title: Divergence of host range and biological properties between natural isolate and full-length infectious cDNA clone of the Beet mild yellowing virus 2ITB.
abstract: Plant infection by poleroviruses is restricted to phloem tissues, preventing any classical leaf rub inoculation with viral RNA or virions. Efficient virus inoculation to plants is achieved by viruliferous aphids that acquire the virus by feeding on infected plants. The use of promoter-driven infectious cDNA is an alternative means to infect plants and allows reverse genetic studies to be performed. Using Beet mild yellowing virus isolate 2ITB (BMYV-2ITB), we produced a full-length infectious cDNA clone of the virus (named BMYV-EK) placed under the control of the T7 RNA polymerase and the Cauliflower mosaic virus 35S promoters. Infectivity of the engineered BMYV-EK virus was assayed in different plant species and compared with that of the original virus. We showed that in vitro- or in planta-derived transcripts were infectious in protoplasts and in whole plants. Importantly, the natural aphid vector Myzus persicae efficiently transmitted the viral progeny produced in infected plants. By comparing agroinoculation and aphid infection in a host range assay, we showed that the engineered BMYV-EK virus displayed a similar host range to BMYV-2ITB, except for Nicotiana benthamiana, which proved to be resistant to systemic infection with BMYV-EK. Finally, both the BMYV-EK P0 and the full-length clone were able to strongly interfere with post-transcriptional gene silencing.
authors.affiliation: Institut de Biologie Moléculaire des Plantes, Laboratoire Propre du CNRS (UPR 2357) Conventionné avec l'Université de Strasbourg, 12 rue du Général Zimmer, 67084, Strasbourg, France; SESVanderHave, Industriepark 15, Soldatenplein Z2, 3300, Tienen, Belgium.
authors.forename: Elodie
authors.initials: E
authors.lastname: Klein
authors.forename: Véronique
authors.initials: V
authors.lastname: Brault
authors.forename: Delphine
authors.initials: D
authors.lastname: Klein
authors.forename: Guy
authors.initials: G
authors.lastname: Weyens
authors.forename: Marc
authors.initials: M
authors.lastname: Lefèbvre
authors.forename: Véronique
authors.initials: V
authors.lastname: Ziegler-Graff
authors.forename: David
authors.initials: D
authors.lastname: Gilmer
mesh_terms: D000818:Animals; D001042:Aphids / Q000821:virology; D017360:Arabidopsis / Q000821:virology; D001483:Base Sequence; D027461:Beta vulgaris / Q000821:virology*; D002999:Clone Cells; D018076:DNA, Complementary / Q000235:genetics*; D020868:Gene Silencing; D016679:Genome, Viral / Q000235:genetics; D058507:Host Specificity; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000235:genetics* / Q000302:isolation & purification*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011523:Protoplasts / Q000821:virology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger