WIĘSYK, Aneta; CANDRESSE, Thierry; ZAGORSKI, Wlodzimierz; GORA-SOCHAKA, Anna

doi:10.1099/vir.0.026286-0

Métadonnées

Afficher la notice complète

Licence d’utilisation du document

WIĘSYK, Aneta
Institute of Biochemistry and Biophysics

CANDRESSE, Thierry
Biologie du fruit et pathologie [BFP]

ZAGORSKI, Wlodzimierz
Institute of Biochemistry and Biophysics

Langue

Article de revue

Ce document a été publié dans

Journal of General Virology. 2011, vol. 92, n° 2, p. 457-466

Microbiology Society

Résumé en anglais

In an effort to study sequence space allowing the recovery of viable potato spindle tuber viroid (PSTVd) variants we have developed an in vivo selection (Selex) method to produce and bulk-inoculate by agroinfiltration large PSTVd cDNA banks in which a short stretch of the genome is mutagenized to saturation. This technique was applied to two highly conserved 6 nt-long regions of the PSTVd genome, the left terminal loop (TL bank) and part of the polypurine stretch in the upper strand of pre-melting loop 1 (PM1 bank). In each case, PSTVd accumulation was observed in a large fraction of bank-inoculated tomato plants. Characterization of the progeny molecules showed the recovery of the parental PSTVd sequence in 89 % (TL bank) and 18 % (PM1 bank) of the analysed plants. In addition, viable and genetically stable PSTVd variants with mutations outside of the known natural variability of PSTVd were recovered in both cases, although at different rates. In the case of the TL region, mutations were recovered at five of the six mutagenized positions (357, 358, 359, 1 and 3 of the genome) while for the PM1 region mutations were recovered at all six targeted positions (50–55), providing significant new insight on the plasticity of the PSTVd genome.< Réduire

Mots clés

VIROID

EVOLUTION

VARIANT

Mots clés en anglais

MUTAGENESE

SELEX

TERMINAL HAIRPIN

POLYPURINE TRACT

VM DOMAIN