Chloroplast envelopes as affected by light or dark pretreatment of pea plants

Glutaraldehyde fixation in 0.33 M sorbitol without any buffer reveals changes in the staining properties of the envelopes of chloroplasts of pea plants kept in the light or in the dark prior to fixation. After dark pretreatment the outer double membrane of the chloroplast does not adsorb heavy metals, resulting in a "white" unstained rim instead of the usual membrane. All other membranes of the cell, including chloroplast grana, are not affected and stain normally. Light pretreatment of the plants allows the usual staining of the outer membrane of the chloroplats. Fixation carried out in the medium usually used to isolate intact CO2 fixing chloroplasts (sorbitol+buffer+ions) reverses the above process and results in unstained envelopes of chloroplasts from preilluminated leaves, while the envelopes of chloroplasts from leaves kept in the dark stain normally. Glutaraldehyde-fixed chloroplats isolated from preilluminated leaves show a very basic isoelectric point during electrofocusing, while fixed chloroplasts from predarkened tissue exhibit an isoelectric point at about pH 7.