S.A., Ronald H. BrlanskyUniversity of Florida Citrus Research and Education Center, Lake Alfred, FL 33850, U.S.A., and John S. HartungUSDA, Agricultural Research Service, Molecular Plant Pathology Laboratory, Beltsville, MD 20705, U.S.A.AffiliationsAuthors and Affiliations Avijit Roy1 † Guillermo Leon M.2 Schyler Nunziata3 Chellappan Padmanabhan3 Yazmín Rivera3 Ronald H. Brlansky4 John S. Hartung1 1USDA, Agricultural Research Service, Molecular Plant Pathology Laboratory, Beltsville, MD 20705, U.S.A. 2AGROSAVIA; Centro de Investigación La Libertad; Villavicencio, Meta, Colombia 3USDA, Animal Plant Health Inspection Service; Plant Protection and Quarantine, Science and Technology, Plant Pathogen Confirmatory Diagnostics Laboratory, Laurel, MD 20708, U.S.A. 4University of Florida Citrus Research and Education Center, Lake Alfred, FL 33850, U.S.A. Published Online:1 Jul 2023https://doi.org/10.1094/PDIS-09-22-2267-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articlePassiflora edulis, commonly known as passion fruit, is a vine species of passionflower native to South America. In Colombia, yellow passion fruit (P. edulis f. flavicarpa) is the most important in terms of net production and local consumption. Recently, two Brevipalpus-transmitted cileviruses, passion fruit green spot virus (PfGSV) and hibiscus strain of citrus leprosis virus C2 (CiLV-C2H), were detected in passion fruit in Brazil and Hawaii, respectively (Olmedo-Velarde et al. 2022; Ramos-González et al. 2020). CiLV-C2H infects both citrus and hibiscus in Colombia (Roy et al. 2015, 2018), but there was no report of PfGSV elsewhere apart from Brazil and Paraguay (da Costa-Rodrigues et al. 2022). Apart from emerging begomovirus diseases, five major viruses are known to infect passion fruit in Colombia: soybean mosaic virus (SMV), cowpea aphid-borne mosaic virus, passion fruit yellow mosaic virus, cucumber mosaic virus, and a tentative Gulupa bacilliform badnavirus A (Cardona et al. 2022). Current findings of CiLV-C2H in passion fruit and PfGSV in hibiscus motivated us to investigate the possibility of cilevirus infection in passion fruit in Colombia. During surveys, along with healthy yellow passion fruit leaves, five symptomatic plant samples from Meta and three from Casanare were collected and sent to the Molecular Plant Pathology Laboratory in Beltsville, MD, under APHIS permit. Samples from Meta showed leaf mottling, rugose mosaic, and leaf distortion, whereas leaf variegation, chlorotic spots, yellowing, green spots in senescent leaves and green vein banding were observed in the Casanare samples. Total RNA was extracted using RNeasy Plant Mini Kit (Qiagen, U.S.A.). To determine the potential cilevirus infection in these samples, three PfGSV specific primer pairs (Ramos-González et al. 2020) and one CiLV-C2 generic primer pair (Olmedo-Velarde et al. 2021) were used in RT-PCR assays. All five samples from Meta failed to produce either PfGSV or generic CiLV-C2 amplicons, but all three Casanare samples amplified 321, 244, and 299 nts of PfGSV-RNA1 and -RNA2 amplicons using C13F/C13R, C6F/C6R, and C8F/C8R primers, respectively. Bidirectional amplicon sequencing followed by BLASTn analysis revealed ≥99% nt identity with the PfGSV-RNA1 (MK804173) and -RNA2 (MK804174) genome sequences. An optimized Ribo-depleted library preparation protocol was used to prepare two cDNA libraries using the RNA extracts of PfGSV suspected positive (Casanare) and negative (Meta) samples (Padmanabhan et al. 2023). HTS libraries of Casanare and Meta samples resulted in 22.7 to 29.5 million raw reads, respectively. After adapter trimming and filtering, clean reads were mapped to the Arabidopsis thaliana reference genome and unmapped reads were de novo assembled (Padmanabhan et al. 2023). BLASTn analysis of the assembled contigs identified one to three contigs corresponding to PfGSV-RNA1 and -RNA2, respectively, from the Casanare sample and three contigs of SMV were identified in the Meta sample. No other virus sequence was obtained from either library. Assembled contigs of the Casanare sample covered 99.33% of the RNA1 and 94.42% of the RNA2 genome, with read depths of 64,474 and 119,549, respectively. Both RNA-1 (OP564895) and −2 (OP564896) segments of the Casanare isolate shared 99% nt identity with PfGSV isolate (MK804173 to 74). Meta sample contigs (OP564897) covered >99% of the SMV genome, which shared >99% nt identity with the Colombian SMV isolates (KY249378, MW655827). Our discovery identified PfGSV in Colombia; the first-time outside Brazil and Paraguay. The findings of PfGSV in passion fruit increase the potential threat and possibility of PfGSV movement via Brevipalpus sp. from passion fruit to other hosts including Citrus spp.The author(s) declare no conflict of interest.References:Cardona, D., et al. 2022. Arch. Phytopathol. Pflanzenschutz 55:1394. https://doi.org/10.1080/03235408.2022.2098588 Crossref, ISI, Google Scholarda Costa-Rodrigues, M., et al. 2022. New Dis. Rep. 45:e12080. https://doi.org/10.1002/ndr2.12080 Crossref, Google ScholarOlmedo-Velarde, A., et al. 2021. Front. Microbiol. 12:660237. 10.3389/fmicb.2021.660237 Crossref, ISI, Google ScholarOlmedo-Velarde, A., et al. 2022. Plant Dis. 106:2539. https://doi.org/10.1094/PDIS-10-21-2314-PDN Link, ISI, Google ScholarPadmanabhan, C., et al. 2023. Front. Plant Sci. 13:1058847. https://doi.org/10.3389/fpls.2022.1058847 Crossref, Google ScholarRamos-González, P. L., et al. 2020. Front. 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Plant Dis. 102:1675. https://doi.org/10.1094/PDIS-01-18-0150-PDN Link, ISI, Google ScholarFunding: Funding was provided by USDA-ARS in house project NP303-8042-22200-319-000D and Citrus Health Response Program (CHRP) Project No. PPQ-013258.The author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 107, No. 7 July 2023SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Article History Issue Date: 26 Jul 2023 Published: 1 Jul 2023 First Look: 5 Dec 2022 Accepted: 28 Nov 2022 Page: 2270 Information© 2023 The American Phytopathological SocietyFundingUSDA-ARSGrant/Award Number: NP303-8042-22200-319-000DCitrus Health Response ProgramGrant/Award Number: PPQ-013258KeywordsBrevipalpus-transmitted virusescilevirushigh-throughput sequencingKitaviridaepassion fruitpassion fruit green spot virussoybean mosaic virusThe author(s) declare no conflict of interest.PDF download