title: Construction of Agrobacterium tumefaciens-mediated tomato black ring virus infectious cDNA clones.
abstract: Tomato black ring virus (TBRV, genus Nepovirus) infects a wide range of economically important plants such as tomato, potato, tobacco and cucumber. Here, a successful construction of infectious full-length cDNA clones of the TBRV genomic RNAs (RNA1 and RNA2) is reported for the first time. The engineered constructs consisting of PCR-amplified DNAs were cloned into binary vector pJL89 immediately downstream of a double cauliflower mosaic virus (CaMV) 35S promoter, and upstream of the hepatitis delta virus (HDV) ribozyme and nopaline synthase terminator (NOS). The symptoms induced on plants agroinoculated with both constructs were indistinguishable from those caused by the wild-type virus. The infectivity of obtained clones was verified by reinoculation to Nicotiana tabacum cv. Xanthi, Chenopodium quinoa and Cucumis sativus. The presence of viral particles and RNA was confirmed by electron microscopy and reverse transcription polymerase chain reaction, respectively. Constructed full-length infectious cDNA clones will serve as an excellent tool to study virus-host-vector interactions.
authors.affiliation: Institute of Plant Protection-National Research Institute, ul. Wł. Węgorka 20, 60-318 Poznań, Poland. Electronic address: a.zarzynska@iorpib.poznan.pl.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Zarzyńska-Nowak
authors.affiliation: Plant Pathology Department, University of California Davis, Davis, CA 95616, USA. Electronic address: ferriol_inm@gva.es.
authors.forename: Inmaculada
authors.initials: I
authors.lastname: Ferriol
authors.affiliation: Plant Pathology Department, University of California Davis, Davis, CA 95616, USA. Electronic address: bwfalk@ucdavis.edu.
authors.forename: Bryce W
authors.initials: BW
authors.lastname: Falk
authors.affiliation: Institute of Plant Protection-National Research Institute, ul. Wł. Węgorka 20, 60-318 Poznań, Poland. Electronic address: n.borodynko@iorpib.poznan.pl.
authors.forename: Natasza
authors.initials: N
authors.lastname: Borodynko-Filas
authors.affiliation: Institute of Plant Protection-National Research Institute, ul. Wł. Węgorka 20, 60-318 Poznań, Poland. Electronic address: b.hasiow@iorpib.poznan.pl.
authors.forename: Beata
authors.initials: B
authors.lastname: Hasiów-Jaroszewska
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000378:metabolism; D000594:Amino Acid Oxidoreductases / Q000235:genetics / Q000378:metabolism; D017796:Caulimovirus / Q000235:genetics / Q000378:metabolism; D027465:Chenopodium quinoa / Q000821:virology; D002999:Clone Cells; D003001:Cloning, Molecular / Q000379:methods*; D018553:Cucumis sativus / Q000821:virology; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000378:metabolism; D018551:Solanum lycopersicum / Q000821:virology*; D017860:Nepovirus / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000821:virology
chemical_list: D018076:DNA, Complementary; D016337:RNA, Catalytic; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase
keywords: TBRV; Tomato black ring virus; agroinfiltration; cDNA infectious clones