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Imunohistochemical Localization of -Amylase in Cotyledons of Vigna mungo Seedlings ¹

Department of Biology, Tokyo Metropolitan University, Setagaya-ku, Tokyo 158, Japan

We studied the localization of -amylase with indirect fluorescence microscopy in transversely sectioned cotyledons of Vigna mungo seedlings. Tissue sections were fixed in periodate-lysine-paraformaldehyde and treated with anti--amylase immunoglobulin G followed by fluorescein isothiocyanate labeled goat anti-rabbit immunoglobulin G. -Amylase appeared in the cells farthest from vascular bundles on the second day of growth and appeared gradually closer to the vascular bundles as growth progressed. The pattern of -amylase appearance was similar in detached cotyledons, indicating that attachment of the embryonic axis has no effect on this pattern. However, in attached cotyledons, -amylase disappeared from the regions where starch grains had been digested, but in detached cotyledons there was no disappearance of -amylase, and digestion was slower than in intact cotyledons.