An immunofluorescent cytochemical technique applying micromanipulation to detect microtubules in plant tissues inoculated with fungal spores

Abstract

An immunofluorescent cytochemical technique to detect microtubules was developed to examine the involvement of microtubules in incipient responses of barley coleoptile cells to fungal attacks. Infiltration of antibodies into target cells was promoted by scratching cell walls of chemically-fixed coleoptile cells with a micromanipulator. In uninoculated control cells, cortical microtubules were arranged obliquely or transversely to the longitudinal axis of the cells. On the other hand, in coleoptile cells which had been fixed 8–10 h after inoculation with a nonpathogen, Erysiphe pisi, microtubules gathered in coleoptile cells beneath mature appressoria of the fungus. When coleoptiles had been fixed 12 h after inoculation, many of the microtubules gathered around an incipient, small papilla, giving a network appearance. The present technique would be helpful for studying the role of microtubules in host cell responses to fungal attack. Key words: microtubules, immunofluorescent cytochemistry, barley coleoptile, Erysiphe pisi.