Abstract: The network structure of cytoplasmic filaments of 10 nm in diameter was detected from callus cells of Arabidopsis thaliana (L.) Heynh by selective extraction combined with whole mount electron microscopy. Western blot analysis showed that the major filament components were 6 polypeptides, which reacted with keratin monoclonal antibody of AE1 or AE3 respectively. By indirect immunofluorescence technique, the AE1 and AE3-reactive antigens were localized throughout the cytoplasm in a diffused pattern. The 10 nm-plant filaments could be reassembled in vitro. These results demonstrated that keratin-like intermediate filaments exist in the cytoplasm of A. thaliana. Using conservative sequence of animal IF genes as primer, a cDNA fragment was further cloned from this model material by RT-PCR, which might shed more light on molecular characterization of IF existence in higher plant.

Key words: Arabidopsis thaliana, Intermediate filament, Selective extraction, Western blot, Assembly in vitro, Gene cloning