The objective of the study was to develop methods for the specific
and rapid identification of Tuber spp. to aid ecological and
taxonomical investigations of Tuber spp. and ectomycorrhizal fungi
in general. Antisera raised against Tuber melanosporum,
T. magnatum, Boletus edulis and Tricholoma matsutake
and monoclonal antibodies produced against T. melanosporum were
used to
develop various immunological assays such as ELISA, Dot-blot and Western
blot. A simple squash Dot-blot was used to visualize
mycorrhizal infection of inoculated host plants grown in tube cultures
and experimental truffières.
Most antisera reacted strongly with homologous antigens from ascomata
or mycelia and mycorrhizal root-tips. Their specificity
allowed differentiation between Tuber spp. and other ectomycorrhizal
fungi. Differentiation among Tuber spp. was possible by
Western blot but not by ELISA or Dot-blot assay. The results suggested
that specificity of the antisera was confined to genus level.
In two cases specificity was increased by absorption with cross-reactive
antigens.
Eleven monoclonal antibodies were produced against T. melanosporum.
Two were directed against proteinaceous material, the
others probably against various carbohydrate determinants. Unfortunately
they could not be used for differentially recognizing
T. melanosporum as they all cross-reacted at least with extracts
of four other ectomycorrhizal fungi.