Journal of Cell Science, Vol 3, 55-64, Copyright © 1968 by Company of Biologists

Submitted on July 5, 1967

Further Ultrastructural Observations on Polysaccharide Localization in Plant Cells

¹ Electron Microscope Unit, John Curtin School of Medical Research, Australian National University Canberra, Australia

Standard periodic acid/Schiff (PAS) techniques have not shown the existence of aldehyde groups in sections of glutaraldehyde-fixed, Araldite-embedded root-tip tissue; peroxidation of such sections resulted in a typical PAS staining pattern. Permanganate-fixed root tips also gave a weak PAS reaction which was intensified by prior peroxidation of the sections. At the ultrastructural level, silver hexamine was used to detect aldehyde groups produced in polysaccharide by permanganate and/or periodate oxidation. Golgi vesicles and slime material in root-cap cells always reacted strongly; the cell wall proper was less reactive. A marked increase in the stainability of the vesicles was evident, the further removed they were from Golgi bodies. This also occurred in root epidermal cells. In both these types of cells, smallersized vesicles and/or the contents of reticulate Golgi cisternae showed evidence of histochemical staining. In meristematic root tip cells, vesicles closely apposed to Golgi bodies did not stain convincingly, though cell walls stained readily. During cell-plate formation, however, both smaller (possibly Golgi) and larger vesicles (phragmoplasts) stained strongly. The walls of permanganate-fixed sieve-tube cells also stained quite strongly, but callose did not unless the tissue block had been treated with periodate before being embedded. In glutaraldehyde-fixed xylem cells, older wall thickenings reacted very strongly even when the sections had been blocked with iodoacetate and bisulphite (which rendered the rest of the section unreactive). If similar sections of younger xylem cells were peroxidized after such blocking reactions, the primary cell wall and the wall thickenings stained, as did many of the Golgi vesicles. The results are related to other experimental observations, both ultrastructural and histochemical, on plant cells.

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