Characterization of the Cell Wall Microdomain Surrounding Plasmodesmata in Apple Fruit

In fleshy fruits ripening is generally associated with a loss in tissue firmness resulting from depolymerization of wall components and separation of adjacent cells. In the regions of the wall that contain plasmodesmata, the usual sequences of ripening events, i.e. depolymerization of the middle lamellae and splitting of the walls, are not observed. In the present study we attempted to characterize in apple (Malus domestica Borkh.) fruit the structural microdomain of the cell wall that surrounds the plasmodesmata by in muro visualization of the cell wall components. Anionic sites of galacturonic acids were labeled with cationic gold. Low-esterified homogalacturonans were labeled with the monoclonal antibody JIM 5. In addition, a polyclonal antibody directed toward [beta](1->3)-glucopyranose was used to target callose in situ. The results indicated that the plasmodesmata-wall complexes were surrounded by a pectic microdomain. This domain was composed of low-esterified homogalacturonans that were not involved in calcium cross-bridging but were probably surrounded by a cationic environment. These structural features may result in the prevention of normal cell wall separation in regions containing plasmodesmata. However, observations by low-temperature scanning electron microscopy suggested that splitting of these walls ruptured the plasmodesmata and ultimately resulted in the spatial separation of adjacent cells.