Starch degradation in cells is closely associated with cereal seed germination, photosynthesis in leaves, carbohydrate storage in tuberous roots, and fleshy fruit development. α-Amylase is considered as one of the key enzymes catalyzing starch breakdown, but up to date its role in starch breakdown in living cells remains unclear because the enzyme was often shown extrachloroplastic in living cells. The present experiment showed that α-amylase activity was progressively increasing concomitantly with the decreasing starch concentrations during the development of apple (Malus domestica Borkh cv. Starkrimson) fruit. The apparent amount of α-amylase assessed by Western blotting also increased during the fruit development, which is consistent with the seasonal changes in the enzyme activity. The enzyme subcellular-localization studies via immunogold electron-microscopy technique showed that α-amylase visualized by gold particles was predominantly located in plastids, but the gold particles were scarcely found in other subcellular compartments. A high density of the enzyme was observed at the periphery of starch granules during the middle and late developmental stages. These data proved that the enzyme is compartmented in its functional sites in the living cells of the fruit. The predominantly plastid-distributed pattern of α-amylase in cells was shown unchanged throughout the fruit development. The density of gold particles (α-amylase) in plastids was increasing during the fruit development, which is consistent with the results of Western blotting. So it is considered that α-amylase is involved in starch hydrolysis in plastids of the fruit cells.
苹果果实发育过程中α
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