title: Recombination-based generation of the agroinfectious clones of Peanut stunt virus.
abstract: Full-length cDNA clones of Peanut stunt virus strain P (PSV-P) were constructed and introduced into Nicotiana benthamiana plants via Agrobacterium tumefaciens. The cDNA fragments corresponding to three PSV genomic RNAs and satellite RNA were cloned into pGreen binary vector between Cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator employing seamless recombinational cloning system. The plasmids were delivered into A. tumefaciens, followed by infiltration of hosts plants. The typical symptoms on systemic leaves of infected plants similar to those of wild-type PSV-P were observed. The presence of the virus was confirmed by means of RT-PCR and Western blotting. Re-inoculation to N. benthamiana, Phaseolus vulgaris, and Pisum sativum resulted in analogous results. Generation of infectious clones of PSV-P enables studies on virus-host interaction as well as revealing viral genes functions.
authors.affiliation: Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection - National Research Institute, Władysława Węgorka 20 St, 60-318, Poznań, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Wrzesińska
authors.affiliation: Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection - National Research Institute, Władysława Węgorka 20 St, 60-318, Poznań, Poland.
authors.forename: Przemysław
authors.initials: P
authors.lastname: Wieczorek
authors.affiliation: Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection - National Research Institute, Władysława Węgorka 20 St, 60-318, Poznań, Poland. Electronic address: olaob@o2.pl.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Obrępalska-Stęplowska
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D000594:Amino Acid Oxidoreductases / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D018532:Pisum sativum / Q000821:virology; D027805:Phaseolus / Q000821:virology; D010935:Plant Diseases / Q000821:virology; D018515:Plant Leaves / Q000821:virology; D010942:Plant Viruses / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000235:genetics; D011995:Recombination, Genetic; D013728:Terminator Regions, Genetic; D014026:Nicotiana / Q000821:virology
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase
keywords: Agrobacterium tumefaciens; Isothermal recombination; Peanut stunt virus; Seamless recombination; Viral infectious clones; cDNA