The distribution, structure and functional state of plasmodesmata were investigated to gain a clearer understanding of the sucrose transport pathway to the storage parenchyma cells in stem tissue in Saccharum officinarum var. NCo376. Evidence from structural studies on sugarcane stems by electron microscopy indicated that there are numerous plasmodesmata from the vascular bundles through to the storage parenchyma cells in mature stem tissue. Our studies, supported by fluorescence microscopy and iontophoresis, indicate that there are functional plasmodesmata in the phloemunloading pathway from transport phloem tissue to the bundle sheath in Saccharum, which could support symplasmic transport; plasmodesmata outside of the sheath cells in the storage parenchyma appear to be constricted by sphincter-like structures within their neck regions. Staining with Aniline Blue revealed evidence of large callose deposits, which co-localized with plasmodesmatal aggregates in the walls of the storage parenchyma cells. This suggests that the sucrose transport into, and accumulation by, storage parenchyma of mature stem tissue is under apoplasmic control.