CHV1 infection of the filamentous ascomycete Cryphonectria parasitica, the causal agent of chestnut blight, results in hypovirulent phenotype. Previous studies showed that virus replication is associated with host membrane vesicles. Heavy water-Ficoll gradients were used to separate subsets of vesicles from the microsomal fraction of virus infected and uninfected strains of C. parasitica. Vesicle accumulation was found to occur in viral infected strains over time whereas in healthy mycelia the amount of these vesicles dramatically decreases after an early proliferation. A subset of coated vesicles was found to contain viral dsRNA and proteins reacting to CHV1 helicase and polymerase antibodies. This coated vesicle fraction from infected strains contains an enriched protein band that in Western blots reacts with anticlathrin heavy chain antibodies. The finding that clathrin accumulates in hypovirulent strains of C. parasitica prompted us to clone the clathrin heavy chain gene and the middle component of its adapter complex 1 to investigate their role in this important plant pathogen.
Characterization of vesicle subsets in CHV1 infected C. parasitica
Turina M;
2001
Abstract
CHV1 infection of the filamentous ascomycete Cryphonectria parasitica, the causal agent of chestnut blight, results in hypovirulent phenotype. Previous studies showed that virus replication is associated with host membrane vesicles. Heavy water-Ficoll gradients were used to separate subsets of vesicles from the microsomal fraction of virus infected and uninfected strains of C. parasitica. Vesicle accumulation was found to occur in viral infected strains over time whereas in healthy mycelia the amount of these vesicles dramatically decreases after an early proliferation. A subset of coated vesicles was found to contain viral dsRNA and proteins reacting to CHV1 helicase and polymerase antibodies. This coated vesicle fraction from infected strains contains an enriched protein band that in Western blots reacts with anticlathrin heavy chain antibodies. The finding that clathrin accumulates in hypovirulent strains of C. parasitica prompted us to clone the clathrin heavy chain gene and the middle component of its adapter complex 1 to investigate their role in this important plant pathogen.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
