• Journal of Plant Biology
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• v.35 no.2
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• pp.99-106
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• 1992

The endosperm protein, vicilin, of ginseng (Panax ginseng C.A. Meyer) was purified by ammonium sulfate precipitaion, gel permeation and ion exchange column chromatography. Vicilin is a glycoprotein composed of 2 subunits with molecular masses of 55,000 (large subunit) and 44,000 (small subunit). The anti-vicilin antibody was raised in rabbit, and purified by DEAE Affi-Gel Blue affinity chromatography. The endosperm cells of the seed were reacted with this anti-vicilin antibody and colloidal gold conjugated secondary antibody. Gold particles were labelled on the elaborating granules of Golgi complex, electron-dense granules and protein bodies in the endosperm cells. These results indicated that the vicilin, which was synthesized in rough endoplasmic reticulum and transported to Golgi, was elaborated in saccules of the Golgi and then transported into protein bodies by electron-dense granules.anules.

• Journal of Ginseng Research
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• v.19 no.3
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• pp.267-274
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• 1995

Vicilin and legumin, the storage Proteins of seed, were Purified from ginseng (Panax ginseng C.A. Meyer) endosperm cells. They were immunized in rabbits, and antibodies were raised respectively. Using these two antibodies, double immunogold labeling of vicilin and legumin was carried out to determine the gap of synthetic time and the transport pattern of vicilin and legumin in the ginseng endosperm cells. Vicilin and legumin were synthesized at the same time at early embryo developmental stage. They were secreted from the Golgi bodies and accumulated into the small vacuoles. As the endosperm cells developed, vicilin and legumin localized in the small vacuoles were gradually transported toward the large central vacuole where they were stored. Protein bodies were derived from the vacuoles filled with proteins and distributed in the endosperm cells of mature red seed. Protein bodies were various in size from 1 to 8 ${\mu}{\textrm}{m}$ in which vicilin and legumin were mixed each other. The number of small particles labeled on the vicilin was greater than that of large particles labeled on the legumin in the protein bodies indicating that the amount of vicilin is higher than that of legumin in the protein bodies.

• Journal of Plant Biology
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• v.39 no.2
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• pp.123-126
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• 1996

The pattern of seed storage protein, vicilin, deposition and site of intracellular localization was examined in cotyledon cells of pea (Pisum sativum) seed using the immunocytochemical methods. The vicilin was confined to the cisternae fo the rough endoplasmic reticulum and dictyosome as well as protein granules newly formed in rough endoplasmic reticulum. Vacuolar protein deposites and protein bodies were also labelled by gold particles. After small protein bodies were formed in the rough endoplasmic reticulum, they were transported to large protein bodies and then fused together. Electron dense protein granule, elaborated in the dictyosome, appears to be transported from dictyosome to protein body. A few unlabelled protein granules seem to be accumulated in other type of proteins than vicilin.

• Applied Microscopy
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• v.29 no.4
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• pp.499-509
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• 1999

Accumulations of the storage proteins in protein storage vacuole and the differentiation of protein bodies from protein-filled ER in developing pea cotyledons have been investigated using conventional and immunoelectron microscopy. To improve the fixation quality, single cells separated enzymatically from sliced cotyledons were used. At early stages of seed development osmiophilic protein accumulates in rER lumen were observed quite often. This protein-filled ER cisternae were differentiated into cytoplasmic protein bodies at late stage by the process called terminal dilations which have been considered a principal route of the formation of cytoplasmic protein bodies somewhat later in seed maturation. Immunocytochemical labellings of the vicilin and legumin show that presence of vicilin on both of the cytoplasmic PB and PD, but limited presence of legumin only on the cytoplasmic PB at intermediate stage of seed development. Immunogold labellings of Bip, ER retention protein, were observed on the inner periphery of protein deposits in protein storage vacuole. This result was regarded that Bip can recognize and retrieve misfolded protein during active accumulation of storage protein to the PD in PSV.

• Applied Microscopy
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• v.31 no.2
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• pp.199-206
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• 2001

In 1980s, the fragmentation or subdivision of protein deposits at the periphery of protein storage vacuole was suggested as the only route of PB development in pea cotyledon cells. Since then, other independant processes such as terminal dilation , transformation and de novo development have been discussed as alternative routes for PB development, and today, these multiple mechanisms of PB development are accepted as a result of active investigations. For analysis of the protein accumulations in the ER cisternae during seed development, immunocytochemical gold labellings were applyed on the single cells separated by enzymatic digestion from cotyledon tissue. Anti-legumin labellings at the early stage, and anti-vicilin labellings at the intermediate stage were observed on the protein-filled ER. The $\alpha-Tip$, which is the ER retention protein, was labelled somewhat at late stage, and PPase, a sort of tonoplast membrane protein, was labelled at early stage.

• Applied Microscopy
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• v.24 no.4
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• pp.86-97
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• 1994

Protein bodies in the endosperm cells of mature red ginseng (Panax ginseng C.A. Meyer) were distributed evenly in the cytoplasm and their size varied from 1 to $8{\mu}m$. Three types of protein bodies were detected and they are spherical or egg-shaped ones containing homogeneous matrix only, spherical ones containing globoids, and irregular shaped ones. Protein bodies degraded in two patterns, one is to start the degration of the body from the surface toward the center, while the other is that the body was broken evenly and then degraded gradually. After degradation, only the limiting membrane remained, that causes the body to be empty. The limiting membranes fused with each other to form a large vacuole. Vicilin and legumin decreased in the endosperm cells as the protein bodies degraded gradually whereas they increased in the umbiliform layers.