Mapping the determinant for necrotic response in Ourmia melon virus infection of Nicotiana benthamiana reveals the plasticity of agro-infectious clones and suggests a new standard for evaluating \"infectivity\" of cDNA clones: derived infectious sequence. (Abstract/Poster in atti di convegno)

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  • Mapping the determinant for necrotic response in Ourmia melon virus infection of Nicotiana benthamiana reveals the plasticity of agro-infectious clones and suggests a new standard for evaluating \"infectivity\" of cDNA clones: derived infectious sequence. (Abstract/Poster in atti di convegno) (literal)
Anno
  • 2013-01-01T00:00:00+01:00 (literal)
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  • Turina M., Rossi, M., Margaria P. , Ciuffo M. (2013)
    Mapping the determinant for necrotic response in Ourmia melon virus infection of Nicotiana benthamiana reveals the plasticity of agro-infectious clones and suggests a new standard for evaluating "infectivity" of cDNA clones: derived infectious sequence.
    in EMBO Workshop: Green viruses, from gene to landscape, Hyeres-les-Palmeiers, France, 7-11 September
    (literal)
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  • Turina M., Rossi, M., Margaria P. , Ciuffo M. (literal)
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Titolo
  • Mapping the determinant for necrotic response in Ourmia melon virus infection of Nicotiana benthamiana reveals the plasticity of agro-infectious clones and suggests a new standard for evaluating \"infectivity\" of cDNA clones: derived infectious sequence. (literal)
Abstract
  • We have recently published the assembly of an Ourmia melon virus (OuMV) agroinfectious clones able to reproduce necrotic symptoms on Nicotiana benthamiana similar to those obtained inoculating the original virus. During the cloning process we also serendipitously originated a set of clones corresponding to the three RNAs which originated a systemic viral infection only displaying mosaic-mottling later fully recovering in newly emerging leaves. We originated reassortant viruses to identify the virus segment responsible for eliciting necrotic response, and showed that the RNA1, coding for the viral RdRP, carried the determinant for the necrotic response. Then a full length sequence comparison with a wild-type clone for RNA1 revealed a 1 nt deletion in position 600 in a stretch of six cytosine residues in the coding region of the RdRP and 16 nt mutations distributed along the same RNA corresponding to three amino acids changes. A combination of chimeric cDNA clones and site directed mutagenesis demonstrated that the sole responsible for the necrotic response is a 1 nt mutation that results in 1 amino acid change (D to N position 364). Surprisingly some of the chimeric constructs were not infectious. We showed that the progeny RNA that accumulates in the plant is different from that of the two original cDNA, in particular replacing the cytosine deletion present in the mild segment at position 600 and mutating a C residue into G in position 2792 in the 3' UTR region of the necrotic allele. The chimeric clones combining the two variants were not infectious, whereas clones carrying only one of the different mutations could be corrected likely at the transcription step (either in vitro or in vivo in a T7 derived clone). Implications of such discovery are discussed and compared to results obtained inserting a 1 nt deletion in a commonly used agro-infectious clone derived from TMV. (literal)
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