Scanning Electron Microscopic Study of Modified Chloroplasts in Senescing Broccoli Florets

Structural changes in chloroplasts of broccoli (Brassica oleracea L., Italica group) florets during senescence were examined using light microscopy, scanning electron microscopy (SEM) with freeze-fracture technique, and transmission electron microscopy (TEM) to better understand the process of chloroplast degradation, particularly at the advanced stage of senescence. Light microscopy revealed that chloroplasts, which initially were intact and green, became obscure in shape, and their color faded during senescence. Small, colored particles appeared in cells as the florets approached the final stage of senescence and became full- to dark-yellow in color. Scanning electron microscopy showed that stroma thylakoids in the chloroplast initially were parallel to each other and grana thylakoids were tightly stacked. As senescence advanced, the grana thylakoids degenerated and formed globules. The globules became larger by aggregation as senescence progressed, and the large globules, called “thylakoid plexus,” formed numerous vesicles. The vesicles ultimately were expelled into the cytosol, and the light microscope revealed many colored particles in the senescent cells. These results indicate that the degradation of chloroplasts in broccoli florets progresses systematically, with the final product being colored particles, which are visible in yellow broccoli sepal cells.