Fine structure and X-ray microanalysis of a red macrophyte cultured under cadmium stress

Thalli of the red alga Audouinella saviana were exposed to 600 microM Cd2+ (LC50), 1000 microM Cd2+ and 1500 microM Cd2+ (final concentrations) for 5, 10 and 15 days (each dose) by adding cadmium nitrate to the culture medium. Untreated thalli were set in triplicate as controls for each experiment. Ultrastructural modifications due to cadmium ad/absorption were observed by TEM/SEM electron microscopy. SEM-EDS X-ray microanalysis, definining the accumulation sites, was performed on cryoprepared samples. TEM studies showed striking changes in the plasmalemma of treated algae, which became irregular and convoluted. Electron-transparent exocytic vesicles, possibly related to cell wall polysaccharide synthesis, were observed. The appearance of ribosomes and Golgi bodies, not significantly present in the cytoplasms of untreated cells, suggested enhanced protein and carbohydrate biosynthesis. The cell walls lost their initial grooves and became smooth and thick. More or less electron-dense vesicular systems were formed. Electron-dense sphaeroids occurred in the plasmalemma-cell wall interface, in the cell wall itself and in nearby vesicular membrane systems. Many small vacuoles containing large metal complexes were formed. Complexes were then sequestered into a large vacuole. SEM observations demonstrated that the cell wall and the membrane systems were the most involved in the defense responses. EDS-X-ray microanalysis confirmed the presence of cadmium in these compartments. Chloroplasts, where no Cd2+ signal was detected, were the least affected organelles, showing only a partial disorganization after lengthy exposure to high Cd2+ concentrations.