Abstract:
‘Mushroom Virus X’ (MVX) disease of the cultivated mushroom Agaricus bisporus
first arose in UK during the 1990’s. This disease resulted in devastating crop losses in
the UK and gradually became more widespread (e.g. Netherlands and Eire). Up to
twenty-six, non-encapsidated, double stranded RNA (dsRNA) elements have been
found to be associated with diseased mushrooms, and these are believed to be the result
of a complex of viruses. Although considerable data has accumulated on the symptoms
of infection, aetiological sources, epidemiology and molecular characterization of the
MVX dsRNA elements are limited. Research described in this thesis focused principally
on sequence characterization of a frequently occurring dsRNA element (
MVX
14.4),
which was shown to be a novel Endornavirus. Assigned ‘Agaricus bisporus
endornavirus 1’ (AbEV1), this represents the first endornavirus known to infect edible
mushrooms. AbEV1 is the first MVX element to be fully sequenced. Putative domains
for RNA-dependent RNA polymerase (RdRp), helicase and glycosyltransferase were
identified and used in comparisons with other viruses. Characterization of an AbEV1-
type dsRNA found in a culture sample derived from a wild Agaricus bisporus collection
indicates a possible source of the MVX dsRNA infections. Epidemiological studies
were used to demonstrate that the AbEV1 dsRNA was transmissible both vertically
through spores and horizontally by mycelial anastomosis between infected donor and
MVX free acceptor strains. As a first step in the effort to understand the role of
AbEV1in MVX infections and to investigate possible host defence mechanisms,
dsRNA hairpin sequences were introduced into A. bisporus by Agrobacterium-mediated
transformation. Both helicase and RdRp sequences were able to confer resistance to the
uptake of MVX dsRNA elements in transformants. These observations suggest that
homology-dependent gene silencing pathway(s) may be present in A. bisporus and
represent a residual antiviral defence mechanism. Advances and approaches developed
in this project open new opportunities to characterize the other dsRNA elements from
the MVX complex, to further our understanding of mycovirus infections and host
responses, and to investigate the origins of infectious dsRNA elements.
