Seeds of the orchid species, Spiranthes sinensis (Pers.)
Ames, were sterilized and germinated in vitro with the
symbiotic fungus Ceratobasidium cornigerum (Bourdot) Rogers. Colonized
embryos developed into protocorms
and these were examined for changes in microtubule arrays, after initial
invasion of fungal hyphae into embryos
and during peloton formation and degradation. Methods utilized to detect
microtubules included immunofluorescence combined with laser scanning confocal
microscopy, conventional transmission electron microscopy
combined with morphometric analysis, and immunogold labelling. Microtubules
were regularly found in close
association with intracellular hyphae and degraded hyphal masses. Cortical
microtubules disappear during peloton
formation but reappear in cells that show fungal lysis. With conventional
transmission electron microscopy and
immunogold labelling the microtubules associated with fungal hyphae and
degenerated hyphal masses were
located close to the perifungal membrane that separates fungal hyphae from
protocorm cytoplasm.