{"pk":41458,"title":"In situ localization of citrus exocortis viroid RNA using an optimized RNAscope™ assay","subtitle":null,"abstract":"Due to their small size, locating pathogenic RNAs, such as viroids, in plant tissue and cell organelles has been difficult. Viroids are small circular single-stranded RNA plant pathogens that reduce plant growth, vigor, and yield in economically important crops such as potato, tomato, hops and citrus. Viroid infections in plants are largely diagnosed by dot blot hybridization, polyacrylamide gel electrophoresis (PAGE) or gels, or real-time quantitative polymerase chain reaction (qPCR). Because traditional plant \nin situ\n hybridization studies for viroids are often limited by the lack of signal amplification and binding specificity due to the small target sequence, we examined the use of RNAscope™ (Advanced Cell Diagnostics Inc., Newark, CA). This\n in situ\n hybridization method increases the detection by amplifying the signal of a single target, to detect the cellular distribution of citrus exocortis viroid (CEVd) with higher sensitivity and specificity. We found that after optimization, CEVd was localized in nuclei of infected cells as clearly distinguishable punctate red dots visible with light microscopy at low magnification, suggesting that the RNAscope™ can be used to study viroids \nin situ\n.","language":"en","license":{"name":"Creative Commons Attribution 4.0","short_name":"CC BY 4.0","text":"Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.\n\nNo additional restrictions — You may not apply legal terms or technological measures that legally restrict others from doing anything the license permits.","url":"https://creativecommons.org/licenses/by/4.0"},"keywords":[{"word":"Viroid, Exocortis, CEVd, RNAscope, Citrus, Plant Pathogen"}],"section":"Articles","is_remote":true,"remote_url":"https://escholarship.org/uc/item/93h2n20h","frozenauthors":[{"first_name":"Daniel","middle_name":"","last_name":"Stanton","name_suffix":"","institution":"Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA","department":"None"},{"first_name":"Scott","middle_name":"J.","last_name":"Harper","name_suffix":"","institution":"Department of Plant Pathology, Washington State University, Prosser, WA, USA.","department":"None"},{"first_name":"Sarah","middle_name":"J.","last_name":"Cowell","name_suffix":"","institution":"Department of Plant Pathology, Washington State University, Prosser, WA, USA.","department":"None"},{"first_name":"Ron","middle_name":"","last_name":"Brlansky","name_suffix":"","institution":"Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA","department":"None"}],"date_submitted":"2021-05-15T03:50:16Z","date_accepted":"2021-05-15T03:50:16Z","date_published":"2021-09-27T07:00:00Z","render_galley":null,"galleys":[{"label":"","type":"pdf","path":"https://journalpub.escholarship.org/iocv_journalcitruspathology/article/41458/galley/31036/download/"}]}