Intracellular distribution of subcellular organelles revealed by antibody against xyloglucan during cell cycle in tobacco BY-2 cells

Immunofluorescence microscopy using an antibody against xyloglucan (XG) rev ealed its dynamics during the cell cycle. In interphase tobacco BY-2 cells, punctate and scattered fluorescence was observed throughout the cytoplasm. Colocalization of such signals with cortical microtubules (MTs) was clearl y observed on the membrane ghosts. They were also associated and accumulate d on MT bundles of the preprophase band. Treatment of protoplasts with cyto chalasin B prior to the preparation of the ghosts had no effect on the patt ern of anti-XG staining, while treatment with propyzamide caused the disapp earance of the staining. These results suggest an association of Golgi appa ratus and/or Golgi-derived vesicles with MTs. In metaphase cells, the stain ing was dispersed in the cytoplasm, except in the area occupied by the meta phase spindle. During anaphase, a broad fluorescence band appeared between daughter chromosomes and gradually concentrated at the equatorial plane bef ore formation of the phragmoplast. At telophase, a bright Line of fluoresce nce appeared at the equatorial plane corresponding to the position of the c ell plate. The length of the line increased as cytokinesis proceeded. Thus, we showed that immunofluorescence microscopy using anti-XG antibody can be considered as a powerful tool for the analysis of Golgi apparatus and Golg i-derived vesicles containing XG.