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Published online 27 August 2001. doi:10.1083/jcb.200105096
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© The Rockefeller University Press, 0021-9525/2001/9/973 $5.00
The Journal of Cell Biology, Volume 154, Number 5, September 3, 2001 973-982


Article

Cotyledon cells of Vigna mungo seedlings use at least two distinct autophagic machineries for degradation of starch granules and cellular components

Kiminori Toyooka1,2, Takashi Okamoto1 and Takao Minamikawa1

1 Department of Biological Sciences, Tokyo Metropolitan University, Tokyo, 192-0397 Japan
2 Department of Infectious Diseases and Tropical Medicine, International Medical Center of Japan, Tokyo, 162-8655 Japan

Address correspondence to Dr. Takashi Okamoto, Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo, 192-0397 Japan. Tel.: 81-426-77-2562. Fax: 81-426-77-2559. E-mail: okamoto-takashi{at}c.metro-u.ac.jp

{alpha}-Amylase is expressed in cotyledons of germinated Vigna mungo seeds and is responsible for the degradation of starch that is stored in the starch granule (SG). Immunocytochemical analysis of the cotyledon cells with anti–{alpha}-amylase antibody showed that {alpha}-amylase is transported to protein storage vacuole (PSV) and lytic vacuole (LV), which is converted from PSV by hydrolysis of storage proteins. To observe the insertion/degradation processes of SG into/in the inside of vacuoles, ultrastructural analyses of the cotyledon cells were conducted. The results revealed that SG is inserted into LV through autophagic function of LV and subsequently degraded by vacuolar {alpha}-amylase. The autophagy for SG was structurally similar to micropexophagy detected in yeast cells. In addition to the autophagic process for SG, autophagosome-mediated autophagy for cytoplasm and mitochondria was detected in the cotyledon cells. When the embryo axes were removed from seeds and the detached cotyledons were incubated, the autophagosome-mediated autophagy was observed, but the autophagic process for the degradation of SG was not detected, suggesting that these two autophagic processes were mediated by different cellular mechanisms. The two distinct autophagic processes were thought to be involved in the breakdown of SG and cell components in the cells of germinated cotyledon.

Key Words: {alpha}-amylase; autophagy; intracellular transport; vacuole; senescence


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