Articles
CHARACTERIZATION OF STOCKY PRUNE VIRUS, A NEW NEPOVIRUS DETECTED IN FRENCH PRUNES
Article number
472_18
Pages
175 – 182
Language
Abstract
A virus was transmitted to Chenopodium murale from GF305 indicator peach seedlings grafted with prunes (Prune d’Ente) showing symptoms of the stocky prune disease (Desvignes, 1990) a disease suspected to be caused by a soil-borne virus.
A purification protocol was developed, based on the use of bentonite for plant extract clarification.
The purified virus sedimented as a single peak in sucrose gradients and showed spherical, nepovirus-like particles upon electron microscope observation.
Analysis of purified particles indicated the presence of two single stranded RNAs of approximately 7.5 and 3.7 kb while double stranded RNAs of equivalent size were detected in infected C. murale. Analysis of the protein content of the purified particles revealed a complex pattern: three major bands of approximately 24.5, 23.5 and 23.2 kDa were consistently observed, together with weaker bands of approximately 52, 41.5 and 40 kDa, which varied in intensity from preparation to preparation.
Partial sequencing of cDNA clones obtained from genomic RNAs revealed significant homology with members of the Comoviridae family.
A 32P-labeled riboprobe prepared from a CDNA clone detected the virus in symptomatic GF305 peach seedlings inoculated with a purified virus preparation.
Taken together, these results indicate that the virus causing the stocky prune disease possesses a set of properties unmatched by previously known nepoviruses.
The name stocky prune virus (StPV) is therefore proposed for it together with its assignment as a tentative member of the nepovirus genus.
A purification protocol was developed, based on the use of bentonite for plant extract clarification.
The purified virus sedimented as a single peak in sucrose gradients and showed spherical, nepovirus-like particles upon electron microscope observation.
Analysis of purified particles indicated the presence of two single stranded RNAs of approximately 7.5 and 3.7 kb while double stranded RNAs of equivalent size were detected in infected C. murale. Analysis of the protein content of the purified particles revealed a complex pattern: three major bands of approximately 24.5, 23.5 and 23.2 kDa were consistently observed, together with weaker bands of approximately 52, 41.5 and 40 kDa, which varied in intensity from preparation to preparation.
Partial sequencing of cDNA clones obtained from genomic RNAs revealed significant homology with members of the Comoviridae family.
A 32P-labeled riboprobe prepared from a CDNA clone detected the virus in symptomatic GF305 peach seedlings inoculated with a purified virus preparation.
Taken together, these results indicate that the virus causing the stocky prune disease possesses a set of properties unmatched by previously known nepoviruses.
The name stocky prune virus (StPV) is therefore proposed for it together with its assignment as a tentative member of the nepovirus genus.
Authors
T. Candresse, R.P. Delbos, O. Le Gall, J. Dunez, J.C. Desvignes
Keywords
etiology, maladie de Brugères, prune d’ente, virus purification, cloning
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